The (Under)Use of Eye-Tracking in Evolutionary Ecology.

To survive and pass on their genes, animals must perform many tasks that affect their fitness, such as mate-choice, foraging, and predator avoidance. The ability to make rapid decisions is dependent on the information that needs to be sampled from the environment and how it is processed. We highlight the need to consider visual attention within sensory ecology and advocate the use of eye-tracking methods to better understand how animals prioritise the sampling of information from their environments prior to making a goal-directed decision. We consider ways in which eye-tracking can be used to determine how animals work within attentional constraints and how environmental pressures may exploit these limitations.

Event-related brain potentials during the visuomotor mental rotation task: The contingent negative variation scales to angle of rotation.

Perceptual judgments about the angular disparity of a character from its standard upright (i.e., mental rotation task) result in a concurrent increase in reaction time (RT) and modulation of the amplitude of the P300 event-related brain potential (ERP). It has therefore been proposed that the P300 represents the neural processes associated with a visual rotation. In turn, the visuomotor mental rotation (VMR) task requires reaching to a location that deviates from a target by a predetermined angle. Although the VMR task exhibits a linear increase in RT with increasing oblique angles of rotation, work has not examined whether the task is supported via a visual rotation analogous to its mental rotation task counterpart. This represents a notable issue because seminal work involving non-human primates has ascribed VMR performance to the motor-related rotation of directionally tuned neurons in the primary motor cortex. Here we examined the concurrent behavioral and ERP characteristics of a standard reaching task and VMR tasks of 35°, 70°, and 105° of rotation. Results showed that the P300 amplitude was larger for the standard compared to each VMR task--an effect independent of the angle of rotation. In turn, the amplitude of the contingent negative variation (CNV)--an ERP related to cognitive and visuomotor integration for movement preparation--was systematically modulated with angle of rotation. Thus, we propose that the CNV represents an ERP correlate related to the cognitive and/or visuomotor transformation demands of increasing the angular separation between a stimulus and a movement goal.

Species with a chemical defence, but not chemical offence, live longer.

Evolutionary hypotheses for ageing generally predict that delayed senescence should evolve in organisms that experience lower extrinsic mortality. Thus, one might expect species that are highly toxic or venomous (i.e. chemically protected) will have longer lifespans than related species that are not likewise protected. This remarkable relationship has been suggested to occur in amphibians and snakes. First, we show that chemical protection is highly conserved in several lineages of amphibians and snakes. Therefore, accounting for phylogenetic autocorrelation is critical when conservatively testing evolutionary hypotheses because species may possess similar longevities and defensive attributes simply through shared ancestry. Herein, we compare maximum longevity of chemically protected and nonprotected species, controlling for potential nonindependence of traits among species using recently available phylogenies. Our analyses confirm that longevity is positively correlated with body size in both groups which is consistent with life-history theory. We also show that maximum lifespan was positively associated with chemical protection in amphibian species but not in snakes. Chemical protection is defensive in amphibians, but primarily offensive (involved in prey capture) in snakes. Thus, we find that although chemical defence in amphibians favours long life, there is no evidence that chemical offence in snakes does the same.

What do I do now? An electroencephalographic investigation of the explore/exploit dilemma.

To maximize reward, we are faced with the dilemma of having to balance the exploration of new response options and the exploitation of previous choices. Here, we sought to determine if the event-related brain potential (ERP) in the P300 time range is sensitive to decisions to explore or exploit within the context of a sequential risk-taking task. Specifically, the task we used required participants to continually explore their options-whether they should "push their luck" and keep gambling or "take the money and run" and collect their winnings. Our behavioral analysis yielded two distinct distributions of response times: a larger group of short-decision times and a smaller group of long-decision times. Interestingly, these data suggest that participants adopted one of two modes of control on any given trial: a mode where they quickly decided to keep gambling (i.e. exploit), and a mode where they deliberated whether to the take the money they had already won or continue gambling (i.e. explore). Importantly, we found that the amplitude of the ERP in the P300 time range was larger for explorative decisions than for exploitative decisions and, further, was correlated with decision time. Our results are consistent with a recent theoretical account that links changes in ERP amplitude in the P300 time range with phasic activity of the locus coeruleus-norepinephrine system and decisions to engage in exploratory behavior.

A comparative analysis of senescence in adult damselflies and dragonflies (Odonata).

Any population whose members are subject to extrinsic mortality should exhibit an increase in mortality with age. Nevertheless, the prevailing opinion is that populations of adult damselflies and dragonflies do not exhibit such senescence. Here, we challenge this contention by fitting a range of demographic models to the data on which these earlier conclusions were based. We show that a model with an exponential increase in age-related mortality (Gompertz) generally provides a more parsimonious fit than alternative models including age-independent mortality, indicating that many odonates do indeed senesce. Controlling for phylogeny, a comparison of the daily mortality of 35 odonate species indicates that although male and female mortalities are positively correlated, mortality tends to be higher in males of those species that exhibit territoriality. Hence, we show for the first time that territoriality may impose a survivorship cost on males, once the underlying phylogenetic relationships are accounted for.

Empirical evidence of senescence in adult damselflies (Odonata: Zygoptera).

1. Age-dependent increases in mortality have been documented in a variety of species of insect under laboratory conditions. However, while strong statistical evidence has been presented for senescence in vertebrate populations in the wild, we know little about the rate and shape of senescence in wild populations of insects. 2. Odonates (damselflies and dragonflies) provide excellent candidate species for evaluating demographic senescence as they are large enough to be marked individually and they are easily re-sighted without recapture. The prevailing opinion - based entirely on qualitative examination of the declines in log numbers alive with time since marking - is that odonates exhibit age-independent daily survivorship. 3. Here, we examine mark-recapture data on the Azure Damselfly Coenagrion puella over two consecutive seasons. For the first time, we evaluate and compare the fit of quantitative models that not only account for weather-dependent daily variation in daily re-sighting rates, but also age-dependent variation in daily survivorship. 4. Models with age-dependent declines in daily survivorship provide a more parsimonious explanation for the data than similar models without these age-dependent effects. In general, models in which mortality increases in an exponential (Gompertz) fashion explain the mark-recapture sequences more efficiently than a range of alternative models, including those in which mortality increases as a power function (Weibull) or reaches a plateau (logistic). These results are indicative of a general senescent decline in physiological functioning, which is particularly marked after 15 days as a mature adult. 5. Weather (temperature, sun and precipitation) and initial mite load influenced the probability of daily re-sighting. Weather and mite load also influenced daily survivorship, but their effects differed between seasons. 6. Overall, fitting models with age as an explicit covariate demonstrates that odonates do indeed senesce. This contradicts previously held assumptions that Odonata do not exhibit age-dependent survivorship in the wild.

Neuropeptide Y-immunoreactive intracardiac neurones, granule-containing cells and nerves associated with ganglia and blood vessels in rat and guinea-pig heart.

Intrinsic neuropeptide Y-containing neurones in rat and guinea-pig hearts were studied at the ultrastructural level by the pre-embedding peroxidase-antiperoxidase immunocytochemical technique. Intracardiac neuronal cell bodies were often weakly or moderately immunostained, and the labelling was usually pronounced in the Golgi complex, multivesicular bodies, some cisterns of granular endoplasmic reticulum and large granular vesicles. Neuropeptide Y-immunoreactive nerve fibres were also observed in association with intracardiac neurones. A subpopulation of neuropeptide Y-immunoreactive granule-containing cells in the rat heart are described for the first time and were very heavily labelled; other granule-containing cells were non-immunoreactive, but were contacted by neuropeptide Y-containing nerves. Preterminal regions of nerve fibres that were located in nerve bundles were only weakly neuropeptide Y-immunoreactive, in contrast to the heavy labelling observed in varicosities that contained many synaptic vesicles. Many neuropeptide Y-immunoreactive nerve fibres were associated with the coronary vasculature and were particularly prominent in the walls of small arteries and arterioles where labelled nerve varicosities were present close to the smooth muscle cells. Immunoreactive nerves were also seen in the myocardium, usually near to capillaries. In axonal varicosities, the central core of large granular vesicles was immunolabelled, and electron-dense immunoreactive material outlined the membranes of small and large clear vesicles. The significance of neuropeptide Y-immunoreactive intracardiac neurones and granule-containing cells and the origin of associated labelled nerve fibres in the heart are discussed.

Olestra, a nonabsorbed, noncaloric replacement for dietary fat: a review.

Olestra has been shown to be safe for its intended use by extensive testing in animals and in humans. It is not digested or absorbed and has no effect on the structure or physiology of the GI tract, the only organ of the body that it contacts. Olestra can interfere with the absorption of other lipophilic substances from the GI tract. The interference occurs because a portion of those molecules that are sufficiently lipophilic partition into the nonabsorbed olestra and is carried out of the body. Whether olestra will interfere with the absorption of a specific molecule can be predicted from the octanol-water partition coefficient of the molecule, a parameter that can be measured or calculated from a knowledge of the structure of the molecule. Olestra does not affect the absorption or efficacy of oral drugs because, in general, they are not sufficiently lipophilic to partition into the olestra. Olestra does not affect the absorption of water-soluble micronutrients or the absorption and utilization of macronutrients. Olestra can reduce the absorption of the fat-soluble vitamins when olestra foods and the vitamins are coingested. These effects can be offset by adding specific amounts of the vitamins to foods made with olestra. Other than the carotenoids and vitamins A and E, olestra does not affect the absorption of potentially beneficial components of fruits and vegetables. The effects on the vitamins can be offset by adding the vitamins to olestra foods. The reduction in the absorption of carotenoids will be less than 6-10% when olestra snacks are eaten under free-living dietary patterns. Any effect this reduction has on vitamin A status can be offset by addition of vitamin A to the foods. The absorption of flavonoids, polyphenols, and most other phytochemicals in fruits and vegetables, which have been shown to provide beneficial health effects, will not be affected by olestra because they are not sufficiently lipophilic. Individuals consuming large quantities of olestra may experience mild or moderate common GI symptoms such as loose or soft stools, gas, or nausea, symptoms similar to those experienced with certain other foods or changed dietary habits. When olestra snack foods are eaten under free-living dietary patterns, the symptoms are not different from those experienced when eating full-fat snack products, in either incidence or severity. When they are experienced, the symptoms resolve in 1-2 days, but may recur. They do not worsen with continued or increased olestra consumption and pose no health risk to the consumer. Olestra products will carry an information label alerting consumers to the possibility of GI symptoms. Olestra foods provide an additional option to those individuals who want or need to lower their total energy intake and body weight. These individuals will find it easier to change dietary habits and to maintain healthful nutritional practices when they use olestra foods. For those who want or need to reduce fat intake but not lose weight, olestra foods can reduce fat intake without affecting energy. Because olestra foods have taste and other organoleptic properties that are similar to those of full-fat foods, individuals will find it easier to switch to low-fat diets.

Heme oxygenase-2 and nitric oxide synthase in guinea-pig intracardiac neurones.

There is increasing evidence that carbon monoxide (CO), like nitric oxide (NO), may be a neuronal messenger molecule. This study investigated the expression of heme oxygenase-2 (HO-2), the enzyme responsible for the synthesis of CO, by intracardiac neurones. Many, if not all newborn guinea-pig intracardiac neurones in culture were HO-2-immunoreactive. Furthermore, double labelling showed that a relatively small subpopulation of these neurones also expressed NO synthase/nicotinamide dinucleotide phosphate (NADPH)-diaphorase (NOS/NADPH-d) activity. These findings suggest that intracardiac neurones can synthesize CO and that CO may be fundamental to their function. Comparison of the proportions of intracardiac neurones that contain HO-2 with those that express NOS/NADPH-d activity also indicates that CO may be more important than NO in the intrinsic neuronal control of the heart.

Causes of death associated with psychiatric illness.

BACKGROUND: A prospective cohort analysis of mortality, among entrants to a population-based psychiatric case register, was undertaken to identify specific causes of death responsible for the increased risk of mortality previously reported in this large group of unselected patients. METHODS: The analysis was based on a study population of 16,871 cases, aged 15-89 years, from Worcester and Kidderminster Health Districts, entering the case register between 1974 and 1984 and generating a total of 85,073 patient-years (PYR) of observation. The underlying cause of death was coded to the relevant revision of the International Classification of Diseases (ICD). Numbers of deaths observed in the study population were compared with the number of deaths expected on the basis of mortality rates for England and Wales. Comparisons were made for eight main causes of death, aggregated at Chapter level of the ICD, and 11 categories of psychiatric diagnoses. Two indices of mortality were used for evaluation: relative risk (RR) = observed deaths/expected deaths; and excess mortality rate (EMR) = (observed-expected deaths)/PYR. RESULTS: RRs were significantly raised for accidents, including suicides, as anticipated, and for various main causes of death. The increased risk of accidental death was found across the majority of the 11 psychiatric diagnostic groups although the EMRs were low at less than 5/1000 PYR. Deaths from respiratory disorders gave rise to the highest RRs after accidental deaths, and were responsible for substantial excess mortality among in-patients and patients with psychotic illnesses (especially dementia). The largest numbers of deaths of both sexes were due to diseases of the circulatory system, with a 40 per cent excess of observed over expected values in the whole series. The excess was due mainly to deaths of in-patients and of patients with psychotic diagnoses. No excess of deaths owing to neoplasms was found for either in-patients or out-patient groups. CONCLUSIONS: The findings that psychiatric illness is associated with an increased risk of death from "natural' causes and that the level of risk was related to the severity and to the diagnostic category of the illness have implications for patterns of care and use of resources.

Nitric oxide synthase-containing neurones and nerve fibres within cardiac ganglia of rat and guinea-pig: an electron-microscopic immunocytochemical study.

The nitric oxide synthase-immunoreactivity and NADPH-diaphorase activity of intracardiac neurones in the rat and guinea-pig was studied at the ultrastructural level. While some nitric oxide synthase-containing intracardiac neurones were very heavily labelled, with electron-dense immunoprecipitate distributed throughout the neuronal cell bodies and their processes, most of the labelled neurones exhibited a lighter and more patchy distribution of nitric oxide synthase-immunoreactive material. Synapses made by nitric oxide synthase-negative nerve fibres with labelled intracardiac neurones were seen. Conversely, many nitric oxide synthase-containing nerve fibres that made synaptic contacts with unlabelled intracardiac neurones were also observed. Some small granule-containing cells were nitric oxide synthase-immunoreactive and were associated with unlabelled nerve terminals, while non-immunoreactive small granule-containing cells that were innervated by nitric oxide synthase-immunoreactive nerves were also seen. Small patches of osmiophilic electron-dense material were observed in the cytoplasm of NADPH-diaphorase-positive intracardiac neurones. This is the first description of the ultrastructural distribution of nitric oxide synthase-immunoreactivity and NADPH-diaphorase activity in a subpopulation of intracardiac neurones of rat and guinea-pig heart and provides further evidence in support of a role for nitric oxide in the local control of the heart by intrinsic neurones.

Ultrastructural investigation of nitric oxide synthase-immunoreactive nerves associated with coronary blood vessels of rat and guinea-pig.

Ultrastructural investigation of nitric oxide synthase-immunoreactive nerves closely associated with blood vessels in rat and guinea-pig hearts revealed many labelled nerve fibres in the walls of the main branches of the coronary arteries, and in arterioles, capillaries and post-capillary venules. The number of nitric oxide synthase-containing nerve fibres associated with different vessels, even those of the same calibre, varied. Terminal regions of nitric oxide synthase-immunoreactive fibres were observed in the endocardium and myocardium. Nitric oxide synthase-labelled fibres displayed electron-dense immunoproduct in both varicose and intervaricose regions. Immunoreactive axonal varicosities contained both small and large synaptic vesicles. The characteristics of the nitric oxide synthase-immunoreactive nerve fibres observed in the heart and the possibility that these fibres represent the processes of intracardiac neurones and/or sensory neurones of extrinsic origin are discussed.

A subpopulation of intracardiac neurons from the guinea pig heart expresses substance P binding sites.

The distribution of binding sites for substance P labeled with [125I]-Bolton-Hunter-reagent was studied in a mixed cell culture preparation from newborn guinea-pig atria and interatrial septum. A relatively small subpopulation of intracardiac neurons expressed substance P binding sites. These neurons exhibited a range of densities of labeling and could be heavily, moderately or lightly labeled with autoradiographic grains. In most cases, the autoradiographic grains were restricted to the neuronal cell body and more proximal regions of the neurites in culture. Intracardiac neurons expressing substance P binding sites were seen in close association with unlabeled neurons. The density of labeling and the distribution of autoradiographic grains over individual intracardiac neurons did not appear to be related to whether they were mono- or binucleate or their associated cell types. The possibility that the substance P binding sites demonstrated here represent functional receptors on intracardiac neurons and their potential role in the heart is discussed.

NADPH diaphorase and nitric oxide synthase are expressed by the majority of intramural neurons in the neonatal guinea pig urinary bladder.

The distribution of neurons expressing NADPH diaphorase activity was examined histochemically in whole mount preparations of the neonatal guinea pig urinary bladder. NADPH diaphorase positive neurons were abundant in the intramural ganglia in both the detrusor and trigone regions of the bladder. Labelled nerve fibres were found in the ganglion interconnectives and in smooth muscle bundles. Mucosal epithelial cells and endothelial cells lining the blood vessels supplying the bladder were also found to express NADPH diaphorase activity. In order to verify that NADPH diaphorase activity represented the presence of nitric oxide synthase in bladder neurons, a well characterised tissue culture preparation was employed. This also provided an opportunity to estimate the proportion of the total population of bladder neurons which expressed NADPH diaphorase activity. Using a combination of histochemical and immunohistochemical techniques, NADPH diaphorase positive neurons were found to constitute approximately 90% of the total neuronal population, which was identified by labelling with an antiserum to the general neuronal marker protein gene product 9.5. Almost all neurons (99%) which expressed NADPH diaphorase activity in culture were also found to be immunoreactive for nitric oxide synthase. These findings indicate that nitric oxide may play a role in the neural control of bladder function, and this possibility is discussed.

Secretion of atrial and brain natriuretic peptides from human cardiac atrial explants in culture: effect of dynorphin.

We have previously developed a method for maintaining human cardiac explants in culture under serum-free conditions, for the assessment of cardiac endocrine function and myocardial growth factors. In order to assess the local role of dynorphin in the human heart, we studied the effects of dynorphin on the secretion of atrial natriuretic peptide and brain natriuretic peptide by human cardiac atrial explants. Dynorphin did not affect the basal secretion of brain natriuretic peptide, but clearly enhanced the release of atrial natriuretic peptide from the human cardiac explants in culture. The atrial content of brain natriuretic peptide was not significantly reduced, whereas the atrial content of atrial natriuretic peptide in cultured explants was reduced two-fold in the presence of dynorphin. These findings indicate that dynorphin may have a direct stimulatory effect on the release of atrial natriuretic peptide, but not brain natriuretic peptide, from human cardiac atria.

Co-expression of four muscarinic receptor genes by the intrinsic neurons of the rat and guinea-pig heart.

Expression of the messenger RNAs encoding the five different muscarinic acetylcholine receptor subtypes was examined in intracardiac neurons from the rat and guinea-pig heart by in situ hybridization techniques. Newborn guinea-pig intracardiac neurons were studied in dissociated cell culture preparations employing both 35S- and digoxigenin-labelled oligonucleotide probes specific for the m1, m2, m3, m4 or m5 muscarinic receptor messenger RNAs. When 35S-tailed oligonucleotides were used, all intracardiac neurons in culture were found to express m1, m2, m3 and m4, but not m5 messenger RNAs. However after hybridization with digoxigenin-tailed probes, only m1 and m2 transcripts were detected. This may reflect differences in the sensitivity of the two techniques. Further to these experiments, intracardiac ganglia in sections of adult rat heart were studied employing m1-, m2-, m3- or m4-specific, 35S-labelled oligonucleotides, and again, all intracardiac neurons expressed messenger RNA for each of these four muscarinic receptor subtypes. Atrial myocytes in culture were only labelled by [35S]- and digoxigenin-tailed m2 oligonucleotides. No other heart cell type seen expressed messenger RNA for any of the muscarinic receptors. The expression of four different muscarinic receptor transcripts by intrinsic neurons of the heart provides the molecular basis for the diverse muscarinic actions observed in these and other autonomic ganglia.

Expression of insulin-like growth factor-1 by human ventricular explants in culture.

There is increasing evidence from animal studies for autocrine and paracrine systems in the heart. As the local role of these peptide systems in the human heart is not clear, we assessed cardiac explants in culture as a model for the study of growth factor expression by the heart in humans. Human right ventricular septal biopsies were maintained as explants for up to 9 days in serum-supplemented medium. Insulin-like growth factor-1 (IGF-1) expression was measured by radioimmunoassay and results corrected for tissue protein content. IGF-1 was synthesised by ventricular explants but not secreted into the culture medium. This culture model has the potential to provide insight into the regulation of and response to growth factor expression by the heart in humans.

Distribution of intracardiac neurones and nerve terminals that contain a marker for nitric oxide, NADPH-diaphorase, in the guinea-pig heart.

There is strong evidence that NADPH-diaphorase can be used as a marker for neurones that employ nitric oxide as a messenger molecule. In the present study, the NADPH-diaphorase activity of intracardiac neurones and nerve terminals in whole-mount stretch preparations and sections of the newborn and adult guinea-pig atria and interatrial septum has been examined histochemically. Together with epicardial, endothelial and endocardial cells, which displayed some NADPH-diaphorase staining, a subpopulation of intracardiac neurones exhibited moderate-heavy labelling for NADPH-diaphorase, while the majority of neurones were only lightly stained or negative. Intracardiac ganglia containing positive neuronal cell bodies were located between the epicardial cells and atrial myocytes in four main regions: in association with the superior and inferior vena cavae, the points of entry of the pulmonary veins, and within the interatrial septum. Nerve terminals exhibiting NADPH-diaphorase activity were seen throughout the atrial tissue, forming basket-like endings around intracardiac neuronal cell bodies; varicose terminals were also observed on atrial myocytes and other non-neuronal structures. A proportion of the nerve fibres was clearly of intrinsic origin, other terminals may well have originated from neuronal cell bodies present outside the heart.

Expression of NADPH-diaphorase activity by guinea-pig paratracheal neurones.

Pharmacological evidence suggests that nitric oxide (NO) is involved in non-adrenergic, non-cholinergic, nerve mediated responses seen in guinea-pig trachealis muscle. The synthetic enzyme for NO (NO synthase) has recently been shown to be responsible for neuronal NADPH-diaphorase activity. Therefore, to determine whether intrinsic paratracheal neurones could be a source of NO in the trachea, expression of NADPH-diaphorase activity was examined histochemically using whole mount preparations of the tracheal plexus. Many paratracheal neurones were found to express moderate to high levels of NADPH-diaphorase activity and are thus likely to be a source of NO in this tissue. This observation provides further evidence that NO is involved in the regulation of relaxation in airway smooth muscle.

Colocalization of nitric oxide synthase and NADPH-diaphorase in the myenteric plexus of the rat gut.

The pattern of distribution and colocalization of nitric oxide-synthase (NOS) and NADPH-diaphorase in the myenteric plexus of whole-mount preparations of the antrum, duodenum, ileum, caecum, proximal colon and distal colon of the rat were investigated using immunohistochemical and histochemical staining techniques. Almost all the myenteric neurons that were NOS-positive in all regions of the gut examined were also stained for NADPH-diaphorase. However, in the stomach, duodenum and ileum, only a few of the NOS-positive nerve fibres in the tertiary and secondary plexuses and circular muscle layer were also stained for NADPH-diaphorase, whereas in the caecum and distal colon almost all the NOS-positive nerve fibres were also stained for NADPH-diaphorase. The results in the present study are consistent with the view that nitric oxide (NO) has a mediating role in gastrointestinal neurotransmission.