RNF185 Is a Novel E3 Ligase of Endoplasmic Reticulum-associated Degradation (ERAD) That Targets Cystic Fibrosis Transmembrane Conductance Regulator (CFTR)

In the endoplasmic reticulum (ER), misfolded or improperly assembled proteins are exported to the cytoplasm and degraded by the ubiquitin-proteasome pathway through a process called ER-associated degradation (ERAD). ER-associated E3 ligases, which coordinate substrate recognition, export, and proteasome targeting, are key components of ERAD. Cystic fibrosis transmembrane conductance regulator (CFTR) is one ERAD substrate targeted to co-translational degradation by the E3 ligase RNF5/RMA1. RNF185 is a RING domain-containing polypeptide homologous to RNF5. We show that RNF185 controls the stability of CFTR and of the CFTRΔF508 mutant in a RING- and proteasome-dependent manner but does not control that of other classical ERAD model substrates. Reciprocally, its silencing stabilizes CFTR proteins. Turnover analyses indicate that, as RNF5, RNF185 targets CFTR to co-translational degradation. Importantly, however, simultaneous depletion of RNF5 and RNF185 profoundly blocks CFTRΔF508 degradation not only during translation but also after synthesis is complete. Our data thus identify RNF185 and RNF5 as a novel E3 ligase module that is central to the control of CFTR degradation.     

Influence of MDR1 C1236T polymorphism on lopinavir plasma concentration and virological response in HIV-1-infected children

Background

Variability in MDR1 and PXR has been associated with differences in drug plasma levels and response to antiretroviral therapy. We investigated whether polymorphisms in MDR1 (T-129C, C1236T and C3435T) and PXR (C63396T) affect lopinavir plasma concentration and the virological or immunological response to HAART in HIV-1-infected children.

Methods

Genotypes were identified in 100 blood donors and 38 HIV-1-infected children. All children received HAART with lopinavir boosted with ritonavir (LPV/r) at the time of LPV plasma level quantification, before (C_trough) and between 1 and 2 h after (C_post-dose) the administration of the next dose of drug. CD4⁺ T-cell counts and plasma viral load were analyzed before and after the initiation of LPV/r.

Results

MDR1 1236T, MDR1 3435T and PXR 63396T alleles showed a frequency of ~ 50% while the MDR1 -129C allele only reached 5%. Children heterozygotes 1236CT showed a significantly lower LPV C_post-dose than homozygotes 1236TT (median C_post-dose = 3.04 μg/ml and 6.50 μg/ml, respectively; p = 0.016). Children heterozygotes 1236CT also had a lower decrease of viral load after 36 weeks of LPV/r exposure compared with homozygotes 1236CC (median viral load changes = − 0.50 log₁₀ copies/ml and − 2.08 log₁₀ copies/ml, respectively; p = 0.047). No effect on the immunological response was observed for polymorphisms of MDR1 or PXR.

Conclusions

Our results suggest that the MDR1 C1236T SNP significantly reduces LPV plasma concentration affecting the virological response to HAART. Heterozygotes 1236CT might have an altered level of P-gp expression/activity in enterocytes and CD4⁺ T lymphocytes that limits the absorption of LPV leading to an impaired virological suppression.     

The role of cutaneous feedback for anticipatory grip force adjustments during object movements and externally imposed variation of the direction of gravity

Grip force adjustments to changes of object loading induced by external changes of the direction of gravity during discrete arm movements with a grasped object were analyzed during normal and anesthetized finger sensibility. Two subjects were seated upright in a rotatable chair and rotated backwards into a horizontal position during discrete movements with a hand-held instrumented object. The movement direction varied from vertical to horizontal inducing corresponding changes in the direction of gravity, but the orientation of the movement in relation to the body remained unaffected. During discrete vertical movements a maximum of load force occurs early in upward and late in downward movements; during horizontal movements two load force peaks result from both acceleratory and deceleratory phases of the movement. During performance with normal finger sensibility grip force was modulated in parallel with fluctuations of load force during vertical and horizontal movements. The grip force profile adopted to the varying load force profile during the transition from the vertical to the horizontal position. The maximum grip force occurred at the same time of maximum load force irrespective of the movement plane. During both subjects' first experience of digital anesthesia the object slipped from the grasp during rotation to the horizontal plane. During the following trials with anesthetized fingers subjects substantially increased their grip forces, resulting in elevated force ratios between maximum grip and load force. However, grip force was still modulated with the movement-induced load fluctuations and maximum grip force coincided with maximum load force during vertical and horizontal movements. This implies that the elevated force ratio between maximum grip and load force does not alter the feedforward system of grip force control. Cutaneous afferent information from the grasping digits seems to be important for the economic scaling of the grip force magnitude according to the actual loading conditions and for reactive grip force adjustments in response to load perturbations. However, it plays a subordinate role for the precise anticipatory temporal coupling between grip and load forces during voluntary object manipulation.     

基于控制模型的药物动力学静注问题的研究

在新药临床的研究中,以静脉注射方式给药时,药物进入全身循环快速分布到体内的各个组织,通过将机体作为单室模型对血药浓度进行控制,可以从血药浓度的变化得到组织药物水平的状态、本文针对药物动力学的特点,利用控制模型,给出了对其稳定性分析的方法．     

AIDS Research and Its Role in China＇s AIDS Prevention and Control Policies

By the end of 2005, the estimated number of HIV infected people in China was 650,000. The seriousness of the epidemic calls for effective control measures to tackle the problems in order to avoid the tragedy in Africa from happening in China. ＂Prevention First＂ is the cornerstone of the country＇s health policy. On 2003 World AIDS Day, Premier Jiabao Wen announced a new national AIDS control policy, ＂Four Frees and One Care＂. This policy clearly shows that the Chinese government has once again taken full responsibility to solve public health problems and has profound impact far beyond the AIDS field. In early 2006, the central government put scientific and technology innovation as a national priority and set the target to build an innovative China by year 2020. Since then, the government has been increasing investment in science and technology with major emphasis on both infectious diseases control and new drug research and development. For the first time, development of 100 new drugs and control of major infectious diseases （AIDS, HBV, TB and other emerging infectious diseases） have been selected as national key scientific projects. China＇s best minds in related fields will be pooled to work together in order to remove the technical barriers blocking efficient control of the major infectious disease in China. Knowledge on molecular epidemiology, immunology, pathogenesis, HAART, as well as HIVDR strains will certainly provide urgently needed scientific information for China＇s AIDS control program. Only evidence-based strategy from good research will provide long-term effective control of AIDS.     

The potential of greenhouse sinks to underwrite improved land management

The current agricultural systems of broad areas of Australia are unsustainable, with large projected increases in salinization, decreases in water quality, wind erosion, and losses of biodiversity. It is well known that these problems can be partially resolved by farmland reforestation; however, a major issue is financing the scale of activity required. The international response to global warming, the United Nations Framework Convention on Climate Change and its Kyoto Protocol, includes provisions that enable greenhouse sinks (sequestration of carbon in soils and vegetation) to be used by parties to fulfil their obligations. The Kyoto Protocol also allows for trading in emission reductions, and this opens the possibility that investment in carbon sinks may help underwrite broader natural resource management objectives. This paper examines the possibilities for improved land management in Western Australia arising from the development of carbon sinks by considering: (a) the likelihood of a carbon market developing and the likely depth of that market as a result of current national and international policies, (b) the data available to provide estimates on different types of sinks, and (c) the likely benefits of wide-scale sink investment.It was estimated that the total amount of carbon that could be sequestered by reforesting 16.8 Mha of cleared farmland is 2200 Mt CO₂-e, and between 290 and 1170 Mt CO₂-e by destocking 94.8 Mha of rangelands. There were insufficient data to produce estimates of sequestration following changes in tillage practice in cropping systems or the revegetation of already salinized land. We conclude that carbon sinks are only likely to become profitable as a broad-scale stand-alone enterprise when carbon prices reach AUD$15/t CO₂-e, with this threshold value varying with carbon yield and project costs. Below this price, their value can be significant as an adjunct to reforestation schemes that are aimed at providing other products (wood, pulp, bioenergy) and land and water conservation benefits. Irrespective of this, carbon sinks provide an opportunity to both sequester carbon in a least-cost fashion and improve soil and watershed management.     

Quality control and integration of genotypes from two calling pipelines for whole genome sequence data in the Alzheimer's disease sequencing project

The Alzheimer's Disease Sequencing Project (ADSP) performed whole genome sequencing (WGS) of 584 subjects from 111 multiplex families at three sequencing centers. Genotype calling of single nucleotide variants (SNVs) and insertion-deletion variants (indels) was performed centrally using GATK-HaplotypeCaller and Atlas V2. The ADSP Quality Control (QC) Working Group applied QC protocols to project-level variant call format files (VCFs) from each pipeline, and developed and implemented a novel protocol, termed “consensus calling,” to combine genotype calls from both pipelines into a single high-quality set. QC was applied to autosomal bi-allelic SNVs and indels, and included pipeline-recommended QC filters, variant-level QC, and sample-level QC. Low-quality variants or genotypes were excluded, and sample outliers were noted. Quality was assessed by examining Mendelian inconsistencies (MIs) among 67 parent-offspring pairs, and MIs were used to establish additional genotype-specific filters for GATK calls. After QC, 578 subjects remained. Pipeline-specific QC excluded ~12.0% of GATK and 14.5% of Atlas SNVs. Between pipelines, ~91% of SNV genotypes across all QCed variants were concordant; 4.23% and 4.56% of genotypes were exclusive to Atlas or GATK, respectively; the remaining ~0.01% of discordant genotypes were excluded. For indels, variant-level QC excluded ~36.8% of GATK and 35.3% of Atlas indels. Between pipelines, ~55.6% of indel genotypes were concordant; while 10.3% and 28.3% were exclusive to Atlas or GATK, respectively; and ~0.29% of discordant genotypes were. The final WGS consensus dataset contains 27,896,774 SNVs and 3,133,926 indels and is publicly available.     

An application of scissored-pair control moment gyroscopes in a design of wearable balance assistance device for the elderly

Impaired balance control ability and degraded functional mobility increases the risk of falling in elderly people. The elderly show more postural sway when standing compared with young people. A sway fall occurs when the center of gravity moves outside the limit of stability. In order to reduce the fall risk from the excessive sway, this study presents the design of wearable balance assistance device for the elderly. Scissored-pair control moment gyroscopes were selected as a torque actuator. A two-axis inclination sensor was used to detect the inclined angle of the wearer’s body. The direction of sway was calculated from the detected inclined angle. The designed device weighs 8.2 kg with a height of 32 cm × width of 40 cm × depth of 22 cm. A multi-segment model of a standing human was used to investigate the device’s performance for balance recovery. According to the simulations, balance recovery in any direction was successfully accomplished with the appropriate initial angle. The relationship between the effective initial angle and detected inclined angle was subsequently established. The stability provided by activation of the device was able to limit the unstable user’s sway boundary. The designed device shows promise for use as a balance assistance device for the elderly.