Strokes cause severe impairment of hand function because of the spasticity in the affected upper extremities. Proper spasticity evaluation is critical to facilitate neural plasticity for rehabilitation after stroke. However, existing methods for measuring spasticity, e.g. Modified Ashworth Scale (MAS), highly depends on clinicians’ experiences, which are subjective and lacks quantitative details. Here, we introduce the first rehabilitation actuator that objectively reflects the condition of post-stroke finger spasticity. The actuator is 3D printed with soft materials. By considering the finger and the actuator together, the spasticity, i.e. stiffness, in finger is obtained from the pressure–angle relationship. The method is validated by simulations using finite element analysis (FEA) and experiments on mannequin fingers. Furthermore, it is examined on four stroke subjects and four healthy subjects. Results show the finger stiffness increases significantly from healthy subjects to stroke subjects, particularly those with high MAS score. For patients with the same MAS score, stiffness variation can be a few times. With this soft actuator, a hand rehabilitation robot that may tell the therapeutic progress during the rehabilitation training is readily available. Stroke has been the leading cause of disability. Every 40 s there will be a new stroke case . Finger Various simple mechatronic devices are designed for standalone finger joint stiffness measurement after stroke . During Recently, robotic hands have been proposed as wearable devices that facilitate movement of each individual finger in both flexion and extension direction. Susanto et al. have introduced the hand exoskeleton robot for active individual finger control via joint moment sensing . TherapeIn this study, we propose the 3D printed soft-elastic composite actuator (SECA) that adopted to our soft robotic hand  for faciStroke causes severe impairment of hand function. Spasticity and motor recovery are both related to neural plasticity after stroke. For rehabilitation planning, the level of finger spasticity that varies over different hand function tasks must be accurately evaluated. Unfortunately, existing spasticity indicators lack reliability, objectivity, consistency, and quantitative stiffness details. Moreover, there exists no hand rehabilitation devices that help objectively evaluate finger spasticity. Here, we introduce the first soft actuator for hand rehabilitation that quantifies spasticity of impaired fingers, i.e. stiffness, in real time. The method is validated with phantom fingers and eight subjects. With the stiffness information, optimal training tasks may be offered by our soft robotic system according to the varying spasticity conditions in fingers. Training outcome may also be improved by indicating the timely therapeutic progress, i.e. the decrease of joint stiffness, to patients for motivating them to achieve better hand function improvement.The 3D printed SECA  presenteMetacarpophalangeal and PIP segments on the SECA correspond to actuation of the MCP and PIP joints of the human finger. When pressurized, the fiber wrapping around the actuator surface suppresses radial expansion, and the torque compensating layer further eliminates any axial elongation at the bottom, leaving only the upper section of the actuator to be elongated for bending the fingers. When depressurized, the torque compensating layer provides an assistive bending moment for the extension of fingers to the initial position . The SEC® – WACKER Chemie AG, to directly 3D print the SECA. In the design of our 3D printed SECA, we select ACEO Silicone GP Shore A 30, an available 3D printed silicone rubber offered by ACEO® – WACKER Chemie AG, as the printing material for the elastomeric actuator body. The 3D printed silicone rubber offers sufficient elongation (450%) and tensile strength (6 MPa) to support large deformation and withstand high input pressure without creating any rupture, while the hardness (Shore A 30) is comparable to the silicone rubber of In this study, we introduce the use of the latest industrial silicone 3D printer  developed by ACEOIn a bending state, assume that the energy loss to the surroundings is negligible, work done by the input air pressure in the chambers is equal to the bending strain energy stored in the elastomer body and torque compensating layer . TherefoWP, WA, and WL denote the work done by input air pressure and the bending strain energy stored in the 3D printed actuator body and the torque compensating layer, respectively.in which Because the SECA bends due to the work done created by the internal pressure acting on the chamber, we equate this work done .P is the input pressure, θ is the bending angle, ΔV is the increase of volume  of the finger joints. Excluding the dynamics associated with the fingers, energy stored in the joints is given by:where θk is the joint stiffness (where tiffness . To trea0) due to the muscle tone naturally presented in finger flexors  being larger than that of finger extensors  (0]) from flexed position would stretch the flexor muscles, creating resistance at the joints to oppose the movement. For stroke patients, strong resistance would be induced against extension due to the excess tone in finger flexors, called hypertonia  . Passivepertonia . In thisSince the major design consideration is for stroke rehabilitation, only the joint stiffness upon extending the fingers is of our interest. Further flexion of the fingers after resting angle , accuracy of the results would be significantly affected. Therefore, it is crucial to define the possible ranges of MCP and PIP joint angles and input pressures for the joint stiffness equation (Eqs 12) as:Near the singularity , and therefore the SECA is not further actuated and influenced by the singularity in the model.where γ is an empirical coefficient chosen to be 0.7 to avoid reaching singularity. In the bending state of the 3D printed SECA, cutoff pressure is defined as soon as the measured MCP or PIP joint angle exceeds its upper limit  as the bending angle of the 3D printed SECA, we can conclude the correct description of energy distribution inside the SECA based on the observed matching between model, FEM, and experiment results  and low stiffness  torsion springs at the PIP and MCP joint positions are designed to model the impaired fingers  Clinical Research Ethics Committee (Ref. ID: NCT03286309). Baseline semi-circular SECA is selected due to the larger ROM generated on the impaired fingers during the measurement of MCP and PIP joint stiffness . In handFour stroke subjects who have demonstrated weak hand strength with different levels of finger flexor spasticity are recruited . Four heA brief physical examination is performed on the subjects . The degQuantification of MCP and PIP joint stiffness is conducted , S3. γ =In this article, we have demonstrated the methodology of applying the 3D printed SECA to finger stiffness evaluation using the analytical models. The accuracy of the models is validated both in free space and on model fingers. Preliminary results showing the joint stiffness of stroke and healthy subjects are obtained using the models, which are supportive to existing clinical measures. In the future, our proposed method can be generalized to a rehabilitation robotic hand that reflects the stiffness of each finger in real time. With the stiffness information, optimal training tasks may be planned for each stroke individual depending on the current finger spasticity condition. Therapeutic progress may also be indicated in detail to motivate patients for achieving better improvement during rehabilitation training.All datasets generated for this study are included in the article/The studies involving human participants were reviewed and approved by the Joint Chinese University of Hong Kong-New Territories East Cluster (CUHK-NTEC) Clinical Research Ethics Committee (Ref. ID: NCT03286309). The patients/participants provided their written informed consent to participate in this study.HH is the first author who developed the mathematical theories, designed the experimental devices and experimental protocols, implemented the data collection software, performed the experiments, analyzed the experimental results, and participated in the manuscript preparation. ZT and XS performed the experiments, contributed to the discussions and analysis, and participated in the manuscript revisions. KT and ZL are the corresponding authors who supervised the project, contributed to the discussions and analysis, and participated in the manuscript revisions. All authors read and approved the submitted version of the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Dear Sir,Reconstruction of total upper eyelid defects pose a considerable challenge, requiring precise lamellar reconstruction to achieve proper eyelid function and aesthetics. Reconstruction techniques often require multiple surgeries, with blocking of the visual axis for varying periods of time before achieving the final result.. The composite graft is sutured to the medial and lateral canthal remnants using the same sutures and the stump of the levator aponeurosis is sutured to the superior edge of the composite graft. If the levator aponeurosis remnant is too short to be attached without causing lagophthalmos, an additional HPG can be placed superiorly. When needed, a mucous membrane graft is harvested from the lower lip and sutured in the superior fornix using a 7–0 polyglactin suture.First, we address the posterior lamellar defect: A full-thickness tarso-conjunctival graft is harvested from the contralateral upper eyelid, leaving at least 4 mm of tarsal plate height to minimize donor site morbidity. This is used to construct the central part of the posterior lamella, and will directly overlie the cornea. We used a graft sized approximately 6 mm X 15 mm. Next, the extent of the remaining defect on each side of the central graft is measured and an appropriately sized HPG is harvested. An oral denture is used for one week to aid healing. The full surface of the HPG is thinned to 70% thickness thereby removing the keratinized mucosa, using a diamond burr . Good eyelid-globe apposition, which can be problematic with HPG grafts and lead to corneal irritation,Using this novel approach, patients were treated in a single sitting, avoiding temporary visual axis occlusion. The creation of a long, pliable, concave posterior scaffold helped avoiding additional surgical manoeuvres and contributed to creating a tension-free, aesthetically pleasing and functioning eyelid.,The part of the composite graft that is in direct contact with the cornea is the tarso-conjunctival graft which provides a like-for-like substitute, contributing to the low rates of ocular surface complications which can sometimes be seen in these patients.Disadvantages include three surgical sites and the need for enough anterior lamella allowing an advancement flap.In summary, this technique adds to the surgeon's armamentarium when tackling a total upper eyelid defect, regaining eyelid function and offering a good aesthetic outcome.None declared.


Editorial It was a great honour when the International Society for Myriapodology  accepted our proposal to organize the 18th International Congress of Myriapodology at the Hungarian Natural History Museum in Budapest, between August 25–31, 2019. We had a successful meeting: 92 scientific participants (plus 13 accompanying persons) from 32 countries, all around the world from Brazil and Uruguay through Europe and South Africa to China and Australia. During the four scientific days we had 48 lectures (including four invited plenary presentations – one each morning) and 55 posters, which covered a wide range of our field, myriapodology, from the zoological subjects of morphology, anatomy, taxonomy, systematics, physiology, biogeography and nature conservation. We also had a joint full day excursion to the nearby Buda Hills, where participants could encounter some representatives of the local soil fauna. In the name of the organizing committee we would like to thank our 16 Hungarian colleagues who helped to make the congress really enjoyable.From the presentations, we are presenting here 11 scientific papers in this Special Issue of Zookeys. We are grateful to Pensoft Publishers and to the editorial board of Zookeys for providing the opportunity to publish a cross-section of the 18th International Congress of Myriapodology.Dr. Zoltán Korsós and Dr. László DányiGuest editors


Q0 exhibited the difference between comb and bottlebrush via no low-frequency Q0 peak of bottlebrush corresponding to backbone relaxation, which is probably related to the stretched backbone conformation in bottlebrush.Linear and nonlinear rheological properties of model comb polystyrenes (PS) with loosely to densely grafted architectures were measured under small and medium amplitude oscillatory shear (SAOS and MAOS) flow. This comb PS set had the same length of backbone and branches but varied in the number of branches from 3 to 120 branches. Linear viscoelastic properties of the comb PS were compared with the hierarchical model predictions. The model underpredicted zero-shear viscosity and backbone plateau modulus of densely branched comb with 60 or 120 branches because the model does not include the effect of side chain crowding. First- and third-harmonic nonlinearities reflected the hierarchy in the relaxation motion of comb structures. Notably, the low-frequency plateau values of first-harmonic MAOS moduli scaled with  Understanding the rheological properties of branched polymers is essential for all forms of production and processing in the polymer industry because the processability of polymers is highly affected by the degree of long-chain branches (LCB) . A brancQ0 was plotted as a function of excitation frequency  generates an oscillatory shear stress response σ(t). The obtained stress response can be decomposed into higher harmonic contributions via Fourier transformation as follows . The cubic amplitude scaling term represents medium amplitude oscillatory shear (MAOS) response, characterized by four MAOS moduli . The first-harmonic MAOS moduli [ω) and ω)] affect asymptotic deviations in average  elasticity or viscosity by causing rotations of elastic or viscous Lissajous curves ω→0. Backbone Rouse time τR,bb was calculated based on the hypothesis suggested by Lentzakis, Vlassopoulos, Read, Lee, Chang, Driva and Hadjichristidis [Mbb,end = Mw,bb/(Nbr + 1), which is the same as the molecular weight of backbone segments between branch points. Then, the effective backbone molecular weight is Mw,bb – 2xcMbb,end, where xc is the fractional length of the backbone-end portion varied by branch size. If branches are longer than backbone ends , xc is 1 because backbone ends have fully relaxed on the branch relaxation timescale. Otherwise , xc is the ratio of molecular weight of the branch to the molecular weight of the backbone end part . Finally, Me)(Mw). Using the calculated τR,bb was calculated from the formula based on the tube theory [τR,bb = τL/. PS290 exhibited the maximum of Q0, G*|) near the terminal relaxation time, as reported in earlier studies [Q0 displayed two local peaks reflecting characteristics of hierarchical relaxation. This implies that third-harmonic MAOS reflects the architectural features of polymers more sensitively than first-harmonic MAOS material function. For PS290-3-44, the peaks of Q0, G*| were observed near τR,bb. The additional peaks of Q0 and τL, which confirms that the low-frequency peaks are due to the remaining effective backbone chains.In  studies ,36,37. TG*|). However, two peaks were more evident in the master curves of third-harmonic properties. For DC PS, the high-frequency peaks of Q0 and τbr. In contrast, the low-frequency peaks of Q0 and τL. All MAOS parameters of DC PS exhibited the local minimum between τR,bb and τbr when their absolute values were used. In addition, featured sign changes of Q0. However, it remains an important question why the stretched backbone conformation makes an additional contribution only to the first-harmonic MAOS moduli and not the third-harmonic MAOS moduli. More systematic approaches would be necessary to answer this question.The first-harmonic MAOS moduli of DC and LB exhibited two local peaks when they were normalized by SAOS complex modulus  in LC regime (Zs > 3), η0 ~ Zs < 3), η0 ~ Zs < 1) and η0 ~ Zs < 0.2). The plateau modulus of the diluted backbone scaled as GN,bb ~ We investigated linear and nonlinear oscillatory shear (SAOS and MAOS) rheology of model comb PS melts, where the number of branches along the backbone was controlled while the branch length was kept. The model PS used was classified into three conformational regimes: loosely grafted comb (LC), densely grafted comb (DC) and loosely grafted bottlebrush (LB). In each regime, scaling of zero-shear viscosity rmations .0 and GN,bb of the experimental data and the model predictions were plotted as functions of Mw. The model failed to predict η0 and GN,bb of PS290-60-44 and PS290-120-44 because the model excludes the effect of side chain crowding in the densely branched topologies.Linear viscoelastic properties of model comb PS set were compared with predictions of the hierarchical model. For PS290-3-44, deviations between the experimental data and the model originated from the statistical distribution of the number and location of branches. Thus, we discussed the effect and importance of this distribution on relaxation processes. For quantitative comparison, ηMw, the high-frequency plateaus were constant whereas the low-frequency plateaus scaled with Q0 and its viscous part Q0 peak corresponding to the backbone part of LB disappeared. The stretched backbone conformation in LB might be related to this no low-frequency Q0 peak. Comparison of MAOS data with characteristic relaxation times  supported the discussion on changes in the number of peaks of relative nonlinearities.We presented the first-harmonic MAOS moduli of comb and bottlebrush polymers for the first time. The first-harmonic MAOS data reflected the hierarchy of relaxation processes in comb topologies well. DC and LB PS exhibited two plateaus in The findings from the first- and third-harmonic MAOS rheology of LC PS indicate that third-harmonic MAOS reflects structural features of different topologies better than first-harmonic MAOS. PS290-3-44 (LC) has a hierarchical relaxation process due to the averagely three branches. However, first-harmonic MAOS moduli displayed one broad peak while third-harmonic MAOS properties exhibited two peaks clearly. Nevertheless, it is necessary to characterize both first- and third-harmonic MAOS properties of polymers with complex architectures because PS290-120-44 (LB) reflected the feature of hierarchical relaxation in first-harmonic MAOS but the stretched conformation of backbone in third-harmonic MAOS.


This paper presents a novel method of calculating desynchronization between transmitters working in a single frequency digital video broadcasting-terrestrial (DVB-T) network. The described method can be a useful tool for enhancing passive radar operations and improving passive coherent location (PCL) sensors to correct their measurements of target localization. The paper presents the problem of localizing DVB-T transmitters utilized by passive radars, and proposes a novel method based on Time Difference of Arrival (TDoA) techniques to solve the problem. The proposed technique has been validated using real signals collected by a PCL sensor receiver. The details of the experiment and extensive result analysis are also contained in this article. The idea of using non-cooperative transmitters as an illuminator of opportunity (IO) for a passive radar receiver is not a new one. The history of passive radar goes back to the 1930s, when Robert Watson-Watt made the first demonstration of this technology utilizing a BBC radio transmitter as an illuminator of opportunity [Nowadays, after nearly three decades, passive radar technology is in a stage of maturity. It provides many applications, from air surveillance ,3,4 to tThere are several scientific works in the literature that focus on problems of passive localization systems utilizing SFNs ,12,13. AIn previous publications , the autIn this article, an analysis of the origin of all the detections is made. Furthermore, the impact of transmitter desynchronization on these detections is explored. Finally, a novel method of calculating the desynchronization between transmitters working in an SFN based on detected TDoA peak positions is presented. The authors believe that this method is a solid addition to any passive radar operating in an SFN, and will be a triggering factor for further studies of the impact of SFN transmitter desynchronization on PCL applications. In the second chapter of this article, passive radar geometry is described. The third chapter provides a brief description of an SFN. In the next chapter, the problem of localizing transmitters used in passive radars is formulated. The next two chapters provide a concise explanation of TDoA. In the seventh chapter, the experiment scenario and setup are presented. In the eighth chapter, the results of the experiment are shown. The next two chapters present the proposed method and impact of measured desynchronization on the passive radar localization of the target. The last chapter concludes this article.Passive radar operates on a different basis from active radar. In traditional active radar, the signal used for illumination is generated by the transmitting part of the radar system. Most commonly, a single antenna is used for both transmission and reception, switching between those two states. This configuration is described as monostatic. In a passive solution, the transmitter, or in this case the IO, works independently of the radar system. This creates a receiver–transmitter (Rx–Tx) pair working in a bistatic configuration.The independence of the exploited transmitter forces each receiver to gather two signals simultaneously: the reference signal gathered from the directional antenna facing the IO; and the measurement signal composed of all echo signals see . Both siTarget localization precision is highly dependent on the accuracy of the transmitter and receiver position data.An SFN is a network in which all transmitters broadcast the same signal simultaneously. In this article, the DVB-T SFN is at the center of interest. When designing a DVB-T SFN, there are many variables to consider. One of which is the ineliminable problem of interference caused by multiple transmitters broadcasting at the same time in the same frequency. It can be experienced by a DVB-T network user at nearly every position. To prevent any signal loss or intersymbol interference, DVB-T uses Orthogonal Frequency-Division Multiplexing (OFDM) with Guard Interval (GI), preventing data loss in transmission. The values of GI vary between 7μs and 224μs for an 8 MHz channel. This method solves the problem of the multipath for the user, therefore no strict synchronization of transmitters in an SFN, nor precise knowledge of its value is needed . HoweverAs mentioned in previous sections, the precise localization of illuminators of opportunity in passive radars is of utmost importance. Any inaccuracy is propagated onto target localization. If the positions of the ellipses’ focal points (Rx and Tx) are uncertain, the whole ellipse position may vary. In extreme cases, when two or more ellipses cross on sections nearly parallel to each other, a slight ellipse shift may result in a disproportionately large target localization error, or the rejection of the measurement.There are many databases provided by broadcast infrastructure operators, government agencies, and noncommercial websites, although transmitters’ positions are frequently inaccurate, out of date, or simply missing. In order to validate data acquired from external sources, it is possible to use a TDoA-based algorithm . Since pn  and the emitter position  can be described asTDoA is a technique used in multilateration localization. Multiple receiver stations gather signals from a single emitter. Considering two-dimensional space, the distance between the nth station position Rxc is the speed of light.Measuring the TDoA between two stations gives information about the difference of distances between the emitter and corresponding stations:The TDoA from single-pair receivers forms a hyperbola on which the source of emission is located see a. By addFor any number of receivers, N, greater than one, it is possible to create a set of N-1 equations and form N-1 hyperbolae b.n and Rxm) is calculated by cross-correlating the acquired continuous signals . Cross-correlation is a mathematical tool used in signal processing to analyze similarities between two signals in the function of time delay (τ). For continuous signals, cross-correlation can be written asThe exact value of the TDoA between two stations  is well known, the calculation of the exact time of occurrence for all four peaks is possible. Writing distance between point A and point B asAssuming that the precise position of transmitters , data from each pair of sensors are cross-correlated. Then, based on a priori knowledge of the localization of emitters, the maxima of the cross-correlation functions are identified. The peaks corresponding to the possible locations of emitters are marked as TDoA peaks. The rest of the peaks are classified as ghost peaks. The next step is to calculate the emitters’ precise localization using measured TDoA peaks. After this step, 3D positions of all emitters are determined. Based on the estimated Tx’s positions and known Rx position, the values of ghost peaks are estimated. Finally, through the comparison of the calculated and measured values of the ghost peaks, the estimated value of the desynchronization between emitters can be found. In the next section, the validation of the proposed algorithm using real measurements is presented.To validate the claim that the synchronization accuracy of two transmitters working in an SFN can be calculated using only passive radar receivers, an experiment was conducted. Four surveillance stations imitating passive radar receivers were placed in different locations in Warsaw and Pruszków, Poland . Each ofThe receivers used a GPS signal to achieve 50 ns (RMS 1-Sigma) synchronization accuracy and 10-m position accuracy. All were set to collect data on the 690 MHz central frequency and 10 MHz bandwidth, which covers the entire 48th DVB-T channel.The stations were controlled remotely via an internet connection provided by a PC. All the data collected by the receivers were transferred to a central processing station.Two DVB-T transmitters operating in an SFN were chosen as illuminators. Both transmitters were located in Warsaw, Poland. The main parameters of both transmitters are shown in It is important to mention here that the precise localization of both radio transmitter towers is ambiguous. Several sources from on-line databases provided different information see . Taking Choosing Wikipedia.org as a source of the transmitters’ positions, and gathered GPS data for the receivers’ positions, all distances were calculated. All previous calculations (5) were expanded by a z-factor to match a three-dimensional scenario. The results of these calculations, recalculated from time units into distance units and rounded to one meter, are presented in A series of measurements were conducted with the described system. After initial signal processing, cross-correlation was calculated for each pair of receivers  is not precise. While it is not a problem for a commercial user, it may be a source of significant localization error for a passive radar system.The described experiment proved not only that the chosen DVB-T transmitters were desynchronized by 3.4 μs, but also that the synchronization error of two transmitters working in an SFN can be calculated using only passive radar receivers, without any additional hardware systems. Such a method is recommended, especially when a priori knowledge of transmitters’ localizations is not reliable or inaccurate. This additional information about desynchronization can be used to alter the localization algorithms of passive radars, and possibly further improve their precision and detection probability, which the authors intend to implement in a future PCL processing chain.


In order to valorise food by-products into healthy and sustainable products, extrusion technology can be used. Thereby, a high expansion rate is often a targeted product property. Rapeseed press cake (RPC) is a protein- and fibre-rich side product of oil pressing. Although there is detailed knowledge about the expansion mechanism of starch, only a few studies describe the influence of press cake addition on the expansion and the physical quality of the extruded products. This study assessed the effect of RPC inclusion on the physical and technofunctional properties of starch-containing directly expanded products. The effect of starch type (native and waxy), RPC level , extrusion moisture content  and barrel temperature (20–140 °C) on expansion, hardness, water absorption, and solubility of the extrudates and extruder response was evaluated. At temperatures above 120 °C, 70 g/100 g of RPC increased the sectional and volumetric expansion of extrudates, irrespective of starch type. Since expansion correlates with the rheological properties of the melt, RPC and RPC/starch blends were investigated pre- and postextrusion in a closed cavity rheometer at extrusion-like conditions. It was shown that with increasing RPC level the complex viscosity |ƞ*| of extruded starch/RPC blends increased, which could be linked to expansion behaviour.  The cultivation and processing of rapeseed—primarily for the production of rapeseed oil—increased worldwide from 50.6 mt in 2007 to 76.2 mt in 2017 ,4,5,6,7.Up until now there has only been limited use of RPC. This has mostly been for animal and not human nutrition due to the fact that RPC contains high contents of antinutritional factors (ANFs) such as phenolic compounds, phytic acid and glucosinolates ,12,13. HLow moisture extrusion is one technology used for texturising starch-based and protein-based biopolymers into food products having an expanded porous structure. There is detailed knowledge about the expansion mechanism of several starch types, but only two studies describe the influence of oilseed press cake addition on the expansion and the physical quality of the extruded products. To our knowledge, there are no studies investigating the impact of RPC on food products. For instance, it has been reported recently that the addition of RPC to starch-based multicomponent products  resulted in inferior physical quality of the extrudates ,19. In pWhen fruit press cakes were added to a starch matrix, lower sectional expansion of extrudates was found in previous studies ,21,22,23It can be assumed that with the addition of a high-protein and high-fibre ingredient such as RPC to starch, the rheological properties of the melt are highly influenced due to the reaction behaviour of RPC components at extrusion temperatures. When the rheological properties change due to RPC addition to starch, an alteration of the expansion mechanism  is expected compared to starch expansion. Additionally, the starch type might have an impact on rheological properties and therefore on expansion due to starch type-related physicochemical and functional properties. Rheological material properties  influence the time of water vapor pressure underrun in the die and determine the direction of expansion . Expansion in turn governs physical product characteristics like specific hardness, wherefore it can be assumed that with different expansion mechanisms, the physical product properties of starch or starch/RPC blends are affected by RPC inclusion. Furthermore, it can be assumed that the technofunctional product properties of starch and starch/RPC blends like water absorption and solubility depend on the addition of RPC and extrusion conditions and indicate underlaying physicochemical reactions of starch and RPC during extrusion. To enable the design of food products including RPC and starch, a systematic evaluation of the impact of RPC on expansion, rheological and technofunctional properties is required. In this study, we have evaluated the impact of the RPC content, starch type and extrusion temperature on the expansion, physical quality and extruder response of extruded starch/RPC blends. In order to draw a correlation between expansion mechanisms and rheological properties, the effect of RPC addition on the reaction behaviour and the complex viscosity of starch/RPC blends was investigated at extrusion-like conditions in a novel closed cavity rheometer. With the investigation of technofunctional properties of starch and starch/RPC blends, indications regarding physicochemical changes in starch and RPC during extrusion processing were collected. A list of abbreviations is provided in TM 100) were kindly provided by Avebe . The amylose/amylopectin ratio of the starches were 25/75 for PS and 1/99 for WPS. Cold-pressed 00 type RPC, rapeseed peel (RP) and rapeseed oil (RO) were kindly provided by Teutoburger Ölmühle . The rapeseed used for RPC production was dehulled before deoiling. For all experiments, RPC and RP were milled to a particle size of < 0.5 mm as described by Martin and Osen . Potato w/w), respectively, to achieve rapeseed protein contents of 5, 16 and 27 g/100 g in the final product. RP and RO were added to the mixtures containing 50 and 70 g/100 g starch to achieve constant fibre and lipid contents in all the mixtures. PS and WPS were mixed with RPC, RP and RO in proportions of 70/10/10/10, 50/40/5/5 and 30/70/0/0  for 60 min. The mixtures were incubated at 20 °C for at least 8 h. Before extrusion, the dry matter in the mixtures was analysed in duplicate as laid down by legislation . The starch/RPC mixtures were extruded as laid down in  with theAs soon as steady-state operating conditions were reached, samples were taken. The average die pressure (bar) and product temperature in front of the die (°C) during the sample-taking period were recorded using CSpro medium software . The specific mechanical energy input (SME) was calculated as described in a number of recent studies ,31.The chemical compositions of the starches, RPC, RP, starch/RPC mixtures and all extruded samples were analysed. Regarding the latter, the extrudates were ground after drying to < 0.5 mm particle size using a knife mill . The dry matter (DM) was measured as laid down in the German Food Act . The proThe particle size distribution of the RPC, RP and starches was determined using a Malvern laser diffraction particle size analyser  as described by Osen and Toelstede . AnalyseSectional and longitudinal expansion indices (SEI and LEI) were calculated according to Alvarez-Martinez and Kondury  and CarvE and diameter dE before being placed on the measuring platform of a texture analyser  equipped with a 50 kg measuring cell. A cylindrical punch (SMS P/25L) compressed the samples to 30% deformation at a test speed of 8 mm s−1 until breakage. The applied force (Fmax) on the lateral surface of the extrudate strands is the specific hardness (N/mm2).For texture analyses, 10 randomly chosen extrudate strands were measured with a digital calliper to determine their length lSample) was placed in a 50 mL centrifuge tube (mCentr) and mixed with 40 mL demineralised water. After 10 min stirring with a reagent shaker , the samples were centrifuged for 10 min at 1000 relative centrifugal force (RCF) and 20 °C using a 6K15 centrifuge . The supernatant was decanted into a tared aluminium dish (mAlu), dried in an oven at 105 °C for 18 h  and then weighed after drying (mAlu + Solids). The residual sediment in the centrifuge tube was also weighed (mCentr + Sed). The water absorption index (WAI) was calculated using Equation (2).The water absorption was determined following the method of 56-20.01 with slight modification . For thiThe water solubility index (WSI) was calculated according to Equation (3) and described the amount of dry solids in the supernatant relative to the original sample mass . The samThe impact of the thermal treatment on structure formation in the starches, RPC and starch/RPC mixtures were investigated using a closed cavity rheometer  as described by Quevedo and Jandt . Before −1). The impact of thermal treatment on the reaction onset temperature was evaluated by temperature sweep analyses in a range from 30 to 140 °C .All experiments were at least performed in duplicate and were given as mean values with standard errors. The effects of starch/RPC ratio and process conditions were determined by one-way analysis of variance (ANOVA) with a significance threshold of The composition of the RPC, RP, PS, WPS and RPC/starch mixtures is summarised in Brassica napus L. variety our supplier processes. Furthermore, peeling the rapeseed before deoiling results in a comparatively low fibre content, increasing the RPC protein content. However, the protein content of the RPC that was used for this work lies within the range of that of rapeseed meals previously studied (34.4–40.6 g/100 g) [The data show that RPC had a protein content of 38.2 g/100 g. Previous studies reported lower RPC protein contents (18.6–30.0 g/100 g) ,7. The cThe RP obtained from the dehulling of the rapeseed had a lower protein content and higher lipid content than RPC (15.7 g/100 g protein and 25.5 g/100 g lipid). The fibre content of RPC (4.7 g/100 g) was much lower than the fibre content of the RP (29.4 g/100 g). In the RPC/starch mixtures, the protein content increases from 5.4 to 26.7 g/100 g with increasing RPC content, whereas the starch content decreases from 69.0 to 29.6 g/100 g. The lipid and fibre contents of the RPC/starch mixtures were kept constant by adding small amounts of RO and RP. PS and WPS had the smaller particle sizes than RPC and RP. As expansion phenomena is linked to flash evaporation at > 100 °C barrel temperature, the expansion indices of samples extruded at such temperatures are discussed here. The increase in temperature has mainly led to an increase in SEI, LEI and VEI in PS samples containing 70 g/100 g RPC. LEI and VEI were lowest for 40 g/100 g RPC samples extruded at 100 °C. In WPS samples, a barrel temperature of 100 °C resulted in higher SEI and VEI regardless of RPC content, but did not have a large impact on LEI. It is expected that the addition of protein- and fibre-rich biopolymers such as press cakes to starch influences the rheological properties of the melt , which fThe observed decrease in SEI due to 40 g/100 g RPC addition in WPS samples in our study is in accordance to previous studies that found lower sectional expansion of extrudates when fruit press cakes or other by-products were added to a starch matrix ,21,22,23The RPC used in this study contains 35.67 ± 5.19 g/100 g total dietary fibre, whereof 88.3% are insoluble and 11.7% are soluble. The impact of extrusion on the dietary fibre content of RPC will be part of future studies. Since with RPC addition, the amount of insoluble fibre in the blends, which act mainly as filler particles, increases markedly, the decrease in SEI in sample WPS40/RPC50 might be explained. However, the high SEI and VEI of samples with 70 g/100 g do not agree with this hypothesis. It can be assumed that high SEI and VEI in RPC-rich samples are driven by swelling expansion mechanisms more than by sudden water evaporation and may be associated with the presence of soluble fibres and an overall high water binding and holding capacity of RPC compared to starch. Soluble fibre fractions have been shown to exhibit large expansion volumes due to their greater water-binding capacity, which enables more nucleation sites for water vapor to develop at die exit ,50. ManyUnfortunately, the extrusion of 70PS/10RPC resulted in severe extruder clogging, wherefore no coherent extrudate strains were generated and the direct measurement of expansion indices was not possible. This means that the impact of starch type on the expansion of samples containing 10 g/100 g RPC cannot be compared here. However, the increase in RPC from 40 to 70 g/100 g increased the SEI and VEI of both PS and WPS based samples. At 140 °C barrel temperature, PS samples containing 70 g/100 g RPC showed a higher SEI compared to WPS-based samples. This effect cannot be explained with our results, but might be due to a low gelatinisation temperature, large granule size and favourable amylose content of PS compared to WPS that allowed bubble growth and extrudate fixation and enhanced the sectional expansion . The specific hardness of the samples is shown in Furthermore, although extruded PS was softer than WPS, no difference in specific hardness was seen between the two starch types when RPC was added, which was also seen for the correlation between starch type and expansion. The hardest extrudates were found for PS and WPS extruded at a moisture content of 29 g/100 g and 120 or 100 °C barrel temperature, respectively. RPC contains a number of components that might have a large impact on the specific hardness of the extrudates, such as proteins, fibres, sugars and lipids. The results suggest that one reason for a decrease in hardness with increasing RPC content in our study might be the relatively high protein content of RPC. The work of Zhu and Shukri  showed tIn starch-rich samples, the temperature-dependent WAI increase was greater than in RPC-rich samples. In PS and WPS samples extruded at 140 °C, the WAI decreased with increasing RPC content, whereas extruded PS and WPS as single materials exhibited low WAI.Water absorption is dependent on the availability of hydrophilic groups that are able to bind water, on capillary forces and on the porosity of the matrix . The high WAI of unextruded RPC-rich samples may be attributed to the water binding ability of fibre components in the RPC, as has been investigated in earlier studies . FurtherThe WSI of the PS and WPS mixtures increased with increasing RPC content at all barrel temperatures and at both moisture contents . The WSIAlthough PS and WPS were not soluble at room temperature (WSI of PS = 0.5 and WSI of WPS = 0.75), the extrusion process markedly increased the WSI of the samples.The starch type and moisture content during the extrusion process did not have a large impact on the WSI of samples. For starch-based matrices, the water solubility (WSI) is often correlated to the extent of degradation of the molecular components of the starch. In multicomponent systems, the WSI indicates the amount of components  that can be solubilised after mechanical breakdown resulting from, e.g., high shear forces in the extruder ,60. A higher RPC content in the formulations leads to an increase in rapeseed protein from 5 g/100 g in the 70PS/10RPC and 70WPS/10RPC mixtures to 26.7 g/100 g in the 30PS/70RPC and 30WPS/70RPC mixtures. The main protein fractions of rapeseed—cruciferin and napin—are soluble at room temperature at neutral pH ,62. ThisBrassica napus canola) are the 11S globulin cruciferin and the 1.7–2S albumin napin, representing, respectively, 60% and 20% of the total accumulated protein [The complex modulus |G*| of the starches, RPC and their mixtures were measured as samples were heated in the CCR rheometer . The val protein . It has  protein . Due to  protein ,67,68,69 protein . This isBetween 100 and 120 °C, the complex modulus |G*| of RPC increases indicating structure formation induced by aggregation reactions. At temperatures above 120 °C, the complex modulus |G*| decreases. There are two possible explanations for this. The aggregation reactions might be complete and |G*| may decrease as a consequence of higher molecular mobility . AlternaThe PS and WPS as single ingredients both showed a steep and constant decrease in |G*| as the temperature increased from 30 to 70 °C. At > 70 °C, |G*| decreased less steeply and at > 105 °C abruptly. In contrast to WPS with constant |G*| in the 120–140 °C region, PS showed a marked increase in |G*| starting at 120 °C. Overall, the complex modulus throughout temperatures between 30 and 110 °C was higher for PS compared to WPS. This can be attributed to the higher amylose content of PS (25 g/100 g amylose) compared to WPS (1 g/100 g amylose). A high amylose content can lead to increased entanglements between the linear amylose chains when starchy melts are heated resulting in a higher viscosity .Pre and TM, PS/RPC70 is extruded at 100 °C exhibited the highest |ƞ*|, followed by PS/RPC70 extruded at 140 °C and PS/RPC40 extruded at 120 °C. PS extruded at 120 °C exhibited the lowest |ƞ*|. The |ƞ*| of PS/RPC70 extruded at 100 °C increased over time, when TPre and TM was 100 °C, which indicates that aggregation reactions in RPC that were induced by extrusion are not completed. PS/RPC70 extruded at 140 °C only shows a slight curve slope, wherefore we assume that a higher TB during extrusion results in a higher degree of aggregation reactions. The comparatively low |ƞ*| of PS extruded at 120 °C and the small impact an increase in TPre and TM (100 or 120 °C) indicates that the starch was gelatinized during extrusion and constitutes an amorphous melt during rheological analysis. Pre and TM were set to the corresponding TB to which the samples were applied to during extrusion. |ƞ*| decreased in the order PS/RPC70 extruded at 140 °C, PS/RPC40 extruded at 120 °C and PS extruded at 120 °C. A high viscosity during extrusion can be associated with a larger SEI, since high viscosity results in high pressure at the die, wherefore the vapour pressure is undergoing later during expansion and the matrix can facilitate its structure during bubble growth. A low viscosity can in contrast be associated with a more pronounced LEI. In our study, for PS samples, the SEI increases and LEI decreased with an increasing RPC content. These observations can be linked to |ƞ*| in rheological analyses and indicate that a closed cavity rheometer can be used as a tool to find indications regarding the expansion properties of biopolymers. To mimic the rheological properties of the melt at the moment of die exit, TThe extruder response is shown in Starch might cause higher viscous dissipation and therefore increase the product temperature due to its high shear viscosity at low temperatures as described, i.e., by Ye and Hu . The pressure at the die is a result of the melt viscosity and is, in turn, influenced by shear and the thermal input in the extruder. Rheological investigations showed, e.g., that the 70WPS/10RPC sample had a dynamic viscosity of 179.5 kPas when treated at 80 °C for 1 min, whereas the 30WPS/70RPC sample had a dynamic viscosity of 54.5 kPas (data not shown). The higher viscosity of the extruded 70WPS/10RPC melt leads to higher die pressure.The aim of this study was to investigate the impact of rapeseed press cake (RPC) concentration on expansion, specific hardness, water binding and solubility, rheological properties and extruder response using a suitable model system. To correlate expansion behaviour with the rheological properties of the melt, a specific rheometer simulating extrusion conditions was used to evaluate the rheological properties of the extruded blends. Starch was chosen as the basis ingredient and the press cake content was varied far beyond those concentrations used in previous studies. Results gained in a specific rheometer show that the addition of RPC had a large effect on the complex viscosity of starch/RPC blends when treated at extrusion like conditions. The RPC increased the complex viscosity of the starch/RPC blends, which could be linked to a more pronounced sectional and decreased longitudinal expansion of the samples. It was possible to extrude starch/RPC mixtures containing up to 70 g/100 g RPC at barrel temperatures of 100–140 °C and moisture contents of 24 or 29 g/100 g. The physical properties of the extrudates were highly dependent on barrel temperature and RPC content, whereas starch type and moisture content did not have a large impact on expansion or texture in the range investigated. At temperatures above 120 °C, 70 g/100 g of RPC increased the sectional and volumetric expansion of extrudates, irrespective of starch type. At the same temperatures and RPC inclusion level, the longitudinal expansion decreased when RPC was added to potato starch and slightly increased when blends contained RPC and waxy potato starch. Moreover, 40 and 70 g/100 g RPC decreased the hardness and water absorption of the extrudates and increased the water solubility. Especially the extrusion of native potato starch/RPC at a high RPC content and high barrel temperatures resulted in a high expansion.In this study, we were able to correlate expansion behaviour and rheological properties using a closed cavity rheometer. However, expansion properties were only physically investigated > 10 s after die exit, whereas it is known from previous studies, that expansion comprises several stages of initial growth and shrinkage. To give a more detailed causal link between material properties and expansion behaviour during all expansion phases, digital imaging at the die exit should be used and the rheological investigations should be extended to the viscous and elastic modulus of the blends under thermomechanical treatment.


Faba bean protein has good functionalities, but it is little used in the food industry. This study identified a challenge from unfavourable starch gelation when utilizing faba bean for producing protein-based emulsion gel foods, and developed processing methods to overcome that. Two types of protein-based emulsion gel foods, namely yogurt and tofu analogue products, were prepared. The processing methods in this study involved steps of thermal pre-treatment of the beans, dehulling, milling, adding plant oil, homogenization, prevention of starch gelation, and inducing protein gelation. Two methods for preventing starch gelation were studied, namely starch removal and hydrolysis. The gel texture, water-holding capacity, and structural properties of the gel products were evaluated. Both starch-gelation prevention methods produced yogurt and tofu analogue products having typical emulsion gel properties. Hydrolysis of starch was favourable for producing the yogurt analogue, because the hydrolysate compounds improved the gel strength and viscosity. Moreover, it utilized the whole flour, meaning all the nutrients from the cotyledon were used and no side-stream was created. In contrast, starch removal was slightly better than hydrolysis for producing the tofu analogue, because the hydrolysate lowered the gel strength and water-holding capacity of the products. It is both possible and ecologically sustainable to utilize whole faba bean flour for making emulsion gel products. Vicia faba L.) is an important plant protein source, being rich in protein (29% on a dry weight basis 23] with The solid contents in all the analogue gels were analysed according to  with modg at room temperature for 30 min. The aqueous solution that was not held by the gel and moved to the top of the gel was carefully collected and air-dried at 95 °C for 8 h. The amount of the evaporated water was regarded as actual released water (Wr). The total amount of water in sample (Wt) was calculated from the total solid content. The water-holding capacity (WHC) was calculated as the percentage of the water held in the gel structure (Wt-Wr) in the total amount of water (Wt). WHC was measured in three replicates.The water-holding capacities of all the analogue gels were analysed as described by  with sli−1 to 100 s−1, kept at 100 s−1 for 10 s, and finally decreased to 0.3 s−1.The viscosity of the water extract and the water solution (1.8% soluble protein content), the emulsions made from them and the yogurt analogues were measured using a ThermoHaake RheoStress 600 rheometer  equipped with cone-and-plate geometry , in three replicates. In order to measure the shear-rate-shear-stress curve of the samples, the shear rate was increased from 0.3 sFrequency-sweep analyses were conducted on the gels using the ThermoHaake RheoStress 600 rheometer. The yogurt analogues were analysed using cone-and-plate geometry . The yogurt analogue was loaded onto the plate using a plastic soft straw dropper. The stress-sweep oscillation test was conducted with an increasing stress between 0.5 Pa and 100 Pa and a constant frequency of 1 Hz on the sample to determine the linear viscoelastic region. The gel sample was then reloaded and analysed with frequency-sweep oscillation test with an increasing angular frequency from 0.6 rad/s to 100 rad/s and a constant stress of 1 Pa. The frequency-sweep tests were conducted on three replicates of each yogurt analogue.The textural properties of the tofu analogues were analysed using a texture analyser  equipped with a 36 mm cylindrical probe (SMSP/36R) at room temperature. The analysed gel samples with a weight of 100 g were prepared in cylindrical glass cups with an inner diameter of 79 mm. A single compression test was conducted, and the probe penetrated the sample at a test speed of 2 mm/s until 40% of strain was reached. Six gels of each sample were prepared and measured independently.w/w), at 25 °C for 20 h. Similarly, the cooked emulsions were mixed with 2% SDS + 10 mM DTT. The mixtures were centrifuged at 20,238× g at room temperature for 15 min. The protein content in the supernatant was analysed using Bio-Rad DC protein assay kits as previously described. The protein resolubility was calculated as the percentage of the protein solubilised from the gel by each solution in the protein solubilised from the emulsion by the 2% SDS + 10 mM DTT solution. The measurements were conducted in three independent replicates.Tofu analogues were mixed with one of the following five solutions: Milli-Q water, 2% sodium dodecyl sulfate (SDS), 10 mM dithiothreitol (DTT), 2% SDS + 10 mM DTT, and 8 M urea, at a ratio of 1:10  was used to conduct one-way analysis of variance (ANOVA) with Tukey’s test at a 95% confidence level.With the processes described above, yogurt analogue and tofu analogue were successfully produced from both whole faba bean flour and a starch-free fraction. The flavour of the tofu analogues was bland, while the yogurt analogues were slightly acidic, and both were considered to be palatable by the evaluation panels. The colour and opacity of the faba bean tofu analogues and yogurt analogues were similar to those of soy-based tofu and commercial soy-based yogurt analogue. Furthermore, the intermediate products made during the process, the emulsions, were similar in appearance to soy-based milk analogues.The solid contents of the starch-free and hydrolysate-containing yogurt analogues were 5.6 ± 0.1% and 11.4 ± 1.3% (mean ± standard deviation), respectively, and those of the starch-free and the hydrolysate-containing tofu analogues were 9.0 ± 0.1% and 16.3 ± 0.6%, respectively. The differences in solid contents in the presence of similar protein and fat contents were attributable to the loss of water-insoluble components during starch removal, and the increase in the solid content due to the starch hydrolysate. The tofu analogues had higher solid contents than the yogurt analogues because they had a lower dilution ratio at the step of protein content adjustment during the process.The yogurt analogues produced from the starch-free and the starch-hydrolysate-containing emulsions both had typical solid gel properties, as shown by the viscoelasticity analysis results , where tThe starch-hydrolysate-containing yogurt analogue was stronger than the starch-free one, as it had higher firmness and viscosity revealed by its higher storage modulus and higher loss modulus values . The higThe solutions, emulsions, and gels at different steps of the production of the yogurt analogues all showed shear-thinning properties . The emuThe viscosities of the starch-hydrolysate-containing solutions, emulsions and gels were all higher than those of the starch-free equivalents . The hydThe starch-hydrolysate-containing water solution had a clear hysteresis of viscosity recovery after shear thinning, and its hysteresis gap was larger than that of the starch-free water extract A. In addThe viscosity of the starch-free yogurt analogue gel was 100-fold higher than that of the emulsion, and that difference was larger than that in the starch-hydrolysate-containing system (80-fold increase) B,C. ThisAs shown in the textural analysis results, the firmness properties of the starch-free and starch-hydrolysate-containing tofu analogues were similar in general . The stap < 0.05). On the other hand, the WHC values of the yogurt analogues did not differ significantly . The presence of the starch-hydrolysate compounds was associated with decreased WHC of the gel, although they are usually considered hydrophilic.The water-holding capacity (WHC) of the starch-free tofu analogue  was slightly higher than that of the starch-hydrolysate-containing tofu analogue  than the starch-free tofu analogue. This finding was in agreement with the larger hysteresis gap in the starch-hydrolysate-containing solution than the starch-free solution in the viscosity analysis . This suThe starch-free and starch-hydrolysate-containing water solutions originally had similar pH values, around 6.6 A. As theAs the pH decreased, the protein solubility decreased in both the starch-free and the starch-hydrolysate-containing solutions B becauseIn summary, the starch hydrolysate compounds enhanced the gel strength of the yogurt analogue but reduced that of the tofu analogue. The effect on the yogurt analogue could be associated with the effects on increased hydrogen bonding, solid content, and viscosity. Moreover, starch hydrolysate compounds may form a weak gel that is compatible and significantly beneficial for the yogurt analogue system . On the Thus, the starch-hydrolysis method was superior for producing faba bean yogurt analogues, as it increased yield, improved gel strength and viscosity, utilized all of the flour and its nutrients, and avoided a fractionation step. This method, however, produced inferior tofu analogues due to reductions in gel strength and water-holding capacity, so further development is necessary to improve the gel properties of a starch-hydrolysate-containing tofu analogue.The production of a “whole flour” emulsion gel food, as a new way of utilizing faba bean, was proven as technologically viable. A process comprising cooking, α-amylase hydrolysis, oil addition, homogenization, and lactic acid bacteria fermentation was proven effective and advantageous in producing the whole-bean yogurt analogues where the proteins effectively stabilised the emulsion and formed a typical gel network structure. The starch hydrolysate compounds improved the gel strength and viscosity of the yogurt analogues. The production of faba bean tofu analogues was also successful, but the starch removal was required in order to maximize gel texture and water-holding capacity. The adverse effects of the starch hydrolysate compounds were attributed mainly to their high viscosity and steric hindrance of proteins. It is likely that these processes will have similar outcomes with other starch-containing grain legumes and there are realistic prospects for producing yogurt analogues from whole flour of faba bean and other pulses in the near future. The development of whole-flour tofu analogues from starch-containing pulses requires further work, but starch-free tofu analogues are feasible already.


Nagashima-type palmoplantar keratosis (NPPK) is the most prevalent palmoplantar keratoderma (PPK) in East Asia. Homozygous or compound heterozygous loss-of-function mutations in serpin peptidase inhibitor, clade B , and member 70 (SERPINB7), which encodes members of the serine protease inhibitor superfamily, have been identified as the cause of NPPK. Clinical manifestations of NPPK include well-demarcated erythema, mild to moderate hyperkeratosis on the whole palm, and sole with transgrediens, extending to the dorsal surfaces of the hands and feet, inner wrists, ankles, and the Achilles tendon areas. In this study, we perform a review of relevant clinical cases aimed at elucidating the clinical characteristics, genetic characterization, differential diagnoses, and clinical management of NPPK. A better understanding of the clinical characteristics and pathogenic gene characterization of NPPK will enhance the diagnosis of NPPK, identify related diseases, and inform on the precise therapy and prognosis. Moreover, it will promote the awareness of NPPK in non-Asian regions. Nagashima-type palmoplantar keratosis  was initially described in Japan by Masaji Nagashima in 1977. It is characterized by transgressive and nonprogressive keratosis with an autosomal recessive trait . HomozygNPPK is characterized by “transgrediens,” which refers to hyperkeratosis that extends beyond the palmar margin of the palmoplantar skin, primarily involving the palms, soles, dorsal surfaces of the hands and feet, inner wrist, ankle joint, and the Achilles tendon area , 3. NotaStudies on the etiology of NPPK have taken more than 30 years. Its first English literature report in 2008 elicited increased research focus and rapid progress on this subject , South KSo far, more than 100 unrelated, molecularly diagnosed cases of NPPK associated with 15 distinct pathogenic SERPINB7 mutations in the homozygous or compound heterozygous state have been reported . This inet al. reported a recessive missense mutation of SERPINB7, c.830C>T (p.P277L), in NPPK patients which causes the mislocalization of SERPINB7 to intracellular aggregates. The mutated protein aggregates and is mislocalized within corneocytes [et al. found a novel mutation, c.271delC , in a Chinese NPPK patient, which formed a premature stop codon that truncated a 98-amino acid protein thereby eliminating a critical reactive site loop  [The establishment of the etiology of NPPK has taken a long time . In 2013neocytes . Later, neocytes . In Julyneocytes , while Kneocytes . In 2018331–352) . Accordi331–352) . In addi331–352) . A recenThe etiology of NPPK has been established. However, its pathogenesis has not been clearly elucidated. The SERPINB7 gene, located in 18q21.3, encodes the serine protease inhibitor B subtype 7, which inhibits serine protease and protects cells from protease-mediated damage , 23. AllSERPINB7 is distributed in the epidermis throughout the body, particularly in the stratum granulosum and upper part of the stratum corneum, whereas NPPK is limited to the palms, soles, knees, and elbows. The pathomechanism of NPPK lesions is usually restricted to specific areas that remain unknown. Frequent involvement of the knees and elbows in NPPK imply that chronic exposure to mechanical stress might be involved in the development of NPPK lesions, by precisely inhibiting mechanical stress-induced proteases and protecting keratinocytes or corneocytes from protease-mediated cellular damage , 19. How+ T lymphocytes had infiltrated the diseased tissue [Given that T cells have been shown to be infiltrated in the affected lesions, it is important to focus on the possible role of T cell-mediated inflammation in the pathogenesis of this superficial genetic disease. In 2009, Sakabe et al. performed immunohistochemical staining on a sample from the skin of NPPK patients and found that CD4d tissue . Subsequd tissue . Elsewhed tissue , while Kd tissue . In addiSince its pathogenesis remains elusive, it is crucial to clarify the mode of its inheritance. NPPK is an autosomal recessive disease, mostly originating from unrelated family mating. However, in some cases, the patients show pseudodominance, an autosomal recessive condition which occurs like the autosomal-dominant inheritance model amongst individuals in 2 or more generations of a family , 4, 26. + T cells were in the dermis with hyperkeratosis. Analysis of the infiltrating cells showed that they primarily included CD3+CD4+ T and a few CD3+CD8+ T cells, without CD1a+ and Langerin+ epidermal Langerhans cells, or exocytosis [et al. reported that the hyperkeratosis and thickening of PPK might reflect the severity of inflammation and oxidative stress, which deteriorates the local microenvironment [Histopathology of the diseased skin of NPPK is nonspecific and exhibits similar manifestations with other PPK. It is primarily characterized by hyperkeratosis, hypergranulosis, and incomplete keratosis of the epidermis, hypertrophy of the granular layer and spinous layer, infiltration of a small number of lymphocytes in the superficial layer of the dermis in a high-power view, and a marked increase in the glandular portion of the eccrine sweat glands at the dermis–hypodermis interface, without epidermolysis, viral inclusion, granular degeneration, or epithelial cell abnormalities  2, 11, , 11, 16.ocytosis . Despiteocytosis . Kogame ironment –30. Furtironment .The clinical features of diffuse PPK and recessive inheritance are not unique to NPPK. Thus, it is necessary to distinguish it from other diseases during clinical diagnosis and treatment . Gene seAlthough the NPPK phenotype is milder when compared to that of other hereditary palmoplantar hyperkeratosis and exhibits a nonprogressive disease progression, an effective treatment is still needed when the high allele gene frequency of founder mutation (c. 796C>T) in Chinese and Japanese populations, as well as discomfort and psychological burden of this disease, is considered. A standard therapeutic option for NPPK is currently unavailable. Therefore, the current management approaches for this disease is mainly aimed at reducing hyperkeratosis using topical vitamin D3 and/or topical keratolytic agents, including 5%, 10% salicylic acid, 10% urea, and adapalene , 4, 47. + T lymphocytes [In 2009, Sakabe et al. performed immunohistochemical staining on a simple affected skin of NPPK patients and found that it was infiltrated by CD4phocytes . They apphocytes .Approximately one-third of alleles that cause genetic diseases carry premature termination codon (PTC), which trigger the production of truncated proteins . MoreoveDue to advances in genetic testing technologies in NPPK diagnosis and treatment as well as the accumulating number of related case reports, NPPK has gradually been recognized by scholars across the globe. However, an internationally recognized diagnostic criteria for NPPK are currently lacking. Notably, the possibility of NPPK cannot be ruled out in patients with clinically sporadic, nondestructive, diffuse palmoplantar keratosis. Genetic testing technologies can be used to confirm the diagnosis of NPPK by integrating medical history, clinical manifestations, and laboratory examination of the patients for differential diagnosis. During diagnosis by genetic testing, priority is given to the detection of the founder mutation (c.796C>T), which exhibits the highest frequency among SERPINB7 mutations. Moreover, considering the high allele frequency of the founder mutation in normal people, it is essential to conduct genetic counseling for NPPK patients or NPPK carriers and their partners. Despite the effectiveness of local immunosuppressants and aminoglycoside antibiotic read-through therapies, their long-term effects have not been determined. More studies should be aimed at elucidating the pathogenesis of NPPK and identifying a precise therapy with stable curative effects.


Historically, liver allografts with >30% macrosteatosis (MaS) on donor biopsy have been associated with early allograft dysfunction and worse graft survival; however, successful outcomes have been reported in small cohorts. This study proposes an elevated MaS threshold for organ utilization without detriment to graft survival.The UNOS Standard Transplant Analysis and Research database was evaluated for transplants between 2006–2015. Graft survival up to 1-year was evaluated by Kaplan-Meier (KM) survival analyses, and by univariate and multivariable logistic regression analyses, including donor and recipient characteristics. Odds ratios (OR) with 95% confidence intervals (CI) for risk of graft loss are reported.Thirty-day risk of graft loss was increased with MaS as low as 10–19%  and peaked with MaS 50–59% (2.921 [1.672–5.103]). At 1-year, risk of graft loss remained elevated with MaS 40–49% (1.465 [1.002–2.142]) and MaS 50–59% . Multivariable models were created for Lower and Higher MELD recipients and MaS cutoffs were established. In Lower MELD recipients, organs with ≥50% MaS had increased risk of graft loss at 30 days  and 1-year post-transplant . Higher MELD recipients had increased risk of graft loss at 30 days with allografts showing MaS ≥40% . At 1-year the risk remained increased, but MaS was not significant predictor of graft loss.048 [1.131–3.710], p = 0.0616). In both MELD cohorts, organs with MaS levels below threshold had similar survival to those transplanted without a donor biopsy.In conjunction with recipient selection, organs with MaS up to 50% may be safely used without detriment to outcomes. Currently, over 13,000 candidates are listed for and awaiting liver transplantation nationwide . Despite a steady increase in the number of liver transplants occurring in the United States over the past five years[Hepatic steatosis is common, being reported in 30–51% of donor livers, a numberet al. demonstrated successful outcomes using donor allografts with <30% MaS.[et al. showed an increased risk ratio (RR) of 1.71 for graft loss at 1-year with allografts having >30% MaS.[et al. found increased rates of primary graft dysfunction, but no difference in 60-day or 3-year patient mortality using allografts with severe steatosis.[The use of grafts with mild steatosis is generally accepted, as Kwon <30% MaS. Converse<30% MaS.,12 Spitz>30% MaS. In contrteatosis. Multipleteatosis.–23 TheseAs waitlist registrations continue to rise, ongoing efforts to expand the donor pool are imperative. Using the Organ Procurement and Transplant Network Standard Transplant Analysis and Research (OPTN STAR) database, we hypothesized that liver allografts with increased levels of MaS above 30% may be used safely in LT. Furthermore, we attempted to identify a threshold MaS level for safe transplantation and characterize the donor and recipient factors which contribute to short and long-term graft survival amongst steatotic allografts in the modern transplant era.www.optn.transplant.hrsa.gov). Institutional approval was sought from the Cedars-Sinai Medical Center review board and given the de-identified nature of the data, it was deemed that additional IRB approval was not necessary.The OPTN STAR database contains prospectively collected data on donors and recipients for transplantation of all organs. A retrospective review of liver transplants and potential liver donors between January 1, 2006 and June 30, 2015 was performed. Data were obtained from the OPTN directly and are available to all interested parties through the OPTN , multi-organ or split-liver recipients, donation after circulatory death (DCD) transplants, recipients with prior liver transplant, recipients listed as Status 1A, as well as any recipient without at least 1-year of follow-up. After applying these criteria, 41,347 transplants were available for analysis. Pre-donation liver biopsy was identified in 19,137 transplants, of which 16,306 had information on donor liver steatosis and comprised the study group.All transplants with donor liver biopsy were categorized by volume of MaS identified on biopsy. Seven MaS groups were identified: 0–9% , 10–19% , 20–29% , 30–39% (n = 762), 40–49% (n = 223), 50–59% (n = 150), and ≥60% (n = 142). Organs without a donor liver biopsy were classified as “No Biopsy.” Donor liver biopsy is typically used in the evaluation of questionable donors, while donors without a biopsy are considered acceptable to transplant for selected recipients. Therefore, these donors comprise an “ideal” control group by which to compare outcomes.Donor and recipient characteristics were collected and evaluated, including age, gender, ethnicity, blood type, body mass index (BMI), viral status for Epstein-Barr (EBV), cytomegalovirus (CMV), hepatitis B virus (HBV) and hepatitis C virus (HCV), as well as cold ischemic time (CIT). Donor-specific variables included biopsy results with percent MaS, calculated Donor Risk Index (DRI), history of hypertension, diabetes, prior myocardial infarctions, history of cigarette or drug use, and Center for Disease Control (CDC) classification as high-risk donor. Recipient-specific variables included lab-based Model for End-stage Liver Disease (MELD) score, etiology of liver disease, history of diabetes, prior abdominal surgery, portal vein (PV) thrombosis, prior transjugular intrahepatic portosystemic shunt (TIPS), and need for dialysis or mechanical ventilation at time of transplant.The primary outcome was graft survival following transplantation. Graft survival was defined as the period between transplantation and graft loss necessitating re-transplantation or recipient death, whichever occurred first. Graft survival was assessed for the duration of individual patient follow-up, up to 1-year after transplantation. Patients who were lost to follow-up with a functioning graft were censored.Subgroup analyses were performed with subgroup determination based on recipient MELD score. Two subgroups were created—Lower MELD (score <33) and Higher MELD (score 33 to 40). For easier application in the current clinical scene, grouping determination was based on recently implemented allocation policies, which facilitate local-regional sharing of available allografts amongst recipients with MELD scores of 33 and higher.Statistical analysis was performed in JMP Pro 13.1 . Single variable analyses were performed using Students t-tests and ANOVA where appropriate; multiple variable categorical analyses were performed using Pearson’s chi-squared tests. Kaplan-Meier (KM) survival analyses were performed and were censored for patients with functioning graft at last known follow-up with survival times truncated to 365 days. Logistic regression models were utilized to evaluate graft survival at 30 days, 90 days, and 1-year post-transplant. Univariable analyses identified factors for inclusion in multivariable analyses using a p-value <0.10 as a threshold for inclusion. A manual, stepwise regression analysis was performed using a threshold p-value of <0.05 for significance with the exception of inclusion for factors previously shown to influence graft survival . Odds ratAfter applying exclusion criteria, 41,347 transplants remained for analysis. Donor liver biopsy was performed in 16,306 (39.4%), comprising the biopsy cohort. Select donor and recipient characteristics are presented in Graft survival was compared between recipients of liver allografts with and without donor liver biopsy. Overall, graft survival was lower for biopsied organs than those without a biopsy at 30 days , 90 days , and 1-year  after transplant. Interestingly, amongst allografts that survived 30 days, there was no difference in 90-day survival  and a small but statistically significant difference in 1-year survival .Incidence of graft loss increased with increasing volumes of steatosis . At 30 dDonor and recipient characteristics within each MaS group are presented in Graft MaS was significantly associated with graft survival at all time points in multivariable analyses . CompareKM survival analyses showed no significant difference across MaS groups for 1-year graft survival in Lower MELD recipients Click here for additional data file.S2 TableComprehensive evaluation of demographic and medical characteristics of all donors and recipients, categorized by percent MaS on biopsy. MaS groups are compared.(DOCX)Click here for additional data file.S3 TableMultivariable regression analyses for 30-Day (A), 90-Day (B), and 1-Year (C) graft loss.(DOCX)Click here for additional data file.S4 TableMultivariable regression analyses for 30-Day (A), 90-Day (B), and 1-Year (C) graft loss amongst recipients with MELD Score <33. Factors known to influence allograft outcomes forced into models .(DOCX)Click here for additional data file.S5 TableMultivariable regression analyses for 30-Day (A), 90-Day (B), and 1-Year (C) graft loss amongst recipients with MELD Scores 33 and higher. Factors known to influence allograft outcomes forced into models .(DOCX)Click here for additional data file.


Asian/Pacific Island Nursing Journal would like to thank the following people who acted as peer-reviewers for the journal from January 2019 through December 2019.The Editor-in-Chief of Patricia AlpertAmnatsatsue KwanjaiBalakrishnan UmamaheswarBooranasuksakul UraipronNafanua BraginskyByon Ha DoChen Wei-TiChad CrossAlona Dalusung-AngostaFelicitas dela CruzRaymond FolenFaye GaryArtemio GonzalesMargaret Hattori-UchimaMargaret HeitkemperNaoko HikitaDianne IshidaIra Kantrowitz-GordonMerle Kataoka-YahiroKarina KatigbakMasashi KawanoJennifer KawiMay KealohaHeeyoung LeeDongmei LeeWen-Wen LeeRebecca LorenzNada LukkahataiMay-Ying LyJennifer NeversAngelina NguyenNa-Jin ParkAndrew ReyesKarol RichardsonReiko SakashitaTodd SetoSetyowatiLillian Tom-OrmeTsai Hsiu-MinNancy WoodsJoyce YangYonezawa Kaori


Patient specific finite element (FE) modeling of the pediatric spine is an important challenge which offers to revolutionize the treatment of pediatric spinal pathologies, for example adolescent idiopathic scoliosis (AIS). In particular, modeling of the intervertebral disc (IVD) is a unique challenge due to its structural and mechanical complexity. This is compounded by limited ability to non-invasively interrogate key mechanical parameters of a patient's IVD. In this work, we seek to better understand the link between mechanical properties and mechanical behavior of patient specific FE models of the pediatric lumbar spine. A parametric study of IVD parameter was conducted, coupled with insights from current knowledge of the pediatric IVD. In particular, the combined effects of parameters was investigated. Recommendations are made toward areas of importance in patient specific FE modeling of the pediatric IVD. In particular, collagen fiber bundles of the IVD are found to dominate IVD mechanical behavior and are thus recommended as an area of primary focus for patient specific FE models. In addition, areas requiring further experimental research are identified. This work provides a valuable building block toward the development of patient specific models of the pediatric spine. Development of patient specific spine models is of increased interest in the treatment of pediatric spinal pathologies. Such models aim to revolutionize clinical practice by providing practitioners with detailed predictions of a patient's spinal biomechanics. For example, such models are already showing promise in predicting outcomes of corrective interventions for AIS . Further, focus will be placed on the annulus fibrosus (AF) as this is the main source of variance in FE IVD models  can be treated as an incompressible fluid Fagan et al., While, most IVD research focuses on the case of healthy adults and/or degenerative IVDs, this work places a special focus on pediatric IVDs—an overlooked area. It thus becomes important to build an understanding of the biomechanics of the IVD, focusing on those parameters relevant for mechanical behavior. There however is a paucity of studies on those parameters relevant for modeling of pediatric IVDs. Below, we aim to highlight key findings, insights and hypothesis of the pediatric IVD biomechanics relevant for FE modeling. Focus will be placed on the AF fiber angle, AF fiber stiffness and AF ground matrix stiffness, as these are the key variables in such FE models. In addition, while the NP is not of focus in this study, development changes will be briefly addressed. Focus will not be placed on the geometry of the IVD as in patient specific models, these are derived from medical imaging. Finally, focus is not placed on the aged or degenerate IVD as these deviate from the healthy case and are outside of the scope of this research  fiber angle observations of fetuses , proteoglycans (65% of dry weight), collagen (20% of dry weight), elastic fibers and other proteins Bogduk, . Due to Based upon the above investigation, the following conclusions can be drawn. First, from an early age  the IVD exhibits the structure of an adult IVD  dataset of the Visible Man . The IVD geometry was identified from CT by manually extracting keypoints of the superior and inferior surface using an in-house MATLAB code. These keypoints were then imported into an in-house Python code, which defined the IVD geometry based upon a parametric description of endplate geometry  and model complexity. Introduction of higher fidelity representations increases the model complexity. In this work, we employ well-established methods of representing the AF ground matrix, AF collagen fibers and NP. Through this we seek to balance the needs of fidelity and complexity.E with a cross-section selected such that the volume of the collagen fibers was 25% that of the AF . In this, the motion of the superior endplate is tracked relative to the inferior endplate and at each instant the center of rotation is found. In this work, we take guidance from Schmidt et al.  who explC10 and ground matrix C01.The objective of this work is to understand the combined influence of IVD parameters on predicted rotational stiffness. To achieve this, the study was split into three studies. In the first study the influence of individual properties on the IVD was explored, through this the properties which had a significant contribution to the IVD stiffness were identified. In the second study, the combined influence of these significant properties was investigated. In the final study, the independence/convolution of combined parameters was explored. As previously discussed, the parameters which are studied are those relating to the AF as these dictate the stiffness of the IVD. The parameters which are explored are fiber angle, fiber stiffness, ground matrix In the first study, parameters of the IVD were varied individually. A large range of IVD parameters are found in literature Galante, , thus to2 = 81 simulations, likewise if four parameters were deemed significant, this would require 94 = 6561 simulations. This is why the first stage sought to identify those parameters which were significant, thus reducing the total number of simulations required.In the second study, the combined effect of IVD parameters was investigated. In this, parameters were varied together across a full range of their potential combinations. For example, if two parameters were deemed significant, then this would result in a total of 9A results in a stiffness increase of 50% while a change in parameter B results in a stiffness increase of 20%. If the effects of there parameters are independent, then changing both A and B together would result in an overall stiffness change of 80% . If the influence of parameters was convoluted , then the combined influence would be other than 80%. Understanding independence of parameters on the overall IVD stiffness is important because, if independent, the full effect of a parameter can be understood from the parametric study presented in study one. However, if parameters are convoluted, then the effect of one parameters is dependent on the other parameters and thus a broader awareness must be maintained when specifying parameters in a patient specific FE model.Study three explored if the effect of changing one parameter is independent of changes in other parameters. By way of explanation, assume in the first parametric study that a change in parameter Pθ, Pk, PC10 be the percentage change in IVD stiffness caused by a change in fiber angle, fiber stiffness, and ground matrix C10, respectively . Likewise, let Pk,C10θ, be the percentage change in IVD stiffness from combined changes of fiber angle, fiber stiffness and ground matrix C10 . If these parameters are independent then:In a more generalized form, let 2) was calculated.In study three, the independence of parameters was tested by using Equation (1) to determine the expected combined effects of parameters from study 1, assuming independence. These were then compared to the effects predicted in study 2, and the coefficient of determination . Thus, in patient specific modeling, high-fidelity representation of the collagen fibers is imperative. Likewise, in efforts to calibrate a model to match experimental findings, fiber angle should be the first target.The behavior under flexion, extension and lateral bending were similar, as these directions are all rotations about axes tangential to the transverse plane. As such, for simplicity, discussion here will focus on flexion, but the overall conclusions are transferable to extension and lateral bending. Under flexion, the fiber angle has the greatest impact, especially for positive increases in fiber angle, where a 50% in fiber angle resulted in a 113% increase in IVD stiffness. Conversely, for a −50% change in fiber angle, the IVD stiffness is reduce by 36%. Interestingly, the second most significant parameter is ground matrix CC01 parameter was of negligible significance. In the first parametric study, the maximum effect of C01 was 1.5% for axial rotation, 6.8% for flexion, 7.3% for extension, and 7.2% for lateral bending. By way of explanation, the larger C10 term of the Mooney-Rivlin model dominates the lesser C01 term, such that changes in C01 have little impact on the overall stiffness of the ground matrix, and thus the overall IVD. Both experimental and modeling studies have shown the strains experienced in the AF are in the range of milli-strains , however as the fiber angle increases or decreases, the convolution increases. This is particularly apparent for large fiber angles, where the convolution is greatest (indicated by an R2 of 0.68). This reinforces the discussion from section 5.3, in which it was observed that increases in fiber angle, magnify the effect of changes in fiber stiffness.However, under flexion, extension and later bending, a degree of dependence is observed between the various parameters, indicated by the locus of data points not sitting on the dashed line. This shows that there is a degree of convolution between the effect of different parameters on IVD stiffness. Interestingly, the degree of convolution is governed by the fiber angle. For a Δθ = 0%, the impact of different parameters follows Equation (1) . This is relevant to patient specific modeling of the IVD as this demonstrates the independence/convolution of IVD parameters under different loadings. In the case of axial rotation, parameters of fiber angle and fiber stiffness are independent. This means that a complete understanding of the effect of these parameters, can be achieved by studying the results of the first parametric study . HoweverBased upon these findings, we propose the following. (1) For patient specific models of the IVD, primary focus should be placed on accurate representation of fiber parameters. Specifically, for models focused on axial rotation both fiber angle and stiffness should be given equal attention, for other motions fiber angle be given greater attention. (2) Non-invasive interrogation of fiber angle for individual patients is a current challenge, thus new techniques need to be developed to incorporate these patient specific fiber parameters. Ultrasonography has been demonstrated as one technique for interrogating parameters such as lamellar number and thickness (both which affect fiber stiffness)  concluded that fiber angles in the pediatric IVD likely match those of the adult. Thus, improved fidelity of fiber angle in patient specific models will improve the overall quality of predictions. Here, the (Michalek, With respect to fiber stiffness, Galante  demonstrConsiderations such as these are of increased significance when dealing with specific pediatric pathologies. Generally, little information is available on IVD parameters in such patients. The discussion above acts as further guidance to required areas of focus in such modeling. For example, in AIS, quantitative variation in the size and orientation of collagen fiber bundles has been observed in opposite sides of the AF (Roberts et al., In any FE model of the IVD certain limitations are inherent and should be noted. First, this study used geometry from a single IVD, from which general conclusions were drawn. For this, it is argued that any effects of patient specific geometries can be extracted from medical imaging, thus focusing on a single, well-studied geometry will aid in translatability of results. Second, this study focused on a single constitutive model for all components, balancing model fidelity and complexity. For example, higher fidelity models can be considered which incorporate non-linear collagen fiber properties (Haut and Little, This work has investigated the influence of various mechanical parameters on the stiffness behavior of the IVD under various loading conditions. Notably, while other studies have investigated the individual influence of individual parameters (Fagan et al., The original contributions presented in the study are included in the article/JL developed the initial IVD model. EP conducted the parametric study, results analysis and wrote the manuscript. PP and JL reviewed the manuscript and provided valuable inputs. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.