Development of a system to analyze oral frailty associated with Alzheimer's disease using a mouse model.

The rapid aging of the population makes the detection and prevention of frailty increasingly important. Oral frailty has been proposed as a novel frailty phenotype and is defined as a decrease in oral function coexisting with a decline in cognitive and physical functions. Oral frailty has received particular attention in relation to Alzheimer's disease (AD). However, the pathomechanisms of oral frailty related to AD remain unknown. It is assumed that the mesencephalic trigeminal nucleus (Vmes), which controls mastication, is affected by AD pathology, and as a result, masticatory function may be impaired. To investigate this possibility, we included male 3 × Tg-AD mice and their non-transgenic counterpart (NonTg) of 3-4 months of age in the present study. Immunohistochemistry revealed amyloid-ß deposition and excessive tau phosphorylation in the Vmes of 3 × Tg-AD mice. Furthermore, vesicular glutamate transporter 1-immunopositive axon varicosities, which are derived from Vmes neurons, were significantly reduced in the trigeminal motor nucleus of 3 × Tg-AD mice. To investigate whether the AD pathology observed in the Vmes affects masticatory function, we analyzed electromyography of the masseter muscle during feeding. The 3 × Tg-AD mice showed a significant delay in masticatory rhythm compared to NonTg mice. Furthermore, we developed a system to simultaneously record bite force and electromyography of masseter, and devised a new method to estimate bite force during food chewing in mice. Since the muscle activity of the masseter showed a high correlation with bite force, it could be accurately estimated from the muscle activity. The estimated bite force of 3 × Tg-AD mice eating sunflower seeds was predominantly smaller than that of NonTg mice. However, there was no difference in masseter weight or muscle fiber cross-sectional area between the two groups, suggesting that the decreased bite force and delayed mastication rhythm observed in 3 × Tg-AD mice were not due to abnormality of the masseter. In conclusion, the decreased masticatory function observed in 3 × Tg-AD mice was most likely caused by AD pathology in the Vmes. Thus, novel quantitative analyses of masticatory function using the mouse model of AD enabled a comprehensive understanding of oral frailty pathogenesis.

Effects of sex, age, choice of surgical orthodontic treatment, and skeletal pattern on the psychological assessments of orthodontic patients.

We aimed to examine the effects of sex, age, choice of surgical orthodontic treatment, and skeletal pattern on psychological assessment scores of orthodontic patients before edgewise treatment. They completed the State-Trait Anxiety Inventory (STAI), Beck Depression Inventory (BDI)-II, and the psychological domain of the World Health Organization Quality of Life 26 (Psych-QOL) for assessment of anxiety, depression, and body image, respectively. No significant effects on psychological assessment scores due to sex or age differences were found. Surgical orthodontic treatment patients and patients with skeletal Class III had significantly higher STAI-Trait and/or BDI-II scores and lower Psych-QOL score. Based on the linear mixed-effects model, the choice of surgical orthodontic treatment had a significant effect on the STAI-Trait, BDI-II, and Psych-QOL scores. No significant interaction effect was found between the choice of surgical orthodontic treatment and the skeletal pattern by ANB angle. Patients with skeletal Class I or III who chose surgical orthodontic treatment had higher STAI-Trait and/or BDI-II scores and/or lower body image score, respectively. These results suggest that patients who chose surgical orthodontic treatment, particularly those with skeletal Class I and III, may be more prone to experience anxiety and depression and have body image dissatisfaction.

Red rice koji extract alleviates hyperglycemia by increasing glucose uptake and glucose transporter type 4 levels in skeletal muscle in two diabetic mouse models.

BACKGROUND: Red rice koji (RRK), prepared by growing Monascus species on steamed rice, has been reported to lower blood glucose levels in diabetic animal models. However, the action mechanism is not yet completely understood. OBJECTIVE: The objective of this study was to examine the mechanism underlying the hypoglycemic action of RRK extract in two diabetic animal models: the insulin-deficiency mice, where the insulin deficiency was induced by streptozotocin (STZ), and insulin-resistance mice, where the insulin resistance was induced by a high-fat diet (HFD). DESIGN: Low (12.5 mg/kg body weight [BW]) and high (50.0 mg/kg BW) doses of RRK extract were orally administered to the mice for 10 successive days (0.25 mL/day/mouse). The protein expression levels of glucose transporter type 4 (GLUT4) in the skeletal muscle and glucose transporter type 2 (GLUT2) in the liver were measured. Blood glucose (BG) levels of STZ-treated mice in insulin tolerance test (ITT) and BG and insulin levels of HFD-fed mice in intraperitoneal glucose tolerance test (IPGTT) were investigated. RESULTS: In the STZ-treated mice, oral administration of RRK extract lowered BG levels and food intake but increased plasma 1,5-anhydroglucitol level. Moreover, the RRK extract lowered the BG levels of STZ-treated mice as measured by ITT. In the HFD-fed mice, we confirmed that the orally administered RRK extract lowered the BG and the homeostasis model assessment index for insulin resistance. Furthermore, the RRK extract lowered the BG and insulin levels of HFD-fed mice in IPGTT. Regarding the protein levels of GLUT, the orally administered RRK extract increased the GLUT4 level in the skeletal muscle; however, the RRK extract did not alter the GLUT2 level in the liver of either the STZ-treated or the HFD-fed mice. DISCUSSION: Our study demonstrates that RRK extract can improve impaired glucose tolerance in mouse models of diabetes by enhancing GLUT4 expression in skeletal muscle. CONCLUSION: These results suggest that RRK extract could potentially be a functional food for the treatment of diabetes mellitus.

Lavender Oil Reduces Depressive Mood in Healthy Individuals and Enhances the Activity of Single Oxytocin Neurons of the Hypothalamus Isolated from Mice: A Preliminary Study.

BACKGROUND: The aim of the present study was to assess the effects of lavender oil inhalation on blood pressure, pulse measurements, cortisol levels, depressive mood, and anxiety in healthy male adults. The mechanism was investigated by the action on oxytocin single neurons in the hypothalamus of rodents. METHODS: The participants (n = 7) were aged 20-40 years. After randomisation, they received an inhaled dose of lavender oil or distilled water for 20 min. They received the other treatment after a washout period of one week. We assessed the outcomes using the Self-Rating Depression Scale, State-Trait Anxiety Inventory, and self-rated unidimensional Visual Analogue Scale for depression; anxiety; and hunger, thirst, and appetite, respectively. Blood pressure, pulse rate, and cortisol concentration in the peripheral blood were assessed before and after inhalation. In the rodent study (n = 4), oxytocin single neurons were isolated from the mouse hypothalamus. Intracellular Ca2+ concentration in the oxytocin neurons isolated from the hypothalamus was measured following direct administration of lavender oil. RESULTS: Seven participants completed the study. Lavender inhalation decreased Self-Rating Depression Scale score and systolic and diastolic blood pressure. Ex vivo administration of lavender oil increased intracellular Ca2+ concentration in the hypothalamic oxytocin neurons. CONCLUSIONS: Lavender oil might be a useful therapy for stress relief, and its mechanism of action may include activation of the central oxytocin neurons.

Helicobacter pylori Vacuolating Cytotoxin A Causes Anorexia and Anxiety via Hypothalamic Urocortin 1 in Mice.

Helicobacter pylori (Hp) infection is related to the pathogenesis of chronic gastric disorders and extragastric diseases. Here, we examined the anorexigenic and anxiogenic effects of Hp vacuolating cytotoxin A (VacA) through activation of hypothalamic urocortin1 (Ucn1). VacA was detected in the hypothalamus after peripheral administration and increased Ucn1 mRNA expression and c-Fos-positive cells in the hypothalamus but not in the nucleus tractus solitarius. c-Fos and Ucn1-double positive cells were detected. CRF1 and CRF2 receptor antagonists suppressed VacA-induced anxiety and anorexia, respectively. VacA activated single paraventricular nucleus neurons and A7r5 cells; this activation was inhibited by phospholipase C (PLC) and protein kinase C (PKC) inhibitors. VacA causes anorexia and anxiety through the intracellular PLC-PKC pathway, migrates across the blood-brain barrier, and activates the Ucn1-CRF receptor axis.

Rubiscolin-6 activates opioid receptors to enhance glucose uptake in skeletal muscle.

Rubiscolin-6 is an opioid peptide derived from plant ribulose bisphosphate carboxylase/oxygenase (Rubisco). It has been demonstrated that opioid receptors could control glucose homeostasis in skeletal muscle independent of insulin action. Therefore, Rubiscolin-6 may be involved in the control of glucose metabolism. In the present study, we investigated the effect of rubiscolin-6 on glucose uptake in skeletal muscle. Rubiscolin-6-induced glucose uptake was measured using the fluorescent indicator 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino]-2-deoxyglucose (2-NBDG) in L6 and C2C12 cell lines. The protein expressions of glucose transporter 4 (GLUT4) and AMP-activated protein kinase (AMPK) in L6 cells were observed by Western blotting. The in vivo effects of rubiscolin-6 were characterized in streptozotocin (STZ)-induced diabetic rats. Rubiscolin-6 induced a concentration-dependent increase in glucose uptake levels. The increase of phospho-AMPK (pAMPK) and GLUT4 expressions were also observed in L6 and C2C12 cells. Effects of rubiscolin-6 were blocked by opioid receptor antagonists and/or associated signals inhibitors. Moreover, Rubiscolin-6 produced a dose-dependent reduction of blood glucose and increased GLUT4 expression in STZ-induced diabetic rats. In conclusion, rubiscolin-6 increases glucose uptake, potentially via an activation of AMPK to enhance GLUT4 translocation after binding to opioid receptors in skeletal muscle.

Evaluation of miniscrew stability using an automatic embedding auxiliary skeletal anchorage device.

OBJECTIVE: To clarify the in vivo effect of an automatic embedding device on miniscrew stability. MATERIALS AND METHODS: 42 miniscrews were implanted into rabbit femurs. The miniscrews with the novel auxiliary device formed the auxiliary group (n = 11 at 4 weeks; n = 11 at 8 weeks) and the miniscrews without the auxiliary device formed the nonauxiliary control group (n = 9 at 4 weeks; n = 11 at 8 weeks). Cortical bone thickness, distance from the cortical bone surface to the miniscrew head, and implantation depth of the spike were measured using micro-computed tomography. The mechanical retention force was evaluated by measuring the displacement of the miniscrew head after it was loaded perpendicular to its long axis. In the lateral displacement test, effects of the auxiliary (with vs without auxiliary), and time (4 vs 8 weeks) were assessed using the Brunner-Langer nonparametric analysis of longitudinal data in factorial experiments. RESULTS: The mean implantation depth of the spike in the auxiliary group at 4 and 8 weeks was 0.28 mm (median: 0.33; SD: 0.12) and 0.37 mm (median: 0.33; SD: 0.19), respectively. The retention force was approximately 2.0 to 2.8 and 1.6 to 1.8 times greater in the auxiliary group than in the nonauxiliary group at 4 and 8 weeks, respectively. CONCLUSIONS: The auxiliary device improved the mechanical retention force without the need to increase miniscrew length or diameter. This may enable the safe use of miniscrews in difficult areas.

Vagal afferent activation induces salivation and swallowing-like events in anesthetized rats.

The aim of this study was to clarify the effect of vagal afferent activation on salivation and swallowing-like events. Salivation is part of a reflex induced by stimulation of the oral area during feeding or chewing. Recently, we reported that nausea induced by gastroesophageal reflux (GER) activation produced salivation and swallowing in humans. Here, we investigated the ability of visceral sensation to enhance salivation and swallowing in rodents in order to inform the mechanism of GER-mediated stomatognathic activation. First, we administered LiCl to anesthetized male rats to induce nausea. LiCl significantly increased salivation and increased the activity of the vagal afferent nerve. Next, we simultaneously recorded salivation and swallowing using an electrode attached to the mylohyoid muscle during vagal afferent stimulation in a physiological range of frequencies. Vagal afferent stimulation significantly increased salivation and swallowing-like events in a frequency-dependent manner. A muscle relaxant, vecuronium bromide, diminished the swallowing-like response but did not affect salivation. These results indicate that visceral sensation induces salivation and swallowing-like events in anesthetized rodents through vagal afferent activation.

Development of a novel spike-like auxiliary skeletal anchorage device to enhance miniscrew stability.

INTRODUCTION: Miniscrews are frequently used for skeletal anchorage during edgewise treatment, and their clinical use has been verified. However, their disadvantage is an approximately 15% failure rate, which is primarily attributed to the low mechanical stability between the miniscrew and cortical bone and to the miniscrew's close proximity to the dental root. To solve these problems, we developed a novel spike-like auxiliary skeletal anchorage device for use with a miniscrew to increase its stability. METHODS: The retention force was compared between miniscrews with and without the auxiliary skeletal anchorage device at each displacement of the miniscrew. The combined unit was also implanted into the bones of 2 rabbits in vivo, and implantation was visually assessed at 4 weeks postoperatively while the compression force was applied. RESULTS: The retention force of the combined unit was significantly and approximately 3 to 5 times stronger on average than that of the miniscrew alone at each displacement. The spiked portion of the auxiliary anchorage device embedded into the cortical bone of the hind limb at approximately a 0.3-mm depth at 4 weeks postimplantation in both rabbits. CONCLUSIONS: The auxiliary skeletal anchorage device may increase miniscrew stability, allow a shortened miniscrew, and enable 3-dimensional absolute anchorage. Further evaluation of its clinical application is necessary.

Replacement of a first molar and 3 second molars by the mesial inclination of 4 impacted third molars in an adult with a Class II Division 1 malocclusion.

This case report presents the successful replacement of 1 first molar and 3 second molars by the mesial inclination of 4 impacted third molars. A woman, 23 years 6 months old, had a chief complaint of crowding of her anterior teeth and linguoclination of a second molar on the left side. The panoramic radiographic images showed that the maxillary and mandibular third molars on both sides were impacted. Root resorption on the distal surfaces of the maxillary second molars was suspected. The patient was given a diagnosis of Angle Class II Division 1 malocclusion with severe crowding of the anterior teeth and 4 impacted third molars. After we extracted the treated maxillary second premolars and the second molars on both sides, the treated mandibular second premolar and the second molar on the left side, and the root canal-filled mandibular first molar on the right side, the 4 impacted third molars were uprighted and formed part of the posterior functional occlusion. The total active treatment period was 39 months. The maxillary and mandibular third molars on both sides successfully replaced the first and second molars. The replacement of a damaged molar by an impacted third molar is a useful treatment option for using sound teeth.

Hesperidin potentiates ghrelin signaling.

Hesperidin, a flavanone glycoside consisting of the flavone hesperitin bound to the disaccharide rutinose, is found in highly nutritious foods, such as oranges, tangelos, tangerines, grapefruits, and other citrus fruits. Exogenous hesperidin has been shown to influence a wide variety of biological functions, including induction of apoptosis and suppression of proliferation in human cancer cells; inhibition of tumor development in various tissues; and expression of antibacterial, antiviral, and antifungal activities. Previous in vivo studies have revealed that hesperidin may play a role in ghrelin secretion from the stomach through antagonism of the serotonin receptors. Because ghrelin appears to be involved in the pathophysiological mechanisms of several human disorders, hesperidin could be a promising target for the treatment of various diseases. This review addresses studies focused on the orexigenic and prokinetic activities of hesperidin in the context of ghrelin secretion. This article also presents some promising patents of hesperidin.

Association of problem behavior with sleep problems and gastroesophageal reflux symptoms.

BACKGROUND: There are few large-scale epidemiologic studies examining the associations between sleep problems, gastroesophageal reflux disease (GERD) symptoms, lifestyle and food habits and problem behaviors (PB) in adolescents. The aim of this study was to evaluate the associations among these factors in Japanese adolescents. METHODS: A cross-sectional survey of 1840 junior high school students was carried out using questionnaires. The subjects were classified into PB or normal behavior (NB) groups using the Pediatric Symptom Checklist (PSC). The scores of the sleep-related factors, sleep bruxism, lifestyle and food habits, and GERD symptoms were compared. Logistic regression analysis was used to determine the factors related to PB. RESULTS: Mean subject age was 13.3 ± 1.8 years. The PB group had significantly longer sleep latency and higher GERD symptom score (P < 0.001). Furthermore, the PB group was significantly more likely to experience absence of the mother at dinner time, skip breakfast, and have <30 min of conversation among family at dinner time. The PB group had significantly higher frequencies of sleep bruxism, difficulty falling asleep within 30 min, nightmares, feeling of low sleep quality, daytime somnolence, and daytime lack of motivation. Feelings of low sleep quality had the strongest association with PB, with an adjusted odds ratio of 12.88 (95% confidence interval: 8.99-18.46). CONCLUSIONS: PB in adolescents are associated with sleep problems, including sleep bruxism, as well as lifestyle and food habits and GERD symptoms.

The role of ghrelin in patients with functional dyspepsia and its potential clinical relevance (Review).

Functional dyspepsia (FD) is a functional gastrointestinal disorder (FGID). According to the Rome III consensus, FD is divided into 2 subgroups: epigastric pain syndrome (EPS) and postprandial distress syndrome (PDS). Although multiple mechanisms of FD pathogenesis have been suggested, its underlying etiology and pharmacological therapy remain unclear. Ghrelin is a gut-derived peptide found in the stomach. It plays a role in the regulation of gastric motility and appetite. The ghrelin gene encodes 3 molecular forms, acyl ghrelin, des-acyl ghrelin and obestatin. Acyl ghrelin acts as an endogenous ligand for growth hormone secretagogue receptor; furthermore, it is orexigenic, with effects on food intake, energy homeostasis and gastrointestinal motility. Des-acyl ghrelin exerts an opposite effect to acyl ghrelin. Obestatin exerts an inhibitory effect on the motor activity of the antrum and duodenum in fed animals. These peptides exert differential effects on gut motility and food intake. The therapeutic potential of ghrelin has attracted attention due to its varied bioactivities. Certain studies have shown that total ghrelin levels are significantly lower in patients with FD compared with healthy volunteers and that the acyl ghrelin levels of patients with FD are higher compared with healthy volunteers. However, a recent study demonstrated that acyl ghrelin levels in patients with PDS were lower compared with healthy volunteers; the association between FD and other ghrelin family gene products also remains unclear. Although certain studies have demonstrated the beneficial effects of acyl ghrelin administration and its agonist in patients with FD, only a few clinical reports exist. Further studies are required in order to examine the effects of ghrelin on FD.

The role of zinc in the treatment of taste disorders.

In the 1990s the number of patients diagnosed with taste disorders in the USA and Japan was over one million people each year, and the number is increasing annually. Taste disorders are caused by several factors such as genetic disease, head trauma, structural changes, glossodynia, cancer, change of lifestyle, and more. The role of zinc in the treatment of taste disorders has been studied since the oral administration of zinc by patients was reported to improve their taste disorders. Carbonic anhydrase (CA), a zinc metalloenzyme, has also been studied in association with taste disorders, since the regulation of serum CA levels was shown to influence the effect of orally administrated zinc in the treatment of taste disorders. Zinc is an essential trace element that contributes to the active center of approximately 300 enzymes. Studies have revealed that zinc is involved in various physiological functions. Moreover, some medications have been shown to induce a zinc deficiency, which has been associated with a variety of clinical conditions. Hence, since the relationship between taste disorder and serum zinc concentration has been discussed for long time, taste disorder may be useful in diagnosing zinc deficiency. Moreover, it appears that medicines of the zinc-containing supplement type contribute to the treatment of taste disorders caused by zinc deficiency. Orally administered zinc has been shown to directly stimulate food intake via neuropeptide in the hypothalamus. Therefore, zinc administration may potentially be used to treat taste disorders, as well as several other diseases by stimulating feeding. The article presents some promising patents on the role of zinc in the treatment of taste disorders.

The roles of salivary secretion, brain-gut peptides, and oral hygiene in obesity.

Obesity has a prevalence of 15-30% among European and American populations. It is an incurable chronic disease associated with considerable mortality and co-morbidity. The co-morbidity risk can be reduced substantially by a moderate weight loss of 5-15%. Notably, additional weight gain exacerbates the morbidity of any concurrent disease. Obesity is also recognized as the basis for metabolic syndrome. Recent research has shown that adipocytes secrete various hormones and cytokines that contribute to obesity. Leptin is an adipostatic hormone that acts on receptors in the hypothalamus to suppress food intake and increase energy consumption. Reduced sensitivity to this molecule can trigger the onset of obesity. Neuropeptides such as leptin also affect salivary secretion. Various neuropeptides have been identified in saliva; the associated receptors are located in the salivary glands or in the nerves innervating the salivary glands. Obesity is associated with hyposalivation and thereby related to several aspects of oral health, such as caries and periodontitis. Hyposalivation is a severe morbidity that can lead to a precipitous decline in oral hygiene, which further leads to multifocal dental caries and periodontitis, or even cardiac disorders. In this article, we review the relationship between salivary secretion and neuropeptides known to play a role in obesity.

The role of ghrelin, salivary secretions, and dental care in eating disorders.

Eating disorders, including anorexia and bulimia nervosa, are potentially life-threatening syndromes characterized by severe disturbances in eating behavior. An effective treatment strategy for these conditions remains to be established, as patients with eating disorders tend to suffer from multiple relapses. Because ghrelin was originally discovered in the stomach mucosa, it has been widely studied over the past decade in an effort to uncover its potential roles; these studies have shed light on the mechanism by which ghrelin regulates food intake. Thus, studying ghrelin in the context of eating disorders could improve our understanding of the pathogenesis of eating disorders, possibly resulting in a promising new pharmacological treatment strategy for these patients. In addition, early detection and treatment of eating disorders are critical for ensuring recovery of young patients. Oral symptoms, including mucosal, dental, and saliva abnormalities, are typically observed in the early stages of eating disorders. Although oral care is not directly related to the treatment of eating disorders, knowledge of the oral manifestations of eating disorder patients may aid in early detection, resulting in earlier treatment; thus, oral care might contribute to overall patient management and prognosis. Moreover, ghrelin has also been found in saliva, which may be responsible for oral hygiene and digestion-related functions. This review discusses the pharmacological potential of ghrelin in regulating food-intake and the role of saliva and oral care in young patients with eating disorders.

Patient with oculo-facio-cardio-dental syndrome treated with surgical orthodontics.

Oculo-facio-cardio-dental (OFCD) syndrome is a rare syndrome characterized by ocular, facial, cardiac, and dental disorders. Only about 20 cases have been reported to date. The most prominent of the various features of this syndrome is canine radiculomegaly. Other features include a long and narrow face, a high nasal bridge, a broad and pointed nose, a bifid nose, ear deformity, cleft palate or submucous cleft palate, maxillary growth retardation, a large gonial angle, open apices, delayed eruption, persistent deciduous teeth, extreme overbite, and constricted maxilla. Orthodontic and prosthodontic treatment has been reported for several patients, but surgical-orthodontic treatment for OFCD has not been reported. An 18-year-old woman with skeletal Class III malocclusion and OFCD syndrome was treated with edgewise appliance therapy combined with orthognathic surgery. We applied a light force during the treatment so as not to induce ankylosis. At the end of the surgical and orthodontic treatments, functional occlusion and an improved facial profile were achieved. After the retention period, stomatognathic function was improved. The results of this treatment suggest that surgical-orthodontic treatment is an effective method for improving skeletal disharmony, facial profile, occlusion, and stomatognathic function in patients with OFCD.

Facial frontal morphological changes related to mandibular setback osteotomy using cephalograms.

AIM: The purpose of this study was to examine morphological changes of the mandibular region using frontal cephalograms following bilateral sagittal split osteotomy (BSSO) for mandibular setback. Furthermore, we compared the stability between screw and plate fixation. METHODS: Pre and postoperative frontal cephalograms were taken of 26 patients. Parameters, Angle "A" which is formed by constructed lines along the mandibular ramus, and mandibular width at the angle were calculated. The subjects were analyzed according to the types of bony fixation, whether by screws or plates. RESULTS: Angle "A" and mandibular angular width of total numbers did not widely change at early period following operation. Late postoperatively, mandibular width of total numbers decreased in conjunction with increasing Angle "A". When comparing the parameters between screw and plate groups, the plate group showed a decrease of Angle "A" as an early change. Late postoperatively, the plate group showed an increase of Angle "A" and a decrease of mandibular width, while the screw group had smaller changes. CONCLUSION: We demonstrate postoperative inclination of mandibular lower ramus towards the facial midline. We suggest that the types of rigid fixation have an influence on frontal skeletal morphology, namely that screw fixation has higher stability.

Intermediate fixed coronary artery stenosis at the site of ergonovine-provoked spasm as a predictor of long-term major adverse cardiac events of patients with coronary spastic angina.

BACKGROUND: Organic coronary artery stenosis is a significant prognostic factor in patients with coronary spastic angina (CSA), so the present study was focused on assessing the impact of intermediate fixed stenosis at sites of provoked spasm on the long-term outcomes of CSA patients. METHODS AND RESULTS: CSA patients diagnosed on the basis of ergonovine-provoked spasm were enrolled and the clinical background and long-term prognosis of CSA patients with intermediate fixed stenosis at the site of provoked spasm (with-fixed-stenosis group, n=37) and those without fixed stenosis (without-fixed-stenosis group, n=126) were retrospectively compared. During the follow-up period (average 4.01 years for with-fixed-stenosis, 4.47 years for without-fixed-stenosis), the with-fixed-stenosis group had a significantly lower event-free survival rate, as well as a higher frequency of admission for unstable angina and percutaneous coronary intervention than the without-fixed-stenosis group, whereas the survival rate did not differ significantly between the 2 groups. In the multivariate analysis, intermediate fixed stenosis at the site of provoked spasm was a predictor of long-term major adverse cardiac events (MACE). CONCLUSIONS: Intermediate fixed stenosis at the site of ergonovine-provoked spasm is an independent risk factor for MACE during the long-term period in CSA patients.