Distinct Pause Research Articles

Roaming Pauses in Cormac McCarthy’s The Road

ABSTRACT This article discusses a textual feature in Cormac McCarthy’s novel The Road (2006) that has hitherto been misunderstood by the reception, namely the spaced ellipses between several paragraphs which are considered by notable McCarthy scholars to signal particularly distinct pauses in the textual flow of the novel. The article demonstrates that this is a misperception with significant consequences for the interpretation of the story’s rhythm. Drawing on textual scholars such as Jerome McGann, Bill Brown, John Bryant, and John B. Thompson, the article finally discusses how neutral bibliographical codes occasionally express their own intentions that are in conflict with those of the author.

Pauses in cholinergic interneuron firing exert an inhibitory control on striatal output in vivo.

The cholinergic interneurons (CINs) of the striatum are crucial for normal motor and behavioral functions of the basal ganglia. Striatal CINs exhibit tonic firing punctuated by distinct pauses. Pauses occur in response to motivationally significant events, but their function is unknown. Here we investigated the effects of pauses in CIN firing on spiny projection neurons (SPNs) - the output neurons of the striatum - using in vivo whole cell and juxtacellular recordings in mice. We found that optogenetically-induced pauses in CIN firing inhibited subthreshold membrane potential activity and decreased firing of SPNs. During pauses, SPN membrane potential fluctuations became more hyperpolarized and UP state durations became shorter. In addition, short-term plasticity of corticostriatal inputs was decreased during pauses. Our results indicate that, in vivo, the net effect of the pause in CIN firing on SPNs activity is inhibition and provide a novel mechanism for cholinergic control of striatal output.

Independent and Stochastic Action of DNA Polymerases in the Replisome

It has been assumed that DNA synthesis by the leading- and lagging-strand polymerases in the replisome must be coordinated to avoid the formation of significant gaps in the nascent strands. Using real-time single-molecule analysis, we establish thatleading- and lagging-strand DNA polymerases function independently within a single replisome. Although average rates of DNA synthesis on leading and lagging strands are similar, individual trajectories of both DNA polymerases display stochastically switchable rates of synthesis interspersed with distinct pauses. DNA unwinding by the replicative helicase may continue during such pauses, but a self-governing mechanism, where helicase speed is reduced by ∼80%, permits recoupling of polymerase to helicase. These features imply a more dynamic, kinetically discontinuous replication process, wherein contacts within the replisome are continually broken and reformed. We conclude that the stochastic behavior of replisome components ensures complete DNA duplication without requiring coordination of leading- and lagging-strand synthesis. PAPERCLIP.

Torque Generation of Enterococcus hirae V-ATPase

V-ATPase (V(o)V1) converts the chemical free energy of ATP into an ion-motive force across the cell membrane via mechanical rotation. This energy conversion requires proper interactions between the rotor and stator in V(o)V1 for tight coupling among chemical reaction, torque generation, and ion transport. We developed an Escherichia coli expression system for Enterococcus hirae V(o)V1 (EhV(o)V1) and established a single-molecule rotation assay to measure the torque generated. Recombinant and native EhV(o)V1 exhibited almost identical dependence of ATP hydrolysis activity on sodium ion and ATP concentrations, indicating their functional equivalence. In a single-molecule rotation assay with a low load probe at high ATP concentration, EhV(o)V1 only showed the "clear" state without apparent backward steps, whereas EhV1 showed two states, "clear" and "unclear." Furthermore, EhV(o)V1 showed slower rotation than EhV1 without the three distinct pauses separated by 120° that were observed in EhV1. When using a large probe, EhV(o)V1 showed faster rotation than EhV1, and the torque of EhV(o)V1 estimated from the continuous rotation was nearly double that of EhV1. On the other hand, stepping torque of EhV1 in the clear state was comparable with that of EhV(o)V1. These results indicate that rotor-stator interactions of the V(o) moiety and/or sodium ion transport limit the rotation driven by the V1 moiety, and the rotor-stator interactions in EhV(o)V1 are stabilized by two peripheral stalks to generate a larger torque than that of isolated EhV1. However, the torque value was substantially lower than that of other rotary ATPases, implying the low energy conversion efficiency of EhV(o)V1.

Multiphasic modulation of cholinergic interneurons by nigrostriatal afferents.

The motor and learning functions of the striatum are critically dependent on synaptic transmission from midbrain dopamine neurons and striatal cholinergic interneurons (CINs). Both neural populations alter their discharge in vivo in response to salient sensory stimuli, albeit in opposite directions. Whereas midbrain dopamine neurons respond to salient stimuli with a brief burst of activity, CINs exhibit a distinct pause in firing that is often followed by a period of increased excitability. Although this "pause-rebound" sensory response requires dopaminergic signaling, the precise mechanisms underlying the modulation of CIN firing by dopaminergic afferents remain unclear. Here, we show that phasic activation of nigrostriatal afferents in a mouse striatal slice preparation is sufficient to evoke a pause-rebound response in CINs. Using a combination of optogenetic, electrophysiological, and pharmacological approaches, we demonstrate that synaptically released dopamine inhibits CINs through type 2 dopamine receptors, while another unidentified transmitter mediates the delayed excitation. These findings imply that, in addition to their direct effects on striatal projection neurons, midbrain dopamine neurons indirectly modulate striatal output by dynamically controlling cholinergic tone. In addition, our data suggest that phasic dopaminergic activity may directly participate in the characteristic pause-rebound sensory response that CINs exhibit in vivo in response to salient and conditioned stimuli.

Single-Molecule Tracking of Collagenase on Native Type I Collagen Fibrils Reveals Degradation Mechanism

Collagen, the most abundant human protein, is the principal component of the extracellular matrix and plays important roles in maintaining tissue and organ integrity. Highly resistant to proteolysis, fibrillar collagen is degraded by specific matrix metalloproteases (MMPs). Degradation of fibrillar collagen underlies processes including tissue remodeling, wound healing, and cancer metastasis. However, the mechanism of native collagen fibril degradation remains poorly understood. Here we present the results of high-resolution tracking of individual MMPs degrading type I collagen fibrils. MMP1 exhibits cleavage-dependent biased and hindered diffusion but spends 90% ± 3% of the time in one of at least two distinct pause states. One class of exponentially distributed pauses (class I pauses) occurs randomly along the fibril, whereas a second class of pauses (class II pauses) exhibits multistep escape kinetics and occurs periodically at intervals of 1.3 ± 0.2 μm and 1.5 ± 0.2 μm along the fibril. After these class II pauses, MMP1 moved faster and farther in one direction along the fibril, indicative of biased motion associated with cleavage. Simulations indicate that 5% ± 2% of the class II pauses result in the initiation of processive collagen degradation, which continues for bursts of 15 ± 4 consecutive cleavage events. These findings provide a mechanistic paradigm for type I collagen degradation by MMP1 and establish a general approach to investigate MMP-fibrillar collagen interactions. More generally, this work demonstrates the fundamental role of enzyme-substrate interactions including binding and motion in determining the activity of an enzyme on an extended substrate.

Paediatric training on breastfeeding

Paediatric training on breastfeeding

Separation and estimation of muscle spindle and tension receptor populations by vibration of the biceps muscle in the frog.

Frog spinal cord reflex behaviors have been used to test the idea of spinal primitives. We have suggested a significant role for proprioception in regulation of primitives. However the in vivo behavior of spindle and golgi tendon receptors in frogs in response to vibration are not well described and the proportions of these proprioceptors are not established. In this study, we examine the selectivity of muscle vibration in the spinal frog. The aim of the study was (1) to examine how hindlimb muscle spindles and GTO receptors are activated by muscle vibration and (2) to estimate the relative numbers of GTO receptors and spindle afferents in a selected muscle, for comparison with the mammal. Single muscle afferents from the biceps muscle were identified in the dorsal roots. These were tested in response to biceps vibration, intramuscular stimulation and biceps nerve stimulation. Biceps units were categorized into two types: First, spindle afferents which had a high conduction velocity (approximately 20-30 m/s), responded reliably (were entrained 1:1) to muscle vibration, and exhibited distinct pauses to shortening muscle contractions. Second, golgi tendon organ afferents, which had a lower conduction velocity (approximately 10-20 m/s), responded less reliably to muscle vibration at physiologic muscle lengths, but responded more reliably at extended lengths or with background muscle contraction, and exhibited distinct bursts to shortening muscle contractions. Vibration responses of these units were tested with and without muscle curarization. Ensemble (suction electrode) recordings from the dorsal roots were used to provide rough estimates of the proportions of the two muscle afferent types.

Strychnine- and bicuculline- induced changes in the firing pattern of motoneurones during in vitro fictive locomotion reveal a possible N -methyl-D-aspartic acid (NMDA)-mediated suppression of motor discharge in wallaby (Macropus eugenii) pouch young

Inhibitory pathways in the spinal cord play an important role in establishing the pattern of motor discharge. In the wallaby spinal cord preparation, disruption of glycinergic and gamma-amino butyric acid (GABA)ergic neurotransmission abolished the alternation between antagonistic motor pools during fictive locomotion. A new pattern of motor discharge also appeared when both glycine and GABAA receptors were blocked simultaneously. This discharge pattern was biphasic, characterized by a distinct pause between two bursts of motoneurone firing during each cycle of motor activity. Whole cell patch recordings showed that the second burst of motor discharge was not caused by a separate inward current at a delayed time course. Furthermore, local injection of an N -methyl-D-aspartic acid (NMDA) specific antagonist converted the biphasic discharge to a continuous burst pattern. The result suggests an NMDA-mediated mechanism, which causes a suppression of motoneurone firing when glutamate release from interneurones is enhanced in the absence of glycinergic and GABAergic inhibition.

Egg deposition behavior in the haplodiploid sawfly Athalia rosae ruficornis Jakovlev (Hymenoptera: Symphyta: Tenthredinidae)

The egg deposition behavior of the turnip sawfly, Athalia rosae (Hymenoptera: Symphyta), is described. Both unmated and mated females lay eggs individually inside of fresh young leaves of cruciferous plants. During an oviposition event, females exhibit a distinct pause in abdominal contractions just before the actual egg deposition act. Unmated females show a longer pause (11.31 s on average) than mated females (4.38 s on overall average). By employing an eye color mutation, the sex of the eggs laid by females was ascertained. Females mated once lay mostly fertilized (diploid female) eggs initially but begin to lay a considerable number of unfertilized (haploid male) eggs later in life. The laying of an unfertilized egg is associated with a longer pause (6.98 s on average) than the laying of a fertilized egg (3.76 s on average). These results are in contrast to previous reports on apocritan Hymenoptera, where the presence of a pause or a longer pause during oviposition was associated with the deposition of fertilized eggs rather than unfertilized eggs. The possibility that mated Athalia rosae females control fertilization and its implications for sex allocation strategies are discussed.

Potential Problems Associated with Use of Speech Recognition Products

Commercial speech recognition products are being used increasingly as alternate input devices for computers, particularly by persons with physical disabilities. These discrete speech dictation systems require the user to insert brief but distinct pauses after each spoken word. Anecdotal evidence suggests that some persons using these products experience moderate to severe problems with their voices, such as hoarseness, sore throats, and even complete loss of voice. This preliminary study, which includes data gathered from survey dissemination and clinical studies, indicates that persons with cumulative trauma disorder may be the most susceptible to these voice problems. Also, we hypotihiesized that in using these discrete speech recognition systems, there may be a tendency to maintain constant pitch, volume, and inflection, keeping the vocal tract musculature in a fixed position. Maintaining this fixed position for extended periods may result in muscle fatigue and, eventually, injury to the laryngeal musculature. Further studies are needed, however, to investigate the effects suggested here. In the meantime, we recommend that users become informed about the unnatural speech patterns used with discrete speech recognition systems; learn to use good vocal hygiene, such as performing warm-up and cool-down voice exercises; and use alternate methods of input along with the speech recognition product.

Effects of 3'-deoxynucleoside 5'-triphosphate concentrations on chain termination by nucleoside analogs during human immunodeficiency virus type 1 reverse transcription of minus-strand strong-stop DNA

We have compared the effects of nucleoside analogs in quiescent and phytohemagglutinin (PHA)-stimulated peripheral blood mononuclear cells (PBMC) exposed to human immunodeficiency virus type 1 (HIV-1) with those of their triphosphorylated derivatives in cell-free HIV-1 reverse transcription assays. We observed a substantial decrease in synthesis of early minus-strand proviral DNA products in HIV-1-infected, quiescent PBMC exposed to each of 3'-azido-3'-deoxythymidine (AZT), 2',3'-dideoxyinosine (ddI), and 2',3'-dideoxy-3'-thiacytidine (3TC), in comparison with nontreated, infected controls. In contrast, no such diminution was observed when PHA-stimulated, HIV-1-infected PBMC were treated with the same drugs. This result was attributed to previously reported findings that PHA-stimulated PBMC possessed larger deoxynucleoside triphosphate (dNTP) pools than quiescent cells did. To further investigate this subject, a cell-free HIV-1 reverse transcription reaction involving HIV-1 RNA genomic template, recombinant purified HIV-1 reverse transcriptase, all four dNTPs and either tRNA3Lys or a deoxyoligonucleotide as primer was used to monitor chain termi-nation mediated by 2',3'-dideoxynucleoside triphosphates (ddNTPs) during synthesis of minus-strand strong-stop DNA. Augmented chain termination was observed with decreasing concentrations of both ddNTP and dNTP when the ratio of dNTP to ddNTP was fixed. We also found that both the number and strength of reverse transcription pause sites were increased at low concentrations of dNTPs and when a deoxyoligonucleotide primer was used in place of the cognate primer, tRNA3Lys. Preferential incorporation of ddATP was observed dur-ing reverse transcription opposite a distinct pause site in a short synthetic RNA template. These results con-firm the notion that the antiviral activities of ddNTP are dependent on both cellular dNTP pools and the state of cellular activation. Pausing of HIV-1 reverse transcriptase during reverse transcription, altered by dNTP concentrations, may be a mechanism that controls the position and extent of incorporation of nucleoside analogs.

Some interruption criteria for short high-frequency vacuum arcs

If the contacts of a vacuum interrupter open shortly before a current zero, the transient recovery voltage (TRV) can cause a reignition and reestablish the arc. When the current in a diffuse vacuum arc passes through zero, there is a distinct pause before the TRV builds up (approximately 40 ns for copper). During this pause the gap carries conduction current only with an ion component which depends on dI/dt, varying between 3 A for dI/dt=60 A/ mu s and 60 A for dI/dt=1235 A/ mu s. The ion current subsequently decays in tens or hundreds of nanoseconds. It can be distinguished from the displacement current at this time by varying dV/dt, keeping the other parameters constant. Among the interruption criteria for short high-frequency vacuum arcs, dI/dt prior to current zero and initial dV/dt are the most important. High values of dI/dt are more likely to precipitate reignitions, but breakdowns can occur after lower dI/dt's if the gap has been subjected to a high current for a relatively long time (>100 mu s). >

Mapping and characterization of transcriptional pause sites in the early genetic region of bacteriophage T7

During transcription of DNA templates in vitro, Escherichia coli RNA polymerase pauses at certain sequences before resuming elongation. Previous studies have established that some pausing events are brought about by the formation of RNA hairpin structures in the nascent transcript; however, it is not known whether this is an invariant and causal relationship. We have mapped and characterized almost 200 distinct pause sites located within the early region of bacteriophage T7 DNA using a collection of T7 deletion mutant DNAs and taking advantage of a procedure that permits synchronous transcription from the T7 A1 promoter. The pausing pattern is sensitive both to the overall concentration of nucleotide substrates and to the relative concentrations of the four nucleotides. The apparent K S value for a particular nucleoside triphosphate can vary over a 500-fold range depending on the nucleotide sequence, and pausing at some sites can be induced by modest reductions in substrate concentration. However, pausing is not solely a consequence of substrate limitation. Pausing at certain sites is caused by some feature of the template or of the transcript itself. Substitution of inosine triphosphate (ITP) for GTP during transcription strongly affects the pattern and strength of pausing events, suggesting that base-pairing interactions involving the RNA strand are important for some pausing events. Other pauses are determined by sequences downstream from the elongation site that have not yet been transcribed, and pausing at these sites is generally insensitive to substitution of IMP for GMP in the nascent transcript. Pausing at one particular site on T7 DNA is strongly enhanced by the presence of E. coli gene nusA protein. These results confirm that there are multiple classes of sites that lead to transcriptional pausing, and provide a collection of sites for further study. Using selected pause sites in the early region of T7 DNA, we have tried to evaluate the possible roles of primary sequence, base composition and secondary structure in pausing. Computer analysis was used to compare primary sequences and potential RNA hairpin structures in transcripts for pauses known to share similar biochemical properties. We see no correlation of pause sites with regions of particular base composition or with specific primary sequences. While some pauses are correlated with the potential to form stable RNA hairpins just upstream from the growing point of the RNA chain, there is not a strict one-to-one relationship between predicted RNA hairpins and the location of pause sites. Since at least one pause site seems to be correlated with an RNA hairpin involving non-Watson-Crick base-pairs (adenine-guanine pairs), this lack of correspondence must be partly due to the inadequacies of current rules for predicting RNA secondary structures. However, elements other than RNA secondary structure must also modulate the process of RNA chain elongation.

Characterization of Giemsa dark- and light-band DNA

Using synchronized cultures of V79-8 Chinese hamster lung fibroblasts, we either alternately labeled early- and late-replicating DNA, or substituted one of these with bromodeoxyuridine to separate them in CsCI density gradients or to identify the bromodeoxyuridine-containing chromosome bands by fluorescence microscopy. The Giemsa light R bands were shown to replicate in the first half of S phase, and the dark G bands were shown to replicate in the last half of S phase. S phase was bimodal, with a distinct pause in the rate of DNA synthesis that separated the period of R-band DNA synthesis from that of G-band DNA synthesis. G-band DNA was found to be 3.2% richer in AT than R-band DNA. Surprisingly, G- and R-band DNA appeared equally transcriptionally active in that alternate labels in chromatin were digested with the same kinetics by DNAase I, and in reassociation experiments, total poly(A)+ RNA drove nick-translated G- and R-band DNA probes similarly. G- and R-band DNA also reassociated with identical kinetics, demonstrating that they contain equal proportions of all kinetic-complexity classes of sequences.

Importance of Prolonged Compression during Cardiopulmonary Resuscitation in Man

Standards for external cardiac compression include compression of the chest for 50 per cent of cycle time and a compression rate of 60 per minute.1 Warnings are given against "quick jabs." Animal studies do, in fact, suggest that the duration of prolonged compression is important in effective resuscitation.2 To achieve a duration of 50 per cent during manual external cardiac compression, there must be a distinct pause at maximal compression. Application of this principle to man appears to have been slow, perhaps because of doubts about the applicability of animal data to man. In view of the obvious importance of . . .

XXI: O.F. (Norman) AOI and AVOI, and English Ahoy

(Other words treated here are Old French (Norman) a voiz, avois, voi, aei; English avoy, hoy, ahey, hey; Provençal aei; Portuguese eoi; Italian voi.) THE recent systematic analysis of the Chanson de Roland with reference to the exact positions of the word aoi in the Oxford manuscript seems definitely to reveal that that puzzling word is almost exclusively used only when there is some sort of shift in the narrative or a distinct pause or a break in the story.

STUDIES IN THE PHYSIOLOGY OF THE NERVOUS SYSTEM. IX.: REFLEX TERMINAL PHENOMENA—REBOUND—RHYTHMIC REBOUND AND MOVEMENTS OF PROGRESSION

STUDIES IN THE PHYSIOLOGY OF THE NERVOUS SYSTEM. IX.: REFLEX TERMINAL PHENOMENA—REBOUND—RHYTHMIC REBOUND AND MOVEMENTS OF PROGRESSION