Effect of cerebellar transcranial magnetic stimulation on soleus Ia presynaptic and reciprocal inhibition.

Previously, we reported that cerebellar transcranial magnetic stimulation (C-TMS) facilitates spinal motoneuronal excitability in resting humans. In this study, we aimed to characterize the descending pathway that is responsible for the C-TMS-associated cerebellar spinal facilitation. We evaluated the effect of C-TMS on ipsilateral soleus Ia presynaptic inhibition (PSI) and reciprocal inhibition (RI) because the vestibulospinal and reticulospinal tracts project from the cerebellum to mediate spinal motoneurons via interneurons associated with PSI. PSI and RI were measured with a soleus H-reflex test following operant conditioning using electrical stimulation of the common peroneal nerve. C-TMS was delivered before test tibial nerve stimulation with conditioning-test interstimulus intervals of 110 ms. C-TMS did not generate motor-evoked potentials, and it did not increase electromyography activity in the ipsilateral soleus muscle, indicating that C-TMS does not directly activate the corticospinal tract and motoneurons. However, C-TMS facilitated the ipsilateral soleus H-reflex and reduced the amount of soleus Ia PSI, but not RI. These findings indicate that C-TMS may facilitate the excitability of the spinal motoneuron pool via the vestibulospinal or reticulospinal tracts associated with PSI. Cerebellar spinal facilitation may be useful for assessing the functional connectivity of the cerebellum and vestibular nuclei or reticular formation.

Task dependency of the long-latency facilitatory effect on the soleus H-reflex by cerebellar transcranial magnetic stimulation.

We investigated whether cerebellar transcranial magnetic stimulation (C-TMS) facilitates the excitability of the ipsilateral soleus motoneuron pool in resting humans, and whether the facilitation is modulated by a task that promotes cerebellar activity. A test tibial nerve stimulus evoking the H-reflex from the right soleus muscle was delivered before or after conditioning C-TMS in prone individuals. The amplitude of the H-reflex was significantly increased at conditioning-test interstimulus intervals of 110, 120, and 130 ms. Furthermore, we revealed that this facilitation effect was inhibited while the individuals tapped their right index finger. These findings indicate that C-TMS facilitates spinal motoneuronal excitability with an ∼100 ms latency in resting humans, and that this cerebellar spinal facilitation is modulated by a task that might increase cerebellar activity. Cerebellar spinal facilitation could thus be useful for assessing the excitability of the cerebellum, or the cerebellar output to spinal motoneurons.

Effect of Posture during Trumpet and Marching Euphonium Performance on the Trunk and Lower Limb Musculoskeletal System.

[Purpose] The purpose of the present study was to investigate the effect of trumpet and marching euphonium performance posture on the trunk and lower limb musculoskeletal system. [Subjects] The subjects were 10 female university students. [Methods] Subjects maintained a resting position, a trumpet performance posture, and a marching euphonium performance posture. The angles and muscle activities of the trunk and lower limbs were then measured. [Results] The anterior tilt angle of the trunk decreased significantly in the trumpet and marching euphonium performance postures compared with the resting standing position, as well as in the marching euphonium performance posture compared with the trumpet performance posture. The muscle activity of the cervical paraspinal muscles, upper fibers of the trapezius, and lumbar paraspinal muscles increased significantly in the marching euphonium performance posture compared with the resting standing position, as well as in the marching euphonium performance posture compared with the trumpet performance posture. [Conclusion] The results suggest that the performance position for trumpet and the marching euphonium performance increases the load on the cervical and thoracic musculoskeletal system, which increases with greater instrument weight. However, the same instrument performance postures had no affect on the musculoskeletal system of the lower limbs.

Selective apoptosis of natural killer-cell tumours by l-asparaginase.

We examined the effectiveness of various anti-tumour agents to natural killer (NK)-cell tumour cell lines and samples, which are generally resistant to chemotherapy, using flow cytometric terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labelling (TUNEL) assay. Although NK-YS and NK-92 were highly resistant to various anti-tumour agents, l-asparaginase induced apoptosis in these two NK-cell lines. NK-cell leukaemia/lymphoma and acute lymphoblastic leukaemia (ALL) samples were selectively sensitive to l-asparaginase and to doxorubicin (DXR) respectively. Samples of chronic NK lymphocytosis, an NK-cell disorder with an indolent clinical course, were resistant to both drugs. Our study clearly separated two major categories of NK-cell disorders and ALL according to the sensitivity to DXR and l-asparaginase. We examined asparagine synthetase levels by real-time quantitative polymerase chain reaction (RQ-PCR) and immunostaining in these samples. At least in nasal-type NK-cell lymphoma, there was a good correlation among asparagine synthetase expression, in vitro sensitivity and clinical response to l-asparaginase. In aggressive NK-cell leukaemia, although asparagine synthetase expression was high at both mRNA and protein levels, l-asparaginase induced considerable apoptosis. Furthermore, samples of each disease entity occupied a distinct area in two-dimensional plotting with asparagine synthetase mRNA level (RQ-PCR) and in vitrol-asparaginase sensitivity (TUNEL assay). We confirmed rather specific anti-tumour activity of l-asparaginase against NK-cell tumours in vitro, which provides an experimental background to the clinical use of l-asparaginase for NK-cell tumours.

Establishment of real-time polymerase chain reaction method for quantitative analysis of asparagine synthetase expression.

We established a real-time quantitative PCR (RQ-PCR) with which to measure abundance of the asparagine synthetase (AS) mRNA. The level of AS mRNA paralleled AS enzyme activity, as well as the AS protein level detected by Western blotting and by in situ immunostaining. Cytotoxicity tests in vitro showed that the AS mRNA level also synchronized with cellular resistance to L-asparaginase in cell lines. Cellular levels of AS enzyme activity correlated with resistance to L-asparaginase. These results indicate that the AS mRNA level is an index of resistance to L-asparaginase. RQ-PCR is superior to enzyme assays, Western blotting, and immunostaining in the following ways: less labor and time, accurate and reproducible quantitativity, and broad dynamic range. In addition, RQ-PCR could evaluate differences in L-asparaginase sensitivity although immunostaining could not. And in clinical samples, we analyzed eight pediatric leukemia cases by this RQ-PCR to evaluate whether this method was applicable to clinical laboratories and the expression level of AS mRNA in each case were predictable for the effectiveness of L-asparaginase treatment. Consequently, this method was useful enough in defining candidates for selective therapy that targets an AS deficiency.