Techniques to compensate for these artifacts were applied to a large jellyfish, Cyanea capillata, freely swimming in ocean waters. The bell kinematics were captured by digitizing exumbrella profiles for two full swimming cycles. Magnification was accounted for by tracking a reference point on the ocean floor and by observing the C. capillata exumbrella arclength in order to have a constant scale through the swimming cycles. A linear fit of the top bell section was used to find the body angle with respect to the camera coordinate system. Bell margin trajectories over two swimming cycles confirmed the accuracy of the correction techniques. The corrected profiles were filtered and interpolated to provide a set of time-dependent points along the bell. Discrete models of the exumbrella were used to analyze the bell kinematics. Exumbrella discretization was conducted using three different methods. Fourier series were fitted to the discretized models and subsequently used to analyze the bell kinematics of the C. capillata. The analysis showed that the bell did not deform uniformly over time with different segments lagging behind each other. Looping of the bell trajectory between contraction and relaxation was also present through most of the exumbrella. The bell margin had the largest looping with an outer path during contraction and inner path during relaxation. The subumbrella volume was approximated based on the exumbrella kinematics and was found to increase during contraction.Obtaining accurate kinematic data of animals is essential for many biological studies and bio-inspired engineering. Many animals, however, are either too large or too delicate to transport to controlled environments where accurate kinematic data can be easily obtained. Often,  Jellyfish have a unique propulsion mechanism that is of interest to a diverse group of researchers from disciplines such as biology Cyanea capillata, see C. capillata are usually of reddish-brown or yellowish color C. capillata have one significant difference from most other jellyfish species; they have a segmented bell as opposed to a uniform bell. Segmentation allows flaps surrounding each of the bell segments to fold on itself during relaxation and expand during contraction. A maximum form area is therefore achieved during contraction and minimized during relaxation. Circular muscles in jellyfish subumbrella are typically solely responsible for propulsion. The C. capillata uses a combination of circular and radial muscles to contract its bell and create propulsion C. capillata tentacles originate from the subumbrella and are used to capture prey as they come in contact with the nematocysts on the tentacles, see Aurelia aurita on hydrodynamic performance. Their specimen diameters ranged from 1.57 to 9.51 cm. Herschlag and Miller A. aurita specimen ranging from 0.36 to 10.2 cm in diameter. C. capillata can reach dimensions over an order of magnitude from what has been analyzed and there is currently no hydrodynamic analysis of such large jellyfish.It is important to understand the implications of size on the propulsion mechanism of jellyfish. However, there is very little information available in literature addressing this issue. McHenry and Jed Jellyfish swim in an oscillating fashion which consists of a two phase cycle: contraction and relaxation. Contraction is achieved by muscles located in the subumbrella as shown in Jellyfish bell kinematics vary significantly from other animals. The locomotion is performed by a bell that is nearly uniform in structure, has high flexibility and undergoes large curvature change. This is different than most animals where motion occurs by actuating rigid internal structures about a pivot point such as in a bone and joint mechanisms. The lack of rigid structures demands new kinematics tracking and analysis methods. Flexible structures can be discretized in order to estimate their deformation and motion. Jellyfish bells have previously been discretized for computational fluid dynamics (CFD) applications The ability to analyze in situ video from field recordings is crucial for the study of many organisms which are either too large or too delicate to work within the laboratory environment. In addition, field recordings allow observation of animals in their natural habitat, thus reducing the probability of behavioral alterations and other effects common in laboratory studies such as wall effects C. capillata to analyze its kinematics. Large C. capillata cannot feasibly be analyzed in a controlled environment and as a result, little information is available in the literature on their behavior and swimming kinematics. These techniques will help in deriving accurate kinematics from video recordings obtained in sub-optimal conditions. This provides the ability to extract potentially valuable biological information from existing footage and future recordings.In this study, we present a novel method for obtaining accurate kinematics from in situ underwater recordings. We apply the method to a recording of a large C. capillata was used to elucidate the swimming kinematics of the animal. The video was obtained from the Ocean Footage video library An in situ video of a large C. capillata, the angle that the animal faces the camera is important. The C. capillata consists of eight radial bell segments and two opposing bell segments should be perpendicular to the field of view to extract the desired bell kinematics. (2) The 2D plane must be at a constant distance from the camera during the duration of the analysis. Otherwise, the animal scale changes over time. (3) The animal must not rotate about the x- or y-axis of the camera. Rotation out of plane causes part of the profile information to be lost when projected onto the 2D camera plane. The later criterion cannot be compensated during post-processing if only one view of the motion is available.The kinematic analysis can be visualized to take place on a 2D surface perpendicular to the camera. Traditionally, in order to capture the animal kinematics accurately, several criteria should be met: (1) The animal motion must occur in a 2D plane parallel to the camera plane. For animals that are not axi-symmetric such as the Cyanea capillata bell kinematics is (i) profile tracking of the bell. This can be achieved by tracking the exumbrella profiles since they provide most of the bell kinematics and can be detected from the high contrast formed with the background. The captured exumbrella profiles need to be corrected in order to provide useful kinematic information. The second step is to (ii) body tracking and removing the overall body motion from the bell kinematics. The third step is to (iii) remove any magnification occurring during the video sequence to prevent a change of scale in the kinematics. The fourth step is to (iv) remove body rotation and the fifth step is (v) strategic bell discretization for kinematic analysis.The first step in capturing the C. capillata met criteria (1) and (3) with negligible rotation about the x- and y-axis but did not meet the criterion (2). The footage begins with the jellyfish's bell fully contracted and covers two full cycles. It was filmed at 25 fps with image resolution of 640×360 pixels. Edge detection was performed manually for the entire exumbrella from one margin to another at every 5 frames using ImageJ resulting in profile points In a dynamic environment such as the ocean, it is difficult to achieve all the aforementioned criteria. The selected video of The profile points were post-processed in order to obtain a set of half profiles which can be used for kinematic analysis. In order to do so, the bell apex must be found. The apex is the top point of the exumbrella when looking at the bell from its side and the location where the axial axis meets the exumbrella. To find the apex, two different methods were considered since post-processing for analysis strongly relies on the apex location. The first method consists of measuring the exumbrella arclength with the jellyfish in the fully contracted position and then taking the midpoint of the full arclength as the apex. This method leads to a significant error in the apex position due to the complexity in locating the bell margin in the images. Margin location largely affects the exumbrella arclength and therefore, the apex location. The second method considered and used, consists of tracking a visible point on the bell near the apex and then shifting the point by the distance between it and the apex as measured using the first method for the first frame in the cycle. The profile points were divided into two half-profiles originating at the apex and ending at the margin. The half-profiles were than normalized by the exumbrella arclength of the half-profile in the relaxed position of the first cycle.The chosen image sequence was recorded from a camera that moves relative to the animal. The animal itself was moving relative to the ocean floor. These factors affected the scale of the animal throughout the video and had to be accounted for in order to resolve the bell kinematics. The easiest approach would be to use an animal feature that has a fixed dimension throughout the video and scale relative to it. However, there are no such features on the jellyfish that can be used for this approach. Therefore, two other techniques were used to account for magnification.a) uses the reference in the background to estimate the motion of the camera relative to the background. Significant image magnification occurs in the image sequence which is due to the camera moving relative to the ocean floor. A rock on the ocean floor was tracked and used to estimate the width variation in the image plane resulting from magnification. This is depicted in The first method  uses a varying dimension on the animal at the same position in different swimming cycles. The C. capillata specimen used in this study and other common jellyfish don't have anatomical features of fix length that can be used as a reliable reference point for determining the magnification. The total exumbrella arclength can be used as a size reference but the exumbrella naturally increases in length during contraction and decreases in length during relaxation. This is due to the swimming muscles compressing the subumbrella and thereby causing a slight expansion of the exumbrella Cyanea capillata and other jellyfish species over several swimming cycles. They have shown that the jellyfish swimming kinematics are cyclical and repeatable during straight swimming. We can therefore assume that the bell will have the same length at a given point during a cycle. The most accurate points and easiest to locate in a given cycle are found during a full contraction or full relaxation. This usually corresponds to a minimum and maximum diameter respectively or the geometry right before the relaxation or contraction respectively. The exumbrella arclength in the fully contracted position was used at three different times in the swimming sequence. The exumbrella arclength is calculated using: The second method  highest deformation as a function of arclength and (ii) highest deformation as a function of time. Three discretization methods were explored to locate nodes which meet the two criteria. (1) The curvature method uses a series of exumbrella profile to determine where most of the deformation occurs over time. (2) The anatomical method consists of observing the animal's anatomy to identify where the variations in structure and anatomical features might affect deformation. (3) The node optimization method consists of optimizing the node location for a set of segments by minimizing the error found between the discrete model and natural profiles.A model of the tA is the area of the triangle. The area can be found using Heron's formula: Curvature is calculated using the definition of a circumcircle which is: a circle passing through three vertices F, G and H of a triangle with sides of length f, g and h. The diameter of a circumcircle is:n is the total number of points for a given exumbrella profile. The derivative of curvature with respect to arclength was taken as follows: The curvature of the circumcircle is:C. capillata ranging from 2 to 30 cm in diameter. A specimen laying on its exumbrella with tentacles removed was used to obtain the dimensions of different features. The features of interests were the central disk, circular muscles, radial muscles and flap, see The second method used to determine node locations consisted of analyzing the anatomical features of the natural animal to determine the musculature and joint arrangements. Gladfelter C. capillata exumbrella profile and the exumbrella model, see An error analysis was conducted to determine a segment configuration which best matched the animal's bell kinematics. It also allowed the evaluation of the other two different node detection methods. The area formed between the This gives the total error Jellyfish propulsion is often modeled as a jetting mechanism where thrust is a function of subumbrella volume change C. capillata bell is axi-symmetric which is a crude representation of the bell geometry but gives a good approximation for this analysis.The total exumbrella arclength Selected exumbrella profiles after processing are shown in The margin trajectories for the two cycles overlay well which means the bell is following a similar path during both cycles. In order for this to occur, magnification and rotation had to be properly compensated. Minor variations are seen after processing which can be due to inconsistencies in deformation when the animal is reacting to water currents, minor errors from manual edge detection, the footage not fully meeting criteria (1) and (3) through the two cycles and the assumption of linear motion between the animal and camera.n = 0.5 of n = 0.62 and xn = 0.28 at the bell margin between the relaxed and contracted positions. The flap flares outwards during contraction and is bent inwards during relaxation. This results in an outer trajectory during contraction and inner trajectory during relaxation. The same observation has been made with A. auritaDuring relaxation, the bell goes over the apex as seen at xThe anatomical analysis revealed five important locations which set delimitation for a four-segment model. The 0% location corresponds to the bell apex which is at the center of the central disk. The central disk region is passive and consists of mesoglea and gonads. It is delimited by the coronal joint and circular muscles at 40%. This joint eases deformation at that location when the circular muscles actuate. The radial muscles begin at the outer edge of the circular muscles which is at 64%. This location of actuator transition will be less stiff then the surrounding area due to the lack of muscles. It is therefore also considered as a node location. The flap starts where the radial muscles end at 88%. This is a location of mechanical transition from active to passive. The flap is also more flexible due to its lack of muscles which will result in more compliance during contraction and relaxation. The fifth node is taken as the bell margin 100%, which also marks the end of the flap.Curvature was calculated for each point on each profile and then averaged over the 52 profiles. The average curvature for the right side of the bell is shown in The average curvature curve shows that there are seven main peaks and valleys at 10, 24, 36, 66, 76, 84 and 96% of exumbrella arclength starting at the apex. These peaks and valleys correspond to potential node locations. The points corresponding to these peaks and valleys were found by identifying the points where C. capillata. It is possible that the anatomical ratios measured varied between the two animals analyzed. Discrepancies in the footage processing can also have led to error in curvature analysis. Since a top view of the subumbrella was used to extract the anatomical information, the actual arclength of the subumbrella could not be used which led to error when calculating the arclength percentage location.Most points along the active part of the bell undergo a similar standard deviation. Variation increases significantly near the point at 82%. Comparing with the anatomical measurements, we can see that this point is close to the 88% point which marks the end of radial muscles and the beginning on the flap. The flap corresponds to a passive part of the bell beginning at the end of the swimming muscles up to the bell margin. The flap undergoes large deformation and movement during swimming but areas on the flap should not be considered node locations from a robotic perspective since this deformation can be achieved by a passive material C. capillata, it is desired to reduce the error between the natural profiles and the model. Depending on the application, the best number of segments used to represent the motion might differ. The error analysis technique was used to find which node location on the exumbrella profile minimized error for a given number of segments. This was done for 1 to 6 segments and the results are shown in When modeling the kinematics of the C. capillata profiles are shown in As expected, the error decreases with increasing number of segments. This is illustrated in C. capillata approximated by four segments for any number of cycles using the following equations:Kinematic models of the four-segment optimized model and the four-segment anatomical model were developed using Fourier series. Segment length C. capillata bell allows for an analysis of individual bell sections. The four-segment anatomical Fourier model is used to help gain an understanding of how the different anatomical regions behave during a swimming cycle. The angles formed between each segment are plotted as a function of time in C. capillata was used for this analysis and is also the same cycle used for the Fourier model. The original data was clipped and shifted in time so that the contraction occurs first instead of relaxation. A discrete representation of the Contraction is taken as the period when the active sections of the bell are moving inwards. This occurs when the circular or radial muscles cause the bell to contract. In the four-segment anatomical Fourier model of Aurelia auritaC. capillata kinematics.The bell trajectories at the anatomical node locations during a full swimming cycle are plotted in C. capillata begins to contract. This is due to the bell extending and flattening as contraction begins. When the radial muscles start contracting at 0.6 s, the subumbrella volume then starts increasing. The volume fluctuates slightly while cruising and starts decreasing during relaxation. This goes against the widely accepted jetting model used to quantify jellyfish propulsion The subumbrella volume and bell diameter as a function of time for one full cycle are plotted in Cyanea capillata biomimetic robot named Cyro Many large or delicate marine animals are poorly studied and understood due to our lack in ability to acquire accurate data about them in their natural environment. The ability to use in situ recordings to obtain accurate kinematics provides insight into how these animals interact with their environment as well as potential for growth and reproduction. For jellyfish, swimming not only provides locomotion but also affects prey encounter C. capillata kinematics since the information we were interested in is contained in that plane. This is a result of the animal's muscle structure and geometry. For more complex motions and geometries, the same techniques can be applied to additional view angles and combined to give kinematic information on different planes. This can help quantify motion that occurs in three dimensions such as the oscillation and undulation of a manta ray's pectoral fin Only one plane was analyzed for the C. capillata, the material behavior and mechanics must also be taken into account. For such application, the dimensions given by the anatomical analysis might have better results. This method shows where artificial actuators should be located if they were able to recreate the same deformation assuming they have the same form factor as the natural muscles. A downside of this method is the required analysis of the natural animal's anatomy which may not be feasible. The kinematic models provided can be used as a time dependent representation of the C. capillata bell motion. The four-segment optimized kinematic model best matches the exumbrella kinematic representation for four segments. The four-segment anatomical kinematic model provides the kinematics of the bell at locations where important anatomical features act.The bell segment facing the camera was tracked and compared with the bell apex to see if there was rotation about the x- and y-axis. The results showed that rotation was in the same range as the error from image tracking and was therefore neglected. The three node detection methods presented in this study offer different benefits. The curvature method determines where the areas of highest deformation occur. With this method, there is no need to actually look at the animal's anatomy. Footage of animal kinematics satisfying the three selection criteria is the only information required. The error analysis method provides the best node and segment representation of the exumbrella for a given number of segments. This can be especially useful for modeling the bell in computational applications as a discrete representation. On the other hand, it does not take into account the mechanics of the bell which would be necessary in a robotic application. To reproduce the same kinematics and morphology of the C. capillata bell kinematics. It was found that the bell does not deform uniformly. The radial muscles actuate with a lag period from the circular muscles and also lag behind the circular muscles during relaxation. The passive flap was found to have significant lag behind all the other sections. This leads to looping in the bell margin causing the bell to have an outer trajectory during contraction and inner trajectory during relaxation. The kinematics analysis also revealed that the C. capillata had a cruise time which lasts for 1/5th of its swimming cycle where the bell geometry stays near constant. Rowers such as the A. aurita do not undergo cruise time. Their contraction is followed by an immediate relaxation C. capillata models found in this study coupled with a hydrodynamic analysis will allow for the investigation of these hypotheses.The four-segment anatomical Fourier model revealed important features of the C. capillata as was found in this analysis. According to the jetting model this should result in a negative thrust. A jetting model is therefore inappropriate for representing the propulsion mechanism of C. capillata. The starting and stopping vortex interaction between the contraction and relaxation phase described by Dabiri et al. C. capillata.Rowing jellyfish propulsion has been modeled as a combination of jetting and paddling C. capillata bell kinematics which requires further exploration is the kinematics of individual bell segments. Bell segments have a three-dimensional deformation which comes from the presence of a passive flap that surrounds the circumference of each bell segments. Also, the tentacles and oral arms being a prominent mass portion of the animal will influence the dynamics and therefore kinematics of the animal. The extent to which they contribute to the kinematics will require further investigation.An important aspect of S1 AppendixFourier transform coefficients.(DOCX)Click here for additional data file.


The reduced capacity to form ARs in bot1-1 was associated with altered expression of MAP-encoding genes along AR induction. While the expression of MAP65-4, MAP65-3, AURORA1, AURORA2 and TANGLED, increased in wild-type but not in bot1-1 plants, the expression of MAP65-8 and MDP25 decreased in wild type plants but not in the bot1-1 plant after two days of IBA-treatment. The expression of MOR1 was increased two days after AR induction in wild type and bot1-1 plants. To examine its expression specifically in AR primordium, MOR1 upstream regulatory sequence was isolated and cloned to regulate GFP. Expression of GFP was induced in the primary root tips and lateral roots, in the pericycle of the hypocotyls and in all stages of AR primordium formation. It is concluded that the expression of MAPs is regulated along AR induction and that reduction in KATANIN expression inhibits AR formation and indirectly influences the specific expression of some MAPs.Adventitious roots (AR) are post embryonic lateral organs that differentiate from non-root tissues. The understanding of the molecular mechanism which underlies their differentiation is important because of their central role in vegetative plant propagation. Here it was studied how the expression of different microtubule (MT)-associated proteins (MAPs) is affected during AR induction, and whether expression differences are dependent on MT organization itself. To examine AR formation when MTs are disturbed we used two mutants in the MT severing protein KATANIN. It was found that rate and number of AR primordium formed following IBA induction for three days was reduced in  ARs are roots that differentiate from non-root tissues, as a response to endogenous or exogenous auxin signaling . The tismor1-1 temperature-sensitive mutant to a restrictive temperature, amorphous clusters of dividing cells were formed rather than AR primordium. In these clusters, which were as big as stage V primordium, MTs were mainly randomly oriented, cell wall properties were changed, PIN1 polarization and auxin maxima were impaired and the differentiation of the root epidermal cell layer was inhibited [KATANIN, bot1-1 formed fewer ARs, a plant overexpressing RIC1 that activates KATANIN [bot1-7 as well as ric1 mutant plants made more flowers than wild type plants when auxin transport was blocked [INFLORESCENCE DEFICIENT IN ABSCISSION (IDA) abscission signaling affects LR primordium morphogenesis and differentiation [In a recent study, the role of MTs in AR formation was investigated. It was found that subtle perturbations to MT dynamics can uncouple cell division from cell differentiation during AR induction . It was nhibited . In the  KATANIN , formed  blocked  suggesti blocked , auxin s blocked , blue li blocked  and mech blocked . The lat blocked , 15, whintiation , 16, 17.ntiation . TherefoEucalyptus grandis had found distinct expression patterns in easy to root juvenile cuttings compared to difficult to root mature cuttings [Of note, another recent study, in which the expression of tubulins and MAPs during AR induction in cuttings .MOR1 was not significantly different during AR formation in wild type and bot1-1 plants, and the expression of GFP under its upstream sequence during early stages of AR primordium further confirms the importance of this MAP in AR induction.In the current work we have asked how the expression of different MAPs, is regulated during AR formation and to what extent the loss of MT severing by the KATANIN ATPase influences these possible changes of expression. The expression of All materials were purchased from Sigma (Rehovot Israel) unless otherwise mentioned.Arabidopsis seeds were germinated and transformed as previously described [MOR1 promoter, a fragment containing the upstream sequence of MOR1 starting just before the ATG and including bp 14964640–14966828 from chromosome 2 was isolated by the primers described in MOR1pro:GFP were isolated. Other plants used were wild-type Arabidopsis thaliana ecotype Columbia or WS. KATANIN mutants bot1-1 and 1–7 were from [escribed . To isolere from . ARs werMOR1prom:GFP mean fluorescence was measured from the whole tissue imaged by the FL500 software from roots of 7d old seedlings which were grown on MS or on MS+10 μ M IBA for 16 hr (6d old seedlings which were transferred to IBA containing media over night). Three independent lines  were examined, from each 3 individual plants were measured for mean fluorescence, total of 9 plants). Mean fluorescence was also measured from hypocotyls which were excised from 7d old etiolated seedlings and from those excised similarly after 5d and incubate in the dark in MS +10 μ M IBA for 2d. For the closer examination of AR primordium, cut etiolated hypocotyls were incubated in MS media supplemented with 1% sucrose and 10 μ M K-IBA. Image acquisition was performed from hypocotyls of the line MOR1prom:GFP-7, after 1, 2, and 3d of induction. An Olympus (Hamburg Germany) IX81/FV500 laser-scanning microscope was used to observe fluorescently labeled cells with the following filter sets: for eGFP, 488-nm excitation, and BA505-525 were used; The objectives used were PlanApo UplanApo 10X 0.4 N.A, UplanApo 20X 0.7 N.A.www.Nanostring.com, Seattle, Washington, USA). The primers used for hybridization are listed in Etiolated hypocotyls were excised from 7d old seedlings and incubated in MS + 1% sucrose with or without 10 μ M IBA. Total RNA was extracted from 3–4 biological replications at day 0 (the day of excision) and after 6h, 1, 2, and 3 d. Expression analysis of selected MAPs was performed by Nanostring  or tubulin β4 using (AT5G44340) using DNase treated RNA as template. No PCR product amplification in RNA samples was used as a quality control to ascertain the complete removal of genomic DNA. Quality checked RNA (1 μg) was used for cDNA synthesis using Maxima First strand cDNA synthesis kit as per manufacturer instructions . The amount of 2.5 μl cDNA was used as a template to carry out real time PCR analysis  and quantify MOR1 expression levels relative to HK genes. Primer sequences are listed in δ Ct relative to HK as previously described [RNA was extracted as above. RNA thus extracted was treated with DNase (Thermo Fisher) 1U/μg RNA at 37°C for 30 min followed by addition of 5 mM EDTA  and heat inactivation at 75°C for 10 min. Removal of genomic DNA was confirmed by carrying out PCR amplification of the following housekeeping (HK) genes, the expression of which was found constant under our experimental conditions; escribed .bot1-1 mutant seedlings are hampered in AR formation [KATANIN expression affects AR formation in cut etiolated hypocotyls treated with IBA. Two KATANIN mutants were used: bot1-1, which is in the Columbia ecotype background, and bot1-7, which is in the Wassilewskija ecotype (WS). bot1-1 and bot1-7 hypocotyls had detectable root primordium at that time respectively. In addition, while about 40% of wild-type hypocotyls showed emerged roots, 0% of bot1-1 and 4% of bot1-7 had such roots 3 days after induction. To determine whether the reduced KATANIN expression affected root induction or root emergence, hypocotyls were also examined after 6d. It was found that while 100% of hypocotyls of wild type had roots after 6d, more than 90% of bot1-7 and 40% of bot1-1 had roots at that time, indicating that once formed, root primordium eventually emerged in the mutant plants   , which is in agreement with the nanostring data.Our analysis indicated that  = 0.09) . When cuimordium . MOR1 mRp = 0.2) . This miMOR1prom in initial steps of AR induction we have induced AR in etiolated hypocotyls and imaged them after 1, 2 and 3 days. To follow the expression of GFP under the In this study we have determined that the expression of MAPs is regulated during AR formation and that this regulation is perturbed when the MT severing protein KATANIN is reduced. Thus, while KATANIN can directly influence MTs organization patterns , 24 it mbot1-1 plants, which could lead to loss of coherent auxin transport within AR primordial cells, inappropriate regulation of various MAPs expression by auxin and to inhibition of AR formation.MTs are sensitive to mechanical signals –29 and aMOR1 function inhibited AR formation and over expression of the RIC1 increased AR formation [Taken together with our previous findings that loss of ormation , the curGiven that MTs are involved in cell division , 36 and bot1-1 samples treated with IBA in which fewer AR were formed. Interestingly, all the samples within each of the above groups clustered together, indicating that MAPs expression remodeling during AR induction was influenced by the IBA treatment and the bot1-1 mutation. Down regulation of some MAPs was observed in wild-type plants 6h after IBA application, which is interesting and yet to be explained. Of note, the expression of MAPs that are known to function during cell division such as MAP65-4, MAP65-3, AURORA1, AURORA2 and TANGLED, [bot1-1 plants. Interestingly, the expression of KATANIN itself increased in wild-type plants induced by IBA but not in IBA untreated plants, indicating its importance. The expression of MAP65-8 and MDP25 [bot1-1 mutant after IBA. MAP65 family members affect MT arrays in different ways. It has been found, for example that while MAP65-1 can promote MT polymerization, MAP65-6 does not [in vitro [clasp plants exhibit auxin-related phenotypes, including abundant lateral roots [bot1-1 and wild-type plants suggest that MAP expression changes during AR induction by IBA is MT-organization dependent. It should, however, be taken into consideration that RNA was extracted from the whole hypocotyl and not preferentially from primordium, so it cannot be ruled out that some MAP transcripts are regulated differently within the dividing cells.Expression profiles of MAPs in wild-type plants treated with IBA for 0–3 days, during which time AR formed, were compared to expression profiles of untreated samples in which no AR were formed and to TANGLED, –42, incrnd MDP25  decreasedoes not . In addidoes not . Differein vitro . The flein vitro . In addiin vitro , 48. In in vitro –51, accuin vitro . clasp pal roots , 50 and al roots , suggestMOR1 promoter has been determined in cells of AR primordium. It was found that the promoter of MOR1 is induced in the primary root tips and LR, in the pericycle of the hypocotyls and in AR primordium following IBA treatment of 16-72h. This suggests that the expression of MOR1 is a target of transcription factors that in turn are direct or indirect targets of auxin. Curiously, GFP under the MOR1 promoter accumulated, albeit to lower levels, in the pericycle of hypocotyls after 2-3d of incubation in liquid MS even when no IBA treatment was applied and no AR primordium were formed Click here for additional data file.S1 Table(DOCX)Click here for additional data file.


Allergic rhinitis (AR) is poorly controlled. Treated patients, even those on multiple therapies still experience symptoms. In a UK survey, 70.5% of moderate-to-severe AR patients took ≥2 medications (either Rx or over the counter) in an attempt to achieve better and faster symptom relief.To explore the extent of co-prescribing by UK general practitioners (GPs) during the hay-fever season in patients with seasonal AR (SAR), perennial AR (PAR), and comorbid asthma.A retrospective database study using the Optimum Patient Care Research Database consisting of data extracted from GP records supplemented with patient-reported outcomes from questionnaires. Patients included in the analysis had a recorded AR diagnosis and ≥1 AR therapy scripts during 1st March 2010 to 31st August 2010.In all, 22,381 AR patients were included. Results are summarized in the table.In contrast to previous surveys, these data relate to prescriptions only and show a high level of co-prescribing behavior among UK GPs. There was a significant shift to combination therapy during the season, particularly for PAR patients, with strong co-prescription evident regardless of asthma co-morbidity. These data indicate that (i) UK GPs are aware that current therapy provides insufficient symptom relief, (ii) that AR is a costly disease to treat requiring several GP visits over the season for therapy modification and (iii) there is a need for an AR therapy which provides more complete symptom relief.


One mechanism of resistance of prostate cancer (PCa) to enzalutamide (MDV3100) treatment is the increased expression of AR variants lacking the ligand binding-domain, the best characterized of which is AR-V7. We have previously reported that Phosphatidylinositol-4-phosphate 5-kinase alpha (PIP5Kα), is a lipid kinase that links to CDK1 and AR pathways. The discovery of PIP5Kα inhibitor highlight the potential of PIP5K1α as a drug target in PCa. In this study, we show that AR-V7 expression positively correlates with PIP5K1α in tumor specimens from PCa patients. Overexpression of AR-V7 increases PIP5K1α, promotes rapid growth of PCa in xenograft mice, whereas inhibition of PIP5K1α by its inhibitor ISA-2011B suppresses the growth and invasiveness of xenograft tumors overexpressing AR-V7. PIP5K1α is a key co-factor for both AR-V7 and AR, which are present as protein-protein complexes predominantly in the nucleus of PCa cells. In addition, PIP5K1α and CDK1 influence AR-V7 expression also through AKT-associated mechanism dependent on PTEN-status. ISA-2011B disrupts protein stabilization of AR-V7 which is dependent on PIP5K1α, leading to suppression of invasive growth of AR-V7-high tumors in xenograft mice. Our study suggests that combination of enzalutamide and PIP5K1α may have a significant impact on refining therapeutic strategies to circumvent resistance to antiandrogen therapies. Although considerable progress is being made to improve therapy of PCa, one-third of treated PCa patients will experience disease recurrence and will progress into castration-resistant PCa (CRPC), which are no longer responsive to anti-androgen therapies –4. One mEnzalutamide inhibits recruitment of AR coactivators and prevents AR nuclear localization . AlthougPhosphatidylinositol 4-phosphate 5-kinase alpha (PIP5K1α) and its phospholipid product PIP2 have emerged as a new class of predictive biomarkers and drug targets in prostate cancer . PIP5K1αCDK1 is the most essential kinase in CDK family, and is sufficient to drive cell division cycle in mammalian cells . CDK1 isIn the present study, we have studied the role of AR-V7 and its functional association with PIP5K1α and CDK1 in PCa progression using PCa cell lines and xenograft mouse models. We have further investigated whether the PIP5K1α inhibitor, ISA-2011B might suppress the growth of AR-V7-overexpressing tumors and selectively block the deregulated AR-V7 pathways in xenograft mouse models. Our study identifies novel cooperative mechanisms involving PIP5K1α, AR-V7, CDK1 and AR, which drive tumor progression and contribute to enzalutamide resistance. Our findings provide new information to guide the development of targeted therapeutic combinations for treatment of invasive castration-resistant PCa.p=0.045, Figure p=0.014; Figure p<0.001, Figure p=0.056, Figure p=0.011, Figure p= 0.023, Figure To evaluate clinical importance of AR-V7 and its correlation with PIP5K1α expression in PCa patients, the first sets of tissue microarrays (TMAs) consisting of begin prostate hyperplasia (BPH) (n=48), primary PCa (n=65), and PCa metastatic tissues (n=43) were immuno-stained with antibodies against AR-V7 and PIP5K1α. A second TMA that contained biopsies of BPH and cancer tissues from 48 PCa patients was also examined for AR-V7 and PIP5K1α expression. AR-V7 expression was predominantly nuclear, whereas PIP5K1α expression was present in both cytoplasm and nucleus of cancer cells Figure . PIP5K1αp<0.001; for PIP5K1α, p=0.006, Figure p=0.002, Figure We further characterized the biological consequences of AR-V7 overexpression in PCa cells and the underlying molecular mechanisms. We introduced overexpression of AR-V7 by transfecting non-malignant PNT1A cells, which do not express endogenous AR-V7, with pEGFP-AR-V7 or pEGFP control vectors. Expression of AR-V7 led to an increase in PIP5K1α expression in PNT1A cells, as measured by densitometric quantification of the blots from three independent experiments using Image J , and showed enhanced nuclear PIP5K1α expression  with ISA-2011B or vehicle control. After two weeks of treatment, the mean tumor volumes in mice treated with ISA-2011B was 3.5-fold smaller compared with those treated with vehicle control  and control cells (p=0.001), and showed more pronounced inhibitory effect on cells overexpressing PIP5K1α  and control cells  and in controls  from 48 patients were constructed at Department of Clinical Pathology and Cytology, Skåne University Hospital, Malmö. The second TMA set that contained BPH and PCa tissues from 48 patients was purchased from Pantomics Inc. . The mRNA expression data of PIP5K1A and PTEN, and AR-V7 status was extracted from the dataset in the cBioPortal database, n= 333 tumors from 425 available cases in a prostate cancer patient cohort [https://tcga-data.nci.nih.gov/docs/publications/prad_2015/. IlluminaGA RNASeq and IlluminaHiseq RNASeq were analyzed [Tissue microarrays (TMAs) containing primary PCa (n=17) and metastatic PCa lesions in distant organs such as lymph node, lung, liver, and bone (n=43) from 14 PCa patients, and benign prostatic hyperplasia (BPH) (n=48) t cohort . The detanalyzed . The stuvs. high grade.Immunohistochemistry on tumor tissue microarrays was performed as previously described . The staThe PNT1A cell line was purchased from Sigma Aldrich (Stockholm). LNCaP, 22Rv1and PC-3 were purchased from American Type Culture Collection  and were maintained in phenol red-containing RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS), 1% penicillin-streptomycin-neomycin and 2 mM L-Glutamine . For treatment, cells were grown in phenol red-free RPMI-1640 medium containing 10% charcoal stripped-serum for 24 hours and then were treated with drugs alone or in combination for 24 hours or 48 hours. Enzalutamide at 5 μM or ISA-2011B at 20 μM or 50 μM final concentrations or solvent DMSO 1% was used. For treatment of 22Rv1 cells with MG132, a proteasome inhibitor, cells were treated with MG132 at 1 μM. For combination treatment of MG132 and ISA-2011B, cells were pre-treated with MG132 for 30 min at 1 μM prior to treatment of ISA-2011B.CDK1 or siRNA negative control duplex were purchased . SiRNAs (50nM) were transfected into 1 × 105 PCa cells using Transfection Reagent TransIT-TKO® according to manufacturers’ protocol (Mirus Bio LCC). After introduction of respective siRNA complexes into PCa cells, cells were then collected after 24, 48 and 72 hours post-transfection.pLPS3-EGFP vector containing full-length human PIP5K1α cDNA or control empty vector were used as previously described . pEGFP v6 cells/mouse in 100 μl sterile vehicle solution were implanted subcutaneously into the flank. Tumor diameters were measured using calipers, and volumes were calculated using the equation (a x b2/2), where a and b represent the larger and smaller diameters, respectively. Tumors were grown to mean volume of 100-150 mm3. Xenograft mice with tumors were randomized (n=6 mice per group) and treatment of xenograft mice with vehicle control, ISA-2011B (40 mg/Kg) was initiated, and treatment for every second day was lasted for 15 days. The weights of mice were measured regularly and survival followed. Tumor samples were collected post-mortem and used for immunoblotting and immuno-histochemical analyses.All animal studies were approved by the Swedish Regional Ethical Animal Welfare Committee. Athymic NMRI nude male mice (n=6 per experiment group) aged 8–12 weeks and weighing 25-27 grams each  were used in each experimental setting. 2×103/well were cultured in 96-well plates for 48 hours. MTS incorporation into the DNA was determined by measuring the absorbance on Infinite® M200 multimode microplate reader (Tecan Sunrise™).Proliferation of transfected PNT1A cells overexpressing AR-V7 or control vectors were determined using MTS proliferation assays (Promega Biotech) according to manufacturer's protocol. Cells at 5 × 10The cells or tumor tissues were harvested and lysed in ice-cold RIPA buffer. Protein (20-30 μg) were separated with 12% SDS-PAGE gels and transferred onto nitrocellulose membranes. Signals were visualized using the Enhanced ChemiLuminescence detection system  and documented with an AlphaImager CCD system. Densitometric quantification of immunoblots was performed by the ImageJ Image Analysis Software  and represented as fold change relative to control and was normalized relative to actin or GAPDH bands. The following antibodies were used in this study: monoclonal antibodies against AR-V7 (produced in JL's lab) and AR-V7 , PIP5K1α , Phosphor-473 AKT , p27 , androgen receptor AR (N20) and P27, Cyclin A2 and anti-GAPDH (Santa Cruz Biotechnology Inc. CA), PSA and Ki-67 , MMP-9  and anti β-Actin  cyclin E (Upstate Inc.) and Cdk1 (BD Transduction Lab Inc.) were used. Secondary antibodies: HRP-conjugated anti-mouse IgG and anti-rabbit IgG .2, 10 mM KCl, 0.5 mM DTT, 1 % Triton X-100, 15 % protease inhibitor cocktail Complete Mini, 1 mM PMSF). After the separation steps, the nuclear and cytoplasmic fractions were obtained and subjected to immunoblot analysis. Antibodies against β-tubulin and Lamin B were used to validate the purity of the cytoplasmic and nuclear fractions, respectively. For immunoprecipitation (IP) assay, antibody against PIP5K1α was used to pull down the immunocomplexes, and antibody to IgG  was used as a negative control. 500 μg of freshly prepared protein lysates were incubated with the antibody of interests and 30 μl of G-sepharose beads  for 3 hours at 4°C. The samples were then washed in RIPA buffer and subjected to immunoblot analysis.Subcellular fractionation was prepared as previously described . For obtPCa cells were grown on the glass coverslips in phenol red-free RPMI-1640 medium containing 10% charcoal stripped-serum for 24 hours and were then treated with the indicated drugs for 24 hours. Cells were fixed with 4% paraformaldehyde in PBS. For blocking background staining from nonspecific interactions, Image-iT™ FX signal enhancer  was used. Primary antibodies against Phosphor-473 AKT, AR and AR-V7 were used. The secondary antibodies including rabbit anti-donkey conjugated to Rhodamine  or anti-goat conjugated to FITC antibodies at 1:200 and goat anti-rabbit Alexa Fluor 488 at 1:500  were used. 4′,6-Diamidino-2-phenylindole counterstain  was used to visualize cell nuclei. The slides were detected under an Olympus AX70 fluorescent microscope . The software ACT2U was used .t-test and Spearman rank correlation tests were performed. All outcome variables are representative of at least three independent experiments. All statistical testes were two-sided, and p values less than 0.05 were considered to be statistical significant. Statistical software, Social Sciences software , was used.Tukey-test, ANOVA, Mann-Whitney test, student


Partial androgen insensitivity syndrome (PAIS) typically presents with micropenis, perineoscrotal hypospadias, and a bifid scrotum with descending or undescending testes and gynecomastia at puberty. It is an X-linked recessive disorder resulting from mutations in the androgen receptor (AR) gene. However, AR gene mutations are found in less than a third of PAIS cases. A 16-year-old boy was admitted with complaints of gynecomastia and sparse facial hair. Family history revealed male relatives from maternal side with similar clinical phenotype. His external genitalia were phenotypically male with pubic hair Tanner stage IV, penoscrotal hypospadias, and a bifid scrotum with bilateral atrophic testes. He had elevated gonadotropins with a normal testosterone level. Chromosome analysis revealed a 46,XY karyotype. Due to the family history suggesting a disorder of X-linked trait, PAIS was considered and molecular analysis of AR gene was performed. DNA sequence analysis revealed a novel hemizygous mutation p.T576I (c.1727C>T) in the AR gene. The diagnosis of PAIS is based upon clinical phenotype and laboratory findings and can be confirmed by detection of a defect in the AR gene. An accurate approach including a detailed family history suggesting an X-linked trait is an important clue for a quick diagnosis. Partial androgen insensitivity syndrome typically presents with micropenis, perineoscrotal hypospadias, and a bifid scrotum with descending or undescending testes and gynecomastia at puberty. It is an X-linked recessive disorder resulting from mutations in androgen receptor (AR) gene. In approximately 50% of cases, a mutation in AR gene cannot be detected.DNA sequence analysis revealed a novel hemizygous mutation p.T576I (c.1727C>T) in the AR gene.Androgen insensitivity syndrome (AIS) is the commonest cause of 46,XY disorders of sex development (DSD) and is characterized with defective masculinization of external genitalia in 46,XY individuals despite normal androgen production and metabolism . The estPatients with CAIS usually present with normal female external genitalia and bilateral inguinal hernias. Individuals who are not diagnosed during childhood are detected after puberty because of primary amenorrhea with a blind vagina and absent uterus. On the other hand, depending on the degree of responsiveness of the external genitalia to androgens, findings in PAIS will show a wide spectrum varying from perineoscrotal hypospadias, bifid scrotum, cryptorchidism and pubertal gynecomastia to extreme inadequate virilization appearing as cliteromegaly and labial fusion .This report describes an adolescent boy with PAIS who presented with gynecomastia at puberty and was found to have a novel mutation in the androgen receptor (AR) gene.A 16-year-old boy was admitted to our clinic with complaints of gynecomastia and sparse facial hair. He was born at term with a birth weight of 3000 grams. A detailed history revealed operations for bilateral cryptorchidism and penoscrotal hypospadias at ages one and two years, respectively. There was no parental consanguinity. Family history revealed male relatives from the maternal side with similar clinical phenotype, including gynecomastia, hypospadias, sparse facial hair, and infertility  following the manufacturer’s instructions. The polymerase chain reaction fragments were sequenced by Illumina MiSeq system using V2 chemistry . Sequencing results were analyzed using IGV software (http://www.broadinstitute.org/igv/).A hemizygous mutation p.T576I (c.1727C>T), which had not previously been reported, was identified in the AR gene . AnalysiThe AIS describes a spectrum of disorders where the degree of receptor insensitivity may vary from minimal to complete insensitivity. In case of MAIS, the individual is phenotypically male with sterility, azoospermia, and gynecomastia without genital abnormalities. The other end of the spectrum comprises XY individuals with CAIS who phenotypically appear as tall females with well-developed breasts, blind vagina, and absent or scanty pubic and axillary hair . The typIn the differential diagnosis of PAIS, 5α-reductase deficiency, partial gonadal dysgenesis , and testosterone biosynthesis defects need to be considered. If the karyotype is 46,XY, then serum testosterone and dihydrotestosterone levels are helpful for the differential diagnosis. Normal or elevated levels of testosterone and dihydrotestosterone are essential to exclude androgen biosynthesis defects. Nevertheless, hCG stimulation test may be necessary to differantiate partial gonadal dysgenesis in prepubertal children . Serum aIn cases with AIS, high levels of testosterone, a substrate for aromatase activity, result in substantial amounts of estrogens, which are responsible for breast development at puberty. However, due to functional AR activity, gynecomastia has not been reported in cases with 5α-reductase type 2 deficiency and partial gonadal dysgenesis . In the The hormonal profile is similar in individuals with CAIS and PAIS. At birth, levels of testosterone and LH remain high or slightly above the normal range for males. During puberty, individuals with PAIS maintain normal or slightly elevated testosterone and LH levels . EstradiAIS is an X-linked recessive disorder resulting from mutations in the AR gene . While tGender decision, genitoplasty, timing of gonadectomy (due to cancer risk), hormone replacement therapy, genetic counseling, and psychological support comprise the basis of management in PAIS. The majority of the patients with PAIS are reared as males . When thPatients with PAIS are under the risk of malignancy which mostly are gonadoblastoma or dysgerminoma. Gonadal tumor risk is 0.8-2% during the prepubertal period and rises up to 30% during late adulthood. Risk of malignancy is low before the age of 25 years and more frequent between 30 and 50 years . The risIn conclusion, PAIS constitutes one of the most common causes of 46,XY DSD. The diagnosis is based upon clinical phenotype and laboratory findings, and can be confirmed by detection of a defect in the AR gene. An accurate approach including a detailed family history suggesting an X-linked trait is an important clue for a quick diagnosis.Informed Consent: It was taken.Peer-review: External peer-reviewed.


Burn injury may be associated with long-term rehabilitation and disability, while research studies on the functional performance after injuries, quality of life (QOL), and abilities to return to work of burn patients are limited. These outcomes are related not just to the degree and nature of injuries, but also to the socio-economical background of the society. This study aimed to identify the factors which might affect burn patients’ abilities to reintegrate back to the society based on a sample in mainland China.A retrospective study was conducted to collect data of demographic characteristics, medical data about burn injuries, physical and psychological status, and self-perceived QOL at the initial phase and upon discharge from a rehabilitation hospital, timing of rehabilitation, and duration of rehabilitation intervention. Four hundred fifteen patients with burn injuries were recruited in the study. Multiple linear regression and logistic regression were used to obtain a model to predict the functional abilities and the perceived QOL at discharge and their changes during rehabilitation, as well as the post-injury work status within 6 months after discharge.The functional performance at discharge and its change were significantly predicted by the functional abilities and QOL at the admission, duration of treatment, timing of rehabilitation, payer source, and total body surface area burned. The perceived QOL at discharge and its change were significantly predicted by the baseline QOL at admission and duration of treatment. The significant predictors of work status within 6 months post-discharge included age, education, payer source, total body surface area burned, perceived QOL, and bodily pain at admission.The present study identified a number of factors affecting the rehabilitation outcomes of people with burn injuries. Identification of these predictors may help clinicians assess the rehabilitation potential of burn survivors and assist in resource allocation. Policy makers should ensure that resources are adequate to improve the outcomes based on these factors. Burn injury is a devastating form of injury and also a major global public health issue. There are about 900 inpatients per burn center annually in 39 burn centers located in 17 provinces in mainland China. Nevertheless, there are only limited epidemiological data on burn injuries in mainland China, and it has been reported that the incidence rate of burn injuries peaks in individuals at the working age . TherefoThere are a few studies exploring the predictors of post-burn functional independence, QOL and work status, and gains in these outcomes from burn rehabilitation –5. HowevThe present study employed a longitudinal retrospective design. The data were obtained by the Department of Burn Rehabilitation of the Guangdong Provincial Work Injury Rehabilitation Hospital from January 2009 to December 2014. All patients were referred by the burn surgeons in the acute hospital in the region. Patients admitted to the rehabilitation hospital would receive a multidisciplinary rehabilitation program including passive and active exercises, splinting and positioning, management of edema and pain, training of activities of daily living (ADL), education of patients and families, and psychosocial support. The program was implemented daily for 6 h excluding Sunday. Subject recruited in this study should have a diagnosis of burn injuries, first admission to the hospital, and between 18 and 60 years old. Those patients with multiple admissions to the hospital or had received other therapy prior to admission were excluded from the study. Those with severe complications unrelated to their burn injuries which impaired functional independence, QOL, or work capacity, such as traumatic brain injuries, spinal cord injuries, serious fractures, amputations, or severe infection would be excluded as well. The study was approved by the Human Research Ethics Committee of the Guangdong Provincial Work Injury Rehabilitation Hospital (no. AF/SC-07/2013.01) and The Hong Kong Polytechnic University (no. HSEARS20150708003).The evaluation was measured at admission to the rehabilitation hospital and upon discharge, and at 6 months after discharge (follow-up).The Chinese version of the Modified Barthel Index (MBI) comprises 10 scored activities including personal hygiene, bathing, dressing, feeding, bowel control, bladder control, toilet transfers, stair climbing, wheelchair/chair-bed transfer, and ambulation . Each acThe World Health Organization Quality of Life (WHOQOL)-BREF scale is a generic measure of QOL, including four domains  . The WHOThe demographic characteristics, medical data about burn injuries, physical and psychological status which were recorded at admission, the time from injury till rehabilitation, and the duration of the rehabilitation (days) were also recorded. The demographic characteristics included age, gender, marital status, level of education, type of payer source for the medical care service , self-perceived economic status, and family and company support were measured using a questionnaire, with the response graded as good, fair, and bad. The medical conditions, including the causes of injuries, burnt area, depth, and regions of burns were also documented. The physical status at admission included MBI , pain and itch ), the number of joints with limited range of motion, and the Pittsburgh Sleep Quality Index (PSQI) were measured. The VAS is a tool used to describe a subjective quality such as pain  and itchThe psychological status was measured by the self-rating depression and anxiety scales (SDS and SAS) . The SDSt tests and chi-square tests were used to explore the changes on MBI, WHOQOL-BREF, and RTW in the subjects. The strength of association between the predicting variables and the outcome measures at discharge, as well as the change in outcome measures from admission, were determined by the Spearman’s correlation coefficient. The predictor variables which showed significant associations with the outcome measures were entered into multiple linear regression (for MBI and WHOQOL-BREF) and logistic regression analyses (for RTW) to identify the key predictors for the rehabilitation outcomes. For RTW, the total body surface area (TBSA) burned, which has been identified as a significant predictor in previous studies , indicates that people with better baseline functional independence and QOL for psychological health and smaller TBSA burned would gain higher functional independence at discharge. For the WHOQOL-BREF scores in physical health and environment at discharge, the significant predictor variables included WHOQOL-BREF in the respective domains at admission, duration of treatment, and timing of intervention. The prediction model indicates that earlier and longer period of rehabilitation and higher baseline QOL in physical health and environment may result in higher WHOQOL-BREF scores in those domains at discharge. For WHOQOL-BREF scores in psychological health and social relationships at discharge, each had an additional predictor: WHOQOL-BREF score in environment for the former and WHOQOL-BREF score in psychological health for the latter, in addition to scores at admission, duration of treatment, and timing of rehabilitation.Table r = −0.418 to 0.688) and longer rehabilitation period (r = 0.026 to 0.050) predicted greater changes. In addition, the payer source with higher percentage of expense reimbursed by insurance, higher QOL in psychological health, smaller area of burn, and better family support predicted better improvements in MBI. The change in QOL in physical health can be further predicted by the TBSA burned. The change in QOL in social relationships and environment were both further predicted by QOL in psychological health at admission.For the changes in MBI and WHOQOL-BREF during the treatment period, lower baseline values  –5. If suThe TBSA burned was found to be a negative predictor of functional independence at discharge and its gain during rehabilitation in this study. This finding is inconsistent with that in the previous studies , 5. The In the current study, the likelihood of returning to work within 6 months after discharge was 61 %, which is comparable to other studies conducted in developed countries , 19. In Despite meaningful findings on prediction of rehabilitation outcomes in a population with burn injuries in China, the present study has some limitations. First, missing data could introduce possible bias because different observations might have been resulted if there were no missing data. The payer source and short duration of treatment could be the causes of missing data, which showed a difference in results with that in the cases included. Second, although the facility where all data were collected is the rehabilitation center for burn injury in mainland China, and the categories of cases could be representative to some extent, the reader should still be cautious about the generalizability of the study outcomes to the whole population in mainland China with burn injury. Third, the reliability and validity of the measurements of rehabilitation outcomes used in people with different disabilities have been reported by a number of previous studies , 12; howIn addition to the above limitations, we would like the reader to be cautious of the following when interpreting the results of the present study. First, this study is an assessment of the performance of a single rehabilitation facility in China which admits and treats a casemix with a majority of workplace injury survivors. Second, the collection and analysis of specific time-related data was not planned a priori, which might influence the interpretation of QOL and RTW outcomes as these are related to discharge from the rehabilitation facility, not the date of injury. Furthermore, the influence of payer source (workplace injury) might introduce significant bias and must be considered in the generalization of results. Finally, work-related injury patients receive more in-patient care and rehabilitation staff input and have greater duration of access to in-patient rehabilitation services than other patients included in this study. These might all contribute an impact on the results of the present study.To conclude, higher baseline functional independence and QOL, payer source with higher percentage of expense reimbursed, better family support, earlier and longer duration of rehabilitation, younger age, higher educational level, lower bodily pain, and smaller area of burn predicted better rehabilitation outcome in burn injury patients in mainland China. Identification of predictors may help assess the rehabilitation potential of burn survivors and assist resource allocation. Among these variables, higher baseline self-care performance and QOL, as well as earlier and longer duration of rehabilitation are modifiable factors, and we suggest policy makers to allocate sufficient medical resources to ensure them.FIM, Functional Independence Measure; MBI, Modified Barthel Index; PSQI, Pittsburgh Sleep Quality Index; QOL, quality of life; RTW, return to work; SAS, self-rating anxiety scale; SDS, self-rating depression scale; TBSA, total body surface area; VAS, visual analogue scale; WHO, World Health Organization


Functional magnetic resonance imaging (fMRI), being an indirect measure of brain activity, is mathematically defined as a convolution of the unmeasured latent neural signal and the hemodynamic response function (HRF). The HRF is known to vary across the brain and across individuals, and it is modulated by neural as well as non-neural factors. Three parameters characterize the shape of the HRF, which is obtained by performing deconvolution on resting-state fMRI data: response height, time-to-peak and full-width at half-max. The data provided here, obtained from 47 healthy adults, contains these three HRF parameters at every voxel in the brain, as well as HRF parameters from the default-mode network (DMN). In addition, we have provided functional connectivity (FC) data from the same DMN regions, obtained for two cases: data with deconvolution (HRF variability minimized) and data with no deconvolution (HRF variability corrupted). This would enable researchers to compare regional changes in HRF with corresponding FC differences, to assess the impact of HRF variability on FC. Importantly, the data was obtained in a 7T MRI scanner. While most fMRI studies are conducted at lower field strengths, like 3T, ours is the first study to report HRF data obtained at 7T. FMRI data at ultra-high fields contains larger contributions from small vessels, consequently HRF variability is lower for small vessels at higher field strengths. This implies that findings made from this data would be more conservative than from data acquired at lower fields, such as 3T. Results obtained with this data and further interpretations are available in our recent research study  [1]. This is a valuable dataset for studying HRF variability in conjunction with FC, and for developing the HRF profile in healthy individuals, which would have direct implications for fMRI data analysis, especially resting-state connectivity modeling. This is the first public HRF data at 7T. Specifications TableValue of the data•This dataset provides a characterization of the variability of hemodynamic response function (HRF) across the brain, and across individuals, which is a confounding negative factor in functional magnetic resonance imaging (fMRI) data analysis •This dataset, which also includes comparable functional connectivity (FC) data, is valuable for studying the impact of HRF variability on varieties of fMRI data analyses, including, but not limited to, resting-state FC modeling.•This dataset characterizes voxel-level HRF variability, hence it could be utilized to develop a generalized whole-brain voxel-level HRF template, with applications in fMRI data analysis.•This is the first study to present HRF data obtained in a 7T MRI scanner. With less noisy HRF estimates, findings from this dataset would be more conservative than that acquired at lower fields, such as 3T.1The dataset presented here contains three parameters that characterize the shape of the HRF 22.13, acquisition matrix=96×96, number of slices=45, and number of fMRI volumes=660 (11 min), with whole-brain coverage. A 32-channel head coil was used. All participants provided informed consent; all procedures were approved by the Auburn University Institutional Review Board (IRB).Forty-seven healthy adults participated in the study. Resting-state fMRI data was obtained in a 7T MAGNETOM scanner  using T2* weighted multiband echo-planar imaging (EPI) sequence 2.2The following standard pre-processing steps were performed on the resting-state fMRI data: slice-timing correction (since the data was acquired using a multiband sequence), realignment and unwrapping, coregistering to the anatomical image, de-spiking, normalization to the MNI space, spatial smoothing (8 mm Gaussian kernel), and regressing out nuisance covariates  signal and cerebrospinal fluid (CSF) signal). Finally, temporal band-pass filtering was performed (0.008–0.1 Hz). Pre-processing was carried out in the Matlab© R2013a platform using Statistical Parametric Mapping (SPM12) 2.3The voxel-wise 3D+time fMRI data was utilized to perform temporal hemodynamic deconvolution. Latent neural time series and corresponding HRF parameters were obtained at every voxel through this process. For deconvolution, we used a popular technique developed by Wu et al. Deconvolution provided the estimated HRF at every voxel of the brain, in every participant. It was characterized by three HRF parameters, as noted earlier – response height (RH), time-to-peak (TTP), and full-width at half-max (FWHM)  between corresponding regions. The DMN regions-of-interest (ROIs) were obtained as 10-mm diameter spheres around the DMN centroids as defined in Power et al. Our main findings associated with this dataset, along with further interpretations, are part of our recent research study


This paper proposes an architecture that explores a gap in the spectrum of existing strategies for robot control mode switching in adjustable autonomy. In situations where the environment is reasonably known and/or predictable, pre-planning these control changes could relieve robot operators of the additional task of deciding when and how to switch. Such a strategy provides a clear division of labour between the automation and the human operator(s) before the job even begins, allowing for individual responsibilities to be known ahead of time, limiting confusion and allowing rest breaks to be planned. Assigned Responsibility is a new form of adjustable autonomy-based teleoperation that allows the selective inclusion of automated control elements at key stages of a robot operation plan’s execution. Progression through these stages is controlled by automatic goal accomplishment tracking. An implementation is evaluated through engineering tests and a usability study, demonstrating the viability of this approach and offering insight into its potential applications. The control of robots is a non-trivial task requiring complex perceptions, decisions and actions to be made using limited information. While many teleoperated systems still only employ human operators, the inclusion of automation is now much more common. Automated control rarely completely replaces human operators, because contemporary technology is not yet adequate, but automation can support their work. Mixing of control shows much promise, relieving operators of workload, and allowing the use of teleoperated robots in environments where they could not be previously controlled , is a type of adjustable autonomy that we initially proposed in Small et al.  to allowThe remainder of the paper is structured as follows. Section In some domains, real robot operations can be well-planned in advance. Whether to direct and regulate human workers, autonomous robots or humans using robots, operations plans are currently used in much existing work. In relatively predictable environments such as warehouses  has long been understood to be a source of trouble in HRI  and how changes are triggered . The responsibility for change is a matter of design. Adjustable autonomy systems  while improving operator experience  by designing better hardware and software for the whole system. A particular focus of this work is a special management of automatic LoA switching, reduction of operator confusion and fatigue, and support for the gradual automation of the execution of the full task over time. The concept is based on five observations, or matters of emphasis, which form the core principles of the design:The other dimension of interest here is whether these changes are planned (e.g. when doing a given task the system is set to a particular mode), or dynamic . The distinction is not always clear cut, because a system with planned changes still needs to be able to dynamically respond to errors, but the normal operating procedure for the system is likely to rely on one of these change strategies more than the other. Figure None of these systems use planned, automatic changes in LoA. Miller and Parasuraman , p. 63 dAR can be understood as a scaffolding for a pre-planned adjustable autonomy teleoperation interface. It allows the LoA of a particular robot to change systematically during the execution of an overall plan. This is new because, unlike other adjustable autonomy approaches, these changes occur automatically at specific plan execution points. Figure By having level changes pre-planned and executed automatically, AR necessarily sacrifices some flexibility in the face of changing conditions. This exchange is motivated by four considerations: (1) A predictable schedule of responsibility allows human operators to know ahead of time when they are and when they are not needed, allowing them to plan breaks. (2) The pre-set levels of autonomy should reduce conflicts between operators, as their role at each stage is well-known to them ahead of time. (3) Pre-determined changes provide boundaries for automation to operate in, reducing programming complexity, and facilitating the gradual automation of the whole task. (4) In scenarios where the use of automation is restricted due to laws, ethical considerations, or simple mistrust in the capabilities of that automation, the ability to specify what is and what isn’t automated becomes desirable.The flexibility issues discussed in the previous paragraph are most apparent if the plan is treated as a static object once the task is underway. Automated planning techniques , a plan is represented as a graph tree in which the nodes are the goals , the edges between them are actions and goals are defined as desirable states of the world  of a tree is a vertex of degree 1.We can interpret these theoretical concepts as follows:The root is the overall goal to achieve, each other vertex is a sub-goal.To achieve a particular sub-goal, its children have to be satisfied, in order .By extension, once all sub-goals are satisfied, the root goal is also satisfied.The leaves are the lowest-level goals, and are the only goals satisfiable directly. Satisfying all of the leaf goals, in order, satisfies the root goal.The resulting plan graph possesses the properties of an ordered rooted tree, the details of which are described below, and illustrated in Fig. To allow automated changes in levels of autonomy to happen as planned, an AR system must be able to track progress through the execution of a task to completion.Current research interest in the monitoring of the execution of plans is focused on supporting the improvement of this execution, and the improvement of the monitoring itself  which may be similarly included in the plan.Not all goals are the same, but may have different natures based on their objective and relation to processing. The proposed architecture supports three types of goals previously described in literature:The plan management module is charged with ensuring that all of the components of the AR system are aware of the current state of the task’s achievement. This state record is used to synchronise all of these components, ensuring the LoA of the system is set as planned, the human operator is aware of the current goal, and that the accomplishment monitoring process is checking for the accomplishment of the correct goal. The plan manager relies on the accomplishment monitoring process for updates to the task progress, and is charged with integrating and disseminating those updates to trigger appropriate changes to the human–machine system.By applying the general theories put forward by .The final assignment is set, ensuring both the human operator and the automated control system are aware of the goals for which they are responsible.In AR each of the plan’s sub-goals are explicitly assigned one of these levels before the execution of the plan. In this example assignment strategy, it is assumed that the human operator is capable of effectively controlling the robot and satisfying all of the goals using it, and that the preference is to relinquish most of the control to the automated modes in order to free up the human operator. These assumptions are for explanatory purposes only and are not, of course, taken for granted. An example assignment process is described below. This process would not typically be done by the operator, rather by a manager or robot specialist, in which case making the operator aware of the contents of the plan is a critical step.Automatic maintenance of physical equipment requires a mobile robot to periodically visit a number of worksites. The robot may perform tasks such as photography, gathering sensor data on environmental conditions, physically probe the integrity of surfaces, joints or attachments, remove panels and/or to change out faulty components Mann . To perfThis section describes a modular design for an AR-based control system for a small mobile robot, allowing that robot to perform the task described above under human and automated supervision.In the maintenance scenario described above—and in most robotics applications—a robotic control system must operate over a network; at the minimum the robot itself and a control station. A modular or component-based approach is ideal when designing software that will be distributed over multiple hardware platforms connected over a network. In a modular design, individual components should be interchangeable with other components capable of performing similar tasks. This is particularly important in robotics, where a system might want to control another robot, or use a different planning algorithm, or in the case of goal accomplishment tracking, load up another goal test.For an AR system, the most important reason to choose a modular approach is the drive to progressively automate the execution of tasks. As automation for more tasks is written, it will be added to the overall system. The easiest way to allow this to happen is to make the automated controller modular: when new automation techniques become available, modules implementing them can be created. Adding new automation capabilities to the system then simply becomes a case of making these new modules available to be loaded up during runtime. Performance statistics of each module can be accumulated once deployed. As mentioned in Section II, one of the advantages of having humans controlling the robot during non-automated tasks is that data can be collected on the human accomplishment of these tasks, possibly paving the way for future automation.This design consists of components divided into three groups: operation, execution, and management .Execution The execution group is mostly concerned with executing commands sent by the operation group, and with handling sensors mounted on the robot. It includes the main control loop for the robot, as well as an interface to handle requests for information gathered by the robot (mostly sensor data).Management The management group is in charge of two major tasks: plan management, and progress tracking. The plan management part is tasked with managing the file storing the plan, making the plan and its goals available to modules that request it, and updating the plan with new information . The progress tracker monitors the plan’s execution in the real world using sensor data to keep the plan up to date.Example task-execution timeline A task’s accomplishment can be broken down into two stages: planning and execution, as shown in Fig. To evaluate AR experimentally requires human operators to actively use it in a reasonably realistic task. To do this, a prototype implementation of the AR system has been implemented in hardware and software. Because AR is primarily a matter of software, the hardware is mostly off-the-shelf parts, modified to support the requirements of the software. The experiment uses a Coroware mobile field robot, built on top of a Lynxmotion base with four driven wheels, with an Intel D2700MUD Mini-ITX single-board computer running Linux. In contains two cameras, a modified 5 DoF manipulator arm and is equipped with a variety of infrared proximity detectors, force and bump sensors. It is linked via 802.11g wireless to a laptop and Logitech game controller that serve as a teleoperation station.The hardware is distributed across two physical components: the control station Fig. a, and thThe software was built from scratch for this implementation. It is divided into three modules as pictured in Fig. The communications between the components of the system are mapped out in Fig. An end-to-end usability experiment was performed on the implemented teleoperation system to evaluate AR design on operator workload, preferences, effectiveness and efficiency. A repeated measures design was chosen. This is commonly used in user evaluations of interfaces for its efficient use of each participant to test all of the interfaces. The drawback of this design is the possibility for participants to perform better in later trials as they gain experience with the system. This confounding effect can be mitigated by ensuring participants do not undertake trials in the same order. It is important that the sample size (n) in the experiment provides for sufficient power in the inferential statistics to be used . This experiment used 20 participants.The task consisted of a mockup maintenance route suggestive of an industrial context. It needed to be complex enough to properly test the human-machine system, yet not so complex as to be too difficult and time consuming for the human operators to learn, and for the automation to accomplish. The robot had to be guided through a sequence of four sectors, each containing a valve that had to be closed using the robot’s arm and gripper. Figure To compare AR to direct, unmediated human control, the experiment included that control mode. The robot had to be operated using four different control options throughout each volunteer’s participation. These control modes covered direct teleoperation (Referred to as control option T) as well as a range of configurations of the AR interface. Control options AR-H and AR-A made use of the progress tracking and plan management sections of the architecture, keeping track of the progress through the task of the robot, as well as keeping the operator informed of that progress. However they had the responsibility assignment for all goals set to H in the case of AR-H and A for AR-A. AR-HA resembles a more typical use of AR as originally imagined, with a set of varying responsibility assignments, as pictured in Fig. During the experiment both qualitative and quantitative data were collected, recording both operator impressions and their performances. Subjective data was collected about the workload placed on the operators in each control option using the NASA Task Load Index (TLX) , or that the LoA of that part of the work be brought back down until the automation is improved. The overall comparison between AR and T shows that collectively, the AR options caused significantly more goal failures, and as stated above this difference is caused by the introduction of automation into the control loop. Altogether, the AR options recorded a 11.67% failure rate.These data show that out of all the recorded runs, the AR options were less effective than the T option in this case. AR can, however, operate very similarly to direct teleoperation if needed (as was done with the AR-H option) and therefore potentially achieve similar effectiveness levels. This does diminish some of the benefits of AR, but not all, as will be shown later. It is important to note that this experiment can only test this particular implementation of automation, which is not claimed to be indicative of the most up-to-date automation techniques.As shown in Fig. The time data showed that T was significantly slower (236.4 s mean time) than AR-H (180.4 s mean) and AR-HA (174 s mean), which were in turn significantly slower than AR-A (104.7 s mean). This highlights important differences in all of the modes. In AR-A, being fully automated, the system never had to wait for operator input, which had two consequences: a faster task accomplishment rate, and a higher goal failure rate. In AR-HA, the amount of time the robot spent moving would have been fairly similar to that of AR-A, however the automation spent time waiting for waypoints to be set by the human operator, which delayed it.One of the more surprising differences lies with AR-H and T. With both those options, all of the control task was performed by the human operator. This is reflected in the similar error rates for both options. However, AR-H had a significantly faster accomplishment rate. This difference comes almost entirely from the goal accomplishment tracking during the AR-H runs. With the T option, the operator had to confirm the closure of the valve visually by testing it repeatedly until satisfied. In the AR-H runs, the goal accomplishment tracking was monitoring the strain placed on the wrist by the valve, allowing for a fast detection of the valve’s state, cutting down the number of grasp attempts needed to close it, and thus saving time.These two sets of data tell a different story, with T outclassing AR as a whole on errors made, but causing a significant increase in time taken for task accomplishment. Overall, the AR system resulted in faster task accomplishment across the board (153 s mean), with the ability to trade a higher error rate for more speed or vice versa (by switching from human control to (semi-automated control) if needed, making it more efficient than the direct teleoperation option.The experiments shown in this paper showed that AR is a viable form of robot teleoperation, capable of accomplishing the tasks set out to operators while imposing less workload on these operators when compared to traditional teleoperation approaches. However, this study can only hint at the situations where AR should be deployed over other adjustable autonomy approaches, and several questions remain to be answered. Are planned changes always easier on the human operators? Are automated changes a viable option when they could catch the human operators unaware? Are manual changes just another task human operators would rather avoid? Under what circumstances should the assignment contract be violated in the interests of safety or efficiency?It became apparent during the trials conducted for this research, that even when planned, automated changes in levels of autonomy could be an issue for human operators. During the trials, one goal was to line up the robot with a valve, a goal which was set to the H/A LoA (human control with assistance from the automation). The automation assisted by highlighting the valve in the video feed to indicate to the operator that the robot was detecting it. The next goal was to close the valve, in full automation. When the valve was detected by the progress tracker, the goal would change, and the automation would take over.This drastic change in level of autonomy, even if planned, caused a lot of frustration to the operators, as the goal might be considered accomplished before they were happy with the placement of the robot. Once the automation took over, the human operator was locked out of the control loop, adding to that frustration if the automation was struggling with closing of the valve due to a poor start position. This proved to be so frustrating that some operators tried to hide the valve from the robot’s camera until they were satisfied with their position. A potential solution to this kind of problem would be to make the operator one of the “sensors” for the goal accomplishment tracking, allowing them to report their satisfaction with the robot’s state when ready. Here is an example of the consideration must be paid to the triggers of automated changes in LoA (planned or not) to avoid inadvertently increasing operator workload.In case of errors, or unforeseen changes in the environment, it is likely that the plan as designed before the task started will no longer produce the desired results. Therefore, a strategy for handling divergences from this plan must be decided on. This problem was not addressed in this paper directly, as it dealt only with the core concepts of AR, but potential solutions can be imagined and discussed here. Plan-altering situations can be considered to fit either of two categories: (1) smaller problems that require a correction before the plan can be resumed, and (2) major problems that will prevent the plan from ever completing. An example of the first would be the loss of a tool required for a future goal in the plan. Before the plan can be completed, the tool must either be found again, or another acquired. Once either has happened, the plan can resume from where it left off. A major issue would be the destruction of a piece of equipment that the plan required the robot to repair. The equipment can now no longer be repaired, rendering the plan un-accomplishable.It is likely that the major failures would result in a cancellation of the plan rather than any kind of in-task re-planning. More interestingly, the smaller problems require on-the-spot inclusion of additional steps (or the replacement of current steps), and thus the modification of the existing plan. Who makes these changes is an important question, as keeping operators aware of the plan is a requirement for AR. A low-tech solution could be to give full control of the robot to the human operator when such a problem arises, with the express goal of returning the situation to an acceptable state. Yet this defeats some of the benefits of AR by imposing unexpected tasks on the human operator, and forcing a single control mode to be used. To make use of the full capabilities of AR, some amount of local re-planning must be done, either automatically or manually. This re-planning needs to add goals, and assign them a LoA. Once the re-planning is done, the modifications must be made available to the operator for approval, ensuring all of the actors in the system are aware of their responsibilities before the task resumes. It is possible that for common problems a library of patches to the plan could be created, which could be rapidly deployed when these problems arise. This could somewhat ameliorate the problem of great additional operator workload, as these patches would likely be familiar to operators that use the system often.While full automation of the robots used today in teleoperated tasks may be desirable, it is in many cases currently infeasible. In the immediate future, a mix of automation and manual control seems the most promising avenue for useful robot control in most scenarios. Direct control teleoperation still reigns, as it is hard to beat the reliability of human operators in most scenarios. If automation is still too untrustworthy to be charged with taking over control, the introduction of automation should primarily focus on assisting the human operators. Let automation take over the small simple tasks that still require time and effort from a human operator, freeing them spend more time and effort on more important tasks such as strategic planning and problem-handling. As time passes and automation techniques become more refined and reliable, more tasks can be automated, gradually reducing demands on the human operator.Assigned Responsibility was designed in part to help this process of gradual automation. The breakdown of the overall task into goals and subgoals sets delimitations that can be used to limit the influence of automation to the simple tasks without jeopardising the greater plan. This also provides context for automation research, breaking down a complex task into a series of simpler tasks, that can be automated one by one instead of as a whole. While that is happening, the robot can keep on working, with the non-automated parts of the plan handled by a human operator. This paper has demonstrated that an Assigned Responsibility system can be used to successfully control a robot through a task using a variety of configurations. It is hoped that this contribution will support further work in the skilful mixing of automation and human-based control of remote robots toward optimal performance.