Monolayer cultures of perikarya isolated from postnatal rat cerebellum

Summary Viable cerebellar perikarya of mixed cell type obtained from 7–9-day-old rats were maintained in monolayer culture for up to 12 days. During this time extensive neurite formation and outgrowth occurred. The large majority of the cells developing in culture were tentatively identified as granule neurons. This identification was based on the large number isolated from the starting tissue, and the cells general morphological features in culture such as perikaryal and nuclear size, the bipolar nature of neurite extension, and their migratory behaviour.D.N. Currie was supported by a grant from Hoechst Pharmaceuticals, Ld, U.K.J.C. was supported by an S.R.C. Grant to S.P.R. RoseThe authors would like to thank Kathleen Tear for excellent technical assistance and members of the Brain Research Group for helpful discussions.     

Signs, toy models, and the a priori: from Helmholtz to Wittgenstein

The Marburg neo-Kantians argue that Hermann von Helmholtz’s empiricist account of the a priori does not account for certain knowledge, since it is based on a psychological phenomenon, trust in the regularities of nature. They argue that Helmholtz’s account raises the ‘problem of validity’ (Gültigkeitsproblem): how to establish a warranted claim that observed regularities are based on actual relations. I reconstruct Heinrich Hertz’s and Ludwig Wittgenstein’s Bild theoretic answer to the problem of validity: that scientists and philosophers can depict the necessary a priori constraints on states of affairs in a given system, and can establish whether these relations are actual relations in nature. The analysis of necessity within a system is a lasting contribution of the Bild theory. However, Hertz and Wittgenstein argue that the logical and mathematical sentences of a Bild are rules, tools for constructing relations, and the rules themselves are meaningless outside the theory. Carnap revises the argument for validity by attempting to give semantic rules for translation between frameworks. Russell and Quine object that pragmatics better accounts for the role of a priori reasoning in translating between frameworks. The conclusion of the tale, then, is a partial vindication of Helmholtz’s original account.     

Natural engineering principles of electron tunnelling in biological oxidation-reduction

We have surveyed proteins with known atomic structure whose function involves electron transfer; in these, electrons can travel up to 14 A between redox centres through the protein medium. Transfer over longer distances always involves a chain of cofactors. This redox centre proximity alone is sufficient to allow tunnelling of electrons at rates far faster than the substrate redox reactions it supports. Consequently, there has been no necessity for proteins to evolve optimized routes between redox centres. Instead, simple geometry enables rapid tunnelling to high-energy intermediate states. This greatly simplifies any analysis of redox protein mechanisms and challenges the need to postulate mechanisms of superexchange through redox centres or the maintenance of charge neutrality when investigating electron-transfer reactions. Such tunnelling also allows sequential electron transfer in catalytic sites to surmount radical transition states without involving the movement of hydride ions, as is generally assumed. The 14 A or less spacing of redox centres provides highly robust engineering for electron transfer, and may reflect selection against designs that have proved more vulnerable to mutations during the course of evolution.     

Observation of coherent optical information storage in an atomic medium using halted light pulses

Electromagnetically induced transparency is a quantum interference effect that permits the propagation of light through an otherwise opaque atomic medium; a 'coupling' laser is used to create the interference necessary to allow the transmission of resonant pulses from a 'probe' laser. This technique has been used to slow and spatially compress light pulses by seven orders of magnitude, resulting in their complete localization and containment within an atomic cloud. Here we use electromagnetically induced transparency to bring laser pulses to a complete stop in a magnetically trapped, cold cloud of sodium atoms. Within the spatially localized pulse region, the atoms are in a superposition state determined by the amplitudes and phases of the coupling and probe laser fields. Upon sudden turn-off of the coupling laser, the compressed probe pulse is effectively stopped; coherent information initially contained in the laser fields is 'frozen' in the atomic medium for up to 1 ms. The coupling laser is turned back on at a later time and the probe pulse is regenerated: the stored coherence is read out and transferred back into the radiation field. We present a theoretical model that reveals that the system is self-adjusting to minimize dissipative loss during the 'read' and 'write' operations. We anticipate applications of this phenomenon for quantum information processing.     

Dynamics of Drosophila embryonic patterning network perturbed in space and time using microfluidics

Biochemical networks are perturbed both by fluctuations in environmental conditions and genetic variation. These perturbations must be compensated for, especially when they occur during embryonic pattern formation. Complex chemical reaction networks displaying spatiotemporal dynamics have been controlled and understood by perturbing their environment in space and time. Here, we apply this approach using microfluidics to investigate the robust network in Drosophila melanogaster that compensates for variation in the Bicoid morphogen gradient. We show that the compensation system can counteract the effects of extremely unnatural environmental conditions--a temperature step--in which the anterior and posterior halves of the embryo are developing at different temperatures and thus at different rates. Embryonic patterning was normal under this condition, suggesting that a simple reciprocal gradient system is not the mechanism of compensation. Time-specific reversals of the temperature step narrowed down the critical period for compensation to between 65 and 100 min after onset of embryonic development. The microfluidic technology used here may prove useful to future studies, as it allows spatial and temporal regulation of embryonic development.     

The effects of primary afferent depolarization on excitability fluctuations of Ia terminals within the motor nucleus

Résumé Sur la préparation de chat spinal aigue, les réponses antidromiques des fibres afférentes la dans un noyeau de motoneurones montrent des fluctuations considérables réduites par des volées afférentes qui provoquent une dépolarisation des afférences primaires. Les réponses provoquées par activation directe des motoneurones sont très stables, et leurs fluctuations non affectées par la stimulation afférente. Ces faits suggèrent que la variabilité du réflexe monosynaptique est principalement due aux fluctuations du potentiel de membrane des parties terminales des fibres afférentes Ia.     

Negation scope detection with a conditional random field model

Identifying negation cues and their scope in a text is an important subtask of information extrac-tion that can benefit other natural language processing tasks, including but not limited to medical da-ta mining, relation extraction, question answering and sentiment analysis.The tasks of negation cue and negation scope detection can be treated as sequence labelling problems.In this paper, a system is presented having two components: negation cue detection and negation scope detection.In the first phase, a conditional random field ( CRF) model is trained to detect the negation cues using a lexicon of negation words and some lexical and contextual features.Then, another CRF model is trained to detect the scope of each negation cue identified in the first phase, using basic lexical and contextual features.These two models are trained and tested using the dataset distributed within the*Sem Shared Task 2012 on resolving the scope and focus of negation.Experimental results show that the system outperformed all the systems submitted to this shared task.     

Reversible redox energy coupling in electron transfer chains

Reversibility is a common theme in respiratory and photosynthetic systems that couple electron transfer with a transmembrane proton gradient driving ATP production. This includes the intensely studied cytochrome bc1, which catalyses electron transfer between quinone and cytochrome c. To understand how efficient reversible energy coupling works, here we have progressively inactivated individual cofactors comprising cytochrome bc1. We have resolved millisecond reversibility in all electron-tunnelling steps and coupled proton exchanges, including charge-separating hydroquinone-quinone catalysis at the Q(o) site, which shows that redox equilibria are relevant on a catalytic timescale. Such rapid reversibility renders popular models based on a semiquinone in Q(o) site catalysis prone to short-circuit failure. Two mechanisms allow reversible function and safely relegate short-circuits to long-distance electron tunnelling on a timescale of seconds: conformational gating of semiquinone for both forward and reverse electron transfer, or concerted two-electron quinone redox chemistry that avoids the semiquinone intermediate altogether.     

Mutations in the gene encoding the 3'-5' DNA exonuclease TREX1 are associated with systemic lupus erythematosus

TREX1 acts in concert with the SET complex in granzyme A-mediated apoptosis, and mutations in TREX1 cause Aicardi-Goutières syndrome and familial chilblain lupus. Here, we report monoallelic frameshift or missense mutations and one 3' UTR variant of TREX1 present in 9/417 individuals with systemic lupus erythematosus but absent in 1,712 controls (P = 4.1 x 10(-7)). We demonstrate that two mutant TREX1 alleles alter subcellular targeting. Our findings implicate TREX1 in the pathogenesis of SLE.