Three-dimensional structure of the channel-forming trans-membrane domain of virus protein "u" (Vpu) from HIV-1

The three-dimensional structure of the channel-forming trans-membrane domain of virus protein "u" (Vpu) of HIV-1 was determined by NMR spectroscopy in micelle and bilayer samples. Vpu(2-30+) is a 36-residue polypeptide that consists of residues 2-30 from the N terminus of Vpu and a six-residue "solubility tag" at its C terminus that facilitates the isolation, purification, and sample preparation of this highly hydrophobic minimal channel-forming domain. Nearly all of the resonances in the two-dimensional 1H/15N HSQC spectrum of uniformly 15N labeled Vpu(2-30+) in micelles are superimposable on those from the corresponding residues in the spectrum of full-length Vpu, which indicates that the structure of the trans-membrane domain is not strongly affected by the presence of the cytoplasmic domain at its C terminus. The two-dimensional 1H/15N PISEMA spectrum of Vpu(2-30+) in lipid bilayers aligned between glass plates has been fully resolved and assigned. The "wheel-like" pattern of resonances in the spectrum is characteristic of a slightly tilted membrane-spanning helix. Experiments were also performed on weakly aligned micelle samples to measure residual dipolar couplings and chemical shift anisotropies. The analysis of the PISA wheels and Dipolar Waves obtained from both weakly and completely aligned samples show that Vpu(2-30+) has a trans-membrane alpha-helix spanning residues 8-25 with an average tilt of 13 degrees. The helix is kinked slightly at Ile17, which results in tilts of 12 degrees for residues 8-16 and 15 degrees for residues 17-25. A structural fit to the experimental solid-state NMR data results in a three-dimensional structure with precision equivalent to an RMSD of 0.4 A. Vpu(2-30+) exists mainly as an oligomer on PFO-PAGE and forms ion-channels, a most frequent conductance of 96(+/- 6) pS in lipid bilayers. The structural features of the trans-membrane domain are determinants of the ion-channel activity that may be associated with the protein's role in facilitating the budding of new virus particles from infected cells.     

Glenohumeral joint dynamics and shoulder muscle activity during geared manual wheelchair propulsion on carpeted floor in individuals with spinal cord injury

This study investigated the effects of using geared wheels on glenohumeral joint dynamics and shoulder muscle activity during manual wheelchair propulsion. Seven veterans with spinal cord injury propelled their wheelchairs equipped with geared wheels over a carpeted floor in low gear (1.5:1) and standard gear (1:1) conditions. Hand-rim kinetics, glenohumeral joint dynamics, and muscle activity were measured using a custom instrumented geared wheel, motion analysis, and surface electromyography. Findings indicated that the propulsion speed and stroke distance decreased significantly during the low gear condition. The peak hand-rim resultant force and propulsive moment, as well as the peak glenohumeral inferior force and flexion moment, were significantly less during the low gear condition. The peak and integrated muscle activity of the anterior deltoid and pectoralis major decreased significantly, while the normalized integrated muscle activity (muscle activity per stroke distance) was not significantly different between the two conditions. Propulsion on carpeted floor in the low gear condition was accompanied by a reduced perception of effort. The notable decrease in the peak shoulder loading and muscle activity suggests that usage of geared wheels may be beneficial for wheelchair users to enhance independent mobility in their homes and communities while decreasing their shoulder demands.     

郑州仰韶时期石纺轮的类型与加工技术

石纺轮是仰韶时期重要的石质工具之一,分析其形态、制作技术对认识这一阶段的石器工业具有重要意义。本文对郑州地区仰韶时期石纺轮开展类型学研究、数理统计和微痕分析后发现,石纺轮制作和纺织品生产中存在较高的专业化水平。郑州地区应存在两种不同类型的石纺轮,器物形制与生产技术间存在紧密关联,分别对应管钻和打制-实心钻两种技术体系,这两种技术体系在郑州地区仰韶时期各遗址内均有发现,表明不同遗址间的石纺轮加工技术水平并无明显差异。     

Ultrasound elastographic assessment of plantar fascia in runners using rearfoot strike and forefoot strike

Forefoot strike is increasingly being adopted by runners because it can better attenuate impact than rearfoot strike. However, forefoot strike may overload the plantar fascia and alter the plantar fascia elasticity. This study aimed to use ultrasound elastography to investigate and compare shear wave elasticity of the plantar fascia between rearfoot strikers and forefoot strikers. A total of 35 participants (21 rearfoot strikers and 14 forefoot strikers), who were free of lower limb injuries and diseases, were recruited from a local running club. Individual foot strike patterns were identified through the measured plantar pressure during treadmill running. The B-Mode ultrasound images and shear wave elastographic images of the plantar fascia were collected from each runner. Two independent investigators reviewed the images and examined the plantar fascia qualitatively and quantitatively. The results demonstrated an overall good agreement between the investigators in the image review outcomes (ICC:0.96–0.98, κ: 0.89). There were no significant differences in the fascial thickness (p = 0.50) and hypoechogenicity on the gray-scale images (p = 0.54) between the two groups. Shear wave elastography showed that forefoot strikers exhibited reduced plantar fascia elasticity compared to rearfoot strikers (p = 0.01, Cohen’s d = 0.91). A less elastic fascial tissue was more easily strained under loading. Tissue overstrain is frequently related to the incidence of plantar fasciitis. While further study is needed for firm conclusions, runners using forefoot strike were encouraged to enhance their foot strength for better protection of the plantar fascia.     

Impact of local temperature increase on the early development of biofilm-associated ciliate communities

Indications of global climate change and associated unusual temperature fluctuations have become increasingly obvious over the past few decades. Consequently, the relevance of temperature increases for ecological communities and for whole ecosystems is one of the major challenges of current ecological research. One approach to investigating the effects of increasing temperatures on communities is the use of fast-growing microbial communities. Here we introduce a river bypass system in which we tested the effect of temperature increases (0, 2, 4, 6°C above the long-term average) on both the colonization speed and the carrying capacity of biofilm-associated ciliate communities under different seasonal scenarios. We further investigated interactions of temperature and resource availability by cross-manipulations in order to test the hypothesis that temperature-mediated effects will be strongest in environments that are not resource-limited. Strong seasonal differences in both tested parameters occurred under natural conditions (no resource addition), and the effects of temperature increase at a given time were relatively low. However, increasing temperature can significantly accelerate the colonization speed and reduce the carrying capacity in particular seasons. These effects were strongest in winter. Simultaneous manipulation of temperature and of resource availability amplified the response to temperature increase, adumbrating strong interactive control of populations by temperature and resource availability. Our results show that the response of communities to local temperature increases strongly depends on the seasonal setting, the resource availability and the stage of succession (early colonization speed vs. carrying capacity). Priority programme of the German Research Foundation—contribution 11.     

Structural characterization of the pore forming protein TatAd of the twin-arginine translocase in membranes by solid-state N-NMR

The transmembrane protein TatA is the pore forming unit of the twin-arginine translocase (Tat), which has the unique ability of transporting folded proteins across the cell membrane. This ATP-independent protein export pathway is a recently discovered alternative to the general secretory (Sec) system of bacteria. To obtain insight in the translocation mechanism, the structure and alignment in the membrane of the well-folded segments 2-45 of TatA_d from Bacillus subtilis was studied here. Using solid-state NMR in bicelles containing anionic lipids, the topology and orientation of TatA_d was determined in an environment mimicking the bacterial membrane. A wheel-like pattern, characteristic for a tilted transmembrane helix, was observed in ¹⁵N chemical shift /¹⁵N-¹H dipolar coupling correlation NMR spectra. Analysis of this PISA wheel revealed a 14-16 residue long N-terminal membrane-spanning helix which is tilted by 17° with respect to the membrane normal. In addition, comparison of uniformly and selectively ¹⁵N-labeled TatA_2-45 samples allowed determination of the helix polarity angle.     

自愿转轮运动对长爪沙鼠身体组成的影响

为研究自愿转轮运动对长爪沙鼠(Meriones unguiculatus)体重的影响,在以前工作的基础上,分析了自愿转轮运动及8周后体重、胴体重和体水,以及心、肝、脾、肺、肾、性腺(睾丸或卵巢)、消化道、腓肠肌、比目鱼肌、肾周脂肪垫和肠系膜脂肪垫等器官及组织的重量变化。结果发现,自愿转轮运动条件下长爪沙鼠胴体湿重、体水和心、肝、脾、肾、腓肠肌、消化道等器官重量增加。自愿转轮运动对比目鱼肌和器官脂肪垫重量的影响存在性别差异。自愿转轮运动使雄性长爪沙鼠器官的脂肪垫重量增加,但雌性降低;对雄性长爪沙鼠比目鱼肌重量没有影响,但雌性增加。以上结果表明,自愿转轮运动促进了长爪沙鼠的体重增长,改变了长爪沙鼠的身体组成。内脏器官和体水重量的增加是体重增加的主要原因。     

Cuprizone demyelination of the corpus callosum in mice correlates with altered social interaction and impaired bilateral sensorimotor coordination

For studies of remyelination in demyelinating diseases, the cuprizone model of CC (corpus callosum) demyelination has experimental advantages that include overall size, proximity to neural stem cells of the subventricular zone, and correlation with a lesion predilection site in multiple sclerosis. In addition, cuprizone treatment can be ended to allow more direct analysis of remyelination than with viral or autoimmune models. However, CC demyelination lacks a useful functional correlate in rodents for longitudinal analysis throughout the course of demyelination and remyelination. In the present study, we tested two distinct behavioural measurements in mice fed 0.2% cuprizone. Running on a ‘complex'' wheel with varied rung intervals requires integration between cerebral hemispheres for rapid bilateral sensorimotor coordination. Maximum running velocity on the ‘complex'' wheel decreased during acute (6 week) and chronic (12 week) cuprizone demyelination. Running velocity on the complex wheel distinguished treated (for 6 weeks) from non-treated mice, even after a 6-week recovery period for spontaneous remyelination. A second behavioural assessment was a resident–intruder test of social interaction. The frequency of interactive behaviours increased among resident mice after acute or chronic demyelination. Differences in both sensorimotor coordination and social interaction correlated with demonstrated CC demyelination. The wheel assay is applicable for longitudinal studies. The resident–intruder assay provides a complementary assessment of a distinct modality at a specific time point. These behavioural measurements are sufficiently robust for small cohorts as a non-invasive assessment of demyelination to facilitate analysis of subsequent remyelination. These measurements may also identify CC involvement in other mouse models of central nervous system injuries and disorders.     

Effects of low-pass filter combinations on lower extremity joint moments in distance running

Inverse dynamics is a standard tool in biomechanics, which requires low-pass filtering of external force and kinematic signals. Unmatched filtering procedures are reported to affect joint moment amplitudes in high impact movements, like landing or cutting, but are also common in the analysis of distance running. We analyzed the effects of cut-off frequencies in 94 rearfoot runners at a speed of 3.5 m/s. Additionally, we investigated whether the evaluation of footwear interventions is affected by the choice of cut-off frequencies. We performed 3D inverse dynamics for the hip, knee and ankle joints using different low-pass filter cut-off frequency combinations for a recursive fourth-order Butterworth filter. We observed fluctuations of joint moment curves in the first half of stance, which were most pronounced for the most unmatched cut-off frequency combination (kinematics: 10 Hz; ground reaction forces (GRFs): 100 Hz) and for more proximal joints. Peak sagittal plane hip joint moments were altered by 94% on average. We observed a change in the ranking of subjects based on joint moment amplitude. We found significant (p < 0.001) footwear by cut-off frequency combination interaction effects for most peak joint moments. These findings highlight the importance of cut-off frequency choice in the analysis of joint moments and the assessment of footwear interventions in distance running. Based on our results, we propose to use matched cut-off frequencies around 20 Hz in order to avoid large artificial fluctuations in joint moment curves while at the same time avoiding a severe removal of physiological high-frequency signal content from the GRF signals.     

dendroTools: R package for studying linear and nonlinear responses between tree-rings and daily environmental data

We introduce in this paper the dendroTools R package for studying the statistical relationships between tree-ring parameters and daily environmental data. The core function of the package is daily_response(), which works by sliding a moving window through daily environmental data and calculating statistical metrics with one or more tree ring proxies. Possible metrics are correlation coefficient, coefficient of determination and adjusted coefficient of determination. In addition to linear regression, it is possible to use a nonlinear artificial neural network with the Bayesian regularization training algorithm (brnn). dendroTools provides the opportunity to use daily climate data and robust nonlinear functions for the analysis of climate-growth relationships. Models should thus be better adapted to the real (continuous) growth of trees and should gain in predictive capabilities. The dendroTools R package is freely available in the CRAN repository. The functionality of the package is demonstrated on two examples, one using a mean vessel area (MVA) chronology and one a traditional tree-ring width (TRW).