No Association between the Polymorphism rs6943555 in the AUTS2 Gene and Personality Traits in Japanese University Students.

OBJECTIVE: The autism susceptibility candidate 2 (AUTS2) gene has been implicated in multiple neurological disorders. Several recent studies have revealed that the polymorphism rs6943555 in the AUTS2 gene is broadly associated with human mental function and behavior. Therefore, in the present study we investigated whether the polymorphism rs6943555 is associated with human personality traits in Japanese university students. In addition, our previous study reported that the AUTS2 rs6943555-rs9886351 haplotype is associated with alcohol dependence. As a preliminary analysis, we also examined whether the AUTS2 haplotypes are related to personality traits. METHODS: After written informed consent had been obtained from the participants, two AUTS2 polymorphisms were analyzed, and personality was assessed using the Temperament and Character Inventory (TCI) in 190 university students. In addition, in order to exclude the influence of the results for students with mental health problems, we gave the Patient Health Questionnaire-9 (PHQ-9) to all subjects. RESULTS: In all the subjects, there was a main effect of the polymorphism rs6943555 genotype on reward dependence (p=0.038) and cooperativeness (p=0.031), although the significance was lost on Bonferroni correction. Similarly, on analysis that excluded the subjects with PHQ-9 scores≥10, no significant association with any TCI dimension score among the rs6943555 genotypes was seen. There was no effect of the rs6943555-rs9886351 haplotypes on the TCI dimension scores. CONCLUSION: This study suggests that the polymorphism AUTS2 rs6943555 is not associated with personality traits. Further large-scale studies with more subjects using other self-report questionnaires are needed.

Statistical Evaluations of Variations in Dairy Cows' Milk Yields as a Precursor of Earthquakes.

Previous studies have provided quantitative data regarding unusual animal behavior prior to earthquakes; however, few studies include long-term, observational data. Our previous study revealed that the milk yields of dairy cows decreased prior to an extremely large earthquake. To clarify whether the milk yields decrease prior to earthquakes, we examined the relationship between earthquakes of various magnitudes and daily milk yields. The observation period was one year. In the results, cross-correlation analyses revealed a significant negative correlation between earthquake occurrence and milk yields approximately three weeks beforehand. Approximately a week and a half beforehand, a positive correlation was revealed, and the correlation gradually receded to zero as the day of the earthquake approached. Future studies that use data from a longer observation period are needed because this study only considered ten earthquakes and therefore does not have strong statistical power. Additionally, we compared the milk yields with the subionospheric very low frequency/low frequency (VLF/LF) propagation data indicating ionospheric perturbations. The results showed that anomalies of VLF/LF propagation data emerged prior to all of the earthquakes following decreases in milk yields; the milk yields decreased earlier than propagation anomalies. We mention how ultralow frequency magnetic fields are a stimulus that could reduce milk yields. This study suggests that dairy cow milk yields decrease prior to earthquakes, and that they might respond to stimuli emerging earlier than ionospheric perturbations.

Horseback Riding Improves the Ability to Cause the Appropriate Action (Go Reaction) and the Appropriate Self-control (No-Go Reaction) in Children.

BACKGROUND: There are many obvious health benefits to riding, including developing a strong core and legs, but there are also many less obvious benefits, such as increased confidence and introspection. Few studies have addressed the effects of horseback riding on children and the mechanisms underlying how riding affects humans. We examined the effects of horseback riding on the ability to distinguish Go/No-go tasks and solve arithmetic problems in children. METHODS: The subjects were 34 boys and 72 girls, aged 10-12 years old, which were divided into three groups (horse riding, walking, and resting). They were healthy typical children, who performed the Go/No-go tasks and solved the arithmetic problems. The heart rate and heart rate variability of the children, and the three-dimensional acceleration of the children while walking horses, were examined. RESULTS: Riding on a half-breed horse or a pony improved the ability to perform Go/No-go tasks and solve arithmetic problems, possibly through sympathetic activity. Some horses, like the Kiso, might provide a healing effect to children through parasympathetic activity. Statistically significant differences in the three-dimensional acceleration and the autonomic activities were observed among the three horses. The acceleration in the Kiso horse group during walking in hand was significantly different from those involving the other two horses, indicating that the vibrations produced by these horses might modify the autonomic activities. CONCLUSION: The most important beneficial factor of horseback riding for children and for human health appears to be associated with the horse's vibrations, which may differ among horses. Riding some horses may improve the ability of children to respond with an appropriate action depending on the situation (Go reaction) or use self-control appropriately (No-go reaction), possibly through the activation of the sympathetic nervous system.

Increased Feeding Speed Is Associated with Higher Subsequent Sympathetic Activity in Dogs.

Although the domestication process has altered the feeding behavior of dogs, some breeds still demonstrate a remarkable ability to gorge, and will eat exceptionally large quantities of food whenever it is available. Lesions in the ventromedial hypothalamus increase appetite and lead to obesity, suggesting that the autonomic nervous system plays an important role in feeding. Focusing on the autonomic activities closely involved in food intake, we investigated sympathetic activities before and after feeding in dogs. The subjects were 56 healthy dogs of 21 different breeds (29 males and 27 females). Based on feeding habits, the 56 dogs were divided into three groups: Fast (n = 19), Slow (n = 24) and Leftover (n = 13). The feeding speed and the amount of food per mouthful of the Fast dogs were significantly greater than those of the Slow and the Leftover dogs. The plasma norepinephrine level in dogs of the Fast group was significantly increased after feeding, while those in the Slow and Leftover groups were significantly decreased after feeding, compared with the pre-feeding concentrations. The low frequency/high frequency ratio of heart rate variability is a good indicator of sympathetic activity and was also significantly higher in the Fast group than in the other groups. Delayed feeding using automatic feeding equipment decreased the plasma norepinephrine concentration and low frequency/high frequency ratio observed after feeding in dogs of the Fast group. In conclusion, dogs eating rapidly with less chewing, which indicates increased sympathetic activity during feeding, may benefit from delayed feeding. The slow eating may activate the parasympathetic nervous system after feeding, which could enhance the activity of the digestive system.

Analysis of Association between Norepinephrine Transporter Gene Polymorphisms and Personality Traits of NEO-FFI in a Japanese Population.

OBJECTIVE: Norepinephrine is an important chemical messenger that is involved in mood and stress in humans, and is reabsorbed by the norepinephrine transporter (NET). According to Cloninger's theory, the noradrenergic system mediates the personality trait of reward dependence. Thus far, although association studies on NET gene polymorphisms and Cloninger's personality traits have been reported, they yielded inconsistent results. Therefore, in the present study we investigated whether or not the 1287G/A, -182T/C and -3081A/T polymorphisms of the NET gene (SLC6A2) are associated with reward dependence-related traits, as assessed by the five-factor model. METHODS: After written informed consent was obtained from participants, the three NET gene polymorphisms were analyzed by polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP), and personality was assessed by the Neuroticism Extraversion Openness-Five Factor Inventory (NEO-FFI) in 270 Japanese university students. RESULTS: A significant relation was found between the -3081A/T functional promoter polymorphism and NEO-FFI scores: those with the T allele exhibited a lower extraversion (E) score than those without the T allele (Mann-Whitney U-test: z=-3.861, p<0.001). However, there was no correlation between the other NET gene polymorphisms and E score, and no association with other dimensions and these three polymorphisms. CONCLUSION: We conclude that the -3081A/T functional polymorphism in the NET gene may affect the extraversion of reward dependence-related traits, as measured by NEO-FFI. However, we used only the shortened version of NEO-PI-R in this study. Further investigations are necessary using the full version of self-rating personality questionnaires.

Social evolution. Oxytocin-gaze positive loop and the coevolution of human-dog bonds.

Human-like modes of communication, including mutual gaze, in dogs may have been acquired during domestication with humans. We show that gazing behavior from dogs, but not wolves, increased urinary oxytocin concentrations in owners, which consequently facilitated owners' affiliation and increased oxytocin concentration in dogs. Further, nasally administered oxytocin increased gazing behavior in dogs, which in turn increased urinary oxytocin concentrations in owners. These findings support the existence of an interspecies oxytocin-mediated positive loop facilitated and modulated by gazing, which may have supported the coevolution of human-dog bonding by engaging common modes of communicating social attachment.

Experimental Verification of the Effects on Normal Domestic Cats by Feeding Prescription Diet for Decreasing Stress.

The objective of this study was to evaluate the effects of diet on the feline stress response by measuring plasma and urinary cortisol. A study diet was developed with a unique combination of nutrients that supports the management of stressful situations. The specific formulation of the diet included alpha-casozepine, which is believed to have an anxiolytic effect, and tryptophan supplementation. Tryptophan is the precursor for the synthesis of the neurotransmitter serotonin. Twenty-one indoor cats were fed with the study diet (n = 10) or a control diet (n = 11) for 8 weeks, after which physiological responses were evaluated. The study diet significantly increased the ratio of plasma tryptophan to large neutral amino acids and decreased urinary cortisol concentrations after being consumed daily for 8 weeks, but there was no effect on plasma cortisol levels following a stressful event (veterinary examination and blood draw). Further studies, such as behavioral analyses, are needed to clarify the effects of the study diet.

Psychological Evaluation of Animal-assisted Intervention (AAI) Programs Involving Visiting Dogs and Cats for Alcohol Dependents: A Pilot Study.

The purpose of this study was to develop an evaluation method for animal-assisted intervention (AAI) programs involving Mood Check List-Short form.2 (MCL-S.2) and the State-Trait Anxiety Inventory (STAI) for psychiatric daycare of Japanese alcohol. dependents. A total of 36 alcohol dependents completed the study and questionnaires assessing their state. A single session of AAI reduced both subjective and physiological measures of state anxiety (A-State); and this program induced a significant reduction in the anxiety after an AAI program session with the dogs and cats involved in the intervention (p = 0.001). The Wilcoxon t-test showed that there were also significant differences in the "anxiety", "pleasantness", and "relaxation". scores for MCL-S.2 among the alcohol dependents, before and after AAI; a significantly decreased "anxiety" score (p = 0.006), and increased "pleasantness" (p = 0.002) and "relaxation" (p=0.012) scores for MCL-S.2 after AAI. The results of this study indicated that alcohol dependents who experienced a group AAI session-program exhibited significant improvements in their feeling; decreased anxiety, and increased pleasantness and relaxation.

Relationship between alcohol dependence and the DBI rs2276596 (C/A) polymorphism in Japanese.

To facilitate elucidation of the pathogenesis of alcohol dependence, we investigated the relationship between a genetic variant of diazepam biding inhibitor (DBI) C/A polymorphism (rs2276596) and alcohol dependence. We determined the DBI genotypes using a novel method involving PCR-RFLP in healthy controls and alcoholics with a diagnosis of alcohol dependence by ICD-10 (F10.20). There was a significant difference in the rs2276596 polymorphism C/A allele frequency of the DBI gene (P < 0.0001) between alcoholics and healthy controls. The present data suggested that a mutant allele of the DBI was one of the risk factors for alcohol dependence as for the rs2276596 polymorphism.

Dogs' Body Language Relevant to Learning Achievement.

The facial expressions and body postures of dogs can give helpful information about their moods and emotional states. People can more effectively obedience train their dogs if we can identify the mannerisms associated with learning in dogs. The aim of this study was to clarify the dog's body language during operant conditioning to predict achievement in the test that followed by measuring the duration of behaviors. Forty-six untrained dogs (17 males and 26 females) of various breeds were used. Each session consisted of 5 minutes of training with a treat reward followed by 3 minutes of rest and finally an operant conditioning test that consisted of 20 "hand motion" cues. The operant tests were conducted a total of nine times over three consecutive days, and the success numbers were counted. The duration of the dog's behavior, focusing on the dog's eyes, mouth, ears, tail and tail-wagging, was recorded during the operant conditioning sessions before the test. Particular behaviors, including wide-eyes, closed mouth, erect ears, and forward and high tail carriage, without wagging or with short and quick wagging, related to high achievement results. It is concluded that dogs' body language during operant conditioning was related to their success rate.

Unusual Animal Behavior Preceding the 2011 Earthquake off the Pacific Coast of Tohoku, Japan: A Way to Predict the Approach of Large Earthquakes.

Unusual animal behaviors (UABs) have been observed before large earthquakes (EQs), however, their mechanisms are unclear. While information on UABs has been gathered after many EQs, few studies have focused on the ratio of emerged UABs or specific behaviors prior to EQs. On 11 March 2011, an EQ (Mw 9.0) occurred in Japan, which took about twenty thousand lives together with missing and killed persons. We surveyed UABs of pets preceding this EQ using a questionnaire. Additionally, we explored whether dairy cow milk yields varied before this EQ in particular locations. In the results, 236 of 1,259 dog owners and 115 of 703 cat owners observed UABs in their pets, with restless behavior being the most prominent change in both species. Most UABs occurred within one day of the EQ. The UABs showed a precursory relationship with epicentral distance. Interestingly, cow milk yields in a milking facility within 340 km of the epicenter decreased significantly about one week before the EQ. However, cows in facilities farther away showed no significant decreases. Since both the pets' behavior and the dairy cows' milk yields were affected prior to the EQ, with careful observation they could contribute to EQ predictions.

Importance of puppy training for future behavior of the dog.

In this study, we attempted to clarify whether puppy socialization and command training class, which was limited to puppies approximately 4 months of age, prevented behavior problems in dogs. We evaluated the behaviors of 142 dogs with 4 types of training experience by using a behavior test and the Canine Behavior Assessment and Research Questionnaire (C-BARQ). Dogs in the puppy class (PC) group (n=44) attended the class for 1 hr each week for 6 weeks, dogs in the puppy party (PP) group (n=39) attended a 1-hr "puppy party," dogs in the adult class (AC) group (n=27) undertook basic obedience lessons for adolescent/adult dogs for 1 hr each week for 6 weeks, and dogs in the no class (NC) group (n=32) underwent no formal training. The behavior test evaluated each dog's response to commands, owner's recall, separation, a novel stimulus and strangers. The C-BARQ evaluated 15 canine behavioral factors. The behavior test results indicated that the PC and AC groups showed significantly higher response to commands than the PP or NC group. Thus, participation in puppy and adult classes improved the obedience behavior of dogs, regardless of age. Positive response to strangers in the PC group was significantly higher than that in the AC and NC groups and tended to be higher than that in the PP group. Therefore, PC may help prevent canine behavioral problems such as disobedience or fear of strangers.

Evaluation of cerebral activity in the prefrontal cortex in mood [affective] disorders during animal-assisted therapy (AAT) by near-infrared spectroscopy (NIRS): a pilot study.

OBJECTIVE: Previous studies have shown the possibility that animal-assisted therapy (AAT) is useful for promoting the recovery of a patient's psychological, social, and physiological aspect. As a pilot study, we measured the effect that AAT had on cerebral activity using near-infrared spectroscopy (NIRS), and examined whether or not NIRS be used to evaluate the effect of AAT biologically and objectively. METHODS: Two patients with mood [affective] disorders and a healthy subject participated in this study. We performed two AAT and the verbal fluency task (VFT). RESULTS: The NIRS signal during AAT showed great [oxy-Hb] increases in most of the prefrontal cortex (PFC) in the two patients. When the NIRS pattern during AAT was compared with that during VFT, greater or lesser differences were observed between them in all subjects. CONCLUSION: The present study suggested that AAT possibly causes biological and physiological changes in the PFC, and that AAT is useful for inducing the activity of the PFC in patients with depression who have generally been said to exhibit low cerebral activity in the PFC. In addition, the possibility was also suggested that the effect of AAT can be evaluated using NIRS physiologically and objectively.

The seasonal fluctuation of plasma amino acids in aquarium-maintained bottlenose dolphins (Tursiops truncatus).

Although there has been extensive research on plasma amino acid profiles of mammals, there is currently a lack of information on seasonal differences in the concentrations of plasma amino acids specifically in cetaceans. The present study examined the response of the plasma amino acids to seasonal changes in the culture environment after controlling for the effect of sex and age. Significant seasonal changes in plasma carnosine (P=0.012), cystine (P=0.0014), isoleucine (P=0.0042), methionine (P=0.002), ornithine (P=0.0096), and taurine (P=0.032) were observed. These amino acids were mainly related to capacity for exercise, ammonia detoxification, thermoregulation, and osmoregulation. We proposed that optimizing plasma amino acids levels by supplementation of amino acids should be of considerable benefit for aquarium-maintained bottlenose dolphins. This study constitutes a first step towards improving our understanding of the metabolism of aquarium-maintained bottlenose dolphins. We also revealed that the ratio of tryptophan to large neutral amino acids significantly declined (P=0.0076), suggesting reduction in serotonin synthesis in winter and autumn. Although further studies are needed, this finding implied that bottlenose dolphins could produce behavioral changes seasonally by the alteration of serotonin activity. To better understand the metabolic machinery for amino acids that facilitate the adaptation of marine mammals to their environments, it is essential to continue monitoring of and further investigations into relationships between plasma amino acids and specific environmental factors.

Urinary oxytocin as a noninvasive biomarker of positive emotion in dogs.

A reliable assay based on physiological parameters that does not require subjective input from the owners is required to assess positive emotions in dogs. In addition, when viewed from an animal welfare perspective, physiological parameters should be collected in a noninvasive manner. Oxytocin (OT) is a biomarker that may be associated with a calm, relaxed state, and positive emotion. We measured the time-lapse in the concentration of plasma OT relative to urinary OT using a radioimmunoassay with sufficient sensitivity and low variability, and examined the relationship between OT and cortisol. Six dogs were injected with exogenous OT intravenously to increase the blood OT concentration. As a result, the highest concentration of urinary OT occurred 1h after the injection, although there was little change in urinary cortisol. Moreover, to evaluate the influence of stimuli on urinary OT and cortisol, we provided three stimuli of eating food, exercising and stroking, all of which were assumed to inspire a positive emotion in dogs, and significantly increased urinary OT concentrations. Our findings indicate that urinary OT might be useful as a noninvasive and objective biomarker of positive emotion in dogs.

Importance of bringing dogs in contact with children during their socialization period for better behavior.

Much attention has been given to the role that dogs play in the social, physical, and psychological development of children. Furthermore, dogs are now playing a new important role in animal-assisted education (AAE). However, some dogs have difficulty getting along with children. The aim of this study was to demonstrate how dogs' contact with children during and after their socialization period influenced their responses toward children. In total, 31 pet dogs were divided into three groups. Group 1 dogs had been in contact with children during the canine socialization period (between 3 and 12 weeks of age); Group 2 dogs had been in contact with children after this socialization period; Group 3 dogs seldom had contact with children. As soon as a child entered the room, the dog was presented three stimuli by the child over the course of several intervals. The dogs' responses were evaluated by analyzing behavior and heart rates. The dogs that had been in contact with children during their socialization period did not show aggressive behavior or excited behavior toward the child in our test. Moreover, their heart rates were significantly decreased even during the last stimulus, which involved the child running around the dog while calling its name. This study suggested that the dogs having contact with children during socialization period might not show inappropriate responses toward children and could be suitable for taking part in AAE.

Characteristic metabolism of free amino acids in cetacean plasma: cluster analysis and comparison with mice.

From an evolutionary perspective, the ancestors of cetaceans first lived in terrestrial environments prior to adapting to aquatic environments. Whereas anatomical and morphological adaptations to aquatic environments have been well studied, few studies have focused on physiological changes. We focused on plasma amino acid concentrations (aminograms) since they show distinct patterns under various physiological conditions. Plasma and urine aminograms were obtained from bottlenose dolphins, pacific white-sided dolphins, Risso's dolphins, false-killer whales and C57BL/6J and ICR mice. Hierarchical cluster analyses were employed to uncover a multitude of amino acid relationships among different species, which can help us understand the complex interrelations comprising metabolic adaptations. The cetacean aminograms formed a cluster that was markedly distinguishable from the mouse cluster, indicating that cetaceans and terrestrial mammals have quite different metabolic machinery for amino acids. Levels of carnosine and 3-methylhistidine, both of which are antioxidants, were substantially higher in cetaceans. Urea was markedly elevated in cetaceans, whereas the level of urea cycle-related amino acids was lower. Because diving mammals must cope with high rates of reactive oxygen species generation due to alterations in apnea/reoxygenation and ischemia-reperfusion processes, high concentrations of antioxidative amino acids are advantageous. Moreover, shifting the set point of urea cycle may be an adaptation used for body water conservation in the hyperosmotic sea water environment, because urea functions as a major blood osmolyte. Furthermore, since dolphins are kept in many aquariums for observation, the evaluation of these aminograms may provide useful diagnostic indices for the assessment of cetacean health in artificial environments in the future.

The feeding behavior of dogs correlates with their responses to commands.

Motivation is one of the most important factors in dog training. To generate motivation, people use various reinforcer mechanisms. In particular, many pet owners use food because it is simple and convenient. The aim of this study was to investigate the association between dogs' level of interest in food and their responsiveness to commands. Thirty-four dogs were divided into three groups based on their feeding patterns (Fast, Slow, and Leftover). The fast group (n=15) had the highest interest in food and showed a high response to commands when food was used as a reinforcer, rather than praise/stroking. The leftover group (n=6) showed the least interest in food and had a low reaction to commands, regardless of the reinforcer. In contrast, the slow group (n=13) had modest interest in food and maintained the same response to commands regardless of the reinforcer. Results of this study indicate that dogs' feeding patterns are indicative of their level of interest in food, and may be useful in determining the optimal training reinforcer. This can help dog owners improve their relationships with their dogs.

The evaluation of autonomic nervous system activation during learning in rhesus macaques with the analysis of the heart rate variability.

The aims of this study were to measure the activity of the autonomic nervous system using heart rate variability (HRV) during learning tasks and to clarify the relationship between learning to overcome a difficult situation and the autonomic nervous system in monkeys. Two young male monkeys (Macaca mulatta) were given simple discrimination learning tasks (DL) and delayed matching to samples tasks (DMTS); Holter-type electrocardiography was done, and HRV was measured. We defined the frequency bands of HRV in rhesus macaques; the low frequency (LF) was 0.01-0.15 Hz, and the high frequency (HF) was 0.15-0.50 Hz. Based on these frequency bands, the LF/HF ratios during learning tasks were analyzed, and a significant increase in the ratio was found during the tasks. The variances in the HF differed between the DL and DMTS tasks; during DMTS tasks, HF variances had a tendency to increase. Our results indicate that increased sympathetic activity accompanied learning and suggest that the parasympathetic nervous system plays a key role during learning, particularly when difficult tasks are being learned.