FNDC5/irisin mediates the protective effects of Innovative theta-shaking exercise on mouse memory.

As a passive motion and non-invasive treatment, theta-shaking exercise is considered an alternative to traditional active exercise for slowing down brain ageing. Here, we studied the influence of theta-shaking exercise on fibronectin type III domain containing 5/irisin (FNDC5/irisin) in the anterior nucleus of the thalamus, hippocampus, and medial prefrontal cortex (ATN-HPC-MPFC). Further, we assessed memory in senescence-accelerated prone mice (SAMP-10 mice) using a behavioural test to confirm the protective effect of theta-shaking exercise against age-related memory decline. SAMP-10 mice were subjected to theta-shaking exercise for 9-30 weeks. Mice then performed the T-maze test and passive avoidance task. Immunohistochemical analysis and ELISA were used to assess FNDC5/irisin, nerve growth factor (NGF), and neurotrophin 4/5 (NT4/5) expression in the ATN-HPC-MPFC. In the shaking group, FNDC5 was locally upregulated within the hippocampus and MPFC area rather than exhibiting even distribution throughout brain tissue. Irisin levels were generally higher in the control group. Meanwhile, hippocampal NGF levels were significantly higher in the shaking group, with no differences noted in neurotrophin levels. Theta-shaking preserved normal neurons in certain sub-regions. However, no beneficial changes in neuronal density were noted in the ATN. Theta-shaking exercise positively affects memory function in SAMP-10 mice. FNDC5 upregulation and higher levels of NGF, along with the potential involvement of irisin, may have contributed to the preservation of normal neuronal density in the hippocampus and MPFC subregions.

Trapping bond exchange phenomenon revealed for off-stoichiometry cross-linking of phase-separated vitrimer-like materials.

Vitrimer materials combined with nano-phase separated structures have attracted attention, expanding the tuning range of physical properties, such as flow and creep properties. We recently demonstrated a preparation of vitrimer-like materials with phase-separated nanodomains in which dissociative bond exchange via trans-N-alkylation of quaternized pyridine was operated. In this study, we demonstrate a new finding about the bond exchange mechanism: that is, the trapping bond exchange phenomenon. The component polymer is a poly(acrylate) containing pyridine side groups randomly along the chain, which is cross-linked by diiodo molecules via pyridine-iodo quaternization, where the quaternized pyridines are aggregated to form nano-size domains. When the cross-linking reaction is performed at an off-stoichiometric pyridine : iodo ratio (i.e., an excess of pyridine groups), free pyridine groups are located in the matrix phase. Since the bond exchange in the present system progresses in an inter-domain manner, the dissociated unit bearing pendant iodo is trapped by the free pyridine groups in the matrix, which generates other small aggregates. This trapping phenomenon greatly affects the relaxation and creep properties, which are very different from those found in conventional knowledge about vitrimer physics.

Exercise through Shaking Stimuli Suppresses Cancer Growth via the Wnt pathway in ApcMin/+ Mice.

OBJECTIVE: Exercise has been reported to suppress colorectal cancer; however, the mechanism of suppression by exercise and its effect on the Wnt pathway, which is particularly involved in the early stage of carcinogenesis, remain unclear. In this study, we subjected ApcMin/+ mice to exercise by shaking stimuli to investigate the mechanisms of suppressing colorectal cancer, and focused on the Ca2+ pathway, which is one of the ß-catenin-independent Wnt signaling pathways that suppress the accumulation of ß-catenin. METHODS: Mice in the exercise group were subjected to exercise by shaking stimuli for 30 min/session, 6 sessions/ week, for a total of 11 weeks. The number and diameter of intestinal polyps were calculated. Expression analysis of ß-catenin and Pak1 from the intestinal tract and Wnt5a-Pan and Wnt5a-Long from the gastrocnemius muscle was performed by western blotting. The expression of ß-catenin and Wnt5a-Pan was observed by immunohistochemical staining. RESULT: The levels of expression of ß-catenin and Pak1 in the small intestine were low in the exercise group, indicating that exercise suppressed the accumulation of ß-catenin. In the gastrocnemius muscle, the levels of expression of Wnt5a-Pan and Wnt5a-Long were significantly higher in the exercise group (p < 0.05). Histological analysis revealed that the percentage of large polyps was significantly lower in the exercise group than in the control group (p < 0.01), revealing that exercise suppressed the growth of polyps. In addition, the villi/crypt ratio (V/C ratio) was significantly higher in the exercise group, suggesting the suppression of exercise-induced local inflammation in the small intestine. CONCLUSION: We believe that the mechanism of polyp growth suppression is related to the inflammatory and not the Wnt pathway. This study clarified the growth-suppressing effect of a novel exercise method on cancer. We believe that its development and clinical application might open new possibilities for the prevention treatment of colorectal cancer.

Novel shaking exercises for hippocampal and medial prefrontal cortex functioning maintain spatial working memory.

INTRODUCTION: The decline in spatial working memory is one of the earliest signs of normal brain aging. OBJECTIVE: We developed a novel physical exercise method, termed the "shaking exercise," to slow down this process. METHODS: The experimental protocol included administering the shaking exercise for 8-32 weeks in male senescence-accelerated mouse prone 10 (SAMP-10). They were subjected to the T-maze test, followed by immunohistochemical analysis, to assess the influence of the shaking exercise on the M1 muscarinic acetylcholine receptor (CHRM1) and α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptor (AMPAR) of the dorsal hippocampus and medial prefrontal cortex (dHC-mPFC). RESULTS: The T-maze test demonstrated that the shaking group had less hesitation in the face of selecting direction at week 24. In the immunohistochemical analysis, more CHRM1s were in the CA3 subregion and more AMPARs were in the subiculum. CHRM1s and AMPARs were maintained in the CA1 region and the mPFC. The CHRM1s seem to have a positive effect on the AMPAR in the dentate gyrus (DG) region and the CA3 region. In the CA1 region, CHRM1s were negatively correlated with AMPARs. In addition, high-density neurons were expressed in the shaking group in the upstream DG, the middle part and the distal part of CA3, the distal part of CA1, and the mPFC. CONCLUSIONS: Our results raise the possibility that maintenance of the spatial working memory effect observed with the shaking exercise is driven in part by the uneven affection of CHRM1s and AMPARs in the dHC-mPFC circuit system and significantly maintains the neuronal expression in the dHC-mPFC.

Exercise by Shaking Alleviates the Decline in Memory due to Aging: A Study in Mice.

INTRODUCTION: Although exercise can prevent cognitive decline due to aging, few elderly individuals are able to exercise for long. Therefore, an exercise method for older adults that is feasible for a long duration without overexertion is necessary. In this study, we focused on exercise by shaking. This study examined the possibility to prevent the decline in memory through regular and long-term shaking exercise using a senescence-accelerated mouse (SAM) model. Behavioral analysis was conducted, and histological changes in the mouse brain were examined to evaluate whether this stimulation method could become a novel exercise method. MATERIALS AND METHODS: The shaking exercise was applied to SAMP10 mice for 30 min 3 times per week for 25 continuous weeks. Behavioral analysis included a step-through passive avoidance test, whereas the histological analysis involved immunohistochemical staining using the anti-glutamate receptor (α-amino-3-hydroxy-5-methyl-4-isoxazole-propionate receptors [AMPAR]) antibody in the hippocampus. The number and area of nerve cells in the hippocampal regions were measured and compared between groups. RESULTS: Behavioral analysis revealed that the shaking group retained memory longer than the control group, and memory capacity decline was suppressed. Additionally, histological examination showed that the shaking group had a higher number of AMPAR receptor-positive neurons per area in the hippocampal CA1 and CA3 regions than the control group, suggesting that degeneration and shedding of neurons due to aging was suppressed. DISCUSSION/CONCLUSION: We believe that shaking could become an exercise therapy that can reduce the decline in memory with aging and expect its human application in the future.

Maintenance of the Amygdala-Hippocampal Circuit Function with Safe and Feasible Shaking Exercise Therapy in SAMP-10 Mice.

INTRODUCTION: Patients with dementia show reduced adaptive, behavioral, and physiological responses to environmental threats. Physical exercise is expected to delay brain aging, maintain cognitive function and, consequently, help dementia patients face threats and protect themselves skillfully. METHODS: To confirm this, we aimed to investigate the effects of the shaking exercise on the avoidance function in the senescence-accelerated mouse-prone strain-10 (SAMP-10) model at the behavioral and tissue levels. SAMP-10 mice were randomized into 2 groups: a control group and a shaking group. The avoidance response (latency) of the mice was evaluated using a passive avoidance task. The degree of amygdala and hippocampal aging was evaluated based on the brain morphology. Subsequently, the association between avoidance response and the degree of amygdala-hippocampal aging was evaluated. RESULTS: Regarding the passive avoidance task, the shaking group showed a longer latency period than the control group (p < 0.05), even and low intensity staining of ubiquitinated protein, and had a higher number of and larger neurons than those of the control group. The difference between the groups was more significant in the BA region of the amygdala and the CA1 region of the hippocampus (staining degree: p < 0.05, neuron size: p < 0.01, neuron counts: p < 0.01) than in other regions. CONCLUSIONS: The shaking exercise prevents nonfunctional protein (NFP) accumulation, neuron atrophy, and neuron loss; delays the aging of the amygdala and hippocampus; and maintains the function of the amygdala-hippocampal circuit. It thus enhances emotional processing and cognition functions, the memory of threats, the skillful confrontation of threats, and proper self-protection from danger.

Body-Specific Attention to the Hands and Feet in Healthy Adults.

To execute the intended movement, the brain directs attention, called body-specific attention, to the body to obtain information useful for movement. Body-specific attention to the hands has been examined but not to the feet. We aimed to confirm the existence of body-specific attention to the hands and feet, and examine its relation to motor and sensory functions from a behavioral perspective. The study included two groups of 27 right-handed and right-footed healthy adults, respectively. Visual detection tasks were used to measure body-specific attention. We measured reaction times to visual stimuli on or off the self-body and calculated the index of body-specific attention score to subtract the reaction time on self-body from that off one. Participants were classified into low and high attention groups based on each left and right body-specific attention index. For motor functions, Experiment 1 comprised handgrip strength and ball-rotation tasks for the hands, and Experiment 2 comprised toe grip strength involved in postural control for the feet. For sensory functions, the tactile thresholds of the hands and feet were measured. The results showed that, in both hands, the reaction time to visual stimuli on the hand was significantly lesser than that offhand. In the foot, this facilitation effect was observed in the right foot but not the left, which showed the correlation between body-specific attention and the normalized toe gripping force, suggesting that body-specific attention affected postural control. In the hand, the number of rotations of the ball was higher in the high than in the low attention group, regardless of the elaboration exercise difficulty or the left or right hand. However, this relation was not observed in the handgripping task. Thus, body-specific attention to the hand is an important component of elaborate movements. The tactile threshold was higher in the high than in the low attention group, regardless of the side in hand and foot. The results suggested that more body-specific attention is directed to the limbs with lower tactile abilities, supporting the sensory information reaching the brain. Therefore, we suggested that body-specific attention regulates the sensory information to help motor control.

Maintaining Aging Hippocampal Function with Safe and Feasible Shaking Exercise in SAMP10 Mice.

INTRODUCTION: The disabling effects of dementia, an incurable disease with little effect on mortality, affect society far more than many other conditions. OBJECTIVE: The aim of this study was to stop or delay the onset of dementia using low-cost methods such as physical exercise. METHODS: Senescence-accelerated model-prone (SAMP) 10 mice were made to perform a user-friendly shaking exercise for 25 weeks. The motor function and hippocampal functions (learning, spatial cognition) of the mice were evaluated using behavioral experiments. The degree of hippocampal aging was evaluated based on brain morphology. The association between behavioral performance of the mice and the degree of hippocampal aging was then evaluated. RESULTS: The behavioral test results showed that the shaking group had higher motor coordination (p < 0.01) and motor learning (p < 0.05). Significantly higher performances in the learning ability were observed in the shaking group at a middle-period experiment (p < 0.05); the spatial cognitive functions also improved (p < 0.05). The shaking group showed delayed ageing of cells in the dentate gyrus (DG; area: p < 0.01) and cornu Ammonis (CA; area: p < 0.01) regions of the hippocampus. CONCLUSIONS: The shaking exercise enhances the activity of mice and reduces age-associated decreases in learning and spatial cognitive functions. Regarding hippocampal morphology, shaking exercise can prevent non-functional protein accumulation, cell atrophy, and cell loss. Specifically, shaking exercise protects cell growth and regeneration in the DG area and enhances the learning function of the hippocampus. Furthermore, shaking exercise maintained the spatial cognitive function of cells in the CA3 and CA1 regions, and prevented the chronic loss of CA2 transmission that decreased the spatial memory decline in mice.

A novel device to prevent osteoporosis by promoting bone metabolism using a newly developed double-loading stimulation with vibration and shaking.

In Japan, 13 million people have osteoporosis, including approximately 9 hundred thousand people who are bedridden owing to bone fractures from falls. Preventing osteoporosis is considered to be an important and effective way of preventing fall-related fractures. Thus, we developed a novel method of locomotor stimulation and analyzed its effectiveness in mice. Specifically, we created a double-loading device that combines vibration and shaking stimulation. The device was used to continuously stimulate ovariectomy-induced decreased bone density mouse models 30 minutes daily for 10 weeks. We then collected femur samples, created undecalcified tissue slices, calculated parameters using bone histomorphomtry, and conducted comparative testing. BS/TV (bone surface/tissue volume), N.Oc/ES (osteoclast number/eroded surface), Oc.S/ES (osteoclast osteoid surface/eroded surface), Omt (osteoid maturation time), Tb.N (trabecular number), Mlt (mineralization lag time) < (p < 0.01), N.Ob (osteoblast number), N.Ob/TV (osteoblast number/tissue volume), sLS (single labeled suface), N.Mu.Oc/ES (multinucle osteoclast number/eroded surface), and N.Mo.Oc/ES (mononucle osteoclast number/eroded surface) (p < 0.05) were significantly higher in the stimulation group than in the non-stimulation group. In addition, BS/BV (bone surface/bone volume), Tb.Sp (trabecular separation), MAR (mineral apposition rate), Aj.Ar (adjusted apposition rate) (p < 0.01), ES (eroded surface ), ES/BS (eroded surface/bone surface), and BRs.R (bone resorption rate) (p < 0.05) were significantly lower in the stimulation group than in the non-stimulation group. These results suggest that stimulation activated osteoblasts and osteoclasts, thereby leading to highly active bone remodeling. We anticipate that bone mineralization will subsequently occur, suggesting that this stimulation technique is effective in preventing osteoporosis by alleviating sudden bone density loss.

Effect of shaking and vibration stimulation on lumbar vertebrae in ovariectomized mice.

OBJECTIVES: Bone fractures affect the activities of daily living and lower quality of life, so investigating preventative measures is important. We developed novel stimulation equipment that combined a vibration stimulus with a shaking stimulus for preventing osteoporosis (one of the causes of bone fractures). We aimed to investigate the effect of this equipment on ovariectomized mice. METHODS: Oophorectomy of 8-week-old female mice was done. The stimulation group was stimulated for 10 consecutive weeks. RESULTS: The stimulation group showed significantly higher values (p<0.05) for osteoid thickness, osteoid volume-to-bone volume ratio and mineral apposition rate than those in the non-stimulation control group. The stimulation group showed significantly higher values (p<0.05) compared with the non-stimulation for expression of bone morphogenetic protein-2, interleukin-1ß, interleukin-6 and myogenic determination gene in quadriceps femoris muscles (QFMs). CONCLUSIONS: These data suggest that cytokine secretion by QFMs carried a humoral factor throughout the body via the blood and blood vessels and acted on bone and various organs. Development of this stimulation method and its clinical application, new methods for preventing and treating osteoporosis could ensue.

Effectiveness of exercise-induced cytokines in alleviating arthritis symptoms in arthritis model mice.

Recently, health awareness in Japan has been increasing and active exercise is now recommended to prevent lifestyle-related diseases. Cytokine activities have many positive effects in maintaining the health of a number of organs in the body. Myokines are cytokines secreted by skeletal muscles in response to exercise stimulation, and have recently generated much attention. Around 700,000 patients in Japan suffer from rheumatoid arthritis, making it the most prevalent autoimmune disease that requires active prevention and treatment. In the present study, a mouse model of spontaneous arthritis (SKG/Jcl) was subjected to continuous exercise stimulation, starting before the disease onset, to examine the effects of anti-inflammatory and inflammatory cytokine secretion on arthritis. For this stimulation, we developed a device that combines shaking and vibration. The results revealed that exercise stimulation delayed the onset of arthritis and slowed its progression. Thickened articular cartilage and multiple aggregates of chondrocytes were also observed. Further, exercise stimulation increased the expression of IL-6, IL-10, and IL-15, and inhibited TNF-α expression. From these results, we infer that the anti-inflammatory effects of IL-6 and IL-10, which showed increased expression upon exercise stimulation, inhibited the inflammatory activity of TNF-α and possibly delayed the onset of arthritis and slowed its progression. Novel methods for preventing and treating arthritis under clinical settings can be developed on the basis of these findings.