Development of quadrupole susceptibility automatic calculator in sum frequency generation spectroscopy and application to methyl C-H vibrations.

Sum frequency generation (SFG) spectroscopy has been established as a powerful interface probe technique based on the electric dipole approximation, while possible signals of quadrupole and bulk origin have also been known for a long time. In this work, we developed a computational tool, namely, Qsac (quadrupole susceptibility automatic calculator), to evaluate the comprehensive contributions of the dipole/quadrupole and interface/bulk in the arbitrary vibrational bands of SFG spectra. The calculations of relevant susceptibility terms are performed on the basis of the theory of energy representation using quantum chemical calculation and molecular dynamics simulation, which allows for semi-quantitative comparison among these terms on the same footing. We applied the Qsac to the methyl C-H stretching bands of organic molecules and found a general trend that the weak asymmetric bands are more sensitive to the bulk contribution than the symmetric ones. The phases of interface and bulk terms tend to cancel in the asymmetric band, which results in the reduced band intensity in the SFG spectra.

Carboxylic Acid Functionalized Spin-Crossover Iron(II) Grids for Tunable Switching and Hybrid Electrode Fabrication.

Two carboxyl-substituted iron(II) grids, one protonated, [Fe4(HL)4](BF4)4·4MeCN·AcOEt (1), and the other deprotonated, [Fe4(L)4]·DMSO·EtOH (2), where H2L = 4-{4,5-bis[6-(3,5-dimethylpyrazol-1-yl)pyrid-2-yl]-1 H-imidazol-2-yl}benzoic acid, were synthesized. Single-crystal X-ray structure analyses reveal that both complexes have a tetranuclear [2 × 2] grid structure. 1 formed one-dimensional chains through intermolecular hydrogen bonds between the carboxylic acid units of neighboring grids, while 2 formed two-dimensional layers stabilized by π-π-stacking interactions. 1 showed spin transition between the 3HS-1LS and 1.5HS-2.5LS states around 200 K, while 2 showed spin-crossover between the 4LS and 2LS-2HS states above 300 K. A modified indium-tin oxide (ITO) electrode was fabricated by soaking the ITO in a solution of 1. The resultant electrode showed reversible redox waves attributed to the original redox processes of iron(II)/iron(III).

Progressive impairment of cerebellar mGluR signalling and its therapeutic potential for cerebellar ataxia in spinocerebellar ataxia type 1 model mice.

KEY POINTS: Spinocerebellar ataxia type 1 (SCA1) is a progressive neurodegenerative disease caused by a gene defect, leading to movement disorder such as cerebellar ataxia. It remains largely unknown which functional defect contributes to the cerebellar ataxic phenotype in SCA1. In this study, we report progressive dysfunction of metabotropic glutamate receptor (mGluR) signalling, which leads to smaller slow synaptic responses, reduced dendritic Ca2+ signals and impaired synaptic plasticity at cerebellar synapses, in the early disease stage of SCA1 model mice. We also show that enhancement of mGluR signalling by a clinically available drug, baclofen, leads to improvement of motor performance in SCA1 mice. SCA1 is an incurable disease with no effective treatment, and our results may provide mechanistic grounds for targeting mGluRs and a novel drug therapy with baclofen to treat SCA1 patients in the future. ABSTRACT: Spinocerebellar ataxia type 1 (SCA1) is a progressive neurodegenerative disease that presents with cerebellar ataxia and motor learning defects. Previous studies have indicated that the pathology of SCA1, as well as other ataxic diseases, is related to signalling pathways mediated by the metabotropic glutamate receptor type 1 (mGluR1), which is indispensable for proper motor coordination and learning. However, the functional contribution of mGluR signalling to SCA1 pathology is unclear. In the present study, we show that SCA1 model mice develop a functional impairment of mGluR signalling which mediates slow synaptic responses, dendritic Ca2+ signals, and short- and long-term synaptic plasticity at parallel fibre (PF)-Purkinje cell (PC) synapses in a progressive manner from the early disease stage (5 postnatal weeks) prior to PC death. Notably, impairment of mGluR-mediated dendritic Ca2+ signals linearly correlated with a reduction of PC capacitance (cell surface area) in disease progression. Enhancement of mGluR signalling by baclofen, a clinically available GABAB receptor agonist, led to an improvement of motor performance in SCA1 mice and the improvement lasted ∼1 week after a single application of baclofen. Moreover, the restoration of motor performance in baclofen-treated SCA1 mice matched the functional recovery of mGluR-mediated slow synaptic currents and mGluR-dependent short- and long-term synaptic plasticity. These results suggest that impairment of synaptic mGluR cascades is one of the important contributing factors to cerebellar ataxia in early and middle stages of SCA1 pathology, and that modulation of mGluR signalling by baclofen or other clinical interventions may be therapeutic targets to treat SCA1.

Enhanced bioactivity of polyvinylidene chloride films using argon ion bombardment for guided bone regeneration.

Polyvinylidene chloride (PVDC) is a long chain carbon synthetic polymer. The objective of this study was to improve the bioactivity of PVDC films through surface modification using argon (Ar) ion bombardment to create Ar-modified PVDC films (Ar-PVDC) to address the clinical problems of guided bone regeneration (GBR), which is technique-sensitive, and low bone regenerative ability. First, the effects of Ar ion bombardment, a low temperature plasma etching technique widely used in industry, on PVDC film wettability, surface chemistry, and morphology were confirmed. Next, fibroblast-like and osteoblast-like cell attachment and proliferation on Ar-PVDC were assessed. As a preclinical in vivo study, Ar-PVDC was used to cover a critical-sized bone defect on rat calvaria and osteoconductivity was evaluated by micro-computed tomography analysis and histological examinations. We found that the contact angle of PVDC film decreased by 50° because of the production of -OH groups on the PVDC film surface, though surface morphological was unchanged at 30 min after Ar ion bombardment. We demonstrated that cell attachment increased by about 40% and proliferation by more than 140% because of increased wettability, and 2.4 times greater bone regeneration was observed at week 3 with Ar-PVDC compared with untreated PVDC films. These results suggest that Ar ion bombardment modification of PVDC surfaces improves osteoconductivity, indicating its potential to increase bone deposition during GBR.

Lesion in the lateral cerebellum specifically produces overshooting of the toe trajectory in leading forelimb during obstacle avoidance in the rat.

During locomotion, stepping over an obstacle under visual guidance is crucial to continuous safe walking. Studies of the role of the central nervous system in stepping movements have focused on cerebral cortical areas such as the primary motor cortex and posterior parietal cortex. There is speculation that the lateral cerebellum, which has strong anatomical connections with the cerebral cortex, also plays a key role in stepping movements over an obstacle, although this function of the lateral cerebellum has not yet been elucidated. Here we investigated the role of the lateral cerebellum during obstacle avoidance locomotion in rats with a lateral cerebellar lesion. A unilateral lesion in the lateral cerebellum did not affect limb movements during overground locomotion. Importantly, however, the lesioned animals showed overshooting of the toe trajectory specific to the leading forelimb ipsilateral to the lesion when stepping over an obstacle, and the peak toe position, in which the toe is maximally raised during stepping, shifted away from the upper edge of the obstacle. Recordings of EMG activity from elbow flexor and extensor muscles suggested that the overshooting toe trajectory in the ipsilateral leading forelimb possibly resulted from sustained elbow flexion and delayed elbow extension following prolonged activity of the biceps brachii. These results suggest that the lateral cerebellum specifically contributes to generating appropriate toe trajectories in the ipsilateral leading forelimb and to controlling related muscle activities in stepping over an obstacle, especially when accurate control of the distal extremity is achieved under visual guidance.

Dual involvement of G-substrate in motor learning revealed by gene deletion.

In this study, we generated mice lacking the gene for G-substrate, a specific substrate for cGMP-dependent protein kinase uniquely located in cerebellar Purkinje cells, and explored their specific functional deficits. G-substrate-deficient Purkinje cells in slices obtained at postnatal weeks (PWs) 10-15 maintained electrophysiological properties essentially similar to those from WT littermates. Conjunction of parallel fiber stimulation and depolarizing pulses induced long-term depression (LTD) normally. At younger ages, however, LTD attenuated temporarily at PW6 and recovered thereafter. In parallel with LTD, short-term (1 h) adaptation of optokinetic eye movement response (OKR) temporarily diminished at PW6. Young adult G-substrate knockout mice tested at PW12 exhibited no significant differences from their WT littermates in terms of brain structure, general behavior, locomotor behavior on a rotor rod or treadmill, eyeblink conditioning, dynamic characteristics of OKR, or short-term OKR adaptation. One unique change detected was a modest but significant attenuation in the long-term (5 days) adaptation of OKR. The present results support the concept that LTD is causal to short-term adaptation and reveal the dual functional involvement of G-substrate in neuronal mechanisms of the cerebellum for both short-term and long-term adaptation.

Quantitative evaluation of posture control in rats with inferior olive lesions.

Impairment of inferior olivary neurons (IONs) affects whole-body movements and results in abnormal gait and posture. Because IONs are activated by unpredicted motion rather than regular body movements, the postural dysfunction caused by ION lesions is expected to involve factors other than simple loss of feedback control. In this study, we measured the postural movements of rats with pharmacological ION lesions (IO rats) trained to stand on their hindlimbs. The coordination of body segments as well as the distribution and frequency characteristics of center of mass (COM) motion were analyzed. We determined that the lesion altered the peak properties of the power spectrum density of the COM, whereas changes in coordination and COM distribution were minor. To investigate how the observed properties reflected changes in the control system, we constructed a mathematical model of the standing rats and quantitatively identified the control system. We found an increase in linear proportional control and a decrease in differential and nonlinear control in IO rats compared with intact rats. The dystonia-like changes in body stiffness explain the nature of the linear proportional and differential control, and a disorder in the internal model is one possible cause of the decrease in nonlinear control.

Prevention of Hamstring Injuries in Collegiate Sprinters.

BACKGROUND: No studies have been reported on how strength, agility, and flexibility training reduce the occurrence of hamstring injuries in sprinters. Therefore, a program for preventing hamstring injury in these athletes has not been established. PURPOSE: To document the incidence of hamstring injuries during times when different prevention strategies were employed to see whether a particular prevention program reduced their occurrence. STUDY DESIGN: Descriptive epidemiology study. METHODS: The study subjects were a total of 613 collegiate male sprinters trained by the same coach over 24 seasons. Tow training was used throughout the research period as a normal sprint training method. The hamstring injury prevention program evolved over time. From 1988 to 1991 (period 1), prevention focused on strength training alone; from 1992 to 1999 (period 2), a combination of strength and agility training was used; and from 2000 to 2011 (period 3), the program incorporated strength, agility, and flexibility training. The incidence of hamstring injuries was compared for each of the 3 prevention strategies. RESULTS: The incidence of hamstring injuries per athlete-seasons was 137.9 for period 1, 60.6 for period 2, and 6.7 for period 3. A significant difference was observed in the incidence of hamstring injury according to the different prevention programs (χ2(2) = 31.78, P < .001, effect size: Cramer V = 0.23, 1 - ß = 0.999). Residual analysis showed that the number of hamstring injuries for period 1 was significantly greater than the expected value (P < .01), whereas that for period 3 was significantly lower than the expected value (P < .01). CONCLUSION: The incidence of hamstring injuries in sprinters decreased as agility and flexibility were added to strength training.

Postural control during quiet bipedal standing in rats.

The control of bipedal posture in humans is subject to non-ideal conditions such as delayed sensation and heartbeat noise. However, the controller achieves a high level of functionality by utilizing body dynamics dexterously. In order to elucidate the neural mechanism responsible for postural control, the present study made use of an experimental setup involving rats because they have more accessible neural structures. The experimental design requires rats to stand bipedally in order to obtain a water reward placed in a water supplier above them. Their motions can be measured in detail using a motion capture system and a force plate. Rats have the ability to stand bipedally for long durations (over 200 s), allowing for the construction of an experimental environment in which the steady standing motion of rats could be measured. The characteristics of the measured motion were evaluated based on aspects of the rats' intersegmental coordination and power spectrum density (PSD). These characteristics were compared with those of the human bipedal posture. The intersegmental coordination of the standing rats included two components that were similar to that of standing humans: center of mass and trunk motion. The rats' PSD showed a peak at approximately 1.8 Hz and the pattern of the PSD under the peak frequency was similar to that of the human PSD. However, the frequencies were five times higher in rats than in humans. Based on the analysis of the rats' bipedal standing motion, there were some common characteristics between rat and human standing motions. Thus, using standing rats is expected to be a powerful tool to reveal the neural basis of postural control.

Measuring body sway of bipedally standing rat and quantitative evaluation of its postural control.

Human generates very slow (<1 Hz) body sway during standing, and the behavior of this sway is known to be changed characteristically depending on the neural ataxia. In order to investigate the sway mechanism and mechanism of neural ataxia through this sway behavior, the present research proposes an experimental environment of rats under bipedal standing. By the experiment, we succeeded the measurement of six intact rats standing for over 200 seconds without postural supports. Moreover, by comparing measured center of pressure (COP) and that of system model with nonlinear PID control model which is proposed as human standing model, control parameters of rats were numerically evaluated. Evaluated control parameters of rats were close to those of human, i.e., control strategy was considered to be comparable between rats and human.

Effect of inactivation of the intermediate cerebellum on overground locomotion in the rat: a comparative study of the anterior and posterior lobes.

The importance of the cerebellum in control of locomotion is demonstrated by the ataxic gait of cerebellar patients. The intermediate cerebellum contains somatotopical representations for forelimbs and hindlimbs in both anterior and posterior lobes. However, it is not known whether these separate regions have discrete roles in control of limb movements during locomotion. Here we compared the effect of muscimol-induced inactivation of the anterior or posterior intermediate cerebellum on limb movements in walking rats. Inactivation of the anterior intermediate cerebellum had clear effects on limb movements during overground locomotion, resulting in excessive toe elevation and hyperflexion of joints in the swing phase. Inactivation of the posterior region resulted in similar but less pronounced deficits. Postural defects were not present in either group of rats. These findings suggest that the intermediate cerebellum of the anterior lobe has a greater influence on the ability to control limb movements during overground locomotion than the posterior lobe.

Gait modification during approach phase when stepping over an obstacle in rats.

Stepping over obstacles to avoid tripping is an essential component in safe and smooth locomotion. Obstacle avoidance during locomotion is completed in several steps during the approach phase toward the obstacle and stepping over the obstacle. The purpose of this study was to investigate gait modification during the approach phase when stepping over obstacles of different heights in rats. In all four limbs, the toe height when the toe was just above the obstacle increased depending on the obstacle height, leaving a safe margin. However, the horizontal distance between toe and obstacle just prior to stepping over was not influenced by obstacle height. In the fore- and hindlimbs that served as trailing limbs, it was found that the stride length and its related swing phase duration in the final step were significantly shorter than those in both the penultimate step and overground locomotion. These results suggest that adjustment of trailing limb in the final step during the approach phase is important in preparation for the stepping movement over an obstacle.

Characteristics of leading forelimb movements for obstacle avoidance during locomotion in rats.

Walking smoothly and safely often involves stepping over an obstacle. The purpose of this study was to examine forelimb movements and toe trajectories in stepping over an obstacle during overground locomotion in rats. We performed a kinematic analysis of forelimb movements and measured electromyographic (EMG) activities in the biceps and triceps brachii of the forelimbs. We found that mean toe height just above the obstacle was lower in the leading forelimb than in the trailing forelimb. The toe positions of the leading forelimb at maximal elevation over the obstacle (peak toe position) were closer to the upper edge of the obstacle than those of the trailing forelimb. The linear distance between peak toe position and the upper edge of the obstacle was significantly less in the leading forelimb compared to the trailing forelimb. The peak toe position of the leading forelimb spatially corresponds to the transition point from flexion to extension of the elbow joint. This transition appeared to be controlled mainly by an offset of EMG activity of the elbow flexor, the biceps brachii muscle. In contrast, the trailing forelimb appeared to be controlled by the shoulder and wrist joints. These results suggest that the toe trajectory of the leading forelimb is more accurately regulated than that of the trailing forelimb. In addition, the activities of the elbow flexor may in part contribute to the toe trajectory of the leading forelimb.

Characteristics of gait ataxia in δ2 glutamate receptor mutant mice, ho15J.

The cerebellum plays a fundamental, but as yet poorly understood, role in the control of locomotion. Recently, mice with gene mutations or knockouts have been used to investigate various aspects of cerebellar function with regard to locomotion. Although many of the mutant mice exhibit severe gait ataxia, kinematic analyses of limb movements have been performed in only a few cases. Here, we investigated locomotion in ho15J mice that have a mutation of the δ2 glutamate receptor. The cerebellum of ho15J mice shows a severe reduction in the number of parallel fiber-Purkinje synapses compared with wild-type mice. Analysis of hindlimb kinematics during treadmill locomotion showed abnormal hindlimb movements characterized by excessive toe elevation during the swing phase, and by severe hyperflexion of the ankles in ho15J mice. The great trochanter heights in ho15J mice were lower than in wild-type mice throughout the step cycle. However, there were no significant differences in various temporal parameters between ho15J and wild-type mice. We suggest that dysfunction of the cerebellar neuronal circuits underlies the observed characteristic kinematic abnormality of hindlimb movements during locomotion of ho15J mice.

Osteoanagenesis after transplantation of bone marrow-derived mesenchymal stem cells using polyvinylidene chloride film as a scaffold.

The aim of this study was to develop a new cell transplantation technique for osteoanagenesis at bone defect sites. Polyvinylidene chloride (PVDC) film was evaluated because of its good biocompatibility and flexibility. We used this film as both a cell scaffold and a barrier membrane. Initially, the cell compatibility of the PVDC film for fibroblast-like cells and osteoblast-like cells was confirmed. Subsequently, bone marrow cells were obtained from rats and cultured on PVDC films in two kinds of medium. The PVDC films with bone marrow-derived mesenchymal stem cells (MSCs) were then applied to critical-sized bone defects in the calvarial bone of rats. After the transplantation, the surgical sites were dissected out and evaluated by soft X-ray radiography, micro-CT analysis and histological examinations. The bone marrow-derived MSC-transplanted rats showed greater bone regeneration than the control rats. Therefore, PVDC film is considered to be useful as a scaffold for bone regeneration.