Exploring Compassion towards Laboratory Animals in UK- and China-Based Undergraduate Biomedical Sciences Students.

Taking a compassionate approach to the non-human animals used in biomedical research is in line with emerging ideas around a "culture of care". It is important to expose biomedical sciences students to the concept of a culture of care at an early stage and give them opportunities to explore related practices and ideas. However, there is no simple tool to explore biomedical sciences students' attitudes towards laboratory animals. Accordingly, there is little understanding of students' feelings towards these animals, or a means of quantifying potential changes to these feelings. We developed a 12-item questionnaire designed to explore compassion (the Laboratory Animal Compassion Scale; LACS) and used it with UK-based and China-based samples of undergraduate biomedical sciences students. In the same samples, we also explored a harm-benefit analysis task and students' beliefs regarding some mental characteristics of laboratory animals, then drew correlations with the quantitative measure of compassion. Compassion levels were stable across years of study and were not related to students' level of experience of working with laboratory animals. We observed a higher level of compassion in females versus males overall, and a higher level overall in the UK-based versus China-based sample. In a task pitting animal suffering against human wellbeing, students' compassion levels correlated negatively with their acceptance of animal suffering. Compassion levels correlated positively with a belief in animals being conscious and possessing emotions. These data are in line with studies that show compassion is gender- and nationality/culture-dependent, and points to links between compassion, beliefs, and choices.

Measuring oxytocin release in response to gavage: Computational modelling and assay validation.

In the present experiments, we tested the conclusion from previous electrophysiological experiments that gavage of sweet food and systemically applied insulin both stimulate oxytocin secretion. To do so, we measured oxytocin secretion from urethane-anaesthetised male rats, and demonstrated a significant increase in secretion in response to gavage of sweetened condensed milk but not isocaloric cream, and a significant increase in response to intravenous injection of insulin. We compared the measurements made in response to sweetened condensed milk with the predictions from a computational model, which we used to predict plasma concentrations of oxytocin from the published electrophysiological responses of oxytocin cells. The prediction from the computational model was very closely aligned to the levels of oxytocin measured in rats in response to gavage.

Neural connectivity between the hypothalamic supramammillary nucleus and appetite- and motivation-related regions of the rat brain.

The supramammillary nucleus (SuM) has an emerging role in appetite control. We have shown that the rat SuM is activated during hunger or food anticipation, or by ghrelin administration. In the present study, we characterised the connectivity between the SuM and key appetite- and motivation-related nuclei in the rat. In adult wild-type rats, or rats expressing Cre recombinase under the control of the tyrosine hydroxylase (TH) promoter (TH-Cre rats), we used c-Fos immunohistochemistry to visualise and correlate the activation of medial SuM (SuMM) with activation in the lateral hypothalamic area (LH), the dorsomedial hypothalamus (DMH) or the ventral tegmental area (VTA) after voluntary consumption of a high-sugar, high-fat food. To determine neuroanatomical connectivity, we used retrograde and anterograde tracing methods to specifically investigate the neuronal inputs and outputs of the SuMM. After consumption of the food there were positive correlations between c-Fos expression in the SuMM and the LH, DMH and VTA (P = 0.0001, 0.01 and 0.004). Using Fluoro-Ruby as a retrograde tracer, we demonstrate the existence of inputs from the LH, DMH, VTA and ventromedial hypothalamus (VMH) to the SuMM. The SuMM showed reciprocal inputs to the LH and DMH, and we identified a TH-positive output from SuMM to DMH. We co-labelled retrogradely-labelled sections for TH in the VMH, or for TH, orexin and melanin-concentrating hormone in the LH and DMH. However, we did not observe any colocalisation of immunoreactivity with any retrogradely-labelled cells. Viral mapping in TH-Cre rats confirms the existence of a reciprocal SuMM-DMH connection and shows that TH-positive cells project from the SuMM and VTA to the lateral septal area and cingulate cortex, respectively. These data provide evidence for the connectivity of the SuMM to brain regions involved in appetite control, and form the foundation for functional and behavioural studies aiming to further characterise the brain circuitry controlling eating behaviours.

Activation of the rat hypothalamic supramammillary nucleus by food anticipation, food restriction or ghrelin administration.

The circulating orexigenic hormone ghrelin targets many brain areas involved in feeding control and signals via a dedicated receptor, the growth hormone secretagogue receptor 1A. One unexplored target area for ghrelin is the supramammillary nucleus (SuM), a hypothalamic area involved in motivation and reinforcement and also recently linked to metabolic control. Given that ghrelin binds to the SuM, we explored whether SuM cells respond to ghrelin and/or are activated when endogenous ghrelin levels are elevated. We found that peripheral ghrelin injection activates SuM cells in rats, reflected by an increase in the number of cells expressing c-Fos protein in this area, as welll as by the predominantly excitatory response of single SuM cells recorded in in vivo electrophysiological studies. Further c-Fos mapping studies reveal that this area is also activated in rats in situations when circulating ghrelin levels are known to be elevated: in food-restricted rats anticipating the consumption of food and in fed rats anticipating the consumption of an energy-dense food. We also show that intra-SuM injection of ghrelin induces a feeding response in rats suggesting that, if peripheral ghrelin is able to access the SuM, it may have direct effects on this brain region. Collectively, our data demonstrate that the SuM is activated when peripheral ghrelin levels are high, further supporting the emerging role for this brain area in metabolic and feeding control.

High-Sugar, but Not High-Fat, Food Activates Supraoptic Nucleus Neurons in the Male Rat.

Oxytocin is a potent anorexigen and is believed to have a role in satiety signaling. We developed rat models to study the activity of oxytocin neurons in response to voluntary consumption or oral gavage of foods using c-Fos immunohistochemistry and in vivo electrophysiology. Using c-Fos expression as an indirect marker of neural activation, we showed that the percentage of magnocellular oxytocin neurons expressing c-Fos increased with voluntary consumption of sweetened condensed milk (SCM). To model the effect of food in the stomach, we gavaged anesthetized rats with SCM. The percentage of supraoptic nucleus and paraventricular nucleus magnocellular oxytocin-immunoreactive neurons expressing c-Fos increased with SCM gavage but not with gastric distention. To further examine the activity of the supraoptic nucleus, we made in vivo electrophysiological recordings from SON neurons, where anesthetized rats were gavaged with SCM or single cream. Pharmacologically identified oxytocin neurons responded to SCM gavage with a linear, proportional, and sustained increase in firing rate, but cream gavage resulted in a transient reduction in firing rate. Blood glucose increased after SCM gavage but not cream gavage. Plasma osmolarity and plasma sodium were unchanged throughout. We show that in response to high-sugar, but not high-fat, food in the stomach, there is an increase in the activity of oxytocin neurons. This does not appear to be a consequence of stomach distention or changes in osmotic pressure. Our data suggest that the presence of specific foods with different macronutrient profiles in the stomach differentially regulates the activity of oxytocin neurons.

The determinants of food choice.

Health nudge interventions to steer people into healthier lifestyles are increasingly applied by governments worldwide, and it is natural to look to such approaches to improve health by altering what people choose to eat. However, to produce policy recommendations that are likely to be effective, we need to be able to make valid predictions about the consequences of proposed interventions, and for this, we need a better understanding of the determinants of food choice. These determinants include dietary components (e.g. highly palatable foods and alcohol), but also diverse cultural and social pressures, cognitive-affective factors (perceived stress, health attitude, anxiety and depression), and familial, genetic and epigenetic influences on personality characteristics. In addition, our choices are influenced by an array of physiological mechanisms, including signals to the brain from the gastrointestinal tract and adipose tissue, which affect not only our hunger and satiety but also our motivation to eat particular nutrients, and the reward we experience from eating. Thus, to develop the evidence base necessary for effective policies, we need to build bridges across different levels of knowledge and understanding. This requires experimental models that can fill in the gaps in our understanding that are needed to inform policy, translational models that connect mechanistic understanding from laboratory studies to the real life human condition, and formal models that encapsulate scientific knowledge from diverse disciplines, and which embed understanding in a way that enables policy-relevant predictions to be made. Here we review recent developments in these areas.

Homeostatic responses to palatable food consumption in satiated rats.

OBJECTIVE: Energy intake is regulated by overlapping homeostatic and hedonic systems. Consumption of palatable foods has been implicated in weight gain, but this assumes that homeostatic control systems do not accurately detect this hedonically driven energy intake. This study tested this assumption, hypothesizing that satiated rats would reduce their voluntary food intake and maintain a stable body weight after consuming a palatable food. METHODS: Lean rats or rats previously exposed to an obesogenic diet were schedule-fed with fixed or varying amounts of palatable sweetened condensed milk (SCM) daily, and their voluntary energy intake and body weight were monitored. RESULTS: During scheduled feeding of SCM, rats voluntarily reduced bland food consumption and maintained a stable body weight. This behavior was also seen in rats with access to an obesogenic diet and was independent of the predictability of SCM access. However, lean rats offered large amounts of SCM showed an increase in total energy intake. To test whether a nutrient deficiency drove this under-compensatory behavior, SCM was enriched with protein. However, no effect was seen on voluntary energy intake. CONCLUSIONS: In schedule-fed rats, compensatory reductions in voluntary energy intake were seen, but under-compensation was observed if large amounts of SCM were consumed.

Dendritic Release of Neurotransmitters.

Release of neuroactive substances by exocytosis from dendrites is surprisingly widespread and is not confined to a particular class of transmitters: it occurs in multiple brain regions, and includes a range of neuropeptides, classical neurotransmitters, and signaling molecules, such as nitric oxide, carbon monoxide, ATP, and arachidonic acid. This review is focused on hypothalamic neuroendocrine cells that release vasopressin and oxytocin and midbrain neurons that release dopamine. For these two model systems, the stimuli, mechanisms, and physiological functions of dendritic release have been explored in greater detail than is yet available for other neurons and neuroactive substances. © 2017 American Physiological Society. Compr Physiol 7:235-252, 2017.

"Eating addiction", rather than "food addiction", better captures addictive-like eating behavior.

"Food addiction" has become a focus of interest for researchers attempting to explain certain processes and/or behaviors that may contribute to the development of obesity. Although the scientific discussion on "food addiction" is in its nascent stage, it has potentially important implications for treatment and prevention strategies. As such, it is important to critically reflect on the appropriateness of the term "food addiction", which combines the concepts of "substance-based" and behavioral addiction. The currently available evidence for a substance-based food addiction is poor, partly because systematic clinical and translational studies are still at an early stage. We do however view both animal and existing human data as consistent with the existence of addictive eating behavior. Accordingly, we stress that similar to other behaviors eating can become an addiction in thus predisposed individuals under specific environmental circumstances. Here, we introduce current diagnostic and neurobiological concepts of substance-related and non-substance-related addictive disorders, and highlight the similarities and dissimilarities between addiction and overeating. We conclude that "food addiction" is a misnomer because of the ambiguous connotation of a substance-related phenomenon. We instead propose the term "eating addiction" to underscore the behavioral addiction to eating; future research should attempt to define the diagnostic criteria for an eating addiction, for which DSM-5 now offers an umbrella via the introduction on Non-Substance-Related Disorders within the category Substance-Related and Addictive Disorders.

Cepheid variables in the flared outer disk of our galaxy.

Flaring and warping of the disk of the Milky Way have been inferred from observations of atomic hydrogen but stars associated with flaring have not hitherto been reported. In the area beyond the Galactic centre the stars are largely hidden from view by dust, and the kinematic distances of the gas cannot be estimated. Thirty-two possible Cepheid stars (young pulsating variable stars) in the direction of the Galactic bulge were recently identified. With their well-calibrated period-luminosity relationships, Cepheid stars are useful distance indicators. When observations of these stars are made in two colours, so that their distance and reddening can be determined simultaneously, the problems of dust obscuration are minimized. Here we report that five of the candidates are classical Cepheid stars. These five stars are distributed from approximately one to two kiloparsecs above and below the plane of the Galaxy, at radial distances of about 13 to 22 kiloparsecs from the centre. The presence of these relatively young (less than 130 million years old) stars so far from the Galactic plane is puzzling, unless they are in the flared outer disk. If so, they may be associated with the outer molecular arm.

Ghrelin, reward and motivation.

Almost all circulating gut peptides contribute to the control of food intake by signalling satiety. One important exception is ghrelin, the only orexigenic peptide hormone thus far described. Ghrelin secretion increases before meals and behavioural and electrophysiological evidence shows that ghrelin acts in the hypothalamus via homeostatic pathways to signal hunger and increase food intake and adiposity. These findings strongly suggest that ghrelin is a dynamically regulated peripheral hunger signal. However, ghrelin also interacts with the brain reward pathways to increase food intake, alter food preference and enhance food reward. Here we discuss ghrelin's role as an endocrine gut-brain reward signal in relation to homeostatic and hedonic feeding control.

Oxytocin, feeding, and satiety.

Oxytocin neurons have a physiological role in food intake and energy balance. Central administration of oxytocin is powerfully anorexigenic, reducing food intake and meal duration. The central mechanisms underlying this effect of oxytocin have become better understood in the past few years. Parvocellular neurons of the paraventricular nucleus project to the caudal brainstem to regulate feeding via autonomic functions including the gastrointestinal vago-vagal reflex. In contrast, magnocellular neurons of the supraoptic and paraventricular nuclei release oxytocin from their dendrites to diffuse to distant hypothalamic targets involved in satiety. The ventromedial hypothalamus, for example, expresses a high density of oxytocin receptors but does not contain detectable oxytocin nerve fibers. Magnocellular neurons represent targets for the anorexigenic neuropeptide α-melanocyte stimulating hormone. In addition to homeostatic control, oxytocin may also have a role in reward-related feeding. Evidence suggests that oxytocin can selectively suppress sugar intake and that it may have a role in limiting the intake of palatable food by inhibiting the reward pathway.

My brain made me do it, and my gut didn't help.

Decisions about what, when and how much to eat are made by the brain, though these choices can be strongly influenced by the hedonic and rewarding properties of sweet or fatty foods. The rumbling before and the fullness after eating tells us that the gut also has an important role in the initiation and termination of feeding. Gut-derived peptides continually convey homeostatic information to the brain to guide feeding. These circulating signals can also modify the pleasure and reward associated with food.

Neural substrates underlying interactions between appetite stress and reward.

Neurobiological mechanisms that normally control food intake and energy expenditure can be overcome by environmental cues and by stress. Of particular importance is the influence of the mesolimbic reward pathway. In genetically susceptible individuals, problematic over-eating likely reflects a changing balance in the control exerted by homeostatic versus reward circuits that are strongly influenced by environmental factors such as stress. Both stress and activation of the reward pathway have been shown to increase food intake and promote a preference for palatable, high-energy foods. Recent research has focused on the important role of circulating and central neuropeptides that powerfully regulate the brain response to food cues. For example, ghrelin has a potent positive effect on the motivational aspects of food intake, and central oxytocin may be involved in satiety. Thus, the decision to eat, or indeed to over-eat, involves a complex integrated neurobiology that includes brain centres involved in energy balance, reward and stress and their regulation by metabolic and endocrine factors.

Peripheral signals modifying food reward.

The pleasure derived from eating may feel like a simple emotion, but the decision to eat, and perhaps more importantly what to eat, involves central pathways linking energy homeostasis and reward and their regulation by metabolic and endocrine factors. Evidence is mounting that modulation of the hedonic aspects of energy balance is under the control of peripheral neuropeptides conventionally associated with homeostatic appetite control. Here, we describe the significance of reward in feeding, the neural substrates underlying the reward pathway and their modification by peptides released into the circulation from peripheral tissues.

Synaptic plasticity in medial vestibular nucleus neurons: comparison with computational requirements of VOR adaptation.

BACKGROUND: Vestibulo-ocular reflex (VOR) gain adaptation, a longstanding experimental model of cerebellar learning, utilizes sites of plasticity in both cerebellar cortex and brainstem. However, the mechanisms by which the activity of cortical Purkinje cells may guide synaptic plasticity in brainstem vestibular neurons are unclear. Theoretical analyses indicate that vestibular plasticity should depend upon the correlation between Purkinje cell and vestibular afferent inputs, so that, in gain-down learning for example, increased cortical activity should induce long-term depression (LTD) at vestibular synapses. METHODOLOGY/PRINCIPAL FINDINGS: Here we expressed this correlational learning rule in its simplest form, as an anti-Hebbian, heterosynaptic spike-timing dependent plasticity interaction between excitatory (vestibular) and inhibitory (floccular) inputs converging on medial vestibular nucleus (MVN) neurons (input-spike-timing dependent plasticity, iSTDP). To test this rule, we stimulated vestibular afferents to evoke EPSCs in rat MVN neurons in vitro. Control EPSC recordings were followed by an induction protocol where membrane hyperpolarizing pulses, mimicking IPSPs evoked by flocculus inputs, were paired with single vestibular nerve stimuli. A robust LTD developed at vestibular synapses when the afferent EPSPs coincided with membrane hyperpolarization, while EPSPs occurring before or after the simulated IPSPs induced no lasting change. Furthermore, the iSTDP rule also successfully predicted the effects of a complex protocol using EPSP trains designed to mimic classical conditioning. CONCLUSIONS: These results, in strong support of theoretical predictions, suggest that the cerebellum alters the strength of vestibular synapses on MVN neurons through hetero-synaptic, anti-Hebbian iSTDP. Since the iSTDP rule does not depend on post-synaptic firing, it suggests a possible mechanism for VOR adaptation without compromising gaze-holding and VOR performance in vivo.

Follow-up data for patients managed by store and forward telemedicine in developing countries.

There is very little published information about the outcomes of patients treated by telemedicine in developing countries. Over a two-year period, seven medical students from five universities spent their electives at a hospital in Papua New Guinea. They assisted with the review of a total of 44 e-referrals made by local doctors; the referrals resulted in 61 queries in a wide range of specialties. The major categories of these queries were internal medicine, paediatrics and surgery. Follow-up data were obtained in 22 of the 44 cases (50%) after a median period of 13 weeks (interquartile range 3-19). The cases were reviewed by an independent doctor. Telemedicine was considered to have assisted with the diagnosis in all cases (median score 5 on a five-point scale from 1 = not helpful at all to 5 = very good/excellent). The advice to the referring doctor for further action was considered helpful in all except one case (median score 5 on the same scale). The outcome for the patient was considered to be good in 15 of the cases (median score 4 on the same scale). Medical students were able to facilitate e-referrals by relieving the pressure on the local doctor to undertake the necessary clerical and technical work. The students reported a rewarding elective experience. The follow-up data showed that low-cost telemedicine can provide useful advice in a low resource setting.

P2X7 subunit-like immunoreactivity in the nucleus of visceral smooth muscle cells of the guinea pig.

P2X receptors are adenosine 5'-triphosphate (ATP)-gated cation-selective channels. Seven cloned subunits (P2X(1-7)) are found in the plasma membrane of many cells in the brain and periphery. ATP-dependent opening of nuclear membrane channels, which resemble P2X(7) receptors, has been reported previously. Therefore, the aim of this study was to use specific antisera to examine P2X(7) receptor subunit expression in the nucleus of cells in visceral organs. In sections of the guinea pig vas deferens and bladder, widespread nuclear immunoreactivity was observed in smooth muscle, epithelial, and lamina propria cells using two antisera raised against different sequences within the P2X(7) subunit. Nuclear immunostaining was also seen in a variety of cell types in the myocardium, lung, stomach, ileum, and colon of the guinea pig. In all tissues, omission of the primary antisera or preincubation of the primary antisera with the antigenic peptide resulted in the loss of specific immunostaining, as did dilution of the antisera beyond 1:32000 or omitting antigen retrieval. Western blot analysis revealed that the anti-P2X(7) antiserum recognised a protein in membrane preparations from both guinea pig vas deferens nuclei and HEK-293 cells transfected with P2X(7) subunits. Thus, the nuclear expression of P2X(7) receptors appears to be widespread in smooth muscle and epithelial cells of visceral organs.