A subset of sweet-sensing neurons identified by IR56d are necessary and sufficient for fatty acid taste.

Fat represents a calorically potent food source that yields approximately twice the amount of energy as carbohydrates or proteins per unit of mass. The highly palatable taste of free fatty acids (FAs), one of the building blocks of fat, promotes food consumption, activates reward circuitry, and is thought to contribute to hedonic feeding underlying many metabolism-related disorders. Despite a role in the etiology of metabolic diseases, little is known about how dietary fats are detected by the gustatory system to promote feeding. Previously, we showed that a broad population of sugar-sensing taste neurons expressing Gustatory Receptor 64f (Gr64f) is required for reflexive feeding responses to both FAs and sugars. Here, we report a genetic silencing screen to identify specific populations of taste neurons that mediate fatty acid (FA) taste. We find neurons identified by expression of Ionotropic Receptor 56d (IR56d) are necessary and sufficient for reflexive feeding response to FAs. Functional imaging reveals that IR56d-expressing neurons are responsive to short- and medium-chain FAs. Silencing IR56d neurons selectively abolishes FA taste, and their activation is sufficient to drive feeding responses. Analysis of co-expression with Gr64f identifies two subpopulations of IR56d-expressing neurons. While physiological imaging reveals that both populations are responsive to FAs, IR56d/Gr64f neurons are activated by medium-chain FAs and are sufficient for reflexive feeding response to FAs. Moreover, flies can discriminate between sugar and FAs in an aversive taste memory assay, indicating that FA taste is a unique modality in Drosophila. Taken together, these findings localize FA taste within the Drosophila gustatory center and provide an opportunity to investigate discrimination between different categories of appetitive tastants.

Blockchain Evaluation Approaches: State-of-the-Art and Future Perspective.

The present increase of attention toward blockchain-based systems is currently reaching a tipping point with the corporate focus shifting from exploring the technology potential to creating Distributed Ledger Technology (DLT)-based systems. In light of a significant number of already existing blockchain applications driven by the Internet of Things (IoT) evolution, the developers are still facing a lack of tools and instruments for appropriate and efficient performance evaluation and behavior observation of different blockchain architectures. This paper aims at providing a systematic review of current blockchain evaluation approaches and at identifying the corresponding utilization challenges and limitations. First, we outline the main metrics related to the blockchain evaluation. Second, we propose the blockchain modeling and analysis classification based on the critical literature review. Third, we extend the review with publicly accessible industrial tools. Next, we analyze the selected results for each of the proposed classes and outline the corresponding limitations. Finally, we identify current challenges of the blockchain analysis from the system evaluation perspective, as well as provide future perspectives.

Environmental Monitoring with Distributed Mesh Networks: An Overview and Practical Implementation Perspective for Urban Scenario.

Almost inevitable climate change and increasing pollution levels around the world are the most significant drivers for the environmental monitoring evolution. Recent activities in the field of wireless sensor networks have made tremendous progress concerning conventional centralized sensor networks known for decades. However, most systems developed today still face challenges while estimating the trade-off between their flexibility and security. In this work, we provide an overview of the environmental monitoring strategies and applications. We conclude that wireless sensor networks of tomorrow would mostly have a distributed nature. Furthermore, we present the results of the developed secure distributed monitoring framework from both hardware and software perspectives. The developed mechanisms provide an ability for sensors to communicate in both infrastructure and mesh modes. The system allows each sensor node to act as a relay, which increases the system failure resistance and improves the scalability. Moreover, we employ an authentication mechanism to ensure the transparent migration of the nodes between different network segments while maintaining a high level of system security. Finally, we report on the real-life deployment results.

Positioning Information Privacy in Intelligent Transportation Systems: An Overview and Future Perspective.

Today, the Intelligent Transportation Systems (ITS) are already in deep integration phase all over the world. One of the most significant enablers for ITS are vehicle positioning and tracking techniques. Worldwide integration of ITS employing Dedicated Short Range Communications (DSRC) and European standard for vehicular communication, known as ETSI ITS-G5, brings a variety of options to improve the positioning in areas where GPS connectivity is lacking precision. Utilization of the ready infrastructure, next-generation cellular 5G networks, and surrounding electronic devices together with conventional positioning techniques could become the solution to improve the overall ITS operation in vehicle-to-everything (V2X) communication scenario. Nonetheless, effective and secure communication protocols between the vehicle and roadside units should be both analyzed and improved in terms of potential attacks on the transmitted positioning-related data. In particular, said information might be misused or stolen at the infrastructure side conventionally assumed to be trusted. In this paper, we first survey different methods of vehicle positioning, which is followed by an overview of potential attacks on ITS systems. Next, we propose potential improvements allowing mutual authentication between the vehicle and infrastructure aiming at improving positioning data privacy. Finally, we propose a vision on the development and standardization aspects of such systems.

The lateral line confers evolutionarily derived sleep loss in the Mexican cavefish.

Sleep is an essential behavior exhibited by nearly all animals, and disruption of this process is associated with an array of physiological and behavioral deficits. Sleep is defined by changes in sensory gating that reduce sensory input to the brain, but little is known about the neural basis for interactions between sleep and sensory processing. Blind Mexican cavefish comprise an extant surface dwelling form and 29 cave morphs that have independently evolved increased numbers of mechanoreceptive lateral line neuromasts and convergent evolution of sleep loss. Ablation of the lateral line enhanced sleep in the Pachón cavefish population, suggesting that heightened sensory input underlies evolutionarily derived sleep loss. Targeted lateral line ablation and behavioral analysis localized the wake-promoting neuromasts in Pachón cavefish to superficial neuromasts of the trunk and cranial regions. Strikingly, lateral line ablation did not affect sleep in four other cavefish populations, suggesting that distinct neural mechanisms regulate the evolution of sleep loss in independently derived cavefish populations. Cavefish are subject to seasonal changes in food availability, raising the possibility that sensory modulation of sleep is influenced by metabolic state. We found that starvation promotes sleep in Pachón cavefish, and is not enhanced by lateral line ablation, suggesting that functional interactions occur between sensory and metabolic regulation of sleep. Taken together, these findings support a model where sensory processing contributes to evolutionarily derived changes in sleep that are modulated in accordance with food availability.

Gustatory processing and taste memory in Drosophila.

Taste allows animals to discriminate the value and potential toxicity of food prior to ingestion. Many tastants elicit an innate attractive or avoidance response that is modifiable with nutritional state and prior experience. A powerful genetic tool kit, well-characterized gustatory system, and standardized behavioral assays make the fruit fly, Drosophila melanogaster, an excellent system for investigating taste processing and memory. Recent studies have used this system to identify the neural basis for acquired taste preference. These studies have revealed a role for dopamine-mediated plasticity of the mushroom bodies that modulate the threshold of response to appetitive tastants. The identification of neural circuitry regulating taste memory provides a system to study the genetic and physiological processes that govern plasticity within a defined memory circuit.

Drosophila fatty acid taste signals through the PLC pathway in sugar-sensing neurons.

Taste is the primary sensory system for detecting food quality and palatability. Drosophila detects five distinct taste modalities that include sweet, bitter, salt, water, and the taste of carbonation. Of these, sweet-sensing neurons appear to have utility for the detection of nutritionally rich food while bitter-sensing neurons signal toxicity and confer repulsion. Growing evidence in mammals suggests that taste for fatty acids (FAs) signals the presence of dietary lipids and promotes feeding. While flies appear to be attracted to fatty acids, the neural basis for fatty acid detection and attraction are unclear. Here, we demonstrate that a range of FAs are detected by the fly gustatory system and elicit a robust feeding response. Flies lacking olfactory organs respond robustly to FAs, confirming that FA attraction is mediated through the gustatory system. Furthermore, flies detect FAs independent of pH, suggesting the molecular basis for FA taste is not due to acidity. We show that low and medium concentrations of FAs serve as an appetitive signal and they are detected exclusively through the same subset of neurons that sense appetitive sweet substances, including most sugars. In mammals, taste perception of sweet and bitter substances is dependent on phospholipase C (PLC) signaling in specialized taste buds. We find that flies mutant for norpA, a Drosophila ortholog of PLC, fail to respond to FAs. Intriguingly, norpA mutants respond normally to other tastants, including sucrose and yeast. The defect of norpA mutants can be rescued by selectively restoring norpA expression in sweet-sensing neurons, corroborating that FAs signal through sweet-sensing neurons, and suggesting PLC signaling in the gustatory system is specifically involved in FA taste. Taken together, these findings reveal that PLC function in Drosophila sweet-sensing neurons is a conserved molecular signaling pathway that confers attraction to fatty acids.

Distinct genetic architecture underlies the emergence of sleep loss and prey-seeking behavior in the Mexican cavefish.

BACKGROUND: Sleep is characterized by extended periods of quiescence and reduced responsiveness to sensory stimuli. Animals ranging from insects to mammals adapt to environments with limited food by suppressing sleep and enhancing their response to food cues, yet little is known about the genetic and evolutionary relationship between these processes. The blind Mexican cavefish, Astyanax mexicanus is a powerful model for elucidating the genetic mechanisms underlying behavioral evolution. A. mexicanus comprises an extant ancestral-type surface dwelling morph and at least five independently evolved cave populations. Evolutionary convergence on sleep loss and vibration attraction behavior, which is involved in prey seeking, have been documented in cavefish raising the possibility that enhanced sensory responsiveness underlies changes in sleep. RESULTS: We established a system to study sleep and vibration attraction behavior in adult A. mexicanus and used high coverage quantitative trait loci (QTL) mapping to investigate the functional and evolutionary relationship between these traits. Analysis of surface-cave F2 hybrid fish and an outbred cave population indicates that independent genetic factors underlie changes in sleep/locomotor activity and vibration attraction behavior. High-coverage QTL mapping with genotyping-by-sequencing technology identify two novel QTL intervals that associate with locomotor activity and include the narcolepsy-associated tp53 regulating kinase. These QTLs represent the first genomic localization of locomotor activity in cavefish and are distinct from two QTLs previously identified as associating with vibration attraction behavior. CONCLUSIONS: Taken together, these results localize genomic regions underlying sleep/locomotor and sensory changes in cavefish populations and provide evidence that sleep loss evolved independently from enhanced sensory responsiveness.

A Harmonized Perspective on Transportation Management in Smart Cities: The Novel IoT-Driven Environment for Road Traffic Modeling.

The unprecedented growth of today's cities together with increased population mobility are fueling the avalanche in the numbers of vehicles on the roads. This development led to the new challenges for the traffic management, including the mitigation of road congestion, accidents, and air pollution. Over the last decade, researchers have been focusing their efforts on leveraging the recent advances in sensing, communications, and dynamic adaptive technologies to prepare the deployed road traffic management systems (TMS) for resolving these important challenges in future smart cities. However, the existing solutions may still be insufficient to construct a reliable and secure TMS that is capable of handling the anticipated influx of the population and vehicles in urban areas. Along these lines, this work systematically outlines a perspective on a novel modular environment for traffic modeling, which allows to recreate the examined road networks in their full resemblance. Our developed solution is targeted to incorporate the progress in the Internet of Things (IoT) technologies, where low-power, embedded devices integrate as part of a next-generation TMS. To mimic the real traffic conditions, we recreated and evaluated a practical traffic scenario built after a complex road intersection within a large European city.

Genetic dissection of sleep-metabolism interactions in the fruit fly.

Dysregulation of sleep and metabolism has enormous health consequences. Sleep loss is linked to increased appetite and insulin insensitivity, and epidemiological studies link chronic sleep deprivation to obesity-related disorders including type II diabetes and cardiovascular disease. Interactions between sleep and metabolism involve the integration of signaling from brain regions regulating sleep, feeding, and metabolic function. Investigating the relationship between these processes provides a model to address more general questions of how the brain prioritizes homeostatically regulated behaviors. The availability of powerful genetic tools in the fruit fly, Drosophila melanogaster, allows for precise manipulation of neural function in freely behaving animals. There is a strong conservation of genes and neural circuit principles regulating sleep and metabolic function, and genetic screens in fruit flies have been effective in identifying novel regulators of these processes. Here, we review recent findings in the fruit fly that further our understanding of how the brain modulates sleep in accordance with metabolic state.

Multisensory integration of colors and scents: insights from bees and flowers.

Karl von Frisch's studies of bees' color vision and chemical senses opened a window into the perceptual world of a species other than our own. A century of subsequent research on bees' visual and olfactory systems has developed along two productive but independent trajectories, leaving the questions of how and why bees use these two senses in concert largely unexplored. Given current interest in multimodal communication and recently discovered interplay between olfaction and vision in humans and Drosophila, understanding multisensory integration in bees is an opportunity to advance knowledge across fields. Using a classic ethological framework, we formulate proximate and ultimate perspectives on bees' use of multisensory stimuli. We discuss interactions between scent and color in the context of bee cognition and perception, focusing on mechanistic and functional approaches, and we highlight opportunities to further explore the development and evolution of multisensory integration. We argue that although the visual and olfactory worlds of bees are perhaps the best-studied of any non-human species, research focusing on the interactions between these two sensory modalities is vitally needed.

Altered regulation of sleep and feeding contributes to starvation resistance in Drosophila melanogaster.

Animals respond to changes in food availability by adjusting sleep and foraging strategies to optimize their fitness. Wild populations of the fruit fly, Drosophila melanogaster, display highly variable levels of starvation resistance that are dependent on geographic location, food availability and evolutionary history. How behaviors that include sleep and feeding vary in Drosophila with increased starvation resistance is unclear. We have generated starvation-resistant flies through experimental evolution to investigate the relationship between foraging behaviors and starvation resistance. Outbred populations of D. melanogaster were selected for starvation resistance over 60 generations. This selection process resulted in flies with a threefold increase in total lipids that survive up to 18 days without food. We tested starvation-selected (S) flies for sleep and feeding behaviors to determine the effect that selection for starvation resistance has had on foraging behavior. Flies from three replicated starvation-selected populations displayed a dramatic reduction in feeding and prolonged sleep duration compared to fed control (F) populations, suggesting that modified sleep and feeding may contribute to starvation resistance. A prolonged larval developmental period contributes to the elevated energy stores present in starvation-selected flies. By preventing S larvae from feeding longer than F larvae, we were able to reduce energy stores in adult S flies to the levels seen in adult F flies, thus allowing us to control for energy storage levels. However, the reduction of energy stores in S flies fails to generate normal sleep and feeding behavior seen in F flies with similar energy stores. These findings suggest that the behavioral changes observed in S flies are due to genetic regulation of behavior rather than elevated lipid levels. Testing S-F hybrid individuals for both feeding and sleep revealed a lack of correlation between food consumption and sleep duration, indicating further independence in genetic factors underlying the sleep and feeding changes observed in S flies. Taken together, these findings provide evidence that starvation selection results in prolonged sleep and reduced feeding through a mechanism that is independent of elevated energy stores. These findings suggest that changes in both metabolic function and behavior contribute to the increase in starvation resistance seen in flies selected for starvation resistance.

Limited taste discrimination in Drosophila.

In the gustatory systems of mammals and flies, different populations of sensory cells recognize different taste modalities, such that there are cells that respond selectively to sugars and others to bitter compounds. This organization readily allows animals to distinguish compounds of different modalities but may limit the ability to distinguish compounds within one taste modality. Here, we developed a behavioral paradigm in Drosophila melanogaster to evaluate directly the tastes that a fly distinguishes. These studies reveal that flies do not discriminate among different sugars, or among different bitter compounds, based on chemical identity. Instead, flies show a limited ability to distinguish compounds within a modality based on intensity or palatability. Taste associative learning, similar to olfactory learning, requires the mushroom bodies, suggesting fundamental similarities in brain mechanisms underlying behavioral plasticity. Overall, these studies provide insight into the discriminative capacity of the Drosophila gustatory system and the modulation of taste behavior.

Measurement of Taste Memory in Drosophila.

The ability to modify behavior as a result of previous experience allows an organism to adapt to changes in its environment. Even innate behaviors, like feeding initiation, can change if previously associated with a noxious stimulus. Here, we describe a taste memory assay pairing appetitive and bitter tastants, resulting in aversive taste conditioning. By training a fly to associate sweet sucrose applied to the tarsus with bitter quinine applied to the proboscis, flies quickly learn to suppress the reflexive proboscis extension to sucrose, providing a bioassay for behavioral and molecular plasticity. This single-fly taste memory assay may be applied to adult Drosophila of any genetic background and allows for interrogation of the neural circuitry and molecular processes encoding memories while simultaneously measuring behavior. Unlike many other memory assays, this system requires few custom components, and therefore can be easily established in laboratories with minimal expertise in the study of fly behavior.

Innate and Conditioned Taste Processing in Drosophila.

Peripheral detection of tastants allows animals to detect the dietary value of food and its potential toxicity. Many tastants such as sugars and fats elicit reflexive appetitive responses, whereas other foods such as quinine induce aversion. The relative value of food can change in accordance with an animal's internal state and prior experience. Understanding the neural and genetic bases for the detection and response to tastants, as well as how these behaviors change with experience, is central to sensory neuroscience. The presentation of attractive tastants to the proboscis or legs of the fruit fly Drosophila melanogaster induces a robust and reflexive proboscis-extension response (PER). This quantifiable response can be used to study the receptors underlying taste detection, the neural circuits involved in sensory processing, and the musculature required for a simple feeding behavior. Furthermore, we have developed a memory assay pairing appetitive and bitter tastants, resulting in aversive taste conditioning, in which the PER response to attractive tastants is diminished. Unlike many memory assays, this assay does not require specialized equipment and can be readily implemented in teaching and research laboratories. Here, we introduce protocols for studying the PER feeding response and aversive taste memory in Drosophila.

Measurement of Reflexive Feeding Response in Drosophila.

The ability to distinguish between food sources that are good and provide nutrients and those that are potentially dangerous is crucial to the survival of an organism. Here, we describe a taste assay that measures the reflexive feeding response to a given tastant. To examine taste preference for a soluble compound, an appetitive tastant is applied to the proboscis, and the proportion of proboscis extensions are recorded. This single-fly assay may be applied to adult Drosophila of any genetic background and facilities examination of the neural circuitry and molecular processes encoding the reflexive taste response. Furthermore, this assay requires few custom components and therefore can be easily established in laboratories with minimal expertise in the study of fly behavior.

Distinct memories of odor intensity and quality in Drosophila.

Even in a simple Pavlovian memory task an animal may form several associations that can be independently assessed by the appropriate tests. Studying conditioned odor discrimination of the fruit fly Drosophila melanogaster we found that animals store quality and intensity of an odor as separate memory traces. The trace of odor intensity is short-lived, decaying in <3 h. Only the last intensity value is stored. In contrast to odor-quality memory, odor-intensity memory does not require the rutabaga-dependent cAMP signaling pathway. Flies rely on their memory of intensity in a narrow concentration range in which they can generalize intensity. Larger concentration differences they treat like different qualities. This study shows that the perceptual identity of an odor is based on at least three lines of processing in the brain: (i) a memory of odor quality, (ii) a memory of odor intensity, and (iii) a range of intensities (and qualities), in which the odor is generalized.

Excessive energy expenditure due to acute physical restraint disrupts Drosophila motivational feeding response.

To study the behavior of Drosophila, it is often necessary to restrain and mount individual flies. This requires removal from food, additional handling, anesthesia, and physical restraint. We find a strong positive correlation between the length of time flies are mounted and their subsequent reflexive feeding response, where one hour of mounting is the approximate motivational equivalent to ten hours of fasting. In an attempt to explain this correlation, we rule out anesthesia side-effects, handling, additional fasting, and desiccation. We use respirometric and metabolic techniques coupled with behavioral video scoring to assess energy expenditure in mounted and free flies. We isolate a specific behavior capable of exerting large amounts of energy in mounted flies and identify it as an attempt to escape from restraint. We present a model where physical restraint leads to elevated activity and subsequent faster nutrient storage depletion among mounted flies. This ultimately further accelerates starvation and thus increases reflexive feeding response. In addition, we show that the consequences of the physical restraint profoundly alter aerobic activity, energy depletion, taste, and feeding behavior, and suggest that careful consideration is given to the time-sensitive nature of these highly significant effects when conducting behavioral, physiological or imaging experiments that require immobilization.

translin Is Required for Metabolic Regulation of Sleep.

Dysregulation of sleep or feeding has enormous health consequences. In humans, acute sleep loss is associated with increased appetite and insulin insensitivity, while chronically sleep-deprived individuals are more likely to develop obesity, metabolic syndrome, type II diabetes, and cardiovascular disease. Conversely, metabolic state potently modulates sleep and circadian behavior; yet, the molecular basis for sleep-metabolism interactions remains poorly understood. Here, we describe the identification of translin (trsn), a highly conserved RNA/DNA binding protein, as essential for starvation-induced sleep suppression. Strikingly, trsn does not appear to regulate energy stores, free glucose levels, or feeding behavior suggesting the sleep phenotype of trsn mutant flies is not a consequence of general metabolic dysfunction or blunted response to starvation. While broadly expressed in all neurons, trsn is transcriptionally upregulated in the heads of flies in response to starvation. Spatially restricted rescue or targeted knockdown localizes trsn function to neurons that produce the tachykinin family neuropeptide Leucokinin. Manipulation of neural activity in Leucokinin neurons revealed these neurons to be required for starvation-induced sleep suppression. Taken together, these findings establish trsn as an essential integrator of sleep and metabolic state, with implications for understanding the neural mechanism underlying sleep disruption in response to environmental perturbation.

A dopamine-modulated neural circuit regulating aversive taste memory in Drosophila.

Taste memories allow animals to modulate feeding behavior in accordance with past experience and avoid the consumption of potentially harmful food [1]. We have developed a single-fly taste memory assay to functionally interrogate the neural circuitry encoding taste memories [2]. Here, we screen a collection of Split-GAL4 lines that label small populations of neurons associated with the fly memory center-the mushroom bodies (MBs) [3]. Genetic silencing of PPL1 dopamine neurons disrupts conditioned, but not naive, feeding behavior, suggesting these neurons are selectively involved in the conditioned taste response. We identify two PPL1 subpopulations that innervate the MB α lobe and are essential for aversive taste memory. Thermogenetic activation of these dopamine neurons during training induces memory, indicating these neurons are sufficient for the reinforcing properties of bitter tastant to the MBs. Silencing of either the intrinsic MB neurons or the output neurons from the α lobe disrupts taste conditioning. Thermogenetic manipulation of these output neurons alters naive feeding response, suggesting that dopamine neurons modulate the threshold of response to appetitive tastants. Taken together, these findings detail a neural mechanism underlying the formation of taste memory and provide a functional model for dopamine-dependent plasticity in Drosophila.