Nucleobindin 2 inhibits senescence in gastric carcinoma.

Here, we focused on the role of Nucleobindin 2 (NUCB2), a multifunctional protein, in gastric carcinoma (GC) progression. NUCB2 expression was investigated in 150 GC cases (20 non-invasive (pT1) and 130 invasive (pT2/pT3/pT4) tumors) by immunohistochemistry (IHC), and in situ hybridization for detection of the mRNA in 21 cases. Using GC cell lines, we determined whether NUCB2 expression was associated with specific cellular phenotypes. In GC clinical samples, NUCB2 was transcriptionally upregulated when compared to normal tissues. High NUCB2 expression was associated with clinicopathological factors including deep tumor invasion, lymphovascular invasion, lymph node metastasis, and advanced clinical stages, and was a significant independent predictor of unfavorable progression-free survival in 150 non-invasive and invasive GC patients. Similar findings were also evident in 72 invasive GC cases in which patients received post-operative chemotherapy, but not in 58 invasive tumors from patients who did not receive the chemotherapy. In cell lines, NUCB2 knockout inhibited proliferation, susceptibility to apoptosis, and migration capability by inducting cellular senescence; this was consistent with higher proliferation and apoptotic indices in the NUCB2 IHC-high compared to NUCB2 IHC-low GC cases. NUCB2-dependent inhibition of senescence in GC engenders aggressive tumor behavior by modulating proliferation, apoptosis, and migration.

A Complex Interplay between Notch Effectors and ß-Catenin Signaling in Morular Differentiation of Endometrial Carcinoma Cells.

Notch signaling contributes to tissue development and homeostasis, but little is known about its role in morular differentiation of endometrial carcinoma (Em Ca) cells. The current study focused on crosstalk between Notch and ß-catenin signaling in Em Ca with morules. Promoters of hairy and enhancer of split 1 (Hes1) and mastermind-like 2 (MAML2) were activated by Notch intracellular domain 1 but not ß-catenin, and a positive feedback loop between Hes1 and MAML2 was observed. Immunoreactivities for nuclear ß-catenin, Hes1, and MAML2, as well as the interaction between ß-catenin and Hes1 or MAML2, were significantly higher in morular lesions compared with surrounding carcinoma in Em Ca. Inhibition of glycogen synthase kinase-3ß (GSK-3ß) increased expression of total nuclear and cytoplasmic GSK-3ß and its phosphorylated forms, as well as Notch intracellular domain 1, Hes1, and active ß-catenin. GSK-3ß inhibition also decreased proliferation and migration, consistent with the response of cells stably overexpressing Hes1. Finally, the nuclear/cytoplasmic GSK-3ß score was significantly higher in morules compared with surrounding carcinoma in Em Ca, and it was positively correlated with nuclear ß-catenin, Hes1, and MAML2 scores. This complex interplay between Notch effectors and ß-catenin signaling through GSK-3ß inhibition contributes to the establishment and maintenance of ß-catenin-mediated morular differentiation, which is, in turn, associated with reduced proliferation and inhibition of migration in Em Ca.

EBP50 Depletion and Nuclear ß-Catenin Accumulation Engender Aggressive Behavior of Colorectal Carcinoma through Induction of Tumor Budding.

Ezin-radixin-moesin-binding phosphoprotein 50 (EBP50) is a scaffold protein that interacts with several partner molecules including ß-catenin. Here, we examined the crosstalk between EBP50 and nuclear catenin during colorectal carcinoma (CRC) progression. In clinical samples, there were no correlations between the subcellular location of EBP50 and any clinicopathological factors. However, EBP50 expression was significantly lower specifically in the outer areas of tumor lesions, in regions where tumor budding (BD) was observed. Low EBP50 expression was also significantly associated with several unfavorable prognostic factors, suggesting that EBP50 depletion rather than its overexpression or subcellular distribution plays an important role in CRC progression. In CRC cell lines, knockout of EBP50 induced epithelial-mesenchymal transition (EMT)-like features, decreased proliferation, accelerated migration capability, and stabilized nuclear ß-catenin due to disruption of the interaction between EBP50 and ß-catenin at the plasma membrane. In addition, Slug expression was significantly higher in outer lesions, particularly in BD areas, and was positively correlated with nuclear ß-catenin status, consistent with ß-catenin-driven transactivation of the Slug promoter. Together, our data suggest that EBP50 depletion releases ß-catenin from the plasma membrane in outer tumor lesions, allowing ß-catenin to accumulate and translocate to the nucleus, where it transactivates the Slug gene to promote EMT. This in turn triggers tumor budding and contributes to the progression of CRC to a more aggressive phase.

Hierarchical object combination and tool use in the great apes and human children.

Object manipulation can be used as a comparative scale of cognitive development among primates, including humans. Combinatory object manipulation is a precursor of tool-using behavior that indicates material intelligence in primates. However, developmental data on it regarding the great apes other than chimpanzees is insufficient. We conducted a longitudinal investigation of humans and chimpanzees as well as a cross-sectional examination of other great-ape infants (two bonobos, three gorillas, and four orangutans) in captive settings by using two kinds of tasks that required either inserting or stacking combinatory action. The four species of great apes and humans demonstrated both types of combinatory object manipulation during infancy. However, the order of development in different types of combinatory object manipulations varied among the great apes. Furthermore, we applied a nesting-cup task to examine the hierarchical complexity in the combinatory strategies of human children and adult chimpanzees. Both of them exhibited highly hierarchical combinations in the nesting-cup task and employed the subassembly strategy, indicating that an action merge may exist not only in human children but also in adult chimpanzees. The results were discussed with reviews of the tool-use literature from the wild great apes. The early acquisition of an inserting action in the chimpanzees may explain the tool utilization commonality reported in wild chimpanzees. The combinatory object manipulation may have worked as an external enhancer to achieve an additional hierarchical complexity in cognition and behavior, eventually leading humans to develop a language system.

Exploring greetings and leave-takings: communication during arrivals and departures by chimpanzees of the Bossou community, Guinea.

In human fission-fusion societies, ritualized non-linguistic signal exchanges that include gestures, vocalizations, and facial expressions are regularly observed at both arrivals (greetings) and departures (leave-takings). These communicative events play an important role in the formation and maintenance of social relationships. Wild chimpanzees also form large communities that split into smaller fluid parties during daily activities, with individuals moving freely between them. However, in chimpanzees only greetings have been reported. This study explores signal exchanges in the Bossou chimpanzee community during fissions (departures) and fusions (arrivals) given an individual's social rank, kinship, position as traveller or party-member, the level of potential threat, and the party size and presence of mature males. We analysed three time periods (1993-1994; 2003-2004; 2013-2014) during which the composition and social hierarchy of the community varied. We show that the occurrence and form of communication during fission and fusion events are mediated by social factors, including rank, kinship, and party size and composition. Individuals were more likely to communicate during fusions than during fissions, communication was more likely to be produced towards a higher-ranking individual and to non-kin individuals, but the tendency to communicate in general increased with an increase in social rank. The presence of more individuals, and in particular mature males, decreased the likelihood of communication. Communication during fusions supported patterns reported in previous studies on greetings, and our results support the argument that, if present, leave-takings are not a common feature of chimpanzee social interactions. Current methodological difficulties regarding the function of declarative signals hinder our ability to discriminate potential parting rituals within communication before departures. Given similar methodological difficulties, we also provide a note of caution in the interpretation of all signals produced during fusions as 'greetings'.

Automated audiovisual behavior recognition in wild primates.

Large video datasets of wild animal behavior are crucial to produce longitudinal research and accelerate conservation efforts; however, large-scale behavior analyses continue to be severely constrained by time and resources. We present a deep convolutional neural network approach and fully automated pipeline to detect and track two audiovisually distinctive actions in wild chimpanzees: buttress drumming and nut cracking. Using camera trap and direct video recordings, we train action recognition models using audio and visual signatures of both behaviors, attaining high average precision (buttress drumming: 0.87 and nut cracking: 0.85), and demonstrate the potential for behavioral analysis using the automatically parsed video. Our approach produces the first automated audiovisual action recognition of wild primate behavior, setting a milestone for exploiting large datasets in ethology and conservation.

Body Site and Body Orientation Preferences during Social Grooming: A Comparison between Wild and Captive Chimpanzees and Bonobos.

Grooming site preferences have been relatively well studied in monkey species in order to investigate the function of social grooming. They are not only influenced by the amount of ectoparasites, but also by different social variables such as the dominance rank between individuals or their levels of affiliation. However, studies on this topic mainly come from monkey species, with almost no report on great apes. This study aimed to explore whether body site and body orientation preferences during social grooming show species-specific differences (bonobos vs. chimpanzees) and environment-specific differences (captivity vs. wild). Results showed that bonobos groomed the head, the front and faced each other more often than chimpanzees, while chimpanzees groomed the back, anogenitals and more frequently in face-to-back positions. Moreover, captive individuals were found to groom facing one another more often than wild ones, whereas wild individuals groomed the back and in face-to-back positions more. While future studies should expand their scope to include more populations per condition, our preliminary 2 by 2 comparison study highlights the influence of (i) species-specific social differences such as social tolerance, social attention and facial communication, and (ii) socioenvironmental constraints such as risk of predation, spatial crowding and levels of hygiene, that might be the two important factors determining the grooming patterns in two Panspecies.

Object sorting into a two-dimensional array in humans and chimpanzees.

Object-sorting tasks have been used as a means of assessing the cognitive development of humans. In order to investigate cognitive development from a comparative perspective, an object-sorting task was conducted in a longitudinal face-to-face situation involving three juvenile/adolescent chimpanzees (7-9 years old) and 17 children (2-5 years old). The subjects were requested to place nine blocks of different categories (distinguished by three colors and three shapes) into the cells of a box arrayed in a three-by-three pattern. Chimpanzees showed complete or partial categorical sorting in 24-43% of pre-cued trials. The youngest children had difficulty in completing a trial by placing all nine blocks into the box. Humans older than 2 years succeeded in making a one-to-one correspondence by placing a block in each cell, while the end-state pattern remained random. The children gradually increased their rate of categorical sorting, where objects of one category were placed in the same row/column; this tendency peaked at 4 years of age. Above this age, the humans spontaneously shifted their sorting strategy to make a completely even configuration (resulting in a Latin square), which may be more cognitively demanding than categorical sorting. While chimpanzees and older children used both color and shape cues for categorical sorting, younger humans preferred to use shape cues. The results of the present study show fundamental similarities between humans and chimpanzees at the basic level of categorical sorting, which indicates that some autonomous rules are applied during object manipulation.

Investigating the Function of Mutual Grooming in Captive Bonobos (Pan paniscus) and Chimpanzees (Pan troglodytes).

Social grooming is often exchanged between individuals in many primate species. Rates of bidirectional (or simultaneous mutual) grooming vary across primate species, and its function is not yet fully understood. For example, mutual grooming is frequent in chimpanzees but rare in most primate species including wild bonobos. There are, however, no quantitative data available in captive bonobos. Therefore, through the direct comparison between captive bonobos and chimpanzees, this study aimed to (i) compare the frequency of mutual grooming between Pan species, (ii) explore and compare the function of mutual grooming, and (iii) discuss the rarity of this behavior in wild bonobo populations. We tested three hypotheses following the previous literature in wild chimpanzees. The social bonding hypothesis states that mutual grooming facilitates the maintenance of strong dyadic bonds. The immediate investment hypothesis states that it serves to signal willingness to invest in the interaction. The switching hypothesis states that mutual grooming serves no function but only occurs as an overlap to change the direction of unidirectional grooming. Our findings strongly supported the immediate investment hypothesis, but not the others. Grooming bouts that included mutual grooming were longer and more equitable than bouts without, illustrating that captive Pan species use mutual grooming to maximize their short-term benefits and increase the social value of their interaction. Captive bonobos performed mutual grooming in similar proportions and for similar functions as captive and wild chimpanzees do. This contrasts with wild bonobos who engage in this behavior only rarely. We suggest that the differences in patterns of mutual grooming between captive and wild bonobos might be explained by different degrees of kinship or by a potential intraspecies variation.

Social influences on grooming site preferences in wild bonobos (Pan paniscus) at Wamba, DRC.

Social grooming is assumed to have both social and hygienic functions. In monkey species, body site preferences during social grooming are related to both the dominance ranks of participating individuals and strength of social bonds. It is suggested that low-ranking groomees and individuals with weak social bonds expose less vulnerable sites of their body because doing so allows them to avoid eye contact, thus reducing the risk of aggression. This study aimed to investigate the influence of (1) the levels of affiliation (through analyses of sex, grooming frequency, and group membership during intergroup encounters), (2) dominance, (3) kinship, and (4) age on grooming site preferences (head, front, back, anogenital region, and limbs) in wild bonobos. Subjects were two groups of wild bonobos living in Wamba, DR Congo. Body site preferences were documented from video clips. Our results illustrate that the levels of affiliation between individuals and their dominance status do not seem to influence body site preferences, contrasting with results in monkey species. However, the selection of body sites was mostly influenced by age and kinship. Mothers were found to groom less accessible sites on their immature offspring's body more than when grooming a non-related immature, suggesting that mothers invest in their offspring more to prevent diseases linked to ectoparasites. Finally, mature individuals groomed the inaccessible back more often than immature individuals, while the latter focused their grooming on the head and front. Overall, this study suggests hygienic functions of social grooming in wild bonobos.

Chimpanzee face recognition from videos in the wild using deep learning.

Video recording is now ubiquitous in the study of animal behavior, but its analysis on a large scale is prohibited by the time and resources needed to manually process large volumes of data. We present a deep convolutional neural network (CNN) approach that provides a fully automated pipeline for face detection, tracking, and recognition of wild chimpanzees from long-term video records. In a 14-year dataset yielding 10 million face images from 23 individuals over 50 hours of footage, we obtained an overall accuracy of 92.5% for identity recognition and 96.2% for sex recognition. Using the identified faces, we generated co-occurrence matrices to trace changes in the social network structure of an aging population. The tools we developed enable easy processing and annotation of video datasets, including those from other species. Such automated analysis unveils the future potential of large-scale longitudinal video archives to address fundamental questions in behavior and conservation.

Water games by mountain gorillas: implications for behavioral development and flexibility-a case report.

Functions of play, which may be performed solo or in a social context, include motor training and behavioral flexibility. Play is often more common in infancy and the juvenile period, although it also occurs in adults of many species. In contrast to social play, few studies have investigated solitary play. Here, we present new empirical data on solitary water play in a subadult and two adult mountain gorillas in Bwindi Impenetrable National Park, Uganda, observed on three different days between January and February 2018. Focal sampling was used to record the behavior of the individuals interacting with water. Movements included vigorous rotation of the arms, splashing the water, tilting the head, making a play face, and sweeping with the hands to create waves on the water surface. One of the episodes represents the first vigorous display of splashing water ever reported for Bwindi gorillas. Our observations highlight three significant components of mountain gorilla development and behavior: play, behavioral flexibility, and exploration.

Behavioral studies and veterinary management of orangutans at Bukit Merah Orang Utan Island, Perak, Malaysia.

The Bukit Merah Orang Utan Island (OUI) Foundation has been conducting behavioral and veterinary research on orangutans as an attempt at ex situ conservation. Since 2010, the Primate Research Institute, Kyoto University has been collaborating with OUI to promote environmental enrichment and infant rearing by biological mothers in addition to the continuous efforts of refining the veterinary management of the endangered species. In 2011, three Bornean orangutans (Pongo pygmaeus pygmaeus) were released on an island, called BJ Island, adjacent to OUI. This island is approximately 5.6 ha in size, and 635 trees belonging to 102 plant species were identified prior to their release. Behavioral monitoring of the released individuals has been conducted to evaluate their behavioral adaptation to the new environment. Two of the three released orangutans were born in the wild, whereas the youngest individual was born on OUI and expected to learn forest survival strategies from the two older individuals. One of the orangutans was pregnant at the time of release and subsequently gave birth to two male infants on BJ Island. The behavioral monitoring indicated that these orangutans traveled more and spent more time on trees following their release onto BJ Island. However, resting was longer for two females both on OUI and BJ Island when compared to other populations. The orangutans consumed some natural food resources on BJ Island. The release into a more naturalistic environment may help the orangutans to develop more naturalistic behavioral patterns that resemble their wild counterparts.

Mother-infant interactions in captive and wild chimpanzees.

We review studies on mother-infant interactions in chimpanzees, our closest living relatives, in captive and wild environments. Infant cognitive development is formed through mother-infant interactions during the long dependent period, which is approximately 5 years. Patterns of interaction between mothers and infants are different from those observed in adult chimpanzees. Mother-infant interactions are relatively altruistic, although solicitation by infants is almost always required. Active teaching has rarely been reported in chimpanzees; instead, infants socially learn new skills through long-term observation. Case studies describing rearing of a disabled infant by a multiparous but inexperienced chimpanzee, and the carrying of dead infants in the wild chimpanzees of Bossou reveal the strong affectionate bond between mothers and infants. Characteristics of human mother-infant interaction are highlighted through comparisons with chimpanzees.

Carbon and nitrogen stable isotopic offsets between diet and hair/feces in captive chimpanzees.

RATIONALE: Estimation of the stable isotopic offsets between tissue and diet is important for dietary reconstructions. Although stable isotopic studies of chimpanzees (Pan troglodytes) are increasing, the isotopic offsets in chimpanzees have never been studied. In this study, the carbon and nitrogen stable isotopic offset values in hair and feces were measured for 13 captive chimpanzees for the first time. METHODS: All consumed food items and quantities were recorded for each individual for 1 week. Food samples were typically collected three times, hair was collected 3 weeks after the experimental week, and feces were collected ad libitum during the experimental week. The stable carbon and nitrogen isotope ratios were measured using elemental analyzer-isotope ratio mass spectrometry (EA-IRMS). RESULTS: As the results of Monte Carlo analysis, the estimated carbon and nitrogen offsets between the hair and diet were +3.0 to +3.9‰ and +2.8 to +3.7‰, respectively, for the 95% confidence intervals (CIs). The 95% CIs of the carbon and nitrogen offset values between the feces and diet were -1.6 to 0.0‰ and +1.2 to +2.7‰, respectively. CONCLUSIONS: These offset values are generally consistent with those of the other primate species reported in previous studies. However, potential variations in the offset values due to dietary and physiological factors should be studied in detail in the future. Copyright © 2016 John Wiley & Sons, Ltd.

Development and behavior of wild infant-juvenile East Bornean orangutans (Pongo pygmaeus morio) in Danum Valley.

Orangutans have a long period of immaturity and the longest inter-birth interval (IBI) of all mammals, which can be explained by their solitary life style, preventing the mother from rearing two offspring simultaneously (solitary life hypothesis) [corrected]. We collected data on mother-offspring dyads living in a primary lowland forest in Danum Valley, East Borneo in an effort to examine the developmental and behavioral patterns of the subspecies Pongo pygmaeus morio. We analyzed developmental changes in mother-offspring distance, contact, and activity budgets in orangutans ranging from 1 to 7 years of age. The results indicated decreased resting and playing with increasing age, whereas feeding, traveling and social play all increased significantly. Mothers' feeding and traveling time were good predictors of their offspring's feeding and traveling activities. Mother-offspring contact lasted longer in resting contexts; contact during traveling was almost non-existent after 4 years of age. Comparisons with previously published data on the Sumatran species Pongo abelli revealed no fundamental differences in these behavioral measures. However, a shorter association time with the mother after behavioral independence is documented for this East Bornean population in comparison to Sumatran populations. These results are best explained by the solitary life hypothesis, in agreement with previous studies. We suggest that environmental constraints in Bornean forests, as well as a lower population density, should be considered when interpreting the differences between Sumatran and Bornean orangutans in both the period of association with mother and the IBI.

Total Synthesis of the Proposed Structure for Aromin and Its Structural Revision.

This paper describes the first total synthesis of the proposed structure for aromin, an annonaceous acetogenin possessing an unusual bis-THF ring system, and its 4S,7R-isomer. The key steps involve an oxidative cyclization of a couple of terminal-diene alcohols and an intermolecular metathesis of an alkenyl tetrahydrofuran with an enone carrying a tetrahydrofuranyl lactone. The spectral data of both samples did not match those of aromin. Re-examination of the NMR data using the CAST/CNMR Structure Elucidator and chemical derivations suggested that the real structure of aromin should be revised to be a tetrahydropyran acetogenin, montanacin D. Cytotoxicities in human solid tumor cell lines for synthetic samples were also evaluated.

A new method of walking rehabilitation using cognitive tasks in an adult chimpanzee (Pan troglodytes) with a disability: a case study.

There are few studies of long-term care and rehabilitation of animals which acquired physical disabilities in captivity, despite their importance for welfare. An adult male chimpanzee named Reo at the Primate Research Institute of Kyoto University, developed acute myelitis, inflammation of the spinal cord, which resulted in impaired leg function. This report describes a walking rehabilitation system set up in a rehabilitation room where he lives. The rehabilitation apparatus consisted of a touch monitor presenting cognitive tasks and a feeder presenting food rewards at a distance of two meters from the monitor, to encourage him to walk between the monitor and the feeder repeatedly. Initially, Reo did not touch the monitor, therefore we needed adjustment of the apparatus and procedure. After the habituation to the monitor and cognitive tasks, he started to show behaviors of saving food rewards without walking, or stopping participation to the rehabilitation. Finally it took seven phases of the adjustment to determine the final setting; when the monitor automatically displayed trials in 4-h, AM (1000-1200 hours) and PM (1400-1600 hours) sessions through a day, Reo spontaneously walked from the monitor to the feeder to receive rewards, and returned to the monitor to perform the next trial. Comparison of Reo's locomotion in a no-task period and under the final setting revealed that the total travel distance increased from 136.7 to 506.3 m, movement patterns became multiple, and the percentage of walking increased from 1.2 to 27.2 % in PM session. The findings of this case study suggest that cognitive tasks may be a useful way to rehabilitate physically disabled chimpanzees, and thus improve their welfare in captivity.