Contextual response selection refers to the animal emitting different types of responses to the same item depending on the context in the background. Contextual item selection occurs when there are multiple items that need to be chosen in a contextual manner. Finally, when multiple items and multiple contexts are involved, contextual item–response selection takes place whereby the animal either chooses an item or inhibits such a response depending on item–context paired association. The literature suggests that the rhinal cortical regions and the hippocampal formation play key roles in mnemonically categorizing and recognizing contextual representations and the associated items. In addition, it appears that the fronto-striatal cortical loops in connection with the contextual information-processing areas critically control the flexible deployment of adaptive action sets and motor responses for maximizing goals. We suggest that contextual information processing should be investigated in experimental settings where contextual stimuli and resulting behaviors are clearly defined and measurable, considering the dynamic top-down and bottom-up interactions among the neural systems for contextual behavior.Animals including humans engage in goal-directed behavior flexibly in response to items and their background, which is called contextual behavior in this review. Although the concept of context has long been studied, there are differences among researchers in defining and experimenting with the concept. The current review aims to provide a categorical framework within which not only the neural mechanisms of contextual information processing but also the contextual behavior can be studied in more concrete ways. For this purpose, we categorize contextual behavior into three subcategories as follows by considering the types of interactions among context, item, and response:  In the absence of such powerful information that effectively sets up the global mode of operation in the brain, physical stimuli in the ever-changing and complex world should produce insurmountable ambiguity and confusion in neural information processing and resulting behavior. In the current review, we will selectively review prior studies including our own that can speak to how the brain enables an animal to learn and remember to respond appropriately in a contextual manner.Animals including humans recognize their surrounding environment rapidly and with seeming ease. A surrounding environment often influences which information needs to be attentively processed in certain ways and also determines an appropriate action set or behavioral repertoire Figure . That isContext is hardly a scientific term. It is often casually used in layman's conversation to loosely refer to a set of physical and cognitive factors that determine the meaning of an otherwise ambiguous element(s) associated with those factors. When a behavioral response to such an element is influenced by those peripherally related factors, one may call it a contextual behavior. The purpose of the current review is to provide a perspective on how contextual system interacts with different neural systems for response, such as item information-processing systems and motor action systems (see below). Therefore, we would not be able to provide an extensive review of the literature on context, but briefly mentioning some related contents from the literature would nonetheless be helpful.What is context? There are many contexts one may encounter during lifetime and there are as many definitions of the term as well. The concept of contextual stimuli was first brought to learning and memory research as learning theories developed largely within the framework of classical and instrumental conditioning  are largely associated with the physical nature of stimuli . On the other hand, more functional definitions of context were proposed by several researchers Hirsh, . Hirsh , for exaRemoving the hippocampus may result in rigid and inflexible S–R memory systems, which would be equivalent to having only habit memory systems , and it should be directly associated with purposeful behavior. By restricting the scope of definition to external and physical stimuli, we intend not to include non-environmental internal variables such as rule, strategy, sequence, motivation, drug state, linguistic variables, etc., as examples of context  is clearly predefined in relation to context , literature has been focusing heavily on how contextual representation is formed and stored in the brain with less emphasis on how contextual memory is “used” during behavioral choice. We would argue in this review that a more wholistic investigation of neural circuits for contextual behavior requires understanding not only how contextual memory is formed and retrieved, but also how contextual information influences different types of choice behavior. Therefore, contextual behavioral studies will be subdivided into three categories in this review using the number of items and the type of responses associated with multiple contexts as major criteria: n Figure . In the Contextual response selection is required when an animal should choose a particular behavioral response from a set of learned behavioral choices in association with the environmental context. This often occurs when there is no obvious elemental cue in the environment that reliably tells the animal which behavioral choice is appropriate or when the elemental cue present is ambiguous (thus rendering decision making for choice behavior difficult), but the surrounding context does the job.One of the widely spread contextual behavioral testing paradigm, contextual freezing or fear-conditioning, requires contextual response selection. In a typical contextual fear-conditioning experiment  are called a context. As an attempt to improve this situation, we tested previously whether the configuration of specific sets of visual cues can serve as effective contextual stimuli for directing the rat's spatial choice behavior to discrete locations  of the VCRS task was developed later in our laboratory, using touchscreen and LCD monitors  recently in another version of VCRS task . One of the examples that may be considered closer to a rodent contextual behavioral paradigm would be a study done by Wirth et al. , in whicIn human studies, combining virtual reality (VR) with functional magnetic resonance imaging (fMRI) techniques has provided a unique experimental opportunity that allows testing visual contextual behavior in a goal-oriented experimental situation that closely resembles the rodent behavioral paradigm . Human imaging studies provide unique opportunities to observe how different neural networks in the whole brain change activity patterns simultaneously, which is not currently feasible in rodent studies. In a VR environment, a visual context is provided by realistic graphical background images and the visual background continuously changes as a subject responds to it. In this situation, a subject can perform a contextual response selection by indicating right or left turn, for example, by pushing corresponding buttons or by manipulating a joystick when he/she encounters a particular visual scene or context. Many tasks that may belong to this category used a virtual maze in which egocentric responses were critically required because the subject moved through alleys connected by several intersections. However, some of these tasks  made spatial response selection  less efficient and produced some differences in the regions of the brain activated for making behavioral choices as compared to the areas for making responses to contextually rich visual stimuli  in conjunction with a specific context in the animal's background. Context in this case plays a critical role especially when the same item(s) appears in multiple contexts, but is rewarded in a certain context only. Importantly, the same response  should be emitted in all cases (so devoid of the response-selection requirement). The response in this case just needs to be directed toward the correct item considering the environmental context.vice versa for odor B. This was the case when scented objects (cups with scented sand) and contexts were novel, but not for familiar odors and contexts. The perceptual discrimination of contexts or items by themselves was not impaired in the NR1-deleted mice. Komorowski et al. also used the same paradigm in rats and found the neural correlates for representing the item–context paired associations in the hippocampus  to the same object appropriate sometimes but not in other times. For example, Bussey et al.  tested rIn our laboratory, we have verified that the hippocampus, prefrontal cortex (PFC), and PER play critical roles and interact with each other as rats are engaged in contextual item-selection behavior. Specifically, we developed a behavioral paradigm in which different arms of a radial-arm maze were associated with either an object-in-context rule or a response-in-context rule in order to test goal-directed behavior  on the left food well in arm 3 (one of the object-in-context arms) at an early learning stage, the rat appeared to become strongly inclined to adopt a response strategy almost immediately. That is, the rat seemed to make an erroneous association and might “think” that it obtained reward because it turned to the object on its left side in the context (instead of associating the object identity with the context and reward). It appears that this dominance of response bias over object-in-context strategy is an innate bias that all rats show almost without exception, albeit to different degrees among individual rats. During the earlier phase of learning, rats acted as if they were only governed by the response-in-context strategy but they started to show inhibitory behavior immediately in front of a wrong object after a few days of training, which was usually a sure sign that foretold an upcoming performance surge in the task in 2–3 days.In our task, since the maze was surrounded by rich visual cues in the background, rats must pay attention to the visual context in order to choose a contextually correct object or response. It is unlikely that rats were dependent more on path integration in identifying the arms visited because, once exiting the start box, rats usually explored the center stage (frequently showing rearing behavior) before finding and entering an open arm. Despite the seemingly complex task demands with potentially conflicting strategies  associated with different contexts, normal rats pre-trained to displace an object to retrieve a food reward learned the task only in approximately 1 week on average. The learning curve itself is interesting because of its non-linear property. That is, rats typically showed almost 50% chance-level performances for several days (approximately 6–7 days) but suddenly exhibited a jump in performance at around day 7 or so during the acquisition. Such learning curve suggests that the rat experienced an “a-ha” moment suddenly after several days of having tried to figure out what the task demands were. This is likely because a typical source of poor performance during acquisition was the Rats with axon-sparing excitotoxic lesions in the hippocampus were severely impaired in the task described above and their performance stayed almost at chance level throughout 10 days of post-surgical testing , it was not difficult to draw a boundary between pre-learning and post-learning stages in the task. Hippocampal neurons mostly fired in a spatially localized fashion  as reported in prior studies O'Keefe, . As has Similar changes in the firing patterns of neurons in the mPFC across learning support the above conjecture. That is, during the initial learning of the task, the firing patterns of neurons in mPFC were critically modulated by the acquisition of the rule. Specifically, the rats used in the above study were implanted with a multi-electrode recording drive that could record single units  from both hippocampus and mPFC simultaneously  is represented more prominently in the mPFC  are reviewed with the topic of contextual item selection in primates. Constraining the scope of review using the definition of context of this review further reduces the number of studies available for review because this type of context is difficult to implement in primate studies. Specifically, monkeys are often tested using a Wisconsin General Testing Apparatus  may qualify those as proper studies to be reviewed here. That is, an object in the Gaffan's paradigm was a small typographic character that can be touched by the monkey in a computer monitor. A context was a visual image covering the entire computer screen and served as the background of the typographic objects. The background image  was composed of a random number of ellipses and segments of ellipses. The colors, positions, and sizes of elliptical shapes were randomly determined by a computer algorithm. The monkey was required to learn where in the monitor a certain typographic object was rewarded when a particular scene was presented as a contextual background. The Gaffan group showed that lesions produced in the fimbria-fornix , entorhinal cortex (EC), and orbital PFC resulted in learning deficits in the task Gaffan, . DisconnIn humans, only very few studies examined contextual item selection as opposed to the more frequent usage of the contextual response-selection paradigm as described above. Among those, Burgess et al.  requiredSo far, we have categorized contextual behaviors into two categories and those were contextual response-selection behavior and contextual item-selection behavior. There is another type of behavioral tasks in which these two categories were used in a mixed fashion. The need for this category of contextual behavior is related to the type of behavioral task in which only a single stimulus  out of multiple candidate items is presented in a given context per trial and the animal is required to either respond to the stimulus directly (or to a dedicated response unit in the apparatus) or to inhibit the response. So it is similar to contextual response selection, but is different at the same time because different items need to be recognized.For example, Good and Honey used a behavioral paradigm in which the rat was trained to press a lever in response to either tone or light stimulus. Each stimulus, however, resulted in reward only in one context, but not in the other context  and this may the most important source of information the brain uses to set the mode of operation in the face of a specific context. The contextual information from the hippocampus should be useful particularly for those brain areas whose functions are critical for controlling final action or response. The PFC and the STR have long been suggested as key players in this regard. For example, many studies have shown the roles of PFC in flexible choice behavior (Ragozzino et al., The functions of STR and the cortico-striatal networks also have long been investigated (Hikosaka and Isoda, In this review, efforts have been made both implicitly and explicitly to emphasize the importance of understanding contextual behavior with the consideration of final goal-related responses. For example, it is critical to understand how the contextual representation in the hippocampus is used by other regions in the brain such as the mPFC and STR in determining the animal's behavior in response to the context. Because the anatomy and physiology suggest that the circuits are built to influence the information processing among these and other contextual behavioral networks in both feedforward and feedback directions, investigating a particular neural structure in isolation may result in insufficient understanding of the network dynamics. As has been emphasized in this review, using goal-directed tasks with well-defined stimulus control is also critical during the investigative endeavor to answer the critical question of where in the information-processing stream the neural manifestation of context emerges as opposed to the representation of a mere sum of elemental percepts. This theme also echoes in understanding the neural systems that were previously considered as pure perceptual systems (Sobotka et al., The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


The existence of an attentional window—a limited region in visual space at which attention is directed—has been invoked to explain why sudden visual onsets may or may not capture overt or covert attention. Here, we test the hypothesis that observers voluntarily control the size of this attentional window to regulate whether or not environmental signals can capture attention. We have used a novel approach to test this: participants eye-movements were tracked while they performed a search task that required dynamic gaze-shifts. During the search task, abrupt onsets were presented that cued the target positions at different levels of congruency. The participant knew these levels. We determined oculomotor capture efficiency for onsets that appeared at different viewing eccentricities. From these, we could derive the participant's attentional window size as a function of onset congruency. We find that the window was small during the presentation of low-congruency onsets, but increased monotonically in size with an increase in the expected congruency of the onsets. This indicates that the attentional window is under voluntary control and is set according to the expected relevance of environmental signals for the observer's momentary behavioral goals. Moreover, our approach provides a new and exciting method to directly measure the size of the attentional window. In everyday life, our visual system is bombarded with signals that continuously attempt to draw our attention. For example, when on the road, we may need to look for street signs that indicate a change in the traffic situation amidst billboards that try to lure us to come over for a quick “bite”. While visually searching and exploring the world, observers continuously move their eyes, to direct the high-resolution section of their retinae to items of interest. During a fixation –the brief period during which the eyes do not move-a portion of the visual field can be scrutinized. Previous results In addition to attending to the information that is relevant for the task at hand, it may be necessary to also consider other environmental signals that may provide unexpected but important information (onsets), for example those that indicate a dangerous situation. Also, the relevance of such unexpected visual information may vary, so we need to exert some kind of top-down control (e.g. inhibition of distraction) over the extent of attentional capture by unexpected visual onsets. These environmental distracters influence following eye-movements in their turn (bottom-up influence) Previous research suggests that one of the mechanisms by which we dynamically control what information is allowed to capture attention, guide our gaze, and potentially drive our behaviour is an attentional window. The attentional window is a theoretical construct describing the limited region of visual space that attention can be paid to. The size of this region is derived by examining whether or not an event presented in visual space can attract or, in other words, capture, attention or gaze. In the latter case, the presence of a capture is determined by verifying whether an event evokes an eye-movement. This is also the approach we take in the present study.The use of gaze recordings is justified by the large body of evidence that indicates that eye-movements are preceded by shifts of attention Theeuwes In a study by Yantis and Jonides The attentional window size may be dependent on the type of search task: i.e. an inefficient, serial search or a more efficient, parallel search e.g. . Serial Several studies suggest that voluntary control over attentional window size could be used to modulate attentional capture. If so, this could be a mechanism by which observers can regulate whether or not environmental signals will capture attention. A voluntary regulation of attentional window size would imply top-down guided processing of stimuli as opposed to a bottom-up adjustment to visual cues. Even though the idea of automatic bottom-up processing of capture events Here, we test the hypothesis that observers exert voluntary control over attentional capture by setting the size of the attentional window. Moreover, we expect them to do so depending on task-characteristics . In other words, we test the hypothesis that attentional window size (as derived from the likelihood of capture) is modulated by the expected relevance of environmental signals for the ongoing task.Hence, we predict that the attentional window size will increase with onset congruency, even in an inefficient serial search task that is generally assumed to impose a narrow attentional focus. We tested this hypothesis by having participants' perform a search task, in which they could freely move their eyes over the screen (i.e. overt orienting). During the task, we presented abrupt onsets that indicated the position of the target with varying degrees of congruency. Cue congruency was reported to the participants prior to each block of trials. We used overt oculomotor capture (i.e. capture of gaze) as our primary measure of attentional capture. The eccentricities of the captured onsets were used to derive the attentional window size. The search task was designed such that we: (a) enabled participants to exhibit gaze behaviour, (b) could study attentional capture at eccentricities up to 20 degrees, (c) were able to study attentional capture during the search process itself, so as to minimize possible contamination by start up processes. Finally, by keeping stimulus and onset characteristics identical in all experimental conditions, we made sure that changes in attentional window size cannot be attributed to changes in onset saliency This study was approved by the ethical committee of the University Medical Centre Groningen, all participants gave written informed consent and were treated according to the declaration of Helsinki.Fourteen university students participated in the study. Age ranged from 20 to 25 years (mean age 22). Participants had normal or corrected-to-normal vision.Participants were seated in a darkened room at 85 cm from a CRT computer monitor (LaCie). The horizontal size of the monitor screen was 27 degrees of visual angle, and its vertical size was 20 degrees of visual angle. A chinrest stabilized the participant's head. A remote eyetracker  was located below the screen to track eye position during the experiment at a sample rate of 1000 Hz. The experiment was written in MATLAB , using the Psychophysics and Eyelink Toolbox extensions 2, respectively.During a trial, participants searched for a single circle with one opening (Target), amidst 11 circles with two openings (distracters) see . CirclesParticipants were required to report the target's open side (left or right) by pressing as fast as possible either the left or right arrow on the keyboard. During the trial, an abrupt onset (a red circle) appeared over 1 of the 12 search elements and disappeared within 100 ms in a shrinking fashion. The onset was presented randomly between one and three seconds from the start of the trial. This guaranteed that participants had commenced searching before the onset's appearance and allowed for sufficient time to react to the onset before the trial ended. Each trial lasted 4 s irrespective of the occurrence of a response. In between trials, a screen was presented for 1 s, showing only a fixation cross in the centre.In different blocks of trials, four levels of congruency of the abrupt onset  were tested. Congruency is defined as the spatial congruence of the target and onset stimuli. For example, during the 50% congruency block, the abrupt onset appeared on the target's location in half of the trials (congruent trails), whereas the abrupt onset appeared on a distracter's location on the remaining trials . In addition to the four blocks with different onset congruencies, two control conditions were performed. In the first control condition, no onsets were presented. This condition was used to monitor gaze behaviour strictly related to the search task. In the second control condition (‘free’), participants were instructed to freely view the screen while an onset appeared. In this condition, all circles were closed and no target search had to be performed. This condition was administered to measure gaze capturing in the absence of a search task situation.In total, each participant performed four sessions, with each session containing a total of six blocks . Each block contained 40 trials. Participants were offered to take a break in between sessions of up to 20 min. During the first and third session, the order of presentation was: no onset, 0%, 10%, 50%, 100% congruency, and free viewing. This order was reversed during the second and fourth session, to prevent performance bias due to learning or fatigue over conditions. Before the start of each block, a message on the screen informed participants about the upcoming condition .Prior to the experiment, participants were instructed to search for the circle with just one opening during the search task and, when found, to report on which side it was open. They were required to do this as fast and as accurately as possible. Furthermore, participants were instructed to gaze freely over the screen in the free viewing condition and to fixate on the fixation cross in between trials. During instruction, stimulus screens were shown and participants completed a few practice trials so that they understood the task. Finally, participants were instructed to try to search for the target immediately after the start of the trial and not wait for the onset to appear. Participants were instructed that in some trials attending to the onsets could help finding the target, while in other trials attending to the onset would hinder finding the target.To measure whether participants' attention was captured, we analysed eye movements (saccades) and used oculomotor capture as a marker for attentional capture. A saccade was defined by a change in eye position of at least 1.0 deg, at a velocity of at least 30 degrees/s, and an acceleration of 8000 degrees/s/s. We applied the following criteria to classify a saccade as an oculomotor capture: (a) a saccade was made that landed on the location of onset appearance, and (b) it was the first or second saccade after the onset's appearance . A saccade was classified as having been made towards the onset, and thus as a capture, when its end position was closer to the location of the object where the onset appeared than to the location of any other object (i.e. the saccade had to land within a radius of 2.5–2.9 deg of an onset object). We disregarded saccades made earlier than 100 ms after onset appearance , as well as saccades with latencies larger than 1000 ms. In 24% of the trials the first saccade following onset appearance was within 100 ms. In these cases the next saccade for this trial was used,  In 5% of the trials, the first saccade following the onset appearance was later than 1000 ms; these trials were removed from further analysis.The oculomotor capture data can be used to determine the capture efficiency. In combination with logistic regression, the capture data was used to determine the attentional window size. An oculomotor capture was assigned a value of 1; an onset that did not capture gaze (a ‘miss’) was assigned a value of 0. We fitted a standard psychometric function  to these outcomes as a function of retinal eccentricity  The psychometric functions were fitted in MATLAB. Fits were performed for each participant and for each condition. To evaluate the influence of chance in determining the size of the attentional window, data in the condition without an onset was analysed based on “virtual” onsets. For this, for each trial a random timing and onset location was chosen, in a way identical to that in actual onset conditions during the experiment. In all other ways, analysis was identical to the analysis of trials with actual onsets present.Repeated measures ANOVA was used to determine an effect of condition on performance (target identification), reaction time, and saccadic latency (in case of captures in conditions with onsets). The relationship between capture effectiveness and attentional window size was investigated using a t-test for paired samples.F   = 49.5, p<0.001) with higher performance for higher congruencies   = 3.5, p = 0.014) were shorter (determined for correct responses). Congruency affected average saccadic latency   = 17.9, p<0.001), with shorter latencies for higher congruency conditions. Latency in the “free” condition was somewhat comparable to that in the 50% congruency condition.Congruency significantly affected search performance, i.e. the relative number of trials in which the side of the target containing the gap was correctly identified  as our primary measure of attentional capture. Although conditions have been reported in which attentional and oculomotor capture are decoupled, during search for a unique shape (gap) as in our experiment, color singletons elicit both attentional capture and oculomotor capture That a adjustable spatial window of attention regulates the occurrence of automatic capture Our results are consistent with the idea that participants voluntarily modulated the size of a spatial window in order to regulate which onsets did result in a capture, and which ones did not. The incentive given in the present experiment was the known relevance of the onset for indicating the target position. In conditions with identical stimulus displays and onsets, varying the relevance was sufficient to induce participants to change the spatial region within which onsets resulted in captures or not. In that sense, our results are entirely consistent whit the attentional window account and can be considered as a direct demonstration of its existence. Moreover, our approach provides a new method to directly measure the size of the attentional window.Accounts in terms of strategic search modes, such as those proposed by Bacon and Egeth , that predict more or less uniform changes in attentional capture over the full extent of the display, by themselves are insufficient to explain our present pattern of results. However, our results cannot fully indicate which search mode was used within the attentional window. The search task required participants to search for gaps in the O's that made up the stimulus display. This would clearly have to be based on feature search mode. However, onset items were red imploding circles, detection of which in itself may have been based on either feature or singleton search mode. Deciding between the search modes used would require variation in the types of onset presented in our paradigm.Our paradigm differs from that usually taken to study the attentional window. For starters, rather than showing that observers can ignore captures when in a parallel search mode, we made sure that participants were engaged in serial search by using a search task. While the default expectation is that this would result in a narrow window, we found otherwise. Participants could select which onsets captured attention, even to the extent that their spatial window covered the whole visual display-thus mimicking parallel search. Clearly, despite the task, participants still had voluntary control over their attentional deployment.behavioural goals. This conclusion does not exclude that the attentional window can be narrow during serial search tasks. In fact, we show that it is possible when little to no benefit for the search task is expected from examining the onset positions. Narrowing one's attentional window appears to be a deliberate choice, since a reaction to abrupt onsets may distract from systematic scanning of the environment The idea that automatic versus top-down control is guided by the type of search task appears to be an oversimplification Our finding that participants used their prior knowledge – of the congruency rates – to voluntary modulate capture is in line with recent results by Sayim et al. Overall, our current results favour the attentional window account. Moreover, this account may be a good candidate to reconcile the results of previous work . In the latter condition, a complete reliance on onset pop-out would still have been very ineffective.In addition, the proposed top-down regulated attentional window account does not provide a complete account of all attentional capture behaviour. Folk, Remington and Johnston Through measuring attentional capture using gaze tracking, we showed that the attentional window is modulated in accordance with the expected relevance of environmental signals. For this, we used an (ongoing) search task that enabled daily life visual behavior. The more relevant the signals were expected to be, the larger the observers set their attentional window. Thus, attentional window size can be voluntarily modulated in accordance with individual behavioural goals. This account of attentional capture may be applied to improve our ability to guide an observer's attention and gaze to regions of interest, with the ultimate goal of improving information communication.


Experimental results suggest a thickness reduction of few-layer graphene sheets subjected to prolonged CF4 plasma treatment while plasma treatment with short time only leads to fluorine functionalization on the surface layer by formation of covalent bonds. Raman spectra reveal an increase in disorder by physical disruption of the graphene lattice as well as functionalization during the plasma treatment. The F/CF3 adsorption and the lattice distortion produced are proved by theoretical simulation using density functional theory, which also predicts p-type doping and Dirac cone splitting in CF4 plasma-treated graphene sheets that may have potential in future graphene-based micro/nanodevices.Structural changes of few-layer graphene sheets induced by CF81.05.ue; 73.22.Pr; 52.40.Hf. Our results suggest an obvious thickness reduction effect in few-layer graphene sheets treated with CF4 plasma as well as surface fluorine functionalization by formation of covalent bonds between the top graphene layer and the ions. The produced disorder in graphene lattice is well reflected in the Raman spectra, and the corresponding mechanism is studied theoretically. The results presented in this work provide a possible direction to obtain giant single-layer graphene sheets with necessary surface functionalization to realize the p-type doping level and to open the Dirac cone for future graphene-based micro/nanodevices.Graphene is one layer of C atoms, arranged in a hexagonal lattice -3. Since2. The samples were then cleaned by ultrasonication in acetone for 30 s to remove the residual of the scotch tape and unattached graphite pieces. The CF4 plasma treatment was carried out in the reaction chamber of a Jupiter III reactive-ion etching setup. The samples were exposed to 0.8-Torr CF4 under radio-frequency plasma (20 W) with different times ranging from 1 to 5 s with a step of 1 s. In our experiment, the number of layers in the few-layer graphene sheets can be estimated by a combination of optical microscopy and Raman spectroscopy. Raman spectra were taken on a Renishaw inVia micro-Raman spectrometer with the 514-nm line of an Ar+ laser as light source . All the measurements were carried out at room temperature.The few-layer graphene sheets used in the current study were produced by mechanical exfoliation from highly ordered pyrolytic graphite and transferred on a silicon substrate covered with 300-nm SiO4 plasma treatment of different times. For a short plasma treatment  and the small full width at half maximum (FWHM) of the 2D band (26 cm−1) indicate that the graphene sheet measured contains only one layer  [−1 and is connected to transverse optical phonons near the K point which require defects or lattice disorder  for their activation via an inter-valley double-resonance Raman process [−1 is remarkably intensified, suggesting an introduction of lattice disorder in the graphene sheet [−1 which is believed to be a combination of D and D' bands [4 plasma treatment. Thus, the Raman measurement was carried out on a few-layer graphene sheet which contains more than one layer before plasma treatment. The obtained spectrum was plotted in the lower panel of Figure −1 and the ratio of I2D/IG is 1.1, elucidating that this sheet consists of a few layers. After 5 s of plasma treatment, the Raman spectrum in the upper panel of Figure −1 and the ratio of I2D/IG is increased to 2.5. The spectral evolution is different from that in 2 s of plasma treatment and indicates that the few-layer graphene layer is thinned to one or two layers [However, a structural modification or surface functionalization may exist during CFreatment ,16,17, ereatment ,21,22. I,680 cm−1. The intraphene) ,23. Besi process . Therefone sheet ; (2) thene sheet ,25; (3)  2D band  cm−1 indD' bands . These fD' bands  and are D' bands ,16. Howeo layers ,23,28, a4 plasma for different times. The relative intensity of the D band (ID/IG) in the sample treated for 5 s is smaller than that for 2 s, implying that the disorder is even remarkable for shorter treatment. To understand this peculiar behavior, we must first make clear the mechanism of disorder production during plasma treatment. The disorder and the corresponding emergence of D and D' bands in our samples may arise from two processes: (1) physically, the order of pristine C atoms in graphene is disrupted by the CF4 plasma and some of the C atoms may be sputtered out, which is named as ion bombardment effect [n (n = 1 to 3) or F species were only adsorbed on the top graphene layer by formation of covalent bonds, which will become saturated because the number of available active C atoms decreases with time. It is worth noting that the formation of covalent bonds can be evidenced by the upshift of 2D peak from 2,685 to 2,691 cm−1[4 plasma treatment therefore possesses disorder features from both the physical and chemical interactions. The chemical interaction takes place only on the topmost layer, while the physical interaction accumulated with time and a long treatment can remove the top layer , exposing the beneath layer. The emergence and intensification of the D and D' bands in the sample subjected to 5 s of treatment thus originates from the disorder created in this new top layer and should increase gradually. Consequently, the disorder probed by Raman spectroscopy is even smaller in the sample treated by CF4 plasma for 5 s than in the sample treated for 2 s (see Figure The evolution of the D band is the most important concern of many researches. The increase of its intensity is generally considered to be evidence that lattice disorder exists in the graphene sheet . An intet effect ,30; and ,691 cm−1. One mayID/IG is obviously relevant to the thickness of few-layer graphene sheet. For a thicker sheet, the weight of the topmost layer with remarkable disorder is smaller, and thus ID/IG should be correspondingly smaller. Therefore, the ration of ID/IG proves to be an effective Raman factor to compare the thicknesses of plasma-treated few-layer graphene sheets. Therefore, the ID/IG proves to be another effective factor to estimate the thicknesses of plasma-treated few-layer graphene sheets other than I2D/IG.Since the intensity of the D band reflects the disorder in the top layer of the few-layer graphene sheet, the relative intensity of the D band may give us a clue about the thickness of the sample. To prove this, we measured Raman spectra of plasma-thinned few-layer graphene sheets with different thicknesses. The three spectra in Figure 2, and CF3 ions. The energy barrier is calculated by the climbing-image nudged elastic band method [2 can hardly be adsorbed, forming covalent bonds, while F and CF3 ions can be adsorbed. The corresponding adsorption energies are 2.2 and 0.4 eV, respectively. The different absorption behaviors can be understood as follows: Because of the sp3 hybridization nature for C atoms in CF, CF2, and CF3, there are two un-bonded electrons in CF2 and only one in CF3, while the C atoms in graphene are sp2 hybridized and there is only one un-bonded electron for each C atom. Thus, it is easier for CF3 to be adsorbed on top of the C atom via a covalent bond (see Figure 2 to form two covalent bonds with two nearby C atoms in the graphene sheet. We checked different absorption configurations by structure relaxation and find that for all cases, the CF2 will leave away from the graphene, as shown in Figure 2 and graphene is mainly due to the large strain/stress. A similar situation is also noted in the CF case. In addition, the calculated energy barrier for the CF3 adsorbed on the graphene surface is only 0.04 eV, which demonstrates that the CF3 adsorption is energetically favorable. Due to the existence of covalent bonds between F/CF3 and C on the graphene sheet, there is distortion around the adsorption position, which gives birth to the appearance of D and D' bands in the Raman spectra of the plasma-treated samples.Although a detailed experimental investigation is so far difficult, we managed to simulate the ion adsorption during the plasma treatment by employing density functional theory and the generalized gradient approximation of the exchange correlation functional with ultrasoft pseudopotentials ,33 to red method , which e3 with coverage of 5.6 %, as shown in Figure 3 functionalization due to electron transfer from the C atoms in the graphene sheet to F/CF3, indicating that the graphene sheets can be p-type doped by fluorinating with both F and CF3. However, it can be found that F is much more efficient to introduce p-type doping in graphene sheets than CF3. In addition, the adsorption of the F/CF3 on graphene sheets also introduces pinning states around the Fermi levels, as demonstrated in Figure 4 plasma in micro/nanoelectronics.Here, we would like to briefly discuss the potential applications of this fluorine functionalization. We calculated the band structures and densities of states of graphene sheets fluorinated with F and CF4 plasma treatment was studied in detail by optical microscopy and Raman spectroscopy. The experimental results indicate a thickness reduction under prolonged plasma treatment while short treatment leads only to fluorine functionalization on the surface layer. The combination of both physical and chemical reactions in the plasma treatment leads to structural modifications which can be well probed in Raman spectra. Theoretical simulation suggests a F/CF3 functionalization by formation of covalent bonds and also predicts a corresponding p-type doping and Dirac cone opening after the F/CF3 adsorption. Although further characterizations are needed to evaluate the electronic properties of treated samples, the current work of thinning and functionalizing few-layer graphene sheets by CF4 plasma under control represents an integrative pathway to industrial fabrication of graphene-based micro/nanodevices.In conclusion, few-layer graphene sheets were prepared by mechanical exfoliation and the structural evolution during CFThe authors declare that they have no competing interests.CS, GH, FD, and YM designed the study. CS performed the experiments with help from RC. YC and US carried out the theoretical study. CS, GH, YC, and YM contributed in drafting the manuscript. All the authors took part in the discussion of the results, and edited and approved the manuscript.


Integrated temperature-humidity-sensitive thick-film structures based on spinel-type semiconducting ceramics of different chemical compositions and magnesium aluminate ceramics were prepared and studied. It is shown that temperature-sensitive thick-film structures possess good electrophysical characteristics in the region from 298 to 358 K. The change of electrical resistance in integrated thick-film structures is 1 order, but these elements are stable in time and can be successfully used for sensor applications. Nanostructured functional spinel-type ceramics based on magnesium aluminates and mixed transition metal manganites are known to be widely used for temperature and humidity measurement -5. But tA number of important problems connected with hybrid microelectronic circuits, multilayer ceramic circuits, temperature sensors, thermal stabilizers, etc. require such resolution, when not bulk , but only the thick-film performance of electrical components (possessing the possibility to group-technology route) is needed . The wel3[2 additives, it was shown that temperature-sensitive elements in thick-film performance attain additionally good humidity sensitivity [B constant value at the level of 3,000 K, such thick-film elements possess only small humidity sensitivity. This disadvantage occurred because of relatively poor intrinsic pore topology proper to semiconducting mixed transition metal manganites in contrast to dielectric aluminates with the same spinel-type structure.To fabricate the integrated temperature-humidity thick-film sensors, only two principal approaches have been utilized, they being grounded on temperature dependence of electrical resistance for humidity-sensitive thick films and/or on humidity dependence of electrical resistance for temperature-sensitive thick films. The first approach was typically applied to perovsite-type thick films like BaTiO3. Within sitivity . Despite2O4-CuMn2O4-MnCo2O4 concentration triangle has a number of essential advantages, non-available for other ceramic composites. Within the above system, one can prepare the fine-grained semiconductor materials possessing p+-type (Cu0.1Ni0.1Mn1.2Co1.6O4) and p-type of electrical conductivity (Cu0.1Ni0.8Mn1.9Co0.2O4). Prepared thick-film nanostructures involving semiconductor NiMn2O4-CuMn2O4-MnCo2O4 and insulating (i-type) MgAl2O4 spinels can be potentially used as simultaneous thermistors and integrated temperature-humidity sensors with extremely rich range of exploitation properties.The thick-film performance of mixed spinel-type manganites restricted by NiMn2O4-CuMn2O4-MnCo2O4 ceramics are used not only as temperature-sensitive layers but also as conductive layers for humidity-sensitive thick films based on MgAl2O4 ceramics.The aim of this work is to develop the separate temperature- and humidity-sensitive thick-film nanostructures based on spinel-type ceramics, in which the semiconducting thick films based on NiMn0.1Ni0.1Co1.6Mn1.2O4, Cu0.1Ni0.8Co0.2Mn1.9O4, and MgAl2O4 spinel ceramics with optimal structural properties [Previously studied and selected samples of Cuoperties -18 were 0.1Ni0.1Co1.6Mn1.2O4 ceramics were sintered at 1,040°C for 4 h and Cu0.1Ni0.8Co0.2Mn1.9O4 ceramics at 920°C for 8 h, 1,200°C for 1 h, and 920°C for 24 h [0.1Ni0.1Co1.6Mn1.2O4 ceramics  and Cu0.1Ni0.8Co0.2Mn1.9O4 ceramics  with additional NiO phase (10%) [Temperature-sensitive ceramics were prepared by a conventional ceramic processing route using reagent grade cooper carbonate hydroxide and nickel  carbonate hydroxide hydrates . The Cu0for 24 h -23. As ase (10%) .2O4 ceramics were prepared via conventional sintering route as was described in more details elsewhere [Ts = 1,300°C for 5 h.The bulk MgAllsewhere -18. The 0.1Ni0.1Co1.6Mn1.2O4/Cu0.1Ni0.8Co0.2Mn1.9O4-based pastes were prepared by mixing powders of basic ceramics  with ecological glass powders (2.9%) without PbO, inorganic binder Bi2O3 (2.9%), and organic vehicle (21.4%). The next content was used for the preparation of humidity-sensitive thick-film pastes: MgAl2O4-based ceramics (58%), Bi2O3 (4%), ecological glass (8%), and organic vehicle (30%).Temperature-sensitive Cu+-conductive paste was formed on humidity-sensitive i-type layer as conductive layer. Then, these structures were sintered in the furnace. The topological scheme of integrated p-i-p+ thick-film structure is shown in Figure The pastes were printed on alumina substrates  using a manual screen printing device equipped with a steel screen. Then, thick films were sintered in PEO-601-084 furnace at 850°C ,23. The The microstructure of the sintered temperature-sensitive ceramics was probed using an electron microscope JSM-6700 F , cross-sectional morphology of the samples being tested near the surface (0- to 70-μm depth) and chip centers. Scanning electron microscopy (SEM) investigations for bulk humidity-sensitive ceramics and thick-film structures were performed using LEO 982 field emission microscope .The pore size distribution of bulk semiconductor and dielectric ceramics in the region from 2 to 1,000 nm was studied using Hg-porosimetry .R on relative humidity (RH) of the environment. The electrical resistance was measured in the heat and humidity chamber PR-3E  at 20°C in the region of RH = 20% to 99%. The electrodes were attached to connecting cables of M-ohmmeter at fixed current frequency of 500 Hz (with the aim of avoidance of polarization of adsorbed water molecules). In addition, the degradation transformation at 40°С and RH = 95% for 240 h was carried out in order to study sample stability in time. The maximal overall uncertainties in the electrical measurements did not exceed approximately ± (0.02 to 0.04) MΩ in electrical resistance. The confidence interval in RH measuring bar restricted by equipment accuracy was no worse than ±1% and in temperature measuring bar ±0.5°C.The electrical resistance of thermistor thick films was measured using temperature chambers MINI SUBZERO, Tabai ESPEC Corp., Japan, model MC-71 and HPS 222. The humidity sensitivity of thick-film structures was determined by measuring the dependence of electrical resistance 2O4 ceramics, which are used for the preparation of humidity-sensitive thick-film layers, are characterized by tri-modal pore size distributions  of each thick-film layers, such effects correspond to the changes in their sensitivity, in particular, decreasing of sensitivity on i-type thick-film layer, due to diminishing of pores connected with capillary condensation processes  and addi+-conductive temperature-sensitive thick-film elements based on spinel-type NiMn2O4-CuMn2O4-MnCo2O4 ceramics have good electrophysical characteristics. These thick-film elements show linear temperature dependences of resistances  and possess high temperature/humidity sensitivities, they will be positively distinguished not only by wider functionality (simultaneous temperature-humidity sensing) but also by unique functional reliability and stability. In the case under consideration, the main advantages proper to bulk transition-metal manganite ceramics  and humidity-sensitive MgAl+ structures with optimal grain-pore structures, where p+-conductive layers was used as a conductive layer, were obtained and studied. Temperature-sensitive thick-film structures possess good temperature sensitivity in the region from 298 to 358 K. The humidity-sensitive elements possess linear dependence of electrical resistance on relative humidity in semilogarithmic scale with some hysteresis in the range of RH ~ 60% to 99%. After degradation transformation, the hysteresis is minimized due to saturation of some nanopores by water, which provide effective adsorption-desorption processes in elements.Integrated temperature-humidity sensitive thick-film p-i-pThe authors declare that they have no competing interests.HK performed the experiments to study the temperature and humidity effects in thick-film structures and drafted, wrote, and arranged the article. IH proposed an idea of the development of integrated thick-film structures and performed the experiments to obtain bulk spinel ceramics and thick films. OS is a supervisor of the whole work, the results of which are presented in this article. MB supervised the experiments performed by IH. All authors read and approved the final manuscript.


The system integrated three parts: a flexible capacitive humidity microsensor, a flexible resistive temperature microsensor, and a radio frequency (RF) module for signal transmission. The results show that the capacitive humidity microsensor has a high sensitivity of 0.83 pF%RH−1 and the resistive temperature microsensor also exhibits a high sensitivity of 2.94 × 10−3 °C−1. The established RF module transmits the signals from the two microsensors. The transmission distance can reach 4 m and the response time is less than 0.25 s. The performance measurements demonstrate that the maximum power density of the fuel cell with and without these microsensors are 14.76 mW·cm−2 and 15.90 mW·cm−2, with only 7.17% power loss.This study developed portable, non-invasive flexible humidity and temperature microsensors and an  The fuel cell is one possible substitute since it produces no carbon dioxide but only water. Fuel cells can be characterized into several categories  such as et al. [et al. [Several researchers have developed methods for inner flooding detection. Nishikawa et al.  drilled  [et al.  manufactConsequently, this study proposes flexible thin-film humidity and temperature microsensors embedded in PEM fuel cell for the purpose of preventing the above issues. In addition, this work also establishes a wireless module to integrate sensors because it has advantages of portability and dependability. Wireless sensors are generally smaller in size and lighter in weight so that it can be carried more easily than conventional instruments. Hence, the investigation employs wireless module to integrate microsensors.The remaining sections are organized as follows: humidity microsensor, temperature microsensor, and wireless module are introduced first. Then, calibration for sensors, the result of wireless integration and measurement consequence within fuel cell are presented.2.2.1.Humidity microsensors can be categorized into resistive and capacitive types ,8, accor2.2.Temperature microsensors are mainly classified into two kinds: thermocouples and resistive temperature sensors. Due to the reason that the fabrication process of thermocouples is complicated so as to increase cost and that resistive temperature sensor shows fine sensitivity, a resistive type sensor acted as our temperature sensor. 2.3.3.3.1.H = ΔC/Δ%RH [−1 at 70 °C. In addition, the linearity gets better as the temperature increases. The humidity microsensor is wire bonded using a dispenser and is tested inside an environmental chamber (Model HUNGTA-HT-8045A). Capacitance measurements were performed at 1 kHz using a LCR meter (Model Wayne Kerr 4230). Tests for the humidity sensor are conducted at various temperatures from 25 °C up to 70 °C as well as for different relative humidity levels ranging from 35%RH to 95%RH. The capacitance value of the microsensor is always recorded after waiting for 15 min at each measurement condition to assure that the whole volume of the climatic chamber reaches a stable temperature and humidity level.  ΔC/Δ%RH ) increasa is resistance value at temperature a °C, Rb is the resistance value at temperature b °C, ΔT is the temperature difference (=a − b). −3 °C−1. In comparison with other works [In the calibration of the temperature microsensor, it is also characterized in the same environmental chamber. Resistance measurements are made through a digital multi-meter (Model HP-HEWLETT-PACKARD-3478A). According to the reference , the sener works –17 as shn as follows:The measurement results by wireless integration are displayed in 3.2.−1 and humidification temperature of 70 °C. Flooding is observed after steady output voltage for 28 min. Thus, it is necessary to revise the humidification condition for the fuel cell to improve this situation. −2 and 15.90 mW·cm−2, indicating the power loss ratio of 7.17%. Such the power loss ratio is much better than 55% of Lee [The wireless sensing system is implemented to perform internal diagnosis of a fuel cell. % of Lee .4.−1 and the temperature sensor exhibits a sensitivity of 2.94 × 10−3 °C−1. Furthermore, a portable, lightweight wireless RF module is developed to transmit the sensor signals. Such the design enables the diagnosis of a fuel cell out of the lab. The wireless detection distance reaches 4 m and its response time is less than 0.25 s. The performance tests show that the maximum power density of the fuel with and without microsensors are 14.76 mW·cm−2 and 15.90 mW·cm−2. In other words, the power density loss ratio is only 7.17%. With regard to further applications of the wireless sensing system, it can be integrated with humidification control equipment to achieve the formation of closed loop system so as to carry out the aim of in situ gas humidification management.This study has successfully established a distinctive means for measuring the internal status of a PEMFC. A thin film and flexible capacitive humidity microsensor and a resistive temperature microsensor are fabricated using MEMS processing. The humidity sensor can reach a sensitivity of 0.83 pF%RH


The European Alps harbour a unique and species-rich biodiversity, which is increasingly impacted by habitat fragmentation through land-use changes, urbanization and expanding transport infrastructure. In this study, we identified the 50 most important questions relating to the maintenance and restoration of an ecological continuum – the connectedness of ecological processes across many scales including trophic relationship and disturbance processes and hydro-ecological flows in the European Alps. We initiated and implemented a trans-national priority setting exercise, inviting 48 institutions including researchers, conservation practitioners, NGOs, policymakers and administrators from the Alpine region. The exercise was composed of an initial call for pertinent questions, a first online evaluation of the received questions and a final discussion and selection process during a joint workshop. The participating institutions generated 484 initial questions, which were condensed to the 50 most important questions by 16 workshop participants. We suggest new approaches in tackling the issue of an ecological continuum in the Alps by analysing and classifying the characteristics of the resulting questions in a non-prioritized form as well as in a visual conceptualisation of the inter-dependencies among these questions. This priority setting exercise will support research and funding institutions in channelling their capacities and resources towards questions that need to be urgently addressed in order to facilitate significant progress in biodiversity conservation in the European Alps. During the past few years, major efforts have been undertaken to maintain and foster pan-alpine biodiversity. The “Platform Ecological Network” of the Alpine Convention , the “Ecological Continuum Initiative”  and the Alpine Space project “ECONNECT” (www.econnectproject.eu) are three examples of such initiatives that also aim at enhancing ecological connectivity across the Alpine range.The European Alps span eight countries, from the Mediterranean shores of Southern France to Slovenia and link with adjacent mountain ranges such as the Carpathians, Balkans and Apennines. The Alps harbour an extraordinary diversity of habitats, plants and animals including many endemics. They are considered as one of the most important regions for the preservation of biodiversity in Europe ECONNECT is a project funded by the EU within the framework of the European Territorial Cooperation Alpine Space Programme and co-funded by the European Regional Development Fund. The project envisions an enduringly restored and maintained ecological continuum, consisting of inter-connected landscapes and protected areas, across the Alpine Arc region, where biodiversity will be conserved for future generations and the resilience of ecological processes will be enhanced. Employing an integrated and multidisciplinary approach, the project has provided an Alpine-wide overview on areas important for ecological connectivity by integrating quantitative and qualitative information on selected sites and the level of interconnectivity among them Sutherland et al. 2011 In this study, we identified the 50 most important questions relating to the maintenance and restoration of an ecological continuum in the European Alps. We initiated and implemented a trans-national priority setting exercise, inviting researchers, practitioners, NGOs, policy makers and other stakeholders from the Alpine region to participate. The exercise was composed of an initial call for pertinent questions, a first online evaluation of the received questions and a final discussion and selection process during a joint workshop. Furthermore, we suggest new approaches in tackling the issue of an ecological continuum in the Alps by analysing and classifying the characteristics of the resulting questions in a non-prioritized form as well as a visual conceptualisation of the inter-dependencies among these questions.The non-prioritized list of the 50 most important questions concerning an ecological continuum in the Alps is shown in The largest proportion of questions (46%) was attributed to the nature context. The nature context questions are rather evenly distributed between the three subtopics. This is followed by the management context (44%) where by far the largest proportion of questions relates to the legislation, policy and planning needs subtopic (63%). Finally the people context makes up a mere 10% of the total questions. From the 50 questions, by far the largest fraction (60%) was formulated as “how” questions, followed by “what” (26%) and “which” questions (14%). Consequently, most attention was given to transformation processes aiming at practices to improve the current situation in Alpine connectivity.We created a graph in the form of a web to illustrate the “location” of each question within the framework of the nine sub-topics in the nature-, people- and management-context . Each coOverall, 27 questions were assigned to a second context area due to their transdisciplinary nature. The sub-topics 6 “economic, social and political needs” (PC) and 9 “tools, incentives, knowledge” (MC) showed the most inter-relations to other context areas .The aim of this study was to identify and analyse gaps of knowledge with respect to achieving, restoring and maintaining an ecological continuum in the European Alps. In order to identify information gaps, we implemented an alpine wide “priority setting exercise” as previously described by Sutherland et al. http://www.chathamhouse.org/about-us/chathamhouserule) will be adhered to is in our view beneficial to encourage openness and the sharing of information. Furthermore, as previously stated by Wuelser et al. The output of priority setting exercises depends on a number of factors, such as the selection of participants, the number of individuals effectively involved in formulating the questions and on the voting system A further issue that hindered and complicated the exercise is the fact that the Alps are distributed across eight countries, 28 regions, 98 provinces and 5 spoken languages. Participants originated from five alpine countries (in the collection phase complemented by one participant from the UK) and divergent cultural backgrounds. In contrast to most previous studies that were English-language based, this study involved four different languages , which made the succinct formulation of questions particularly difficult. We tried to alleviate the fact that English was a foreign language to all of the participants by consulting with a native speaker during the entire process. Similarly, Braunisch et al. Similarly question 15 “What is an effective set of indicators (for species and habitats) that can be used to evaluate and monitor connectivity at different scales?” is a prerequisite in addressing 3 other questions: namely question 2 “How are corridors best implemented; with clearly spatially defined borders or as functional units integrated in wide ecological continuums?”, question 19 “Are artificially engineered ecological networks a threat or benefit to endemic species?” and question 46 “What methods can be used to record the effectiveness of functional connectivity, or its elements, across different spatial and temporal scales?”. Essential questions like question 15, dealing with indicators and methodological approaches, should be given clear priority, as their answer is a precondition for scientifically sound answers to other, subsequent questions.Solving certain questions in a consecutive order indicates a potential time lag between putting forward a question and actually answering it. This time lag may lead to a change of the initial situation, which would imply that the results of the preceding question are no longer suitable for solving the initial issue. This time lag in connectivity conservation makes it impossible to “come back” to the initial situation after the extended process of research, decision-making and implementation. This clearly indicates that there is an asymmetric drift of the initial problem and its proposed solution due to the general inertia in decision-making processes dealing with environmental issues. As holds true for numerous environmental problems, most notably global climate change, we suggest that the conservation and restoration of an ecological continuum is a “wicked problem” The gaps of knowledge, in conserving and restoring connectivity emphasized in this exercise make evident that these involve a highly dynamic and interconnected process rather than a simplistic and straightforward approach. It appears essential to reconcile the dynamic and complex nature of the problem with the problem solving approaches. Inadequate simplification of the interdependencies will possibly lead to results that are not relevant in forming policy In order to address the complex issue of an Alpine ecological continuum, it appears necessary to apply a forward reasoning approach which identifies possible future scenarios and integrates uncertainties , reflects the sectoral structure of society, governance and administration with respect to environmental problems in general. To overcome this, an integrative transdisciplinary approach is necessary. What appears to be missing in order to find a starting point to address the problem of the Alpine ecological continuum, is a common strategy or vision The visual “chaos” and multi-structural character of our results, shown in the “web of questions” , reflectFor this type of study to provide guidance and contribute towards conservation-action implementation the results must be disseminated accordingly. As has been pointed out previously, bridging the gap of knowledge between research and conservation practice cannot be achieved with unidirectional platforms A core team of five researchers and practitioners initiated and facilitated the entire exercise. The priority setting process was divided into four steps: initiation, assembling and organizing initial questions, pre-workshop voting and workshop voting. The method has been previously described in detail in various publications Initiation. Based on the individual networks of the core team and striving to depict a representative cross-section of the main players in Alpine biodiversity conservation, we invited 48 institutions including researchers, conservation practitioners, NGOs, policymakers and administrators from the Alpine area to participate in this initiative.Collating of initial questions. 25 of the invited 48 institutions (52%) generated 484 initial questions. The core team collated and assigned these questions to twelve main topics for clarity .Pre-workshop voting. In order to reduce the set of questions, we distributed the initial-questions-list to all participants for a first evaluation. Rephrasing and adding of questions was actively encouraged at this stage. In preparation for the workshop phase, participants were then asked to select a maximum of 55 questions from the total of 484 questions submitted. Participants were also asked to only select questions from areas in which they had sufficient knowledge base. A total of 385 questions received at least one vote and an additional 15 questions were proposed at this stage.Workshop. Final discussion and voting took place during a two-day workshop in December 2010 in Switzerland. Unfortunately, only 16 participants were able to attend. The remaining 400 questions assigned to the twelve topic areas  were discussed in four sessions. In each session, three groups worked simultaneously to reduce and rephrase the questions. In the plenary session the participants decided to reduce the questions to 50 instead of 55 due to overlaps. The final 50 questions were selected in a plenary session by majority vote following discussion.The core team ascribed individual questions from the final non-prioritized list of 50 questions, to three interacting context areas according to a slightly modified, previously published, management framework for connectivity conservation: nature-, people- and management contexts. Each context area has 3 interacting sub-topic areas, We subsequently assigned each question to one or more sub-topic area within the individual context areas  [15]. WeAll questions were formulated either as (i) what - asking for information about something, (ii) which - asking about choice and (iii) how – questioning manner, condition or quality in order to further breakdown and classify the outcomes.Finally, we established a simple non-prioritized list that attributes all 50 questions to the three context areas and respective sub-topics  and 2. T


Sarcoidosis is a disease process which predominantly affects the lungs but can involve virtually any organ in the human body. Neurosarcoidosis is a rare manifestation which can present in a variety of ways. There is no single diagnostic test for sarcoidosis; hence, the diagnosis is based on combined clinical, laboratorial, and radiological grounds. We describe a rare case where a patient presented with dysphagia, hoarseness, hearing loss, and unsteadiness. Sarcoidosis is a multisystem granulomatous disorder of unknown cause. Although it can involve virtually any organ system, it shows a predilection for the lungs with up to 90% of cases displaying pulmonary sarcoidosis. This is followed by ocular, skin, lymph node, and salivary gland involvement. Neurosarcoidosis is relatively rare accounting for approximately 5–15% of cases . As the A previously healthy 32-year-old lady presented acutely to the ORL service with a 2-day history of increasing dysphagia, hoarseness of voice, and a blocked sensation in the right ear. She was unable to swallow any solids and experienced choking spells with thin fluids. She also described a recent viral-like illness that lasted for about 2 weeks followed by an episode of left sided facial weakness and asymmetry earlier during the week which seemed to have resolved completely at the time of presentation. On further questioning, she reported vague symptoms of unsteadiness especially while driving her car.On examination, she had a very husky voice associated with difficulty in swallowing her saliva. She had weak palatal muscles and an absent gag reflex. Flexible laryngoscopic examination showed marked hypomobility of the left vocal cord and pooling of secretions in the hypopharynx. Pure tone audiogram showed a mild, high frequency sensorineural hearing loss on the left and a downsloping mild to moderate sensorineural hearing loss on the right side, (as illustrated in Supraclavicular lymph node core biopsy showed granulomatous lymphadenitis. She was diagnosed with sarcoidosis on the grounds of clinical, radiological, and histological findings.She made a full recovery after a course of corticosteroids.The diagnosis of neurosarcoidosis can be difficult to make due to the variability of presentation which includes cranial nerve palsies, meningitis, seizures, and neuropsychiatric symptoms , 2.Multiple cranial neuropathies are uncommon presentations with diverse causes. Tumours in general appear to be the commonest cause accounting for about 30% of cases. This is followed by vascular disease, trauma, and infection. In Keane's 34-year review of 979 cases of multiple cranial palsies, sarcoidosis was the cause in 2 patients only [The most common neurologic manifestation of sarcoidosis is cranial neuropathy secondary to nerve granulomas, raised intracranial pressure, or granulomatous meningitis . The facAlthough the cause of sarcoidosis is unknown, an abnormal immune response to infectious and noninfectious environmental factors appears to be a key factor , 2. ThisThere are various diagnostic criteria suggested for neurosarcoidosis, and these classify cases as definite, probable, and possible neurosarcoidosis. A CNS or extraneural biopsy is usually required for a diagnosis of definite sarcoidosis .Although there are no established treatment guidelines in neurosarcoidosis, corticosteroids are the first drugs of choice and patients usually respond to high doses. Cytotoxic drugs such as methotrexate and azathioprine can be used in patients where corticosteroids are contraindicated or in cases that are resistant to corticosteroids. Surgical intervention is reserved from life-threatening situations with large intracranial or spinal granulomas , 2.Neurosarcoidosis is still a diagnostic dilemma due to its rarity and variable presentation. However, cranial nerve palsies are common presentations in otolaryngology; hence, granulomatous disorders such as sarcoidosis should always be part of the differential diagnosis.


It is widely accepted that orthologous genes have lost or gained introns throughout evolution. However, the specific mechanisms that generate these changes have proved elusive. Introns are known to affect nearly every level of gene expression. Therefore, understanding their mechanism of evolution after their initial fixation in eukaryotes is pertinent to understanding the means by which organisms develop greater regulation and complexity.To investigate possible mechanisms of intron gain and loss, we identified 189 intron gain and 297 intron loss events among 11 Drosophila species. We then investigated these events for signatures of previously proposed mechanisms of intron gain and loss. This work constitutes the first comprehensive study into the specific mechanisms that may generate intron gains and losses in Drosophila. We report evidence of intron gain via transposon insertion; the first intron loss that may have occurred via non-homologous end joining; intron gains via the repair of a double strand break; evidence of intron sliding; and evidence that internal or 5' introns may not frequently be deleted via the self-priming of reverse transcription during mRNA-mediated intron loss. Our data also suggest that the transcription process may promote or result in intron gain.Our findings support the occurrence of intron gain via transposon insertion, repair of double strand breaks, as well as intron loss via non-homologous end joining. Furthermore, our data suggest that intron gain may be enabled by or due to transcription, and we shed further light on the exact mechanism of mRNA-mediated intron loss. Spliceosomal introns, segments of RNA that are excised by the spliceosome during the processing of pre-mRNA in eukaryotes, are found in varying quantities and positions among orthologous genes. By identifying orthologs, aligning gene sequences, and coupling intron absences/presences with known species phylogenies, numerous studies have identified the number of intron gains and losses that have occurred among species throughout evolution -11. HoweAs a deeper understanding of gene expression emerges, it is evident that introns not only increase proteome diversity through their well known role in alternative splicing , but alsReverse Transcriptase-Mediated Intron Loss (referred to as RTMIL in this work)   . The annSTMapper . Ortholoab initio gene finding programs that were used to annotate genes in the 11 Drosophila genomes and the use of well annotated D. melanogaster genes during the annotation of the other 10 Drosophila genomes greatly increased the reliability of these annotations [D. melanogaster may lack experimental validation, annotation errors may exist. Therefore, we applied quality controls to each intron identified as an intron gain in a single species. First, we excluded all intron gains located within a single species that were length 3 n (where "n" is an integer) and did not contain a premature termination codon (PTC) . This criterion was based on a recent study in Drosophila that also used computationally annotated introns to identify intron gains and losses. In that study, 86% of predicted intron gains that were located in a single species and were length 3 n without PTCs were annotation errors as opposed to novel introns [The ability to accurately identify intron gains and losses relies upon accurate gene annotation. The multitude of comparative and otations . HoweverPY participated in the design of the study, performed and conceived analyses, and drafted the manuscript. BK wrote computer programs for data analysis. LZ conceived of the study, participated in its design, wrote computer programs for data analysis and helped to draft the manuscript. All authors read and approved of this version of the manuscript.Supplementary figures. Figures used to provide further information about the alignments and various cases of intron gain/loss events.Click here for fileOrtholog dataset. A matrix of all orthologs used in final analyses. Each ortholog group is listed on one line.Click here for file


Cough is a common symptom in people who develop interstitial lung diseases (ILD). The pathological features of the ILDs are many and varied suggesting that the cause of cough may also vary with each disease. This article reviews what is currently known about cough in sarcoidosis and idiopathic pulmonary fibrosis; two of the commonest ILDs. It also outlines some of the theories which have been proposed to explain why cough develops in these conditions and describes what little is known about how to treat it. Sarcoidosis is a multi-system disorder of unknown aetiology which is characterised histologically by the infiltration of affected tissue by non-caseating granulomatous inflammation. Whilst any part of the body can be involved, sarcoidosis typically affects hilar and mediastinal lymph nodes, the lung interstitium and bronchial epithelum. Less commonly, the upper respiratory tract, pleura and pulmonary vessels may be involved. A number of epidemiological studies have investigated the prevalence of cough in people with sarcoidosis and whilst it appears rare in Japan, it is common in other parts of the world where the prevalence is between 30 and 50% Table  1-3].-3.1-3].Whilst little is known about the mechanisms by which sarcoidosis causes cough , clinicaOne retrospective study from China supports the link between granulomatous inflammation of the airways and cough in sarcoidosis. This study compared respiratory symptoms in 87 patients who had undergone tracheal biopsy and found that significantly more (92%) of those who had granulomatous tracheitis complained of cough compared with those (49%) who had normal tracheal biopsies . This stSarcoidosis of the upper respiratory tract (SURT) occurs in approximately 5% of cases and may variably affect the larynx, epiglottis, arytenoids, the aryepiglottic folds and vocal chords as well as the nasal mucosa and sinuses . There aThe enhanced cough reflex observed in people with saroidosis is likely initiated by sensory fibres of the vagus nerve that innervate the airways and larynx which have become sensitised by granulomatous inflammation. The precise mechanism by which this occurs is uncertain. However, it is interesting that Ricci and co-workers demonstrated that a number of non-neuronal cell types can express neurotrophins and their receptors in normal human lung . These iOccasional, unusual manifestations of sarcoidosis have been described which cause patients to present with the symptom of cough. These include granulomatous infiltration of the pulmonary veins causing pulmonary veno-occlusive disease with engorgement of airway epithelium and sarcoidosis affecting the vagus nerve at the level of the jugular foramen causing vocal cord palsy and cough ,13.There have been no randomised, placebo-controlled clinical trials which have investigated the effect of any pharmacological treatment specifically for cough in sarcoidosis. However, clinical observation suggests that when used in an appropriate clinical setting such as progressive interstitial lung disease, oral corticosteroid therapy has a beneficial effect on cough when it is a troublesome accompanying symptom. One double-blind, placebo-controlled study of inhaled corticosteroids in newly diagnosed patients with pulmonary sarcoidosis reported an improvement in global symptom score which included cough . HoweverIdiopathic pulmonary fibrosis (IPF) is the commonest form of idiopathic interstitial pneumonia. It is a disease of unknown aetiology which develops in the sub-pleural aspects of the posterior, basal parts of the lungs and becomes more extensive over time. On CT scan of the thorax the lungs show reticular opacities, often associated with ‘traction bronchiectasis’ and honeycombing Figure . On histlater, with severe and extensive fibrosis cough, unproductive or with scanty mucoid expectoration, may be troublesome’[It has been known for many years that cough is a clinical feature of IPF. In 1974, Scadding observed: ‘ublesome’. Howeverublesome’. More reublesome’. Interesublesome’. There iublesome’.Superficially it appears anomalous that an interstitial lung disease, which predominantly affects the alveolar walls in the peripheral lung where sensory innervation is relatively sparse , should silent dysphagia’ was a possible cause of ‘pulmonary lesions of obscure origin’. This hypothesis has been given further support by recent objective studies which used 24-hour oesophageal pH monitoring and oesophageal manometry to demonstrate that up to 87% of patients with IPF have evidence of acid reflux which is often asymptomatic [A significant barrier to the methodical study of cough in IPF is that patients frequently have confounding co-morbidities which can cause or exacerbate cough. These include COPD, sinusitis, hypertension treated by angiotensin converting enzyme inhibitors and, most importantly, gasto-oesophageal reflux disease (GORD). The latter is a known cause of chronic cough  and it hptomatic ,27. Inteptomatic .A recent, preliminary study applied the technique of combined oesophageal pH and impedance monitoring, which characterises all reflux episodes regardless of pH, to show that non-acid reflux also occurs in people with IPF . When coThe hypothesis that aspiration of gastric contents is an explanation for both the pathogenesis of IPF and its associated symptom of cough, has been confounded by a number of recent clinical and epidemiological observations. Firstly, that obstructive sleep apnoea (OSA) may be prevalent in people with IPF ,33; secoA different theory for the pathogenesis of cough in IPF proposes that mechanical distortion of the larger airways by progressive interstitial fibrosis, so-called ‘traction bronchiectasis’ Figure  increaseAs with sarcoidosis, there is some evidence for increased expression of neurotrophins in the lungs of people with IPF. It has been demonstrated that induced sputum from such people contains higher levels of NGF and BDNF compared with normal controls  and BALFTreatment of cough in IPF remains problematic for both patients and physicians, particularly in the later stages of the condition when it may be associated with increasingly severe breathlessness. In such cases, palliative therapy using conventional anti-tussive agents such as opiate-derived preparations often proves to be of limited benefit, as does optimal pharmacological treatment of acid reflux. One small, uncontrolled, open-labeled study found that cough sensitivity to capsaicin and cough symptom score was reduced after one month of high-dose oral corticosteroids suggesting that cough in IPF should be amenable to pharmacological therapy . In an oA recent double-blind, two-treatment, two-period crossover trial comparing thalidomide with placebo demonstrated a beneficial treatment effect on cough and quality of life as determined by questionnaires and visual analogue score . Whilst At the present time, physicians are all too often reduced to treating the debilitating cough of IPF with opiates as part of an end-of-life pathway. There is thus an urgent need for better treatments which can only be developed through our better understanding of the cough reflex in IPF.The author declares that he has no competing interests.


Mongolia’s riverine landscape is divided into three watersheds, differing in extent of permafrost, amount of precipitation and in hydrological connectivity between sub-drainages. In order to assess the vulnerability of macroinvertebrate communities to ongoing climate change, we consider the taxonomic and functional structures of stream communities in two major watersheds: The Central Asian Internal Watershed (CAIW) and the Arctic Ocean Watershed (AOW), together covering 86.1% of Mongolia’s surface area. We assess the consequences of the hydrological connectivity between sub-drainages on the nestedness and distinctness of the stream communities. And accordingly, we discuss the expected biotic changes to occur in each watershed as a consequence of climate change.Gamma and beta diversities were higher in the CAIW than the AOW. High community nestedness was also found in the CAIW along with a higher heterogeneity of macroinvertebrate assemblage structure. Assemblages characteristic of cold headwater streams in the CAIW, were typical of the drainages of the Altai Mountain range. Macroinvertebrate guilds of the CAIW streams exhibited traits reflecting a high stability and low resilience capacity for eutrophication. In contrast, the community of the AOW had lower nestedness and a combination of traits reflecting higher stability and a better resilience capacity to disturbances.Higher distinctness of stream communities is due to lower connectivity between the drainages. This was the case of the stream macroinvertebrate communities of the two major Mongolian watersheds, where connectivity of streams between sub-drainages is an important element structuring their communities. Considering differences in the communities’ guild structure, hydrological connectivity and different magnitudes of upcoming impacts of climate change between the two watersheds, respective stream communities will be affected differently. The hitherto different communities will witness an increasing differentiation and divergent adaptations for the upcoming changes. Accordingly, in an increasing awareness to protect Mongolia’s nature, our results encourage adapting conservation planning and management strategies specifically by watershed. Over the last two decades human-caused climate change has emerged as a leading threat to biodiversity conservation -3. The pTo predict the response of communities to climate change, we need first to understand how communities are currently structured and regulated. The structure of a stream invertebrate community is the result of several multiscale filters representing both historical factors and ecological constraints. The historical factors are considered to operate on a larger geographical scale and over geological time periods; while the ecological constraints are considered to operate mainly on the habitat scale and on shorter time scales ,13. The The impact of climate change on stream macroinvertebrate communities has been mainly studied in Europe  and Nort2). The AOW is bounded by Russia to the north, by the Hentei Mountain range to the east and by the Hangai Mountain range to the south and west. The CAIW is also bounded by Russia to the north, and is separated from China by the Gobi Desert to the south  The AOW covers only 20.6% of Mongolia’s surface area yet receives 51% of the annual precipitation of Mongolia while the CAIW covers 65.6% of Mongolia’s surface area and receives only 12% of the annual precipitation .2) The AOW has a highly connected network of sub-drainages that exit Mongolia by draining to the north, providing about 50% of the water inflow to Russia’s Lake Baikal , while the CAIW has 11 disconnected sub-drainages believed to be remnant of Tertiary and Quaternary drainages . Therefo3) The AOW is located at the boundary of the Siberian cryosphere, therefore having the highest permafrost extent in Mongolia Figure B, while http://clade.ansp.org/entomology/mongolia/] during multiple expeditions carried out in Mongolia from 2003 to 2010. Each stream site was sampled once during the 8 years timeframe. Sample sites in the AOW had a median elevation of 1432 m a.s.l.  and sample sites of the CAIW had a median elevation of 2029 m a.s.l. . This difference in altitudes of the stream-sites can be considered as an uncontrolled bias as high altitude streams from the AOW were scarce overall in the watershed and the majority were dry during the sampling expeditions. A recent sharp decrease of the glaciers in the Hangai Mountains may have contributed to the lack of summer-flow in the headwater streams of this mountain range . The frost-free period in Mongolia extends on average from late May to late August; therefore, all the sampling expeditions were carried out in July.The database for this study was obtained from 114 streams (37 from the AOW and 77 from the CAIW) sampled by the Mongolian Aquatic Insect Survey [At all sites the invertebrate communities were sampled following a kick sampling protocol using a 500 μm net mesh size and complying with the US EPA Rapid Bioassessment Protocol . Inverteet al. [Temperature, precipitation and humidity patterns, snow-cover decrease, glaciers melting and permafrost thaw are considered to be the main abiotic elements strongly affected by climate change in Mongolia. A substantial increase of the mean temperature by 1.6°C was observed over a 60-year period in Mongolia (1940-2001) . In addiet al.  found thet al. . In addiet al.  and the et al.  suggestsThese changes will be directly affecting the stream invertebrate communities. For instance, water temperature and thermal regimes regulate invertebrate population growth, population dynamics and overall niche expansion . IncreasFor these reasons, all biological traits deemed to be sensitive to or to describe affinities to temperature, disturbance, substrate structure, trophic status and eutrophication Table  were usehttp://purl.oclc.org/estimates.The total number of taxa per watershed constitutes gamma diversity. Since the numbers of sampled stream-sites per watershed were not equal, richness accumulation curves  were uset-test, (significance threshold 5%), followed by an analysis of nestedness to assess the impact of rare species on the community structure. Nestedness represents the degree to which small assemblages of taxa are subsets to successively larger assemblages and can be viewed as a region-wide outcome of a taxa pool being filtered by site-specific environmental constraints [T) of 0° shows a perfectly nested distribution (rare taxa are in diversity rich sites) and a T of 100° shows a random distribution of the taxa. Matrix temperatures were tested for statistical significance using a Monte-Carlo test (comparison to 999 randomly simulated matrices). Nestedness analysis was performed using NESTEDNESS software [Beta diversity, between stream diversity, was used to quantify the level of distinctness of the stream invertebrate communities at the watershed scale. Beta diversity values between watersheds were compared using straints ,38. To qsoftware .We excluded all rare taxa  and performed a Correspondence Analysis (CA) to describe the structure of the metacommunities. The CA was conducted on a reduced matrix of 100 taxa (59 taxa were removed) and 114 stream-sites. Prior to the CA, data were transformed to logarithmic values (ln(x + 1)) in order to reduce and normalize the variance.et al.[Taxa were assigned to particular biological trait categories Table  accordinet al.. The datBiological traits were available for 69 of the 100 taxa that occurred more than once in our data set. Therefore, the analysis was computed on a matrix of 69 taxa and 114 stream-sites. Ten biological traits described in 43 categories were included in the analysis  were computed using the ade4 package in R .p < 0.001) between the two watersheds. Values were higher at the CAIW having a median value of 124, while the median value was 68 in the AOW. The nestedness analysis showed also that communities of the CAIW had a higher nested structure than communities of the AOW . Therefore, these 3 results show that the CAIW has a higher heterogeneity of its stream communities when compared to the AOW.In total 159 taxa were identified from 114 stream-sites, including 125 different genera  and taxa adapted to intermittent stream flow . The other taxa of this assemblage were mainly trout-stream type taxa, like Suwallia sp., Suwallia teleckojensis, Isoperla sp., and Acentralla sp. While 28 stream-sites out of 77 in the CAIW (≈ 36%) were correlated to this assemblage  reported previously in glacier streams of the Himalaya and Tian-Shan Mountain ranges [The differences in the structure of communities between the two watersheds are the result of geological history and natural variation in the riverine landscape. No dams or channelization has occurred in the Mongolian rivers, so the lower biodiversities (both gamma and beta diversities) of the AOW cannot be attributed to anthropogenic homogenization across drainages [sensu, . A predin ranges . The assHigher beta diversity values and lower nestedness temperature in the CAIW confirm our initial hypothesis that lower sub-drainages connectivity would support higher distinctiveness of communities. The importance of dispersal through the riverine network was also indicated by guild structure analysis. The disconnected sub-drainages of CAIW prevent passive dispersal via the riverine network ; consequently both aquatic and aerial passive dispersals were less relevant to the structure of the communities in this watershed Figure B. In conet al.[A higher connectivity of sub-drainages also means a higher connectivity of vegetated riparian buffers that can provide sheltered habitat corridors for the dispersal of aquatic imagos. In Mongolia, such dispersal routes (i.e. following the network of riparian vegetation) can play an important role due to the structure of the Mongolian landscape. The lack of dense vegetation on the xeric Mongolian short grass steppe and a windy environment may restrict swarming activities of aquatic imagos to the riparian vegetation. This hypothesis corroborates the outcome of Petersen et al. finding The consequences of global warming on the stream communities in Mongolia will be dependent on 1) their current composition, and 2) the magnitude of the thermal and hydrological changes. Macroinvertebrates in streams of the AOW have more traits providing better resistance and resilience to disturbance, and many taxa there are typified by r-selected traits. Correspondingly, the AOW community has more taxa with r strategy, having a higher occurrence of r-selection traits . Small bBeyond the functional differences between communities of the watersheds, remarkable differences in the extent of permafrost  due to local extinction and reduced colonization by better adapted taxa. Conservation planning must incorporate the connected nature of drainages, a key factor for the conservation of freshwater biodiversity  when undThe authors declare that they have no competing interests.AM participated to the field expedition in 2010, compiled data, ran the analysis and wrote the paper. JG served as PI on the two grants supporting this work and led the expeditions of the Mongolian Aquatic Insect Survey. All authors read and approved the final manuscript.