Comprehensive structural model of the mechanochemical cycle of a mitotic motor highlights molecular adaptations in the kinesin family

Kinesins are responsible for a wide variety of microtubule-based, ATP-dependent functions. Their motor domain drives these activities but the molecular adaptations that specify these diverse and essential cellular activities are poorly understood. It has been assumed that the first identified kinesin - the transport motor kinesin-1 – is the mechanistic paradigm for the entire superfamily, but accumulating evidence suggests that this is not the case. To address the deficits in our understanding of the molecular basis of functional divergence within the kinesin superfamily, we studied kinesin-5s, which are essential mitotic motors whose inhibition blocks cell division. Using cryo-electron microscopy and subnanometer resolution structure determination, we have visualised conformations of microtubule-bound human kinesin-5 motor domain at successive steps in its ATPase cycle. Following ATP hydrolysis, nucleotide-dependent conformational changes in the active site are allosterically propagated into rotations of the motor domain and uncurling of the drugbinding loop L5. In addition, the mechanical neck-linker element that is crucial for motor stepping undergoes discrete, ordered displacements. We also observed large reorientations of the motor N-terminus that indicate its importance for kinesin-5 function through control of neck-linker conformation. A kinesin-5 mutant lacking this N-terminus is enzymatically active, and ATP-dependent neck-linker movement and motility is defective although not ablated. All these aspects of kinesin-5 mechanochemistry are distinct from kinesin-1. Our findings directly demonstrate the regulatory role of the kinesin-5 N-terminus in collaboration with the motor’s structured neck-linker, and highlight the multiple adaptations within kinesin motor domains that tune their mechanochemistries according to distinct functional requirements

Supramolecular hierarchy among halogen and hydrogen bond donors in light-induced surface patterning

Halogen bonding, a noncovalent interaction possessing several unique features compared to the more familiar hydrogen bonding, is emerging as a powerful tool in functional materials design. Herein, we unambiguously show that one of these characteristic features, namely high directionality, renders halogen bonding the interaction of choice when developing azobenzene-containing supramolecular polymers for light-induced surface patterning. The study is conducted by using an extensive library of azobenzene molecules that differ only in terms of the bond-donor unit. We introduce a new tetrafluorophenol-containing azobenzene photoswitch capable of forming strong hydrogen bonds, and show that an iodoethynyl-containing azobenzene comes out on top of the supramolecular hierarchy to provide unprecedented photoinduced surface patterning efficiency. Specifically, the iodoethynyl motif seems highly promising in future development of polymeric optical and photoactive materials driven by halogen bonding

Concurrent Conditions and Human Listeriosis, England, 1999–2009

The epidemiology of listeriosis in England and Wales changed during 2001–2008; more patients >60 years of age had bacteremia than in previous years. To investigate these changes, we calculated risk for listeriosis by concurrent condition for non–pregnancy-associated listeriosis cases reported to the national surveillance system in England during 1999–2009. Conditions occurring with L. monocytogenes infection were coded according to the International Classification of Diseases, 10th Revision, and compared with appropriate hospital episode statistics inpatient denominator data to calculate incidence rates/million consultations. Malignancies (especially of the blood), kidney disease, liver disease, diabetes, alcoholism, and age >60 years were associated with an increased risk for listeriosis. Physicians should consider a diagnosis of listeriosis when treating patients who have concurrent conditions. Providing cancer patients, who accounted for one third of cases, with food safety information might help limit additional cases

Structural identification to improve bridge management

This paper presents results from static loads tests performed on the new Langensand Bridge built in Switzerland. A systematic study of over 1000 models subjected to three load cases identifies a set of 11 candidate models through static measurements. Predictions using the set of candidate models are homogenous and show an averaged discrepancy ranging of 4 to 7% from the displacement measurements. All candidate models have values for material proprieties that are close to expected values. This finding confirms that the behaviour of the structure conforms to the design expectations. Comparing the candidate model set to a design model that takes into account only main structural elements shows that the structure has approximately 30% reserve capacity with respect to a typical deflection risk scenario according to Swiss codes. The population of candidate models may be used to understand and predict the behaviour of the full bridge prior to its completion

CMS4SI Structural Identification Approach for Interpreting Measurements

The ageing of existing structures and new innovative designs are increasing the necessity for a greater understanding of structural behaviour. A better understanding would improve effectiveness of activities such as assessing reserve capacity, evaluating load increases and replacement decision making. Identification methodologies are needed to indicate the right behaviour using indirect measurements and behaviour models. This paper proposes a methodology that is able to accommodate multiple explanations while overcoming limitations of other SI approaches. The algorithm is called Candidate Model Search for System Identification (CMS4SI). Metrology guidelines are extended for use in the field of system identification while systematically including uncertainties and their correlations. The CMS4SI approach provides the necessary robustness and simplicity to support decisions related to the identification and understanding of structural behaviour. The approach is evaluated by full scale-testing of the Langensand Bridge. A critical aspect for meaningful identification is the uncertainty associated with model simplifications. The adaptation of clustering techniques and the use of radar plots allow for a convenient visualisation of results involving several parameters. Finally, models that are identified can be used to perform predictions of unmeasured behaviour, thereby supporting infrastructure management

Predicting Usefulness of Measuring Structures during Load Tests

This paper presents an approach for model identifiability that builds upon recent research into measurement data interpretation. The objective of this approach is to determine probabilistically to what degree the number of models able to explain a measured behaviour can be reduced in comparison to the initial solution space. The procedure is intended to be used prior to obtaining measurements from full-scale testing. The new methodology evaluates the probability of occurrence of two performance indices; the expected number of candidate models and the expected parameter range. It allows users, prior to taking measurements, to determine whether or not performing tests is likely to be useful. Since it does not require any intervention on the structure, this method may be used for a fraction of the cost required for full-scale testing. These features are illustrated through a case study, the Langensand Bridge (Switzerland). The methodology is the basis for a new generation of sensor placement techniques that determine to what extent particular sensor and load configurations are useful

HIV-1 selectively targets gut-homing CCR6+CD4+ T cells via mTOR-dependent mechanisms

Gut-associated lymphoid tissues are enriched in CCR6+ Th17-polarized CD4+ T cells that contribute to HIV-1 persistence during antiretroviral therapy (ART). This raises the need for Th17-targeted immunotherapies. In an effort to identify mechanisms governing HIV-1 permissiveness/persistence in gut-homing Th17 cells, we analyzed the transcriptome of CCR6+ versus CCR6- T cells exposed to the gut-homing inducer retinoic acid (RA) and performed functional validations in colon biopsies of HIV-infected individuals receiving ART (HIV+ART). Although both CCR6+ and CCR6- T cells acquired gut-homing markers upon RA exposure, the modulation of unique sets of genes coincided with preferential HIV-1 replication in RA-treated CCR6+ T cells. This molecular signature included the upregulation of HIV-dependency factors acting at entry/postentry levels, such as the CCR5 and PI3K/Akt/mTORC1 signaling pathways. Of note, mTOR expression/phosphorylation was distinctively induced by RA in CCR6+ T cells. Consistently, mTOR inhibitors counteracted the effect of RA on HIV replication in vitro and viral reactivation in CD4+ T cells from HIV+ART individuals via postentry mechanisms independent of CCR5. Finally, CCR6+ versus CCR6- T cells infiltrating the colons of HIV+ART individuals expressed unique molecular signatures, including higher levels of CCR5, integrin β7, and mTOR phosphorylation. Together, our results identify mTOR as a druggable key regulator of HIV permissiveness in gut-homing CCR6+ T cells

Estimation of modelling errors in structural system identification

This paper presents a method for measurement-system design through criteria related to model based structural identification. Using a multi-model approach and results from previous research carried out at EPFL, an improved algorithm is proposed. The algorithm accounts for various types of sensors having different accuracies and taking different kinds of measurements. The algorithm selects sensor types and locations that minimise the number of non-identified candidate models. The results show that the approach provides an alternative to selecting and placing sensors using engineering experience alone, and that a scientific approach based on sensor characteristics and modelling error is feasible. A single span composite bridge is used to illustrate the algorithm. It is shown that adding more than 9 sensors, from a possible set of 34, will not provide further useful information for structural identification

Multivisceral intestinal transplantation: Surgical pathology

We report the diagnostic surgical pathology of two children who underwent multivisceral abdominal transplantation and survived for 1 month and 6 months. There is little relevant literature, and diagnostic criteria for the various clinical possibilities are not established; this is made more complicated by the simultaneous occurrence of more than one process. We based our interpretations on conventional histology, augmented with immunohistology, including HLA staining that distinguished graft from host cells in situ. In some instances functional analysis of T cells propagated from the same biopsies was available and was used to corroborate morphological interpretations. A wide spectrum of changes was encountered. Graft-versus-host disease, a prime concern before surgery, was not seen. Rejection was severe in 1 patient, not present in the other, and both had evidence of lymphoproliferative disease, which was related to Epstein-Barr virus. Bacterial translocation through the gut wall was also a feature in both children. This paper documents and illustrates the various diagnostic possibilities.. © 1989 Informa UK Ltd All rights reserved: reproduction in whole or part not permitted