Synthesis of 1,3-Dibromopyrene as Precursor of 1-, 3-, 6-, and 8-Substituted Long-Axially Symmetric Pyrene Derivatives.

Pyrene derivatives bearing substituents at positions 1, 3, 6, and 8 find numerous applications, as exemplified by their use in lasers, sensors, and bioimaging probes. However, these derivatives typically have point-symmetric or short-axially symmetric structures, whereas long-axially symmetric derivatives remain underexplored because of the difficulty in obtaining their precursor, 1,3-dibromopyrene. To address this problem, we herein synthesized 1,3-dibromopyrene from 1-methoxypyrene in an overall yield (71 % over four steps) considerably exceeding those of existing methods. 1,3-Dibromopyrene was converted into 13OPA, a long-axially symmetric pyrene dye with electron-donor (alkoxy) groups at positions 1 and 3 and electron-acceptor (formyl) groups at positions 6 and 8. 13OPA exhibited photophysical properties distinct from those of its point-symmetric and short-axially symmetric isomers, featuring a broad and strongly redshifted absorption, strong fluorescence with reduced sensitivity to protic solvents, and small dipole moment change upon photoexcitation. The derivatization of 13OPA into a Schiff base and its functionalization via Lewis acid-base pairing were also demonstrated. Thus, our work expands the design scope of pyrene-based molecules, particularly those used as emitters.

Visual snake aversion in Octodon degus and C57BL/6 mice.

Phobia against spiders or snakes is common in humans, and similar phobia-like behaviors have been observed in non-human animals. Visual images of snakes elicit phobia in humans, but sensory modalities that cause snake aversion in non-human animals are not well examined. In this study, we examined visually induced snake aversion in two rodent species. Using a three-compartment experimental chamber, reactions to images of snakes were compared between the diurnal precocious rodent Octodon degus and nocturnal laboratory mice. The snakes whose images were presented do not live in the original habitats of degus or mice. Snake aversion was assessed by presenting snake vs. no-image, snake vs. flower, snake vs. degu, and snake vs. mouse images. The time spent in a compartment with the snake image and with the non-snake images were measured. Degus avoided images of snakes in every tests. In contrast, mice did not display snake aversion. Degus are diurnal animals, i.e., visual information is important for their survival. Since mice are nocturnal, visual information is less important for survival. Such behavioral differences in the two species may explain the difference in visually induced aversion to snakes. A principal component analysis of the stimulus images suggests that elementary cues, such as color, do not explain the differences in the species' aversion to snakes. Finally, snake aversion in degus suggests that aversion is innate, since the animals were born and raised in a laboratory.

Preference and discrimination of facial expressions of humans, rats, and mice by C57 mice.

Social animals likely recognize emotional expressions in other animals. Recent studies suggest that mice can visually perceive emotional expressions of other mice. In the first experiment, we measured the preference of mice for two different facial expressions (a normal facial expression and an expression of negative emotion such as pain) of rats, mice, and humans. Results revealed that mice showed a slight preference for the normal expression over the face expressing pain in the case of rats, but no preference in the case of others. In the second experiment, we trained mice to discriminate between the two facial expressions in an operant chamber with a touch screen. They could discriminate facial expressions of mice and rats, but they did not show discrimination of human facial expressions. Principal component analysis of the images of stimuli reveals negative correlation between pixel-based dissimilarity of training stimuli and the number of sessions to criterion. The mice showed generalization to novel images of the mouse faces with and without pain but did not maintain their discriminative behavior when new rat faces were shown. These results suggest that mice display category discrimination of conspecific facial expressions but not of other species.

Spatial learning in Japanese eels (Anguilla japonica).

Japanese eels (Anguilla japonica) were trained on a Morris-type spatial learning task. There were four tubes in a pool, but the eels could hide in only one of these. The eels learned the position of the open tube, and maintained their performance when the pool was rotated to remove possible intra-maze cues. The eels could not maintain their performance in a dark room, suggesting that spatial learning involved extra-maze visual cues. When the position of the open tube was randomly changed every day, the performance of the eels in finding the open tube did not improve.

Discrimination of artificial starry sky by pigeons.

Night migratory birds use the star compass for their navigation. Here, I examined discrimination of a starry sky by a nonmigratory bird, the pigeon. Four pigeons were trained in an operant chamber to discriminate between an artificial starry sky created using software (Stella Theater Pro) and a black sky without stars. After they learned the task, they received three generalization tests. Test 1 presented a starry sky with different classes of stars (i.e., the number of stars was changed). Test 2 presented a starry sky at different times (i.e., a horizontal shift of the sky). Test 3 presented a starry sky at different latitudes (i.e., a vertical shift of the sky). The pigeons displayed a clear generalization gradient with a peak shift to the sky with more stars in Test 1 and a peak close to the original sky in Test 3, but almost a flat gradient in Test 2. Therefore, pigeons demonstrated stimulus control using the pattern of the stars, and they were more sensitive to the vertical shift than to the horizontal shift.

The whole is equal to the sum of its parts: Pigeons (Columba livia) and crows (Corvus macrorhynchos) do not perceive emergent configurations.

We previously demonstrated that chimpanzees, like humans, showed better accuracy and faster response time in discriminating visual patterns when the patterns were presented in redundant and uninformative contexts than when they were presented alone. In the present study, we examined the effect of redundant context on pattern discrimination in pigeons (Columba livia) and large-billed crows (Corvus macrorhynchos) using the same task and stimuli as those used in our previous study on chimpanzees. Birds were trained to search for an odd target among homogenous distractors. Each stimulus was presented in one of three ways: (1) alone, (2) with identical context that resulted in emergent configuration to chimpanzees (congruent context), or (3) with identical context that did not result in emergent configuration to chimpanzees (incongruent context). In contrast to the facilitative effect of congruent contexts we previously reported in chimpanzees, the same contexts disrupted target localization performance in both pigeons and crows. These results imply that birds, unlike chimpanzees, do not perceive emergent configurations.

Dark-Field Microscopic Detection of Bacteria using Bacteriophage-Immobilized SiO2@AuNP Core-Shell Nanoparticles.

To replace molecular biological and immunological methods, biosensors have recently been developed for the rapid and sensitive detection of bacteria. Among a wide variety of biological materials, bacteriophages have received increasing attention as promising alternatives to antibodies in biosensor applications. Thus, we herein present a rapid and highly selective detection method for pathogenic bacteria, which combines dark-field light scattering imaging with a plasmonic biosensor system. The plasmonic biosensor system employs bacteriophages as the biorecognition element and the aggregation-induced light scattering signal of gold nanoparticle-assembled silica nanospheres as a signal transducer. Using Staphylococcus aureus strain SA27 as a model analyte, we demonstrated that the plasmonic biosensor system detects S. aureus in the presence of excess Escherichia coli in a highly selective manner. After the sample and the S. aureus phage S13'-conjugated plasmon scattering probe were mixed, S. aureus detection was completed within 15-20 min with a detection limit of 8 × 104 colony forming units per milliliter.

Rapid assessment of the dose-response relationship of methamphetamine using the progressive-dosing procedure.

This paper describes a new method to rapidly obtain dose-response curves for a drug with rewarding properties using the conditioned place preference protocol. In the usual single-dosing procedure, different animals receive single, varying doses. Thus, a large number of animals are required to generate a curve. A new procedure, known as progressive dosing, alternates increasing drug doses with saline. In this way, the same animal can receive multiple tests. The dose-response curves of the rewarding effect of methamphetamine in mice were obtained using both single-dosing and progressive-dosing procedures. Although the progressive-dosing curves were not identical to the single-dosing curves, they showed a similar pattern. The progressive-dosing procedure was replicated with a new set of mice to confirm the reliability of the method and a dose-response curve similar to the previous one was obtained. This new method can reduce the number of animals required and shorten the duration of the experiment.

Impaired Pavlovian predictive learning between temporally phasic but not static events in autism-model strain mice.

Autism-spectrum disorder (ASD) is a multi-aspect developmental disorder characterised by various social and non-social behavioural abnormalities. Using BTBR T+ tf mouse strain (BTBR), a promising animal model displaying a number of behavioural and neural characteristics associated with ASD, we tested the hypothesis that at the core of various symptoms of ASD lies a fundamental deficit in predictive learning between events. In five experiments, we conducted a variety of Pavlovian conditioning tasks, some requiring the establishment of associations between temporally phasic events and others involving static events. BTBR mice were impaired in the acquisition of conditioned magazine approach responses with an appetitive unconditioned stimulus (US) (Experiment 1) and conditioned freezing with an electric shock US (Experiment 2). Both of these tasks had temporally phasic conditioned stimuli (CSs). Conversely, these mice showed normal acquisition of conditioned place preference (CPP), whether the US was a systemic injection of methamphetamine (Experiment 3A) or the presence of food (Experiment 3B). Experiment 4 showed normal acquisition of conditioned taste aversion (CTA) to a flavour-taste compound CS, although BTBR mice still exhibited an abnormal stimulus selection when learning for each element of the compound CS was assessed separately. Experiment 5 revealed a weaker latent inhibition of CTA in BTBR mice. The BTBR mouse's impaired predictive learning between phasic events and intact associations between static events are discussed in terms of dysfunctional contingency-based, but not contiguity-based learning, which may accompany abnormal selective attention to relevant cues. We propose that such dysfunctional contingency learning mechanisms may underlie the development of various social and non-social symptoms of ASD.

Preference for and discrimination of videos of conspecific social behavior in mice.

We showed mice videos of three conspecific social behaviors, namely sniffing, copulation, and fighting, in pairwise combinations using iPods and evaluated preference as determined by time spent in front of each iPod. Mice preferred the copulation video to the sniffing video, the fighting video to the sniffing video, and the fighting video to the copulation video. In Experiment 1a, we used a single video clip for each social behavior but used multiple video clips for each social behavior in Experiment 2a. Next, we trained mice to discriminate between the fighting and copulation videos using a conditioned-place-preference-like task in which one video was associated with injection of morphine and the other was not. For half of the subjects, the fighting video was associated with morphine injection, and for the other half, the copulation video was associated with morphine injection. After conditioning, the mice stayed longer in the compartment with the morphine-associated video. When tested with still images obtained from the videos, mice stayed longer in the compartment with still images from the video associated with morphine injection (Experiment 1b). When we trained mice with multiple exemplars, the subjects showed generalization of preference for new video clips never shown during conditioning (Experiment 2b). These results demonstrate that mice had a preference among videos of particular behavior patterns and that they could discriminate these videos as visual category. Although relationship between real social behaviors and their videos is still open question, the preference tests suggest that the mice perceived the videos as meaningful stimuli.

Pigeons (Columba livia) fail to connect dots in learning biological motion.

Biological motion point-light displays provide a powerful method for studying motion perception. Nonhuman animals are capable of discriminating point-light displays, but it remains unknown how they perceive biological motion in these displays. We trained two groups of pigeons to discriminate video stimuli using two different classification rules. The motion-congruent group was trained to discriminate full-detail and corresponding point-light displays of pigeons from full-detail and point-light displays of humans. The motion-incongruent group was trained to discriminate full-detail pigeons and point-light humans from the other displays. Both groups acquired the discrimination. When tested with novel displays, pigeons showed good transfer of learning. Transfer was poorest with the point-light displays in the motion-congruent group. The results indicate that the pigeons failed to make the connection between the full-detail displays and their point-light counterparts even when the common motion was available as a cue.

Air flow cued spatial learning in mice.

Spatial learning experiments in rodents typically employ visual cues that are associated with a goal place, even though it is now well established that they have poor visual acuity. We assessed here the possibility of spatial learning in mice based on an air flow cue in a dry version of the Morris water maze task. A miniature fan was placed at each of the four cardinal points of the circular maze, but only one blew air towards the centre of the maze. The three other fans were blowing towards their own box. The mice were able to learn the task only if the spatial relationship between the air flow cue and the position of the goal place was kept constant across trials. A change of this spatial relationship resulted in an increase in the time to find the goal place. We report here the first evidence of spatial learning relying on an air flow cue.

Common experience modifies the reinforcing properties of methamphetamine-injected cage mates but not morphine-injected cage mates in C57 mice.

The aim of this study was to determine whether previous exposure to a drug affects the social facilitation of conditioned place preference (CPP) for a drug-injected cage mate. Twenty-two male C57/BL6J mice received drug injections (methamphetamine or morphine) and 22 male C57/BL6J mice received saline injections. All 44 mice then received CPP training, during which one compartment of a conventional CPP apparatus was associated with a drug-injected cage mate (stimulus mouse) and the other compartment was associated with a saline-injected cage mate (stimulus mouse). The subject mice did not receive any drug injection during this CPP training. Time spent in the compartment associated with the drug-injected cage mate was measured before and after training. Subject mice that had previously received methamphetamine injections showed an increase in the time spent in the compartment associated with the methamphetamine-injected cage mate after CPP training. This effect was not observed in subject mice that had previously received saline injections. Subject mice did not show an increase in the time spent in the compartment associated with the morphine-injected cage mate irrespective of whether they had previously received morphine or saline injections. Therefore, in agreement with previous reports, common experience with methamphetamine induced reinforcing properties, but that with morphine did not.

Long-term follow-up after non-operative management of biloma due to blunt liver injury.

INTRODUCTION: Some case series have reported that hepatectomy was used to treat major bile leakage and biloma. However, it is unknown whether non-operative management (NOM) can be used to treat these complications. Our hospital uses NOM primarily for blunt liver injuries. This study describes the incidence and treatment of newly developed biloma in hemodynamically stable patients with blunt liver trauma and investigates NOM as a treatment option. METHODS: A retrospective chart review was conducted from January 2006 to May 2012 at a tertiary care hospital in Japan. The primary outcome measures were the incidence of biloma and the number of patients who required operative management. Biloma was defined as a cystic lesion with low density near the site of liver injury on contrast-enhanced abdominal computed tomography. RESULTS: Chart review identified 98 patients (63 males and 35 females). Thirty-five of 98 patients (35.7 % [95 % CI, 26.2-45.2]) developed biloma. Infected biloma in three, of whom one required percutaneous drainage. Hepatectomy was not performed. CONCLUSION: Our data suggest that biloma after NOM of blunt liver injury is common (36 %), but infected biloma is rare. All patients with bilomas were treated using NOM. Most bilomas are self-limited, and NOM is feasible.

Tissue gadolinium deposition in hepatorenally impaired rats exposed to Gd-EOB-DTPA: evaluation with inductively coupled plasma mass spectrometry (ICP-MS).

PURPOSE: This study was undertaken to quantify tissue gadolinium (Gd) deposition in hepatorenally impaired rats exposed to gadolinium ethoxybenzyl diethylenetriamine pentaacetic acid (Gd-EOB-DTPA) by means of inductively coupled plasma mass spectrometry (ICP-MS) and to compare differences in Gd distribution among major organs as possible triggers for nephrogenic systemic fibrosis. MATERIALS AND METHODS: Five hepatorenally impaired rats (5/6-nephrectomized, with carbon-tetrachloride-induced liver fibrosis) were injected with Gd-EOB-DTPA. Histological assessment was conducted and Gd content of the skin, liver, kidneys, lungs, heart, spleen, diaphragm, and femoral muscle was measured by inductively coupled plasma mass spectrometry (ICP-MS) at 7 days after last injection. In addition, five renally impaired rats were injected with Gd-EOB-DTPA and the degree of tissue Gd deposition was compared with that in the hepatorenally impaired rats. RESULTS: ICP-MS analysis revealed significantly higher Gd deposition in the kidneys, spleen, and liver (p = 0.009-0.047) in the hepatorenally impaired group (42.6 ± 20.1, 17.2 ± 6.1, 8.4 ± 3.2 µg/g, respectively) than in the renally impaired group (17.2 ± 7.7, 5.4 ± 2.1, 2.8 ± 0.7 µg/g, respectively); no significant difference was found for other organs. In the hepatorenally impaired group, Gd was predominantly deposited in the kidneys, followed by the spleen, liver, lungs, skin, heart, diaphragm, and femoral muscle. Histopathological investigation revealed hepatic fibrosis in the hepatorenally impaired group. CONCLUSIONS: Compared with renally impaired rats, tissue Gd deposition in hepatorenally impaired rats exposed to Gd-EOB-DTPA was significantly increased in the kidneys, spleen, and liver, probably due to the impairment of the dual excretion pathways of the urinary and biliary systems.

Analysis of visual discrimination in Japanese eel (Anguilla japonica).

Japanese eels were trained to discriminate between a checkerboard panel and a plain gray panel in a circular pool with three pipes. One of the pipes was open, whereas the others were closed. The correct choice of discriminative stimulus was reinforced by entering the pipe. When the panels were displayed vertically (on the wall), the eels successfully learned discrimination, but they were unable to acquire the task when the panels were presented horizontally (on the floor). Enucleation of the retina impaired discrimination, whereas ablation of the olfactory plates did not. In the second experiment, the eels underwent three tests after discriminative training with vertical stimuli displayed. When plain black or white panels were presented instead of a checkerboard panel, the eels could not discriminate. Thus, the discriminative stimulus must have both black and white components. The eels exhibited a generalization gradient along the fines of the checkerboard. Finally, the pallium was damaged by coagulation, and the eels did not maintain the discrimination after the lesions. The behavioral deficits were classified into successful relearning and no relearning. Damage to the dorso-lateral (DL) or dorso-central (DC) pallium was associated with severe impairment (no relearning), although it was not possible to isolate the particular brain area or combination of brain areas which was required. The DL damage probably causes memory deficits, but the deficits caused by the DC damage might be motor or motivational deficits.

Spontaneous tempo production in cockatiels (Nymphicus hollandicus) and jungle crows (Corvus macrorhynchos).

Musical and rhythmical abilities are poorly documented in non-human animals. Most of the existing studies focused on synchronisation performances to external rhythms. In humans, studies demonstrated that rhythmical processing (e. g. rhythm discrimination or synchronisation to external rhythm) is dependent of an individual measure: the individual tempo. It is assessed by asking participants to produce an endogenous isochronous rhythm (known as spontaneous motor tempo) without any specific instructions nor temporal cue. In non-human animal literature, studies describing spontaneous and endogenous production of motor tempo without any temporal clue are rare. This exploratory study aims to describe and compare the spontaneous motor tempo of cockatiels and jungle crows. Data were collected on spontaneous beak drumming behaviours of birds housed in laboratory. Inter beak strokes intervals were calculated from sound tracks of videos. The analyses revealed that inter beak strokes intervals are non-randomly distributed intervals and are isochronous. Recorded spontaneous motor tempos are significantly different among some cockatiels. Since we could only conduct statistical analysis with one corvid, we cannot conclude about this species. Our results suggest that cockatiels and jungle crows have individual tempos, thus encouraging further investigations.

Infrared thermography for non-invasive measurement of social inequality aversion in rodents and potential usefulness for future animal-friendly studies.

Infrared thermography is a method that detects thermal radiation energy and can measure the body surface temperature of animals from a distance. While rectal temperature has traditionally been used to measure animals' core temperature, thermal imaging can avoid the stress and potential rise of body temperature deriving from handling of the animals. Additionally, being non-invasive and contactless, thermal imaging allows free movement of the animals. The validity of this technique as a psychophysiological method has been proven in a series of stress-induced hyperthermia (SIH) studies of mice under social inequality conditions. Restraint in a holder elicits SIH in mice. A restrained mouse surrounded by freely moving cage mates displays increased SIH suggesting that social inequality enhances the stress. Social inequality can be examined also in unrestrained mice, in particular through unequal distribution of food. In this protocol, a food-deprived mouse is given a small piece of cheese, while its cage mate is given a large piece of cheese. This inequity causes SIH, suggesting social inequality aversion in mice. Thus, social inequality in different situations similarly increased SIH. Importantly, in future studies infrared thermography could also be used to evaluate emotional arousal states different from stress (for example to assess reactivity to rewards or in social and sexual preference tests). Moreover, the technique could be used to investigate also cognitive arousal induced by novelty. Indeed, infrared thermography could be a particularly useful tool for animal-friendly studies of cognition and emotion in rodents.

A versatile and fast-sampling rate wearable analog data logger.

We propose a wearable, versatile, and open-source data logger that harvests the capacities of a low-cost microcontroller and enables fast-sampling recording of Analog signals into a microSD card. We describe here the circuit design and an exhaustive list of instructions to build a small, lightweight, and fast sampling rate data logger (up to 5 kHz for simultaneous recording of 3 channels and up to 40 kHz when using a single channel). We provide data analysis instructions, including publicly available scripts to facilitate its replication and customization. As a straightforward proof-of-concept, we tested our device embedded with a three-axial Analog accelerometer and were able to record triple axis acceleration of body movements in high resolution. A Fourier transform followed by a principal component analysis discriminated accurately between body motions of two participants and two types of movement recorded (walking VS running). Our wearable and fast-sampling rate data logger overcomes limits that we identified in previous studies, by being low-cost, capable of fast sampling rate, and easily replicated. Moreover, it can be customized to fit with a wide variety of applications in biomedical research by substituting the three-axial Analog accelerometer with virtually any type of Analog sensors or devices that output Analog signals. •We present a method to build and use a low-cost, fast-sampling rate and wearable Analog data logger, where having an engineering background is not required.•The data logger we present can collect Analog signals from 3 channels simultaneously at 5kHz and up to 40 kHz when using a single channel.•We demonstrate that our data logger can record data from a triple axis Analog accelerometer at 5 kHz, however, signals from virtually any Analog sensor or device that outputs Analog signals can be collected.

Crows cross-modally recognize group members but not non-group members.

Recognizing other individuals by integrating different sensory modalities is a crucial ability of social animals, including humans. Although cross-modal individual recognition has been demonstrated in mammals, the extent of its use by birds remains unknown. Herein, we report the first evidence of cross-modal recognition of group members by a highly social bird, the large-billed crow (Corvus macrorhynchos). A cross-modal expectancy violation paradigm was used to test whether crows were sensitive to identity congruence between visual presentation of a group member and the subsequent playback of a contact call. Crows looked more rapidly and for a longer duration when the visual and auditory stimuli were incongruent than when congruent. Moreover, these responses were not observed with non-group member stimuli. These results indicate that crows spontaneously associate visual and auditory information of group members but not of non-group members, which is a demonstration of cross-modal audiovisual recognition of group members in birds.