Active anti-predator behaviour of red titi monkeys (Plecturocebus cupreus).

Due to their inconspicuous behaviour and colouration, it has been assumed that titi monkeys' main anti-predator behaviour is passive crypsis and hiding. So far, active predator mobbing has been documented only for black-fronted titi monkeys, Callicebus nigrifrons. Here we report for the first time mobbing behaviour of red titi monkeys, Plecturocebus cupreus (previously Callicebus cupreus), as reaction to an ocelot (Leopardus pardalis) and a Boa constrictor. We also report other active anti-predator behaviours, such as alarm calling and approaching, as reactions to tayras (Eira barbara) and raptors. Our observations provide additional evidence for sex differences in anti-predator behaviour, possibly related to the evolution and maintenance of social monogamy.

Transverse forces in skeletal muscle with massage-like loading in a rabbit model.

BACKGROUND: The objective of this study was to quantify the transverse forces in skeletal muscle subjected to constant compressive massage-like loading (MLL) following eccentric exercise (ECC). METHODS: Twenty-eight New Zealand White rabbits were used for this two-part study. For all testing, a customized electromechanical device was utilized to apply a constant compressive force MLL to the tibialis anterior (TA) muscle and the resultant transverse forces were quantified. The device consisted of two stepper motors that were positioned orthogonally to each other and connected to separate sliding tracks. A stainless steel cylindrical massage tip was mounted to a customized two-axis sensor consisting of two strain gauges with which forces along the two axes were measured. First, we determined the effects of tissue loading frequency and compression magnitude on transverse forces in the TA. Following a bout of ECC, sixteen rabbits were randomly assigned to a protocol with MLL frequency of 0.25 Hz or 0.5 Hz at a constant compressive force of 5 N or 10 N. Secondly, we utilized a protocol of 0.5 Hz, 10 N, 15 min MLL that was performed on 4 consecutive days commencing immediately post ECC (n = 6 animals) or 48 hours following ECC (n = 6 animals). Transverse forces were measured during all 4 MLL sessions for the entire 15 min duration for both the immediate and the delayed groups. RESULTS: Both frequency and magnitude of compressive force due to MLL showed an effect on the magnitude of transverse force (p < 0.05 for each parameter). Furthermore, MLL beginning immediately following ECC produced higher transverse forces than MLL delayed by 48 hours with an average 20% difference between the two MLL groups over the four day protocol. Forces were higher in the middle 5 minutes compared to the first 5 minutes for all MLL bouts in both groups. CONCLUSIONS: Frequency and magnitude of MLL and timing for delivery of MLL following ECC affect resultant transverse force values for exercised muscle. The application of our findings to humans receiving massage following exercise remains unknown at this time.

Rapid purification of recombinant histones.

The development of methods to assemble nucleosomes from recombinant histones decades ago has transformed chromatin research. Nevertheless, nucleosome reconstitution remains time consuming to this day, not least because the four individual histones must be purified first. Here, we present a streamlined purification protocol of recombinant histones from bacteria. We termed this method "rapid histone purification" (RHP) as it circumvents isolation of inclusion bodies and thereby cuts out the most time-consuming step of traditional purification protocols. Instead of inclusion body isolation, whole cell extracts are prepared under strongly denaturing conditions that directly solubilize inclusion bodies. By ion exchange chromatography, the histones are purified from the extracts. The protocol has been successfully applied to all four canonical Drosophila and human histones. RHP histones and histones that were purified from isolated inclusion bodies had similar purities. The different purification strategies also did not impact the quality of octamers reconstituted from these histones. We expect that the RHP protocol can be readily applied to the purification of canonical histones from other species as well as the numerous histone variants.

Effects of immediate vs. delayed massage-like loading on skeletal muscle viscoelastic properties following eccentric exercise.

BACKGROUND: This study compared immediate versus delayed massage-like compressive loading on skeletal muscle viscoelastic properties following eccentric exercise. METHODS: Eighteen rabbits were surgically instrumented with peroneal nerve cuffs for stimulation of the tibialis anterior muscle. Rabbits were randomly assigned to a massage loading protocol applied immediately post exercise (n=6), commencing 48h post exercise (n=6), or exercised no-massage control (n=6). Viscoelastic properties were evaluated in vivo by performing a stress-relaxation test pre- and post-exercise and daily pre- and post-massage for four consecutive days of massage loading. A quasi-linear viscoelastic approach modeled the instantaneous elastic response (AG0), fast (g1(p)) and slow (g2(p)) relaxation coefficients, and the corresponding relaxation time constants τ1 and τ2. FINDINGS: Exercise increased AG0 in all groups (P<0.05). After adjusting for the three multiple comparisons, recovery of AG0 was not significant in the immediate (P=0.021) or delayed (P=0.048) group compared to the control group following four days of massage. However, within-day (pre- to post-massage) analysis revealed a decrease in AG0 in both massage groups. Following exercise, g1(p) increased and g2(p) and τ1 decreased for all groups (P<0.05). Exercise had no effect on τ2 (P>0.05). After four days of massage, there was no significant recovery of the relaxation parameters for either massage loading group compared to the control group. INTERPRETATION: Our findings suggest that massage loading following eccentric exercise has a greater effect on reducing muscle stiffness, estimated by AG0, within-day rather than affecting recovery over multiple days. Massage loading also has little effect on the relaxation response.

A mechatronic system for quantitative application and assessment of massage-like actions in small animals.

Massage therapy has a long history and has been widely believed effective in restoring tissue function, relieving pain and stress, and promoting overall well-being. However, the application of massage-like actions and the efficacy of massage are largely based on anecdotal experiences that are difficult to define and measure. This leads to a somewhat limited evidence-based interface of massage therapy with modern medicine. In this study, we introduce a mechatronic device that delivers highly reproducible massage-like mechanical loads to the hind limbs of small animals (rats and rabbits), where various massage-like actions are quantified by the loading parameters (magnitude, frequency and duration) of the compressive and transverse forces on the subject tissues. The effect of massage is measured by the difference in passive viscoelastic properties of the subject tissues before and after mechanical loading, both obtained by the same device. Results show that this device is useful in identifying the loading parameters that are most conducive to a change in tissue mechanical properties, and can determine the range of loading parameters that result in sustained changes in tissue mechanical properties and function. This device presents the first step in our effort for quantifying the application of massage-like actions used clinically and measurement of their efficacy that can readily be combined with various quantitative measures (e.g., active mechanical properties and physiological assays) for determining the therapeutic and mechanistic effects of massage therapies.

Structure and subunit topology of the INO80 chromatin remodeler and its nucleosome complex.

INO80/SWR1 family chromatin remodelers are complexes composed of >15 subunits and molecular masses exceeding 1 MDa. Their important role in transcription and genome maintenance is exchanging the histone variants H2A and H2A.Z. We report the architecture of S. cerevisiae INO80 using an integrative approach of electron microscopy, crosslinking and mass spectrometry. INO80 has an embryo-shaped head-neck-body-foot architecture and shows dynamic open and closed conformations. We can assign an Rvb1/Rvb2 heterododecamer to the head in close contact with the Ino80 Snf2 domain, Ies2, and the Arp5 module at the neck. The high-affinity nucleosome-binding Nhp10 module localizes to the body, whereas the module that contains actin, Arp4, and Arp8 maps to the foot. Structural and biochemical analyses indicate that the nucleosome is bound at the concave surface near the neck, flanked by the Rvb1/2 and Arp8 modules. Our analysis establishes a structural and functional framework for this family of large remodelers.

Massage timing affects postexercise muscle recovery and inflammation in a rabbit model.

PURPOSE: This study compared the effect of immediate versus delayed massage-like compressive loading (MLL) on peak isometric torque recovery and inflammatory cell infiltration after eccentric exercise (EEX). METHODS: Eighteen skeletally mature New Zealand White rabbits were instrumented with peroneal nerve cuffs for the stimulation of hindlimb tibialis anterior muscles. After a bout of EEX, rabbits were randomly assigned to an MLL protocol (0.5 Hz, 10 N, 15 min) that started immediately post-EEX, 48 h post-EXX, or no-MLL control and performed for four consecutive days. A torque-angle (T-Θ) relationship was obtained for 21 joint angles pre- and post-EEX and after four consecutive days of MLL or no-MLL. Muscle wet weights and immunohistochemical sections were obtained after final treatments. RESULTS: EEX produced an average 51% ± 13% decrease in peak isometric torque output. The greatest peak torque recovery occurred with the immediate application of MLL. There were differences in torque recovery between immediate and delayed MLL (P = 0.0012), immediate MLL and control (P < 0.0001), and delayed MLL and control (P = 0.025). Immunohistochemical analysis showed 39.3% and 366.0% differences in the number of RPN3/57 and CD11b-positive cells between immediate (P = 0.71) and delayed MLL (P = 0.12). The area under the T-Θ curve showed a difference for immediate (P < 0.0001) and delayed (P = 0.0051) MLL as compared with control. Exercise produced an average 10° ± 0.2° rightward shift from preexercise peak isometric torque angle. Control, immediate MLL, and delayed MLL produced an average leftward angular shift from the postexercise angle (P = 0.28, P = 0.03, and P = 0.47, respectively). CONCLUSION: Post-EEX, immediate MLL was more beneficial than delayed MLL in restoring muscle function and in modulating inflammatory cell infiltration. These findings invite similar human studies to make definitive conclusions on optimal timing of massage-based therapies.

Dose-dependency of massage-like compressive loading on recovery of active muscle properties following eccentric exercise: rabbit study with clinical relevance.

BACKGROUND: Optimal strategies for massage and its use in athletes have not been conclusively demonstrated. PURPOSE/STUDY DESIGN: Effects of varying duration, frequency and magnitude of massage-like compressive loading (MLL) on recovery of skeletal muscle active properties (torque angle (T-Θ) relationship) following exercise-induced muscle injury were studied. METHODS: Twenty-four New Zealand White rabbits were surgically instrumented with bilateral peroneal nerve cuffs for stimulation of hindlimb tibialis anterior muscles. Following a bout of eccentric exercise (EEX), rabbits were randomly assigned to a MLL protocol of 0.25 or 0.5 Hz at 5 or 10 N for 15 or 30 min. T-Θ was obtained for 21 tibiotarsal joint angles pre- and post-EEX and post 4 consecutive days of MLL. Muscle wet weight and H&E sections were obtained following final treatments. RESULTS: EEX produced an average 61.8%±2.1 decrease in peak isometric torque output. Differences in torque recovery were found between magnitudes (5 and 10 N; p=0.004, n=12) and frequencies (0.25 and 0.5 Hz; p=0.012, n=12), but no difference for durations (15 and 30 min) with the 0.5 Hz, 10 N, 15 min protocol showing greatest recovery 4 days post-EEX. MLL muscle (n=12) wet weight was 3.22±0.18 g, while no MLL tissue (n=9) weighed 3.74±0.22 g (p=0.029). Histological analysis showed a difference in torn fibres between low-parameter and high-parameter MLL (6.5±1.04 vs 0.5±0.29 per 0.59 mm(2), p=0.005). CONCLUSIONS: Results showed a dose-response effect for magnitude and frequency of MLL on recovery of active muscle properties following EEX. Future studies will investigate underlying biological mechanisms for this enhanced recovery of muscle function.

In vivo passive mechanical properties of skeletal muscle improve with massage-like loading following eccentric exercise.

A quasi-linear viscoelasticity (QLV) model was used to study passive time-dependent responses of skeletal muscle to repeated massage-like compressive loading (MLL) following damaging eccentric exercise. Six skeletally mature rabbits were surgically instrumented with bilateral peroneal nerve cuffs for stimulation of the hindlimb tibialis anterior (TA) muscles. Following the eccentric exercise, rabbits were randomly assigned to a four-day MLL protocol mimicking deep effleurage (0.5 Hz, 10 N for 15 min or for 30 min). The contralateral hindlimb served as the exercised, no-MLL control for both MLL conditions. Viscoelastic properties of the muscle pre-exercise, post-exercise on Day 1, and pre- and post-MLL Day 1 through Day 4 were determined with ramp-and-hold tests. The instantaneous elastic response (AG(0)) increased following exercise (p<0.05) and decreased due to both the 15 min and 30 min four-day MLL protocols (p<0.05). Post-four days of MLL the normalized AG(0) decreased from post-exercise (Day 1, 248.5%) to the post-MLL (Day 4, 98.5%) (p<0.05), compared to the no-MLL group (Day 4, 222.0%) (p<0.05). Exercise and four-day MLL showed no acute or cumulative effects on the fast and slow relaxation coefficients (p>0.05). This is the first experimental evidence of the effect of both acute (daily) and cumulative changes in viscoelastic properties of intensely exercised muscle due to ex vivo MLL. It provides a starting point for correlating passive muscle properties with mechanical effects of manual therapies, and may shed light on design and optimization of massage protocols.

Dpb11/TopBP1 plays distinct roles in DNA replication, checkpoint response and homologous recombination.

DPB11/TopBP1 is an essential evolutionarily conserved gene involved in initiation of DNA replication and checkpoint signaling. Here, we show that Saccharomyces cerevisiae Dpb11 forms nuclear foci that localize to sites of DNA damage in G1, S and G2 phase, a recruitment that is conserved for its homologue TopBP1 in Gallus gallus. Damage-induced Dpb11 foci are distinct from Sld3 replication initiation foci. Further, Dpb11 foci are dependent on the checkpoint proteins Mec3 (9-1-1 complex) and Rad24, and require the C-terminal domain of Dpb11. Dpb11 foci are independent of the checkpoint kinases Mec1 and Tel1, and of the checkpoint mediator Rad9. In a site-directed mutagenesis screen, we identify a separation-of-function mutant, dpb11-PF, that is sensitive to DSB-inducing agents yet remains proficient for DNA replication and the S-phase checkpoint at the permissive temperature. The dpb11-PF mutant displays altered rates of heteroallelic and direct-repeat recombination, sensitivity to DSB-inducing drugs as well as delayed kinetics of mating-type switching with a defect in the DNA synthesis step thus implicating Dpb11 in homologous recombination. We conclude that Dpb11/TopBP1 plays distinct roles in replication, checkpoint response and recombination processes, thereby contributing to chromosomal stability.

Influence of Transmural Pressure and Cytoskeletal Structure on NF-κB Activation in Respiratory Epithelial Cells.

Respiratory epithelial cells are exposed to complex mechanical forces which are often modulated during pathological conditions such as Otitis Media and acute lung injury. The transduction of these mechanical forces into altered inflammatory signaling may play an important role in the persistence of disease conditions and inflammation. In this study, we investigated how static and oscillatory pressures altered the activation of NF-κB inflammatory pathways and how changes in the actin cytoskeleton influenced the mechanotransduction of pressure into NF-κB activation. An in vitro system was used to apply static and oscillatory pressures to alveolar epithelial cells cultured at an air-liquid interface. Latrunculin A and Jasplakinolide were used to alter the cytoskeleton and tight-junction structure and ELISA was used to monitor activation of NF-κB. Results indicate that both static and oscillatory pressures can activate NF-κB and that this activation is magnitude-dependent at low oscillation frequencies only. Jasplakinolide treated cells did not exhibit significant changes in normalized NF-κB activation compared to unloaded controls while Latrunculin treated cells exhibited increases in normalized NF-κB activation only at low frequency or static pressures. These results indicate that altering the actin cytoskeleton may be a useful way to mitigate the mechanotransduction of pressure forces into inflammatory signaling.