Electroporation enables the efficient mRNA delivery into the mouse zygotes and facilitates CRISPR/Cas9-based genome editing

Recent use of the CRISPR/Cas9 system has dramatically reduced the time required to produce mutant mice, but the involvement of a time-consuming microinjection step still hampers its application for high-throughput genetic analysis. Here we developed a simple, highly efficient and large-scale genome editing method, in which the RNAs for the CRISPR/Cas9 system are electroporated into zygotes rather than microinjected. We used this method to perform single-stranded oligodeoxynucleotide (ssODN)-mediated knock-in in mouse embryos. This method facilitates large-scale genetic analysis in the mouse

Development of active icosahedron and its application to virtual clay modeling

We have developed an active link mechanism for physical man-machine interaction. We report an active icosahedron consisting of intelligent cylinders and its application to virtual clay modeling. Intelligent pneumatic cylinders are newly developed to realize active link mechanisms. This cylinder aims at a novel cylinder in which various sensors and control devices are built. Active link mechanisms are highly integrated and enhanced by intelligent cylinders. A control system is built for the active icosahedron. In the control system, a key element is a control program implementing drawing of a virtual model on display and controlling of active links. Virtual clays are deformed by the program based on the apex positions converted from cylinder lengths. The active icosahedron realized dynamic interaction with virtual objects in PC, showing the potential of the devices as a haptic interface.</p

Nuclear-localized subtype of end-binding 1 protein regulates spindle organization in Arabidopsis

End-binding 1 (EB1) proteins are evolutionarily conserved plus-end-tracking proteins that localize to growing microtubule plus ends where they regulate microtubule dynamics and interactions with intracellular targets. Animal EB1 proteins have acidic C-terminal tails that might induce an autoinhibitory conformation. Although EB1 proteins with the same structural features occur in plants (EB1a and EB1b in Arabidopsis thaliana), a variant form (EB1c) is present that lacks the characteristic tail. We show that in Arabidopsis the tail region of EB1b, but not of EB1c, inhibits microtubule assembly in vitro. EB1a and EB1b form heterodimers with each other, but not with EB1c. Furthermore, the EB1 genes are expressed in various cell types of Arabidopsis, but the expression of EB1c is particularly strong in the meristematic cells where it is targeted to the nucleus by a nuclear localization signal in the C-terminal tail. Reduced expression of EB1c compromised the alignment of spindle and phragmoplast microtubules and caused frequent lagging of separating chromosomes at anaphase. Roots of the eb1c mutant were hypersensitive to a microtubule-disrupting drug and complete rescue of the mutant phenotype required the tail region of EB1c. These results suggest that a plant-specific EB1 subtype has evolved to function preferentially on the spindle microtubules by accumulating in the prophase nucleus

Endurance training facilitates myoglobin desaturation during muscle contraction in rat skeletal muscle.

At onset of muscle contraction, myoglobin (Mb) immediately releases its bound O2 to the mitochondria. Accordingly, intracellular O2 tension (PmbO2) markedly declines in order to increase muscle O2 uptake (mVO2). However, whether the change in PmbO2 during muscle contraction modulates mVO2 and whether the O2 release rate from Mb increases in endurance-trained muscles remain unclear. The purpose of this study was, therefore, to determine the effect of endurance training on O2 saturation of Mb (SmbO2) and PmbO2 kinetics during muscle contraction. Male Wistar rats were subjected to a 4-week swimming training (Tr group; 6 days per week, 30 min × 4 sets per day) with a weight load of 2% body mass. After the training period, deoxygenated Mb kinetics during muscle contraction were measured using near-infrared spectroscopy under hemoglobin-free medium perfusion. In the Tr group, the VmO2peak significantly increased by 32%. Although the PmbO2 during muscle contraction did not affect the increased mVO2 in endurance-trained muscle, the O2 release rate from Mb increased because of the increased Mb concentration and faster decremental rate in SmbO2 at the maximal twitch tension. These results suggest that the Mb dynamics during muscle contraction are contributing factors to faster VO2 kinetics in endurance-trained muscle

Pneumatic direct-drive stepping motor for robots

A new type of pneumatic stepping motor, named pneumatic nutation motor, was developed. This motor achieves stepping positioning of 720 steps/rotation without any electrical devices or sensors mounted on the servo mechanisms. This makes the motor possible to be used under hazardous conditions such as in water and in strong magnetic fields where conventional electromagnetic motors cannot be used. The motor torque is so big that the motor can be used as a direct motor. In this report, the driving principle and design of this motor are presented. Its characteristics are analyzed experimentally and theoretically. The motors were applied to a parallel linkage mechanism with six degrees of freedom. The mechanism shows that the pneumatic nutation motors can be used as a direct servo motor for robot mechanisms.</p

Intracellular oxygen tension limits muscle contraction-induced change in muscle oxygen consumption under hypoxic conditions during Hb-free perfusion.

Under acute hypoxic conditions, the muscle oxygen uptake (mV˙O2) during exercise is reduced by the restriction in oxygen-supplied volume to the mitochondria within the peripheral tissue. This suggests the existence of a factor restricting the mV˙O2 under hypoxic conditions at the peripheral tissue level. Therefore, this study set out to test the hypothesis that the restriction in mV˙O2 is regulated by the net decrease in intracellular oxygen tension equilibrated with myoglobin oxygen saturation (∆PmbO2) during muscle contraction under hypoxic conditions. The hindlimb of male Wistar rats (8 weeks old, n = 5) was perfused with hemoglobin-free Krebs-Henseleit buffer equilibrated with three different fractions of O2 gas: 95.0%O2, 71.3%O2, and 47.5%O2 The deoxygenated myoglobin (Mb) kinetics during muscle contraction were measured under each oxygen condition with a near-infrared spectroscopy. The ∆[deoxy-Mb] kinetics were converted to oxygen saturation of myoglobin (SmbO2), and the PmbO2 was then calculated based on the SmbO2 and the O2 dissociation curve of the Mb. The SmbO2 and PmbO2 at rest decreased with the decrease in O2 supply, and the muscle contraction caused a further decrease in SmbO2 and PmbO2 under all O2 conditions. The net increase in mV˙O2 from the muscle contraction (∆mV˙O2) gradually decreased as the ∆PmbO2 decreased during muscle contraction. The results of this study suggest that ΔPmbO2 is a key determinant of the ΔmV˙O2

Active link mechanisms for physical man-machine interaction

&lt;p&gt;In this paper, we propose a new type of haptic interface, named active link mechanism. This device realizes physical man-machine interaction (PMI) between machines and persons. Two prototypes were developed to demonstrate the potential of the active link mechanisms. Developed interface devices are an active tetrahedron and an active icosahedron. Nine-DOF micro spherical joints and pressure control pneumatic cylinders were developed to realize the active tetrahedron, while fifteen-DOF micro spherical joints and intelligent pneumatic cylinders were developed for the active icosahedron. The tetrahedron successfully realizes &quot;virtual touch&quot;; the operators feel actions, forces, and shapes of the virtual objects in PC and also move and deform them. Real time PMI is realized by building the developed devices into MSC.Visual-Nastran4D. MSC.VisuaI-Nastran4D is a mechanism analysis software, which can make motion analysis in real time. The active icosahedron also realized dynamic interaction with virtual objects in PC, showing the potential of the devices as a haptic interface.&lt;/p&gt;</p