Interaction Between Motor Domains Can Explain the Complex Dynamics of Heterodimeric Kinesins

Motor proteins are active enzyme molecules that play a crucial role in many biological processes. They transform the chemical energy into the mechanical work and move unidirectionally along rigid cytoskeleton filaments. Single-molecule experiments suggest that motor proteins, consisting of two motor domains, move in a hand-over-hand mechanism when each subunit changes between trailing and leading positions in alternating steps, and these subunits do not interact with each other. However, recent experiments on heterodimeric kinesins suggest that the motion of motor domains is not independent, but rather strongly coupled and coordinated, although the mechanism of these interactions are not known. We propose a simple discrete stochastic model to describe the dynamics of homodimeric and heterodimeric two-headed motor proteins. It is argued that interactions between motor domains modify free energy landscapes of each motor subunit, and motor proteins still move via the hand-over-hand mechanism but with different transitions rates. Our calculations of biophysical properties agree with experimental observations. Several ways to test the theoretical model are proposed.Comment: To appear in New J. Phy

Understanding Mechanochemical Coupling in Kinesins Using First-Passage Time Processes

Kinesins are processive motor proteins that move along microtubules in a stepwise manner, and their motion is powered by the hydrolysis of ATP. Recent experiments have investigated the coupling between the individual steps of single kinesin molecules and ATP hydrolysis, taking explicitly into account forward steps, backward steps and detachments. A theoretical study of mechanochemical coupling in kinesins, which extends the approach used successfully to describe the dynamics of conventional motor proteins, is presented. The possibility of irreversible detachments of kinesins from the microtubules is also explicitly taken into account. Using the method of first- passage times, experimental data on the mechanochemical coupling in kinesins are fully described using the simplest two-state model. It is shown that the dwell times for the kinesin to move one step forward or backward, or to dissociate irreversibly are the same, although the probabilities of these events are different. It is concluded that the current theoretical view, that only the forward motion of the motor protein molecule is coupled to ATP hydrolysis, is consistent with all available experimental observations for kinesins.Comment: Submitted to Biophysical Journa

X-ray detection with Micromegas with background levels below 10−6^{-6} keV−1^{-1}cm−2^{-2}s−1^{-1}

Micromegas detectors are an optimum technological choice for the detection of low energy x-rays. The low background techniques applied to these detectors yielded remarkable background reductions over the years, being the CAST experiment beneficiary of these developments. In this document we report on the latest upgrades towards further background reductions and better understanding of the detectors' response. The upgrades encompass the readout electronics, a new detector design and the implementation of a more efficient cosmic muon veto system. Background levels below 10$^{-6}$keV$^{-1}$cm$^{-2}$s$^{-1}$ have been obtained at sea level for the first time, demonstrating the feasibility of the expectations posed by IAXO, the next generation axion helioscope. Some results obtained with a set of measurements conducted in the x-ray beam of the CAST Detector Laboratory will be also presented and discussed

Generalized exclusion and Hopf algebras

We propose a generalized oscillator algebra at the roots of unity with generalized exclusion and we investigate the braided Hopf structure. We find that there are two solutions: these are the generalized exclusions of the bosonic and fermionic types. We also discuss the covariance properties of these oscillatorsComment: 10 pages, to appear in J. Phys.

Motion-Based Piloted Simulation Evaluation of a Control Allocation Technique to Recover from Pilot Induced Oscillations

This paper describes the maturation of a control allocation technique designed to assist pilots in the recovery from pilot induced oscillations (PIOs). The Control Allocation technique to recover from Pilot Induced Oscillations (CAPIO) is designed to enable next generation high efficiency aircraft designs. Energy efficient next generation aircraft require feedback control strategies that will enable lowering the actuator rate limit requirements for optimal airframe design. One of the common issues flying with actuator rate limits is PIOs caused by the phase lag between the pilot inputs and control surface response. CAPIO utilizes real-time optimization for control allocation to eliminate phase lag in the system caused by control surface rate limiting. System impacts of the control allocator were assessed through a piloted simulation evaluation of a non-linear aircraft simulation in the NASA Ames Vertical Motion Simulator. Results indicate that CAPIO helps reduce oscillatory behavior, including the severity and duration of PIOs, introduced by control surface rate limiting

An optimized single chain TCR scaffold relying on the assembly with the native CD3-complex prevents residual mispairing with endogenous TCRs in human T-cells.

Immunotherapy of cancer envisions the adoptive transfer of T-cells genetically engineered with tumor-specific heterodimeric α/β T-cell receptors (TCRα/β). However, potential mispairing of introduced TCRα/β-chains with endogenous β/α-ones may evoke unpredictable autoimmune reactivities. A novel single chain (sc)TCR format relies on the fusion of the Vα-Linker-Vβ-fragment to the TCR Cβ-domain and coexpression of the TCR Cα-domain capable of recruiting the natural CD3-complex for full and hence, native T-cell signaling. Here, we tested whether such a gp100(280-288)- or p53(264-272) tumor antigen-specific scTCR is still prone to mispairing with TCRα. In a human Jurkat-76 T-cell line lacking endogenous TCRs, surface expression and function of a scTCR could be reconstituted by any cointroduced TCRα-chain indicating mispairing to take place on a molecular basis. In contrast, transduction into human TCRα/β-positive T-cells revealed that mispairing is largely reduced. Competition experiments in Jurkat-76 confirmed the preference of dcTCR to selfpair and to spare scTCR. This also allowed for the generation of dc/scTCR-modified cytomegalovirus/tumor antigen-bispecific T-cells to augment T-cell activation in CMV-infected tumor patients. Residual mispairing was prevented by strenghtening the Vα-Li-Vβ-fragment through the design of a novel disulfide bond between a Vα- and a linker-resident residue close to Vβ. Multimer-stainings, and cytotoxicity-, IFNγ-secretion-, and CFSE-proliferation-assays, the latter towards dendritic cells endogenously processing RNA-electroporated gp100 antigen proved the absence of hybrid scTCR/TCRα-formation without impairing avidity of scTCR/Cα in T-cells. Moreover, a fragile cytomegalovirus pp65(495-503)-specific scTCR modified this way acquired enhanced cytotoxicity. Thus, optimized scTCR/Cα inhibits residual TCR mispairing to accomplish safe adoptive immunotherapy for bulk endogenous TCRα/β-positive T-cells

Comparison of serious inhaler technique errors made by device-naïve patients using three different dry powder inhalers: a randomised, crossover, open-label study

Background: Serious inhaler technique errors can impair drug delivery to the lungs. This randomised, crossover, open-label study evaluated the proportion of patients making predefined serious errors with Pulmojet compared with Diskus and Turbohaler dry powder inhalers. Methods: Patients ≥18 years old with asthma and/or COPD who were current users of an inhaler but naïve to the study devices were assigned to inhaler technique assessment on Pulmojet and either Diskus or Turbohaler in a randomised order. Patients inhaled through empty devices after reading the patient information leaflet. If serious errors potentially affecting dose delivery were recorded, they repeated the inhalations after watching a training video. Inhaler technique was assessed by a trained nurse observer and an electronic inhalation profile recorder. Results: Baseline patient characteristics were similar between randomisation arms for the Pulmojet-Diskus (n = 277) and Pulmojet-Turbohaler (n = 144) comparisons. Non-inferiority in the proportions of patients recording no nurse-observed serious errors was demonstrated for both Pulmojet versus Diskus, and Pulmojet versus Turbohaler; therefore, superiority was tested. Patients were significantly less likely to make ≥1 nurse-observed serious errors using Pulmojet compared with Diskus (odds ratio, 0.31; 95 % CI, 0.19–0.51) or Pulmojet compared with Turbohaler (0.23; 0.12–0.44) after reading the patient information leaflet with additional video instruction, if required. Conclusions These results suggest Pulmojet is easier to learn to use correctly than the Turbohaler or Diskus for current inhaler users switching to a new dry powder inhaler

Genome-Wide Transcriptional Reorganization Associated with Senescence-to-Immortality Switch during Human Hepatocellular Carcinogenesis

Cataloged from PDF version of article.Senescence is a permanent proliferation arrest in response to cell stress such as DNA damage. It contributes strongly to tissue aging and serves as a major barrier against tumor development. Most tumor cells are believed to bypass the senescence barrier (become "immortal") by inactivating growth control genes such as TP53 and CDKN2A. They also reactivate telomerase reverse transcriptase. Senescence-to-immortality transition is accompanied by major phenotypic and biochemical changes mediated by genome-wide transcriptional modifications. This appears to happen during hepatocellular carcinoma (HCC) development in patients with liver cirrhosis, however, the accompanying transcriptional changes are virtually unknown. We investigated genome-wide transcriptional changes related to the senescence-to-immortality switch during hepatocellular carcinogenesis. Initially, we performed transcriptome analysis of senescent and immortal clones of Huh7 HCC cell line, and identified genes with significant differential expression to establish a senescence-related gene list. Through the analysis of senescence-related gene expression in different liver tissues we showed that cirrhosis and HCC display expression patterns compatible with senescent and immortal phenotypes, respectively; dysplasia being a transitional state. Gene set enrichment analysis revealed that cirrhosis/senescence-associated genes were preferentially expressed in non-tumor tissues, less malignant tumors, and differentiated or senescent cells. In contrast, HCC/immortality genes were up-regulated in tumor tissues, or more malignant tumors and progenitor cells. In HCC tumors and immortal cells genes involved in DNA repair, cell cycle, telomere extension and branched chain amino acid metabolism were up-regulated, whereas genes involved in cell signaling, as well as in drug, lipid, retinoid and glycolytic metabolism were down-regulated. Based on these distinctive gene expression features we developed a 15-gene hepatocellular immortality signature test that discriminated HCC from cirrhosis with high accuracy. Our findings demonstrate that senescence bypass plays a central role in hepatocellular carcinogenesis engendering systematic changes in the transcription of genes regulating DNA repair, proliferation, differentiation and metabolism

Color Reflection Invariance and Monopole Condensation in QCD

We review the quantum instability of the Savvidy-Nielsen-Olesen (SNO) vacuum of the one-loop effective action of SU(2) QCD, and point out a critical defect in the calculation of the functional determinant of the gluon loop in the SNO effective action. We prove that the gauge invariance, in particular the color reflection invariance, exclude the unstable tachyonic modes from the gluon loop integral. This guarantees the stability of the magnetic condensation in QCD.Comment: 28 pages, 3 figures, JHEP styl

New solar axion search in CAST with 4^4He filling

The CERN Axion Solar Telescope (CAST) searches for $a\to\gamma$ conversion in the 9 T magnetic field of a refurbished LHC test magnet that can be directed toward the Sun. Two parallel magnet bores can be filled with helium of adjustable pressure to match the X-ray refractive mass $m_\gamma$ to the axion search mass $m_a$. After the vacuum phase (2003--2004), which is optimal for $m_a\lesssim0.02$ eV, we used $^4$He in 2005--2007 to cover the mass range of 0.02--0.39 eV and $^3$He in 2009--2011 to scan from 0.39--1.17 eV. After improving the detectors and shielding, we returned to $^4$He in 2012 to investigate a narrow $m_a$ range around 0.2 eV ("candidate setting" of our earlier search) and 0.39--0.42 eV, the upper axion mass range reachable with $^4$He, to "cross the axion line" for the KSVZ model. We have improved the limit on the axion-photon coupling to $g_{a\gamma}< 1.47\times10^{-10} {\rm GeV}^{-1}$ (95% C.L.), depending on the pressure settings. Since 2013, we have returned to vacuum and aim for a significant increase in sensitivity.Comment: CAST Collaboration 6 pages 3 figure