Variations in hormone concentrations across the menstrual cycle affect human female mate preferences. It has been shown that around the time of ovulation human females prefer more masculine male voices, faces, and bodies while simultaneously preferring less faces that are more feminine. They prefer also displays of male dominance, males with more symmetrical faces, and the scent of males with high levels of body symmetry. The aim of the experiments reported here was to investigate whether there are changes in female preferences for walking gaits across the menstrual cycle.Experiment 1 showed female observers could discriminate between point-light walkers with low and high levels of fluctuating asymmetries in their gaits. Female observers were more sensitive to asymmetries in female gaits than they were for asymmetries in male gaits. Experiment 2 showed that level of gait asymmetry did not affect the abilities of observers to discriminate female from male walkers. Experiment 3 showed that female observers did not change their preference for low and high asymmetry walkers across their menstrual cycles. However, females showed a decreased preference for all female walkers at the time during which it was estimated observers were at peak fertility. That same change in preference was not observed for male walkers.These data suggest female observers may not value gait asymmetry, as a mate selection cue, in the same way that they value asymmetries in faces and bodies. While only “average” gaits were used in these experiments, rather than the gaits of individual walkers, the types of asymmetries in gait tested here were not used in the same way as static cues for judging the apparent healthiness of individuals. Females do discriminate well average female gait asymmetries and do change their preferences for those gaits across their menstrual cycle. Doing so may reflect the operation of processes that equip females with an advantage when competing for mates at times of peak fertility. Variations in hormone concentrations across the menstrual cycle affect human female mate preferences. It has been shown that during the stage of the menstrual cycle during which women are most likely to become pregnant from a single act of sexual intercourse  that they prefer more masculine male voices, faces-6, and bThe relationship between HCR and preferences for more, rather than for less symmetry are interesting. Every human body and face has some level of fluctuating asymmetry (FA), manifesting as deviations from perfect bilateral symmetry. Usually FA is low with individuals accumulating only small asymmetries during development. Indeed, resistance to environmentally induced FA is thought to indicate underlying genetic strength. PerhapsIndividuals with low levels of FA are also going to be more symmetrical in their movements. Those with structural asymmetries will typically move asymmetrically: One leg substantially shorter than the other will lead, for example, to a limp. As such, biological motion (BM) has also become a social cue usefully exploited by both female and male observers. Humans are, for example, very good at discriminating the sex of others,22, emotWith that in mind, it is unsurprising that there is evidence that females use BM as a cue for judging mate potential,13. To dThe experiments described here were conducted using methods approved by the formally constituted Human Research Ethics Committee (HREC) at Southern Cross University . The HREC approval ensures these experiments were conducted in accordance in compliance with the Helsinki Declaration.Experiments 1 and 2: Twenty-six premenopausal females  participated in these two experiments. Participants were students at the University of Southern Cross. All identified themselves as heterosexual and between the ages of 18-36. All had normal or corrected-to-normal vision.Experiment 3: 16 premenopausal females met inclusion criteria on initial testing: They reported to be between the ages of 18-36 years old; had regular monthly menstrual cycles ; were not currently using any form of hormonal contraception or had not been using any hormonal contraception in the last three months; and identified as heterosexual. Of the original 16, four were removed due to irregular menstrual cycles (e.g. menstruating twice in a month) during the course of the experiment. Thus, 12 naturally ovulating women were included in the repeated measures analyses .Custom designed software  was used to create point-light walkers, each composed of 15 point lights. Each point-light represented a major joint on the human body plus the centre of the sternum and hips, FigureWith that in mind, the stimuli used here were based on “averaged” walkers developed by Troje,26. TrojEach point-light walker from the continuum contained an average amount of fluctuating asymmetry , four were male , and one was sexually neutral . Each walker was presented with one level (low or high) of fluctuating asymmetry. Thus, in each block 45 trials presented low FA walkers  and 45 trials presented high FA walkers. Each exposure lasted 3 seconds during which the walker completed approximately 4 steps. The total duration of the experiment on any one occasion was less than 15 minutes. In experiment 1, participants were instructed to indicate, on every trial, whether they thought the gait of the walker onscreen was “symmetrical” or “not symmetrical”. The definition of symmetrical provided was, “Does the motion of one side of the body match or ‘mirror’ the motion of the other side?”. In experiment 2, and using the same paradigm, participants indicated whether they thought the walker was “female” or “male”.Experiment 3: Following previous studies-6, a staParticipants filled out a questionnaire to ensure they met inclusion criteria. If so, they were asked about the date of onset of their last menstrual period. If the participant indicated their onset had already occurred, forward counting from the day of onset allowed testing days to be scheduled for the days estimated most likely to be HCR and LCR. If the participant could not name the day of onset of their last period they were asked to notify the researcher when the next period began. From this information participants were randomly scheduled for their first testing session during either their estimated HCR or estimated LCR depending on which phase they were closest to upon successful completion of the questionnaire (see below). Seven females completed their first testing session during their estimated HCR , and 5 females completed their first session during their estimated LCR . Participants were instructed to indicate (via key press) whether they ‘liked’ the walk of each walker onscreen. Specifically, they were told, ‘regardless of your sexual orientation and regardless of whether you think the walker onscreen is a male or a female, do you like the way they walk?’ Key press was counterbalanced across participants. Participants were then scheduled to come in for the second testing session during their other conception risk phase and complete the same process. Before the start of the second session, participants were required to confirm where they were in their cycle to ensure accurate testing days were captured.Participants in all experiments were instructed to press “M” or “Z”, and the alternatives were counterbalanced across participants in each experiment, to signal symmetrical/non-symmetrical walkers or female/male walkers. No feedback was given as to performance.All experiments were designed as repeated measures studies, with each participant being tested on every condition. All analyses were designed for Repeated Measures ANOVAs with planed orthogonal contrasts. Data were analysed using the PSY programme . Target stimuli for these experiments were the -4 walker, the 0 walker, and the +4 walker, with others providing distractor stimuli to prevent repetition effects on the targets.The challenge in testing gait preferences, as opposed to form or appearance preferences, is to eliminate, as far as possible, interactions between form and motion cues. With that in mind, the stimuli used here were variations of the so-called point-light walkers. PrefereThe aim of Experiment 1 was to test whether female observers could detect asymmetries in the gaits of point-light walkers used here. In particular, it was hypothesised that females would be able to discriminate low levels of FA in gait from high levels of FA.1, 25 = 20.653, p < .000), ambiguous walker  and male walker . Note, however, that none of the walkers tested were judged to be symmetrical 100% of the time, demonstrating that observers were able to discriminate even small levels of FA from gait.Data analyses were conducted on three point-light walkers only , and for two levels of fluctuating asymmetry (low and high). These walkers were isolated to capture responses to unambiguously female gait, gender ambiguous gait, and unambiguously male gait respectively. For each of those walkers mean performances on each of the three stimuli of interest were computed for each individual participant, and then group means for each level of FA were calculated. Mean proportions of “symmetrical” judgments across these three walkers and two levels of FA are represented in Figure1, 25 = 7.053, p = .014), high FA walkers . That is, female observers were more acute detectors of asymmetry in female walkers than in ambiguous or in male walkers .Interestingly, for both low and high FA walkers there were differences between performances according to sex of the walker. For both FA levels, female walkers were judged to be symmetrical less often than either ambiguous or male walkers: Low FA walkers , ambiguous walkers , or male walkers . That is, female walkers were never reported to be male and male walkers were always reported to be male. Interestingly, for both levels of FA, ambiguous walkers were judged to be male more often than female, an example of the reliable male bias reported elsewhere[1, 25 = 11289.063, p = .000). Similarly, the high FA male walker was reported to be “male” more often than the high FA ambiguous and female walkers . Thus, it appears that FA does not affect acuity for sex discrimination for female observers.Mean performances on each of the three stimuli of interest were again computed for each individual participant and then group means for each level of FA calculated. Mean proportions of “male” judgments across these three walkers and two levels of FA are represented in Figurelsewhere. Also, aThe hypothesis that different levels of asymmetry in gait would not affect accurate sex discrimination was supported. Both the low and high FA female walkers were never reported to be ‘male’, and the low and high FA male walkers were always reported to be ‘male’. A “male-bias”, the tendency to perceive sexually ambiguous stimuli as male more often than female was obseThe previous experiments revealed that female observers could detect a difference between gaits low in asymmetry and gaits high in asymmetry and confirmed that ability to successfully discriminate the sex of a walker is not disrupted by high asymmetries. Given that asymmetries were detectable, the sex of the walkers was identifiable, and that symmetry is preferred in structural traits when women are in their HCR phase, the aimMean performances on each of the three stimuli of interest were computed for each individual participant and then group means for each level of FA calculated. Mean proportions of “liking” judgments across the three walkers and two levels of FA when female observers are in their low conception risk phase, and high conception risk phase are represented in Figures1, 11 = 2.570, p > 0.05), of ambiguous walkers , or of male walkers . In other words, these data suggest there are no differences in female preferences for low or high FA gaits in walkers of any sex at estimated LCR.“Liking” judgements at estimated LCR: As highlighted in Figure1, 11 = 0.048, p > 0.05), of ambiguous walkers , or of male walkers . In other words, these data suggest there are no differences in female preferences for low or high FA gaits in walkers of any sex at estimated HCR.“Liking” judgements at estimated HCR: Figure1, 11 = 8.520, p ≤ 0.017; Bonferroni adjusted at α = 0.05, k = 3). Most interestingly, there was no difference in liking judgements across conception risk phases for the ambiguous walker  or male walker . That is, female observers did not differ in their preferences for male gaits as their risk of conception varied. They did, however, show decreased preferences for the gaits of other females as they moved from low conception risk to high.Given that there was no effect of asymmetry on observers’ liking judgements data were collapsed across levels of FA, and a post-hoc one-way repeated measures ANOVA was run to investigate whether there was any differences in the way females evaluated gait across the menstrual cycle independent of asymmetry. FigureThus, in summary, these data do not support the hypothesis that females at high conception risk would prefer male gait that was more symmetrical to gait that was less symmetrical. Instead, these data suggest changes in hormonal state relating to conception risk affect female perceptions of other female’s biological motions rather than their perceptions of male biological motions. Of course, these findings need replicating. This was an unexpected result and should directly be investigated in future work.The general aim of this research was to investigate whether human females experienced changes in their preferences for walking gaits across the menstrual cycle. In particular, it was predicted that female observers would show preferences for male gaits with low levels of FA during the estimated HCR phase of their menstrual cycle compared to judgements made during their estimated LCR phase. To test that hypothesis point-light walkers with low and high levels of FA were constructed and female observers were asked first to discriminate low from high levels of FA, and they could do that reliably (Experiment 1). Indeed, females were more acute observers of asymmetries in the gaits of other females than they were of asymmetries in male gaits. Those same observers were asked also to discriminate female from male walkers with both low and high levels of FA and they could do that reliably also (Experiment 2). Surprisingly, and contrary to predictions, a sub-set of the observers showed no changes in preferences for low and high FA across their menstrual cycles (Experiment 3). That is, conception risk did not predict female preferences for male gaits with low FA: Low FA walkers were never preferred to high FA walkers in either conception risk phase. This finding contrasts the prediction that females would prefer male walks that are more symmetrical at HCR. Interestingly, however, it was found that independent of FA, female observers liked the gaits of other female walkers less during the estimated HCR phase of their menstrual cycle compared to judgements made during their estimated LCR phase. That same variation was not evident in judgements of non-female walkers.While not predicted, that result is not without precedent. It has been shown previously that females at peak fertility prefer less the faces of other females. That is, women show decreased preferences for feminine faces when progesterone levels are low and oestrogen levels are high. Jones aWhat is clear from these data is that the levels of FA in the gaits used in these experiments do not interact with variations in fertility: Even though female observers could discriminate between female gaits showing low and high levels of FA, the decrement in “liking” for the gaits of other females was independent of level of asymmetry being displayed. Similarly, there were no variations observed here in preferences for male gaits with low and high levels of FA across the menstrual cycle. Those observations suggests that while female observers are sensitive to gait asymmetries, especially in other females, they do not use that particular cue in the same way they use structural cues to health or genetic robustness. As noted above, there is a substantial literature indicating females do vary their preferences for faces and other features correlated with health-related traits as fertility varies.It is tempting to speculate that gait asymmetries are not among those. Of course, that argument needs to be tempered with the observation that the walkers used here are not individuals but average walker. Similarly, the asymmetries added to these stimuli are not natural in that the fluctuating asymmetries in our stimuli are based on changes between averages. That means that before stronger conclusions can be drawn it may be necessary to test the effects reported here with real asymmetries in real walkers. Doing so may recruit discrimination mechanisms not sensitive to the manipulations used here.That there were no interaction between FA and fertility seen here could be because symmetry in motion may not be as reliable a cue to the (genetic) characteristics on an individual as symmetry in static, structural traits like faces, bodies, and scents. Symmetry in structural traits does not usually vary, and under normal circumstances, cannot be ‘faked’. ContrasThis study contributes some innovations to research on human mate preferences and the underlying mechanisms that drive such processes. This study indicates that female observers may not value gait asymmetry as a mate selection cue in the same way that they value symmetry in faces and bodies. In summary, these data show female observers are capable of discriminating the sex of individuals on the basis if their gait and that they can do so across different levels of fluctuating asymmetry. These data show too that the magnitudes of those asymmetries do not affect female observers’ preferences for individuals as fertility varies across the menstrual cycle. Thus, asymmetries in the motion cues used here are not used in the same way as static cues for judging the apparent healthiness of individuals, female or male. Interestingly, female observers did changes in their preferences for other females, on the basis of their gaits, across their menstrual cycles. Females showed decreased preferences for other females at times when it was estimated that the observers were at peak fertility. While the neural mechanisms mediating those behaviours may be both unconscious (and currently unknown) they may equip individual females to act in ways that would increase opportunities for mating with strong males at times of peak fertility.Both authors declare that they have no competing interests.NH carried out the data collection and preliminary treatment of all data. vdZ designed the study and with NH performed the statistical analysis. NH and vdZ conceived of the study, and both drafted the manuscript. vdZ supervised the project. All authors read and approved the final manuscript.


Current mobile phones come with several sensors and powerful video cameras. These video cameras can be used to capture good quality scenes, which can be complemented with the information gathered by the sensors also embedded in the phones. For example, the surroundings of a beach recorded by the camera of the mobile phone, jointly with the temperature of the site can let users know via the Internet if the weather is nice enough to swim. In this paper, we present a system that tags the video frames of the video recorded from mobile phones with the data collected by the embedded sensors. The tagged video is uploaded to a video server, which is placed on the Internet and is accessible by any user. The proposed system uses a semantic approach with the stored information in order to make easy and efficient video searches. Our experimental results show that it is possible to tag video frames in real time and send the tagged video to the server with very low packet delay variations. As far as we know there is not any other application developed as the one presented in this paper. In [YouTube and situated the in Google Maps service, to seek videos using a time restriction is presented. This can be done implementing temporal tagging that allows the user to specify queries of concepts like within, contains or overlap [In a semantic sensor web, as defined by Henson in , sensor amio. In  a proof  overlap .In , Henson In this paper we present a sensor-based video tagging system that allows a mobile phone to record videos and tag them with some extra information taken from the embedded sensors. Later, the tagged video is placed on an Internet server, so it can be viewed by other users jointly with the sensed data in order to appreciate better the environment when the video was recorded. Moreover, we have added a semantic approach in the server in order to provide easy and efficient video searches. Experimental results show that videos can be tagged in real time and with a good grade of precision, and the transactions with the Web server can be done with low delays. This facilitates to implement a real time query system directly on the mobile phones in real time.The remainder of the paper is as follows: in Section 2 we present the main concepts germane to mobile video tagging using the sensors, that are included in the current smart mobile phones, and we describe some related work found in the literature. The architecture of the semantic system, the ontologies and the semantic system operation are presented in Section 3. Section 4 is devoted to the analytical model presentation. Section 5 shows the experimental evaluation of our semantic system. Finally, in Section 6, we draw our main conclusions and future work.2.Web Ontology Language to define the ontology and temporal logic to describe the semantic events and reasoning. In [Traditionally, several formats for video tagging have been used related to different specific purposes. Moreover, the video content can be analyzed using ontology. In  soccer dning. In , an origMultimedia Contend Description Interface) helps describe video content and thus enables the user to easily perform many tasks such as find, store and locate a video [New multimedia standards, such as MPEG4 and MPEG7, provide the basic functionalities in order to manipulate and transmit objects and metadata. Concretely, MPEG7  it is possible automatically generate this semantic video indexing . These sA not considered challenge is to take into account the MPEG7 annotation of videos in order to allow Peer-to-Peer communication among them as explained in . SimilarA semantic modeling of video contents retrieval is presented in . In thisIn  a very iIn  a dense There are several ways to include semantic information from sensors. The first step is to consider a set of tags that can describe the values that the sensors provide, and the second step is to implement an ontology that can allow the extraction of knowledge.A good introduction of the reasons for and benefits of annotating the information retrieved from sensors is done in . The autOntoSensor and the CSIRO ontologies represents the current limit of expressive capability for semantic sensors. But, neither current ontology nor a combination of the available ontologies is able to express all the properties required for the capabilities. So, it is needed more work in order to improve this expressivity. Moreover, the early work presented in [In  the authented in  shows thCommonSense system and they propose an architecture for WSNs with semantic web technologies as enablers. Finally, they have tested their system on a fire accident scenario in a building.In  the JenaSemantic Streams framework [The ramework  allows uIn  we presewhere, when, who or hybrids. The prototype uses a camera connected to different sensors to tag the video. Moreover, they show the difficulty of tagging video frames with sensor data. In this paper we explore the applicability of mobile phones for tagging video frames. We take maximum advantage of the mobile phone sensors in order to implement a low cost system. We will show that our proof of concept application is efficient and our experimental results show that real time video tagging (for further semantic search) could be easily used without decreasing the performance of the mobile phone.Finally, in  the auth3.In this Section we first describe the architecture of our semantic system, including a review of the tradeoffs for the design of video tagging in mobile phones. Secondly, we show the knowledge representation and finally the semantic operation of our system.3.1.geotaging each physical object represented in the images that form a video and also obtain useful measurements of the mobile phone’s sensors for each image. All these data are stored in an eXtensible Markup Language (XML) file. Therefore, each recorded video file is associated with tags. The tagged video is stored in a single multimedia container that can be stored locally on the mobile phone or streamed to an Internet video Server. Then, the files can be shared with other users by accessing to the server. The users can also search for videos on that platform (Internet video Server), according to a certain semantic criteria, based on data that are in the tagged files of the shared videos.The implemented mobile application allows users to record videos and tagging them with semantic sensor data. This extra information is associated with each of the images composing a video recorded from a mobile phone, allowing users to exploit this information when sharing tagged videos. Specifically, the process consists in User Interface used to add the needed data in order to begin the recording process and tagging is shown in where he/she is and what kind of object he/she thinks of recording. There are different options for each object type and location. The user must also specify the name of the file to be generated after the recording and tagging. And he/she must indicate the distance between the object and the mobile phone’s camera using a specific value or by choosing a particular approach factor . These data will be used in the tagging process later.The Global Positioning System (GPS) and (4) the tagging process. We describe these four processes next.Once the user has entered the above information using the user interface, then four key processes will run concurrently in order to generate tagged videos: (1) the video recording process, (2) the process of capturing the remaining data necessary for tagging from the sensors of the mobile phone, (3) the process of capturing necessary data from the Secure Digital (SD) card in an MPEG4 file.The video recording process is the responsible for accessing the hardware of the phone’s camera and to capture the images that compose a video. This video is stored on the Accelerometer: It calculates the acceleration applied to the device. Specifically, it provides three values: the acceleration in the depth axis (Z axis) of the mobile phone, on the horizontal axis (X axis) and on the vertical axis (Y axis). These values are obtained by measuring the forces applied to the mobile phone and taking into account the gravity of the Earth. The three acceleration components are in 2m/s.Gyroscope: It measures the angular velocity in each of the axes of the phone, in rad/sec. It is especially useful in gaming applications.Light: It detects the brightness of the screen, providing a value in SI units: lux.Magnetic: It measures the value of the magnetic field in the X, Y, Z axes of the mobile phone in μTesla unit.Orientation: It calculates three values  to detect the mobile phone inclination and the cardinal point that the mobile phone’s camera is pointing.Proximity: It calculates the proximity through the distance in cm between the mobile phone and the detected object. It is very useful in applications that detect human or animal presence in a given area.Gravity: It calculates the components in the Z, X and Y axes of gravity in m/s2.Temperature: It calculates the temperature at which the device is.Rotation vector: It represents the rotation of the device as a combination of an angle and an axis .Linear accelerometer: It indicates the acceleration along each device axis, not including gravity. All values have units of m/s2.Pressure: It calculates atmospheric pressure in mbars unit.The process of capturing the remaining data needed for tagging accesses the sensors available on the mobile phone in order to read the sensed data which are later used to tag the video. An Android mobile phone may have a maximum of 10 sensors, which are listed next :AccelerThe GPS of the mobile phone provides two kinds of values: the geographical coordinates and the altitude of the mobile phone. Finally, the tagging process is responsible for generating a XML file that contains the information of the tagging of each of the images captured during video recording.GPST and the average time it takes for the sensors to provide a value is SenT . In addition, we must take into account the GPS’ reading time and capturing its values (RGPST) and the reading time of the sensors and the time to capture their values (RsenT). Moreover, it is also important to consider the time taken to generate the tagged file, which corresponds to the time of writing all the necessary data for each image (TagT). We must also take into account the total processing time, which corresponds to the reading of data, its capturing, and the subsequent tagging (TotT).In the process defined above there are several factors to consider as they define the operation of the application and more specifically the video tagging. All these factors arise because the tagging should be done for each image of the video. Assuming that the mobile phone’s camera captures 25 frames per second (fps), the tagging process should be done every 40 ms. But this process is subject to the time taken by other processes (the reading of data from sensors and GPS). The time spent in reading the values of the GPS and the sensors may be higher or lower than 40 ms. Then, from now on we will assume that the average time it takes for the mobile phone’s GPS to provide a value is As mentioned above, we have assumed that 25 images must be generated every second (in order the user can appreciate movement). And therefore, tagged every 40 ms. But this is not entirely true and taking into account that the current video cameras use compression methods based on MPEG4, they do not generate 25 fps. Instead, they use other techniques that exploit the temporal information of a frame to generate the next (in the video sequence). Then, another factor that affects the performance is the need to synchronize the video recording and tagging effectively.Finally, it is also important to consider the storage time of the recorded video frames. Although it is not possible to access to the code used for storage in the mobile phone we use an approximation to measure this time.3.2.An ontology is a formal explicit representation of knowledge as a set of concepts within a domain, that is used to describe the domain, its properties, and the relationships between those concepts ,43. It psingle, multiple and hybrid ontology approaches. Our system is based on a hybrid approach  labels the video with a GPS position, and the type of object that is being recorded, for example a flower and the user specifies if it is a rose, a poppy or a margarita. Ontology has its primitive for each flower, but we have a shared vocabulary. When we ask for a red flower, red and flower are shared vocabulary. This vocabulary must be understood by all ontologies in order to let our system be able to answer the question: Is there a video with red flower? Our system should answer with the videos of roses and poppies (red flowers), but not with the videos of margaritas (white flower). This is an easy example to explain which ontology approach is more adequate for our proposal. Now, if we introduce some more labels from the sensors of the mobile phone adequately, the system should be able to answer questions such as: in which coast is a red flower in a sunny day?The main advantage of this approach is that new sources can easily be added without the need of modification in the mappings or in the shared vocabulary. It supports the acquisition and evolution of the ontology. Our system can be used for several applications. This scalability feature lets the system be extended without much effort. The use of a shared vocabulary makes the source ontologies comparable and avoids the disadvantages of multiple ontology approaches. The drawback of hybrid ontology approach is that it cannot be reused easily. It has to be re-developed from scratch, because all source ontologies have to refer to the shared vocabulary.An example of a part of a local ontology of our system is detailed next:VIDEO name>. The video will have a name and this tag is the global one.<Place>. Initially we have programmed for this tag only three attributes: <Hotel>, <Beach>, and <Restaurant> which are qualified with a name. The idea is the mobile user can specify the name of a hotel, a beach or a restaurant. This means the mobile user can be in any of these places or in a beach in which there is a hotel in which there is a restaurant. This tag can be easily extended with more attributes such as <Mountain>, <Supermarket>, <Home>, and many more.<Object>. Initially we have programmed for this tag only three attributes. So, the user can specify the name of: an <Animal>, a <Plant> and a <Monument>. We can add many more such as <Furniture>, <Food>, <Vehicle>, etc.<Tagging of whole video. The idea is to provide a global description of the scene in a particular scenario. We considered the following tags:Each frame of video (or a set of frames depending on the processing power of the mobile phone) is tagged with the values of the sensors the user wants to include. Each sensor has a tag named with the name of the sensor and an attribute that contains its value. Moreover we include the time in which the frame was captured by the video camera and the GPS value. In Taking into account the sensors of an Android mobile phone, we are able to generate the tagging of the video in two different levels:The structure of the tagging file is shown in Up to now we have not added image processing for forms recognition. But our purpose is to extract concepts directly from the images. We have seen two approaches in the related literature that are being studied for our purposes: the first one uses a frame-based image classification , and the3.3.The system operation is as follows : the mobwhere (using location tags), what kind of object (using the object tag), what interval of time (using time tag)… For doing that, the semantic searcher uses the tag file generated when the mobile user recorded the mobile video. This semantic software can be implemented as a proxy or as a plugin that must be connected to the Web services aforementioned. Our proposal is to adapt a typical semantic searcher. When semantic searches are used, the semantic searcher must access to a service in which the mobile phones be registered. This registering software will be in charge to control the mobile phone video availability.Once the mobile video has been stored in an Internet Video Server, any user (mobile or not) can search the video using a typical Web searcher or using a semantic search. Contrary to a typical Web search, a semantic searcher will admit queries like YouTube with a simple click) or even add an image in a location of the Google Maps. This type of procedure provides an easy-to-use system. In the experimental results shown in Section 5 we use our server to test this proposal and we are researching the way to upload the tagged videos to public servers.4.T is given by sensingt is the time required to gather the data from the embedded sensors. It is the time of the most delayed sensor data. The highest time is the time needed to scan the values of the GPS (GPST), as we will see in the next section. So, bearing this in mind, we can affirm that sensingt ≅ GPST, which is around 1 second. processingt is the time required by the device mobile to read the tag from the sensor and to tag the video frames. In this case processingt = RSenT + TagT. According to our measures shown on the next section, the value will be around 50 ms. TXt is the time needed to encapsulate all data into the IP packet in order to send the information through several networks. RXt is the time required to decapsulate the data. Generally TXt is equal to RXt. Nowadays these times are very low, because they depend on features of the end devices and these devices have high performance. propagationt is the time needed to send all the data via the interconnected networks to the Internet server. It depends on RQt (the request time) and the propagation time of each network. Bearing in mind that the propagation time in a network (iDelay) may vary depending on the amount of traffic in the network, we have added the instantaneous efficiency of each network (iφ). Hence, the propagation time, when there are n networks between the mobile phone and the server, is shown in Our first step has been to model the time needed to perform all the processes carried out in our system in order to test if it will provide a good service for the end users. First, it is necessary to assess the time required for the system from the video capturing and tagging until its storage in Internet servers. This time should be analyzed in order to see how much time is required since we started recording a video until the video can be accessed by another user using the Internet. The total time savingt and presentationt to savingt, because presentationt is negligible when the data have been stored on the servers. In order to analyze the parameter savingt we have used an approximation of GI/G/m model [λ and the squared coefficient of the variation of an interarrival time  is defined in Ψ  in the ϕ4  can be estimated using the set of expressions shown in EW can be expressed by EW (M/M/m) is defined in ϕ to the server. Finally, the server processes the request and sends a reply. If this reply is OK, the mobile device will send the tagged video to the server. In this case, the request time is RQ_Meth_At. On the right we show our proposed method (Method B). It allows improving the system in terms of time. The mobile device sends the RQ packet immediately after starting the sensing process. Then, the system follows the regular designed steps. We can observe that request time is shorter. In this case, the request time is given by In sendingt. In this figure we see that the improvement is higher than in t2 ≫ Δt1). According to this data we can affirm that if we use method B, regardless of sendingt, we will obtain a shorter RQt. Moreover, this improvement, Δt, is directly proportional to sendingt.In failed message. In method A, we see that the RQ message is sent after sensing, so the device stores the data until it receives the RSfailed to perform the appropriate task (store the tagged video on a file or delete the tagged video). In method B, the mobile device sends the RQ message when it starts the sensing process, so the system receives the RSfailed before completing the sensing process. In this case, we have considered two options. The first one lets the user to stop the process and delete the data, and the second process lets the user continue sensing and storing the tagged video locally.When the request process is denied, because the server cannot store the data for any reason, Method B continues improving the system. 5.We have performed several experiments in order to test the performance of our mobile application. In particular, we have evaluated the tagging process and the communication performance between the mobile phone and the Server. We have used a HTC Tattoo mobile phone to experimentally show the viability of our proof of concept. The technical characteristics of the mobile phone are presented in 5.1.GPST, RGPST, SenT, RsenT, TagT and TotT. For the values presented in this section we repeated the experiments several times obtaining for all the measurements similar values .In this section we present the experimental results after measuring the next parameters: GPST using a satellite (we do not use the combined method of GPS and WiFi). The minimum time spent in scanning the GPS value was 903 ms and the maximum value was 1,068 ms. The mobile phone scanned different sets of 19 values with similar results. The average value was 1,000 ms. That is, the time needed to scan the GPS value is 1 s. This is a bigger value than 40 ms. This means some images must be tagged with the same GPS value. The RGPST is insignificant compared with the GPST due to its values are in the interval from 1 ms to 7 ms, with an average value of 3.5 ms. In We measured the values of SenT measured in two consecutive reads for the following sensors: temperature, accelerometer, orientation, light, magnetic, pressure and proximity. In addition we also show the time needed to read simultaneously all the sensors. Some interesting points must be highlighted: The major values are concentrated in the interval from 1 ms to 30 ms. That means the latency of the measurement of consecutive values is in the normal case very low. We also show that there are several values in the approximate interval from 100 ms to 120 ms. This means that in several cases the latency is around 115 ms what means that consecutive images must be tagged with the same value of the sensor. Only a few values are greater than 150 ms and lower than 220 ms. When all the sensors are read the same shape of the curve is obtained. But in this case a very high value is found for the 18th measure. The conclusion is that the Android software introduces a variable time to read the sensors which implies to repeat the value of the sensor for several consecutive images.In RsenT are very small in all cases as is shown in TagT are normally ten times higher than the values of RsenT (with singular cases). The values of TotT are always in the interval from 40 ms to 80 ms but a singular value is found for the 8th measure. These values mean that in general the values needed to tag the video frames and store them in the SD card are acceptable.The values The experimental results of tagging in the mobile phone indicated that it is possible to tag mobile videos in the mobile phone in real time. Moreover, the overhead introduced by our tagging software is relatively low.5.2.As mentioned before, the application allows users to share videos tagged with others through a server platform. The video upload from the mobile phone has been tested using a WiFi connection because it is more and more usual to find WiFi hotspots offering WiFi connectivity at no additional cost. Our proposal does not depend on the wireless technology used to send the videos, and it can be used in 3G technology as well, because the upload rate could even be up to about 10 Mbps in some operators. A detailed performance test about the bandwidth needed for video delivery over several wireless technologies is shown in .Taking into account that the user sends a multimedia container file with the tag and the video and taking into account that this multimedia container is approximately 1 MB (considering video files of 10 seconds stored in the mobile phone), one can stay that for a connection with a speed of 11 Mbps, there will be not problem to send the files to the server as regards bandwidth availability.jitter of packets  in communication networks is defined as the difference in point to point delays, among several packets selected from a data stream, where losses are ignored. This definition is taken from the Request For Comments (RFC) 3393 [The delay FC) 3393 , which eFC) 3393 . In factPDV is the packet delay time from the start of transmission at the source, from the first byte of the packet until the last byte is received at the destination. To calculate the PDV, the delay components that do not vary can be overlooked. For example, if the packet size is the same and the transmission time and receiving time in the data buffers do not vary, the contribution of the time (the one the packet takes to cross the network to reach the destination buffer) will only be taken into account.The most important factors that affect the PDV are undoubtedly congestion or channel bit rate . This leads to packet losses or to a relatively large difference between the time of transmission of a packet and the time of receipt. If there are no disconnections, when the delay of packets increases, the PDV will increase. Thus, if a user is downloading a file, the waiting time will increase proportionally to the increase of PDV. On the contrary, if disconnections occur, unless there is an attention mechanism to them, we must forward the video and its tagging file again starting from scratch. In this case, the time the user must wait will increase, but the delay between packets will depend only on the channel conditions. The variation of packet delay grows exponentially as the RSSI decreases its value, which corresponds to a WiFi signal level received less, and therefore slows down the operation of the application .IT: Time spent by the current packet to be transmittedI-1T: Time spent by the previous packet to be transmittedN: Number of packets that are considered in the measurement. They correspond to time interval.We have measured the PDV both communicating only the video and communicating the tagged video in order to make a comparison between the results obtained in both cases. For a given level of RSSI (weak or strong), we have considered the following parameters to measure the communicating time (delay) extreme-to-extreme:The delay  and PDV In The values obtained for delay and PDV indicate that the real time communication of tagged videos is possible and with low PDV for a WiFi (version g) channel. Moreover, the tagging of videos we introduced produced a very low overhead in communications.6.Current mobile smart mobile phones have very powerful video cameras. They also have several sensors that can be used to tag the mobile video. The processing power of current smart mobile phones is very high. This make possible to implement mobile platforms like Android over 1 GHz processors that have up to 256 MB RAM. They also have very high speed wireless communication cards like WiFi (version g).In this paper we have presented an application that records and tags the video frames with data collected by the Android mobile phone’s sensors. Tagged videos are uploaded to an Internet video server and accessible for users. The experimental results show that is possible to tag mobile video frames in real time and using the WiFi wireless card interface of the mobile phone is also possible to make real time transactions with the video Server that hosts the videos.Long Term Evolution) will exhibit transmission speeds of about 50 Mbps. This means that the tests we have implemented using WiFi could be used in these kinds of mobile communication technologies naturally. We modestly think that this is a starting work that has opened the door to new kind of mobile video applications in the close future. We think this means that our solution could be very successful in the near future.The next version of mobile communication (YouTube). Then another mobile user could make a semantic video search in the Internet Video Servers. These searches can be done in Real Time. On the other hand the semantic video searches can also be done in tagged videos stored in the mobile phones. This fact confers to our system the typical characteristics of a semantic wireless sensor web considering the mobile phones as high powerful wireless sensors.As part of the future work we will propose to a Telco operator the possibility to implement our mobile application in their mobile transport network. We also will work in the definition a semantic searching application of mobile semantic videos in the Video Server. Integrating this application with our current mobile application it will be possible a mobile user to take a short video from a determined place, to tag it, to send it to the Internet Video Server (a public one such as


In such interiors, a high-performance method of microbial contaminant reduction need be introduced, especially when large groups of young patients are medically cured. Respecting fungal aerosol, its average indoor concentration (88 cfu/m3) was significantly lower than outdoor level (538 cfu/m3). It confirms that ventilation system provides efficient barrier against this type of biologically active propagules. Among identified micro-organisms, the most prevalent indoors were Gram-positive cocci, which constituted up to 80 % of airborne microflora. As highly adapted to the diverse environments of its human host , they can be easily released in high quantities into the air. The number of people introduced into such subterranean chambers should be in some way limited. The analysis of microclimate parameters revealed that temperature and relative humidity influenced significantly the level of bacterial aerosol only. Hence, a constant control of these parameters should be scrupulously superintended at this type of subterranean premises.Nowadays, an inhalation of naturally generated aerosols has again become a widely practiced method of balneological treatment of various respiratory diseases. The aim of this study was to characterize the microbial aerosol of subterraneotherapy chambers at the Bochnia Salt Mine Health Resort in southern Poland. The measurements were carried out using a 6-stage Andersen impactor over a period of 1 year in both indoor  and outdoor air. The maximum bacterial aerosol concentrations in the chambers reached 11,688 cfu/m In 1958 in Wieliczka, nearby Cracow, Poland, the world’s first salt spa for patients with respiratory diseases was open. The unique presence of dry salt aerosols having both specific concentration and size maintains “clean,” that is, micro-organism- and allergen-free atmosphere. Such elimination and/or attenuation of those immunologically reactive stimulants determines the efficiency of subterraneotherapy and in numerous cases may heal adverse health effects  formed microclimate conditions in the subterranean chambers. Despite the known benefits of such treatments, a subterraneotherapy is implemented in a few health resorts in the world only . According to the Latin maxim “There is broad medical evidence that specific microclimate conditions in subterranean chambers  have a beneficial effect on human health, especially in case of respiratory diseases. The importance of this phenomenon is emphasized by the fact that current environmental changes, particularly the increase in gaseous and particulate pollutant concentrations in the air, reduce the natural immunity of human organism . Both an impact of microclimate parameters  as well as an influence of atmospheric air  on the indoor microbial population were also evaluated.The study was carried out in the Bochnia Salt Mine Health Resort, being a part of the oldest salt mines in Poland. The Bochnia Salt Mine origin dates back to the thirteenth century, and it possesses the oldest functioning shaft in European mining constructed in 1251. Since 1995, a few mine excavations have been converted into a sanatorium and used for therapeutic purposes .3, the Europe’s biggest man-made subterranean room) and Kołdras , located 248 and 176 m below the ground level and housing 250 and 42 beds for patients, respectively. In both chambers, the samples were taken during the treatment courses with patients and in the same premises 24 h after curative treatment without them. This 24 h break before the post-treatment sampling was chosen due to the scheme of exchange between the “old”  and “new”  groups of patients. The patient groups (150–200 people) consisted mostly of children. Majority of the patients (90 %) recruited from among elementary school children; their parents, teachers or chaperons made the remaining 10 % of the persons present in the chambers. Sampling was carried out 24 h after curative treatment of patients in empty chambers. Moreover, the air samples were taken from the ramp, an incline of 139 m long being a part of the corridor connecting Ważyn and Kołdras chambers. In addition, in each of the investigated areas, an indoor background station was established to measure bacterial and fungal aerosol concentrations (IN-B). All these measurements were carried out in the galleries located on the same levels as the studied chambers, through which the air was introduced into them.The bioaerosol measurements were performed over a period of 1 year (from January to December), twice in each of four seasons. The air samples were collected in two subterranean chambers: Ważyn , and filter set (cotton filter) was responsible for a delivery of the atmospheric air into the studied subterranean chambers. The air was forced down into the chambers from the surface level through an old (and not cleaned recently) 210 m long brick downcast shaft (housing a wooden ladder way), and then its movement in the underground spaces was regulated by the barrier walls and a specific location of therapy chambers in the corridor bends. The shaft was unheated. In order to obtain data on the background atmospheric level of microbial contamination (OUT-B), the air was sampled in front of a fan outside the salt mine above the ground. The location of the Bochnia Salt Mine Health Resort and selected sampling points characteristics is presented in Fig. The air samples were collected using a 6-stage Andersen impactor . The sampler was placed at a height of 1.0–1.5 m above the floor or ground level to simulate aspiration from the human breathing zone. A 5 min sampling period was applied for collection of bacterial and fungal aerosols. Samples were taken at a flow rate of 28.3 L/min. All bioaerosol samples were taken in triplicates. Bacteria were collected, on blood trypticase soy agar , and after impactor reloading, fungi were collected on malt extract agar . During sampling, the air temperature and relative humidity (RH) were measured using a hytherograph .3). Bacterial strains were identified to the genus and/or species level by Gram staining, their morphology and, finally, by the biochemical API tests . Fungi were identified according to their morphology using several identification keys  and Mann–Whitney tests  as well as Spearman correlation  using Statistica  version 7.1—2006 . The “3 and from 189 to 2,517 cfu/m3 in indoor and outdoor air, respectively. The highest bacterial aerosol concentrations were observed in the Ważyn chamber  during physical activity of the patients. When the patients were absent, the measured bacterial concentrations in the air were significantly lower than when they were present , reaching the level characteristic for both indoor and outdoor backgrounds. There were no statistically significant differences between the bacterial aerosol concentrations in the outdoor background, indoor background and subterranean chambers after curative treatment without patients . The comparison of bacterial aerosol concentrations between the studied premises revealed the highest differences  between the physical exercise area in the Ważyn chamber  and the Kołdras chamber . Moreover, it was shown that the concentrations of fungi in chambers during treatment courses  and without them were not statistically different (Mann–Whitney test: p > 0.05). The highest indoor fungal aerosol concentration was observed at a physical exercise area of the Ważyn chamber (566 cfu/m3) and inside the Kołdras chamber  ranged from 7 to 566 cfu/m3 Table . Outdoorp > 0.05). Despite the natural (in temperate climatic zone) fluctuations of outdoor air temperature over a year, there were also no differences between the median temperature values measured indoors and those noted in outdoor background.During all bioaerosol measurements, environmental parameters, that is, temperature and relative humidity of the air, were also controlled. Median values and ranges for both these parameters measured indoors and outdoors are presented in Table p < 0.01).Compared to air temperature, the changes in relative humidity within a year in the health resort chambers were much more distinct. The humidity ranged from 25 % in winter to 73 % in summer and the median yearly value in the studied premises reached 56, and 70 % outdoors. Statistically significant differences in the examined premises were observed between summer (median: 70 %) and winter (median: 35 %) RH values only . This relationship was the most significant in the Kołdras chamber (correlation coefficient: R = 0.75 at p < 0.05).The analyses of the influence of microclimatic parameters on the observed bacterial and fungal aerosol concentrations showed that only bacterial aerosol levels were affected by these factors. The correlation analysis revealed that each rise in temperature and relative humidity in the studied chambers resulted in significant increase in bacterial concentration in the air , mesophilic actinomycetes (mainly Streptomyces) and molds (Cladosporium and Penicillium) predominated, which is a typical picture for both outdoor and indoor microflora. Molds (68 %), Gram-positive cocci (17.5 %) and yeast-like fungi (9 %) were the most prevalent in the studied outdoor environment. Gram-positive cocci were the most numerous among all analyzed microbial groups in the studied chambers. Irrespective to the presence of patients their contribution to the total bacterial community indoors reached 78–79 %. Endospore-forming Gram-positive rods were also frequently isolated from the air of studied premises. Their percentage contribution to the total airborne microflora was about 8 %. The presence of yeast-like fungi in the air was 2 %. Qualitative analyses revealed that the most numerous species belong to: among bacteria—Staphylococcus (8 species) and Bacillus (4 species) genera, and among fungi—Penicillium genus (4 species). The conditions inside the studied chambers favor the growth of Staphylococcus and Bacillus species. The bacteria from genus Bacillus were the most numerous in a bedroom in the Ważyn chamber (when the patients were present) but were also frequently isolated from the outdoor air. The air was free from Gram-negative rods in all analyzed subterranean chambers when the patients were present. It shows that there are unfavorable conditions for these micro-organisms survival and lack of any indoor sources of their emission and infiltration. The percentage contribution of Gram-positive cocci to the total number of bacteria in the studied chambers, when the patients were present, ranged from 70 to 96 %, and when the premises were empty—from 82 to 95 %. In this study, mesophilic Gram-positive cocci (mainly Staphylococcus and Micrococcus) predominated among identified micro-organisms of the subterranean chambers. Their percentage contribution to the total microflora ranged from 56 to 86 % and was significantly higher than those noted for outdoor background, where ranged from 24 to 29 % (Mann–Whitney test: p < 0.05). Frequent occurrence of Staphylococcus  in the studied premises was noted. Their percentage contribution to the total bacterial microflora ranged from 8 to 27 %. In the examined premises, their highest concentration was particularly evident in the Kołdras chamber and in the physical activity area of the Ważyn chamber. Their high isolation frequency in the outdoor air (between 24 and 57 %) was also naturally reflected in their high abundance in the analyzed chambers (probably due to the transport through forced ventilation). Among isolated molds, Cladosporium cladosporioides predominated constituting from 42 to 66 % of the total fungal microflora in the studied chambers.Figure 4 and 5.7 × 102 cfu/m3, respectively. It shows that they were significantly higher than the concentrations reported by Dolezal et al.  on bacterial aerosol concentration both in each of the studied seasons and throughout the whole year. The highest bacterial aerosol concentration was observed during physical activity of the patients. The observed differences in bacterial aerosol concentrations between chambers result, interalia, from the fact that the Kołdras chamber is much smaller in terms of the size and bed capacity than the Ważyn chamber. The fact that the Ważyn chamber is adjusted for both the exercise therapy (sports field) and resting (sleeping area) is also important regarding its biological contamination.The presence of patients in subterranean chambers had a significant effect  may have bacteriostatic properties  was the only value that exceeded the admissible level. Hence, a high-performance method of microbial contamination reduction should be introduced in such premises to protect the patients, who have already suffered from respiratory diseases.The obtained results of both indoor and outdoor measurements of bioaerosol concentrations were compared with the Polish proposals for threshold limit values (TLVs), which are 5 × 10ts Górny . In almop > 0.05) for indoor and outdoor backgrounds. This observation is different from those noted by the other authors in numerous “overground” locations where seasonal fluctuations in bioaerosol concentrations are quite typical  promoting a fungal growth, has no significant influence  prevailed also in the outdoor air (where isolation frequency ranged between 24 and 57 %). Hence, forced ventilation was also responsible for delivery of these microbial contaminants to the studied subterranean chambers. Fungal species from Penicillium, Alternaria, Aspergillus and Cladosporium genera were the most often isolated in salt mine chambers, and these results are in a good agreement with the observations conducted by other authors  when patients were present, and physical exercises were carried out in the chambers. These confirm that human and its different activities including physiological  and physical  are the main source of bacterial aerosol in this indoor environment. The air contamination with fungal aerosol in investigated subterranean chambers was lower than outdoors . The results clearly indicate that more attention should be paid to a quality of the air, which is delivered to subterraneotherapy chambers through the shafts and corridors. The microbial analyses of the air in indoor and outdoor environment revealed that Gram-positive cocci , mesophilic actinomycetes (mainly Streptomyces) and molds (Cladosporium and Penicillium) were the predominant micro-organisms. As Gram-positive cocci can be easily released in high quantities from human body into the air, the number of people entering subterranean chambers should be limited. As the level of bacterial aerosol in subterranean chambers was significantly influenced with temperature and relative humidity, a constant control of these parameters should be scrupulously superintended in this type of premises dedicated for therapeutic purposes.The bioaerosol concentrations in subterraneotherapy chambers at the Bochnia Salt Mine Health Resort ranged from 35 to 11,688 cfu/m


Visceral leishmaniasis (VL) causes considerable morbidity and mortality in Ethiopia. Data on the prevalence and associated risk factors on malnutrition among VL patients in Ethiopia are scarce. This study aimed to assess the prevalence of malnutrition and its associated risk factor among VL patients in Northwest Ethiopia.An institution-based cross-sectional study was conducted from June to September 2012 at four leishmaniasis treatment sites in Northwest Ethiopia. Four hundred and three adult VL patients were enrolled in the study. Malnutrition was defined as a body mass index (BMI) ≤ 18.5 kg/m2. The data collected from the VL patients included sex, age, residence, occupation, weight, height, laboratory results . Multivariate logistic regression model was used to determine the strength of association between malnutrition and associated risk factors.Giardia intestinalis and Ascaris lumbircoides were the leading prevalent intestinal parasites. Factors such as age, sex, residence, occupation, HIV status and anemia were not associated with severe malnutrition.Among 403 adult VL patients 385 (95.5%) were malnourished. Twenty eight percent (n = 113), 30.3% (n = 122), and 37.2% (n = 150) were mildly, moderately and severely malnourished, respectively. The prevalence of intestinal parasitic infection was 47.6% (n = 192) and it was associated with malnutrition (P = 0.01). The prevalence of VL-HIV co-infection was 10.4% (n = 42). Hook worm, The prevalence of malnutrition in VL patients was very high and it was associated with intestinal parasitic infections. Therefore, screening of severely malnourished VL patients for intestinal parasitic infections during admission is recommended. Visceral leishmaniasis (VL) is one of the most neglected infectious diseases . It affeMalnutrition is one of the risk factors for the development of VL -8. MalnuAscaris lumbricoides, ongoing blood loss by hookworm infections and/or schistosomiasis . In this study 192 (47.6%) of VL patients were infected with one or more intestinal parasites. Of whom more than half n = 100; 52.1%) were severely malnourished. Forty eight VL patients had multiple intestinal parasitic infections; all were severely malnourished. Malnutrition due to intestinal parasitoses might be due to nutrient demand of parasite itself or blocking of the absorbing surface of the mucosa by adult worms, and ongoing blood loss by hookworm infections and/or schistosomiasis [0; 52.1% VL can be a risk factor for malnutrition and can lead to low BMI. On the other hand, malnutrition can affect the immune system of the host, making people vulnerable to infections. Malnutrition can make a person susceptible to infection and infection also contributes to malnutrition, which causes a vicious cycle .We use only BMI which depends upon weight and the square of height, it ignores basic scaling laws whereby mass increases to the 3rd power of linear dimensions.The prevalence of severe malnutrition and intestinal parasitic infection among adult VL patients was found high and associated. Therefore, screening of severely malnourished VL patients for intestinal parasitic infections is recommended. Possibilities of synchronized nutritional rehabilitation and universal deworming programs with VL treatment should be looked for.BMI: Body mass index; HIV: Human immuno-deficiency virus; OR: Odds ratio; VL: Visceral leishmaniasis.The authors declare that they have no competing interests.BM, ME, YT conceived the study, designed, participated in sample collection, performed laboratory experiments, conducted data analysis and drafted the manuscript for publication. AF, TT, KM reviewed the initial and final drafts of the manuscript. ED treated the patients, reviewed the initial and final drafts of the manuscript. All authors read and approved the final manuscript.


Pan troglodytes) from two separate groups watched videos of familiar and unfamiliar individuals yawning or at rest (control). The chimpanzees yawned more when watching the familiar yawns than the familiar control or the unfamiliar yawns, demonstrating an ingroup-outgroup bias in contagious yawning. These results provide further empirical support that contagious yawning is a measure of empathy, which may be useful for evolutionary biology and mental health.Humans favor others seen as similar to themselves (ingroup) over people seen as different (outgroup), even without explicitly stated bias. Ingroup-outgroup bias extends to involuntary responses, such as empathy for pain. However, empathy biases have not been tested in our close primate relatives. Contagious yawning has been theoretically and empirically linked to empathy. If empathy underlies contagious yawning, we predict that subjects should show an ingroup-outgroup bias by yawning more in response to watching ingroup members yawn than outgroup. Twenty-three chimpanzees ( Homo sapiens) favor other humans seen as belonging to their own group (ingroup) over humans seen as belonging to different social groups (outgroup), even in absence of explicitly stated bias Humans , the closer the social bond between individuals, the more likely they would yawn when the other yawned The link between empathy and contagious yawning has empirical support. Humans who performed better at self-recognition and theory-of-mind, two abilities that contribute to complex empathy, performed more contagious yawning Macaca arctoides) Canis familiaris) Contagious yawning has been documented in five mammalian species: humans Our hypothesis was that if empathy is the mechanism underlying contagious yawning, then contagious yawning should show the same biases as other measures of empathy, specifically the ingroup-outgroup bias. We tested two groups of captive chimpanzees by showing them yawn and control videos of their own group and the strange group. Chimpanzees form communities that are territorial and exclude neighboring individuals and communities Ethics: The experiment presented was approved by the Institutional Animal Care and Use Committee of Emory University (#083-2008Y) and was conducted in accordance with the “Guidelines for the treatment of animals in behavioural research and teaching” by the Animal Behavior Society/Association for the Study of Animal Behaviour and the Weatherall report on “The use of non-human primates in research”. We used voluntary testing to minimize stress on the subjects.Pan troglodytes, age 10–46, 4 m/19 f) housed in two groups of 12 at the Yerkes National Primate Research Center . The 4 males and 19 females ranged in age from 10 to 46. The chimpanzees lived in large outdoor enclosures  with indoor sleeping quarters and could not see each other. Group 1 had an additional indoor testing building. Chimpanzees were tested indoors or outdoors depending upon where they approached the video player. Chimpanzees saw only one stimulus per day between 10.00 h and 13.30 h.The subjects were 23 adult chimpanzees  digital video camera. We obtained yawns from seven individuals from each group, selected one yawn from each individual based on the quality of the segment , and edited each yawn clip to 9 s using iMovie HD . From the same footage we selected 9 s control segments from each of the same individuals at rest. By using the same footage we were able to select control clips with virtually identical viewing angles, postures, background compositions, lighting, etc. We included 1 s of green screen between each clip and assembled them into a yawn video and a control. Each clip was shown once before repeating, and the order within a set was randomized on the condition that the same clip could not be shown consecutively.The videos were presented on an iPod Touch  with a 7.5×5 cm screen. The iPod was presented on its own to a chimpanzee when other individuals were not within view. When more than one individual was within view, we placed the iPod up against an opaque container with an eyehole at the opposite end. The hole was small enough that only one chimpanzee could see through it at a time. Chimpanzees were tested alone, in small groups, or with the entire group based upon their comfort. Some individuals were at ease being alone and were tested closed in a room indoors. Most individuals were more comfortable when in a group. Small groups and individual testing was also needed to ensure that low-ranking individuals could get access to the video player. Regardless of whether an individual was alone or in a group, the small screen of the iPod and the container with the eyehole ensured that we could present the video to one subject at a time.a priori reasons for order effects to occur when the stimuli changed to something novel.Each chimpanzee was exposed to the videos for a total of 20 minutes on one or more days, depending upon the interest and cooperativeness of each individual. Some chimpanzees reached 20 min in one session, but most needed more than one day of testing. Timing for each session started after the chimpanzee first looked at the video. The chimpanzee was then free to watch the video or not as it chose, however the entire 20 min of recording was after observing the video and thus reflects the influence of viewing the video on the chimpanzee. Within each condition, ingroup or outgroup, the order of yawn and control videos was counter balanced (as close as possible due to the odd number of subjects). However, all subjects saw the ingroup videos before switching to the outgroup videos, as the outgroup exposure was a follow-up. Whereas we can envision order effects when the videos were of the same subjects , we cannot envision t-tests unless indicated). Values reported are means ± SEM, and all statistics were two-tailed.A digital video camera recorded each session. We coded each session for the number of yawns by each chimpanzee after the first attention toward the video. We also coded the amount of attention each individual paid to the video in s. We analyzed the results using PASW Statistics 18.0 for Macintosh  according to our previous recommendations t22 = 3.61, p = 0.002, d = 1.05, t22 = 1.40, p = 0.175, d = 0.34, t22 = 2.73, p = 0.012, d = 0.51, The chimpanzees yawned more frequently in response to the ingroup yawn video than to the ingroup control . Fourteen individuals saw clips of themselves yawning in the ingroup video (1 in 7 clips), but these individuals yawned at similar rates (5.21±1.42) as the 9 individuals who did not . There were no relationships between the amount of yawning and the number of sessions (1–5) needed to reach 20 min of exposure .There were no differences in yawning between females  and males  for either stimulus .The subjects paid similar amounts of attention to the yawn and control videos for both conditions (ingroup: The chimpanzees yawned more in response to the familiar yawn video than the familiar control, demonstrating contagious yawning. However, the video of unfamiliar chimpanzees had no detectable effect, as the difference in yawning between the yawn and control videos was nonsignificant. Critically, the chimpanzees yawned more in response to the familiar yawns than the unfamiliar yawns, demonstrating ingroup-outgroup bias. This bias supports the hypothesis that empathy is the mechanism underlying contagious yawning.per se as a mediating factor and supports the idea that social identification with the stimuli influenced the rate of contagion.The link between empathy and contagious yawning is further supported by our data on attention. The chimpanzees actually watched the videos of unfamiliar individuals more than the videos of familiar individuals. They attended more to the unfamiliar yawns, but yawned more to the familiar yawns. This finding rules out attention a priori reasons for an order effect. The attention data show that the chimpanzees did not lose interest in the videos since they watched the outgroup videos more than the ingroup videos. There is no evidence nor are there suggestions in the literature that contagious yawning is transient and fluctuates over time. These same subjects were previously tested and showed contagious yawning Even though all of the ingroup videos were presented before the outgroup videos, we can think of no In contrast to chimpanzees, humans We should also be mindful of social structure, as we may have two different factors at work: familiarity and group membership. Chimpanzees are territorial and aggressive toward neighboring communities Exposed to artificial stimuli that transcend the ingroup-outgroup distinction, chimpanzee yawn contagion shows patterns similar to those of brain imaging studies of empathy. Chimpanzees yawned in response to 3D computer-animated chimpanzees yawning Contagious yawning in humans has not yet been tested for biases, including social closeness


Paenibacillus polymyxa is a bacterium widely used in agriculture, industry, and environmental remediation because it has multiple functions including nitrogen fixation and produces various biologically active compounds. Among these compounds are the antibiotics polymyxins, and the bacterium is currently being reassessed for medical application. However, a lack of genetic tools for manipulation of P. polymyxa has limited our understanding of the biosynthesis of these compounds.P. polymyxa under conditions where homologous recombination is enhanced by denaturation of the suicide plasmid DNA. To test this system, we targeted P. polymyxa α-and β-amylase genes for disruption. Chloramphenicol or erythromycin resistance genes were inserted into the suicide plasmid pGEM7Z-f+ (Promega). To mediate homologous recombination and replacement of the targeted genes with the antibiotic resistance genes nucleotide sequences of the α-and β-amylase genes were cloned into the plasmid flanking the antibiotic resistance genes.To facilitate an understanding of the genetic determinants of the bacterium, we have developed a system for marker exchange mutagenesis directly on competent cells of P. polymyxa E681. We propose that P. polymyxa isogenic mutants could be developed using this system of marker exchange mutagenesis. α-and β-amylase genes provide a useful tool for direct recombinant screening in P. polymyxa.We have created a simple system for targeted gene deletion in  Paenibacillus polymyxa (formerly Bacillus polymyxa), the type species of Paenibacillus, is considered to be a plant growth-promoting rhizobacterium (PGPR) with a broad host plant range P. polymyxa is widespread in the soil and is widely used in agriculture, industry, and environmental remediation because of its multiple functions P. polymyxa have been sequenced by four groups P. polymyxa E681 genome is composed of one circular chromosome of 5.4 Mb P. polymyxa isolates have potential to produce various economically important biologically active compounds; however, a lack of genetic tools available for manipulation of P. polymyxa has hindered our understanding of the biosynthetic steps involved in natural product synthesis. Only two very important antibiotic compounds, fusaricidins and polymyxins, have been successfully mutagenized in the bacterium. This was performed by targeted mutagenesis protocol based on genomic libraries of PKB1 and E681 P. polymyxa corresponding chromosomal loci has not been demonstrated previously probably due to low efficiency of transformation or to the low efficiency of homologous recombination events. Library-based mutagenesis is a costly and time-consuming approach and not realistic when a number of genes have to be mutated simultaneously. In this study α- and β-amylase genes were targeted for inactivation to test the mutagenesis system we developed.The enzyme α-amylase randomly hydrolyzes α-1,4-linkages in both amylose and amylopectin chains. The enzyme β-amylase hydrolyzes alternate β-1,4 linkages from the ends of amylose and amylopectin chains. P. polymyxa E681 cells under conditions where homologous recombination is enhanced by denaturation of the suicide plasmid DNA. Alpha -and -beta-amylase genes were targeted to test the system. The genes provide a useful tool for direct recombinant screening in P. polymyxa.Here we report on successful gene replacement in Paenibacillus polymyxa E681 was isolated from the rhizosphere of winter barley Escherichia coli DH5alpha cells (Invitrogen) acted as host for recombinant plasmids and were cultivated at 37°C on LB agar. Bacillus subtilis JH642  was used for the complementation experiment. B. subtilis and P. polymyxa E681 were grown in half strength Tryptic soy broth (TSB) (Difco) at 30°C. Brain heart Infusion medium (Difco) containing 10% sucrose was used for the transformation of P. polymyxa E681. The extraction of plasmid and chromosomal DNA were performed according to standard procedures P. polymyxa genes by replacement with a chloramphenicol or erythromycin resistance cassette was constructed from the plasmid pGEM7Z-f+ (Promega) which replicates in E. coli but not in P. polymyxa  and was then ligated into pGem7Zf(+) via EcoRI and BamHI cleavage sites resulting in plasmid pGEM7Zf Cat. Subsequently, 760 bp and 870 bp fragments of the flanking regions of the α-amy gene were amplified by PCR from the E681 chromosomal DNA using α-amy-FF – α-amy-FR and α-amy-RF – α-amy RR primer pairs resulting in PCR products A and B respectively. Fragments A and B were then introduced into pGem7Zf cat (+) using the NsiI and BamHI, and XbaI and ApaI restriction sites, respectively, resulting in plasmid pGEM7Za-amy.The cat gene was replaced with the gene encoding erythromycin resistance (erm) using primers EmyF and EmyF. Then, 742 bp and 794 bp fragments of the flanking regions of β-amy gene were amplified by PCR from the E681 chromosomal DNA using β-amy –FF/β-amy -FR and β-amy –RF/β-amy RR primer pairs resulting in PCR products C and D respectively. The fragments were inserted to into pGem7Zf erm NsiI and BamHI, and XbaI and ApaI restriction sites resulting in plasmid pGEM7Zfb-amy.The pGEM7zf Cat P. polymyxa was performed by electroporation. In order to prepare competent cells, a single P. polymyxa E681 colony was picked from 1/2 TSA solid medium and cultured in BHIS broth 600 of 0.5, the bacterial growth was interrupted by placing the culture on ice for 10 min and centrifuged at 5000×g for 10 min at 4°C. After washing the cells twice with cold SM buffer Transformation of 2O, and kept on ice before electroporation. For UV treatment, 18 ml of DNA solution in a droplet on a plate was subjected to 2 50, 100 or 250 mJ cm−2 UV irradiation in a UV chamber (CL1000 UV crosslinker UVP Inc.), and DNA was kept on ice before electroporation. Competent cells were mixed with DNA, transferred to 2 mm cuvettes, and placed on ice for at least 5 min. The sample was pulsed with a voltage of 6.25 kV cm−1, a capacitance of 25 µF, and a resistance of 200 Ω. One ml of 30°C pre-warmed BHIS was added and the cell suspension was incubated at 30°C for 3 h.It is known that homologous recombination may be strongly stimulated by denaturation of double-stranded circular DNA. DNA for electroporation was prepared after slightly modified protocols of Oh and Chater (1997) and Hinds et al (1999) Transformants were selected on respective antibiotics plates and cassette insertion was confirmed by PCR with primers flanking the sequences used in the mutagenesis vector.P. polymyxa was achieved, the complementation was performed in B. subtilis JH642. For this purpose, the α-amylase gene of B. subtilis JH642 was first deleted by using pDG1662 integration plasmid . For the complementation of the α-amylase gene of P. polymyxa E681, the α-amylase gene was amplified with C-α-amyF and C-α-amyR then cloned into pHPS9 , an E.coli/Bacillus shuttle vector, via the NdeI and BamHI restriction sites. For complementation of the β-amylase gene, the full structural gene of β-amylase was amplified with C-β-amyF and C-β-amyR. This fragment was cloned into pHPS9 using the NdeI and BamHI restriction sites. These plasmids carrying amylase genes were introduced into B. subtilis by natural uptake. A fresh single colony cultivated overnight was inoculated in Spizizen’s medium As only partial complementation of amylase genes in 2 2H2O), followed by 300 µl of detection solution (0.01% w/v I2 and 0.1% w/v KI in 1 M HCl). The starch concentration was measured by changes in the optical density at 620 nm.Production of α- and β- amylase was quantified as follows. Each of the strains was inoculated in half strength tryptic soy broth (TSB) containing 1% soluble starch . An aliquot of the culture was removed at each measurement time and centrifuged at 8,000×g for 10 min at 4°C. The supernatant was diluted ten times with distilled water and 150 µl of diluted supernatant was added to 600 µl of reaction buffer  and flat colonies (F type). Similar colony morphology has been observed e.g. in P. aeruginosa. The mechanism of phase variation includes site specific recombination, gene conversion, epigenetic modifications, nested DNA inversions and slipped strand mispairings but could also involve recently reported new mechanisms UvrABC). This has been shown to be an effective recombination substrate owing to the rapid formation of RecA filaments P. polymyxa phase variation remains to be elucidated. Yet it is already clear that on and off switching of phenotype expression is involved when manipulating the bacterial populations.There is considerable interest in P. polymyxa E681 α-amylase and β-amylase genes PPE_02348 and PPE_04705 were created. In addition to testing the system the genes encode enzymes that participate in starch degradation and hence provide a system for direct screening for various heterologous fragments to the genes to P. polymyxa on starch clearing plates. For deletion of the α -amylase gene, its structural gene flanking regions were cloned into a pGEM7Zderivative containing the cat gene which inactivates chloramphenicol. For the β-amylase gene deletion the gene flanking regions were cloned into a pGEM7Z derivative containing the erm gene which encodes erythromycin resistance (P. polymyxa (cat and erm genes) into the α-and β-amylase genes their amylolytic activity was disrupted. β-amylase and double mutant show distinct differences in starch clearance on plate assays. These phenotypes therefore allow the direct screening of recombinant clones. Further, the identification of the recombinant clones can be performed directly on the transformation plates by a simple assay. In contrast to other screening possibilities commonly employed for positive differentiation of recombinants, starch clearance plates provide excellent results at modest costs.In order to demonstrate the functionality of this gene knockout system, deletion mutations of sistance . After tsistance . This issistance . Double polymyxa . Our resP. polymyxa E681 starch degradation in ½ TSB over 60 h (P. polymyxa during late stationary phase. The complementation of amylase genes was attempted in P. polymyxa E681derivatives using pHPS9, an E.coli/Bacillus shuttle vector. The α- amylase gene was amplified with C-α-amyF and C-α-amyR primers and β- amylase with C-β-amyF and C-β-amyR primers. The amplified products were inserted into pHPS9 between NdeI and BamHI restriction sites. Yet only 25% starch degradation with complementation experiment was achieved. It is likely that the p59 promoter driving the expression of cloned amylase genes in pHPS9 had a reduced activity in P. polymyxa. Hence, the complementation experiment with amylase genes was performed in B. subtilis JH642. Initially the JH642 α-amylase gene was deleted and replaced by P. polymyxa α- and β-amylase genes (To confirm disruption of the amylase genes, we studied the dynamics of ver 60 h . The resse genes . Howeverse genes  C and D.P. polymyxa genome may be inferred from sequence similarities, the functions of the majority of the genes are unknown. In this study, we developed a new genetic tool for P. polymyxa that enables efficient and less costly creation of gene knockouts. To our knowledge, this represents the first reported use of gene replacement by homologous recombination based on P. polymyxa cells. Likewise we show that the α- and β-amylase genes provide a valuable genetic marker that enables rapid direct screening of heterologous genes in P. polymyxa cells. Even though this study focused on the specific insertional mutagenesis of α - and β-amylase genes, the technique described could be used more generally for mutagenesis of various genes in P. polymyxa. The isolation of mutants deficient in important functions would be extremely useful in elucidating the genes that contribute to the beneficial effect of P. polymyxa.While the function of many genes in the