Chordoid glioma is a rare and relatively recently defined tumour entity. Worldwide there have only been around 90 cases described until now. A chordoid glioma comprises a low-grade suprasellar neuroepithelial neoplasm originating in the anterior part of the third ventricle, with consistent radiological features on MRI. This lesion should be considered as a differential of third ventricle tumours. The patient described in this paper is quite unique in the sense that despite only partial tumour resection was obtained, the residual tumour was not progressive during several years of follow-up. Preoperative recognition of this disease entity is crucial to modify the treatment approach and improve patient outcome. Chordoid glioma is a rare and relatively recently described low-grade suprasellar neuroepithelial neoplasm originating in the anterior part of the third ventricle. This entity was first named by Brat et al. in 1998 [The name of the tumour is derived from the histological appearance, resembling a chordoma, but with other characteristics pointing towards its glial origin. Most physicians have little knowledge of this tumour entity, which make it a very confusing lesion. It has to be considered as a differential of third ventricle tumours. Though it is a low-grade tumour, the treatment-related morbidity is very high. Knowledge of this disease entity is crucial to better estimate patient prognosis and improve patient outcome by modifying the treatment approach. This case report is quite unique in the sense that despite only partial tumour resection was obtained, the residual tumour was not progressive during several years of follow-up.A 43-year-old man presented at the neurology department complaining of increasing visual disturbances during reading, evolving into bilateral hemianopsia. In his clinical history, the patient underwent bariatric surgery one year before. Further general examination was negative.First, our patient underwent a CT scan of the brain. This showed a voluminous well-circumscribed oval mass on the midline in the third ventricle, with its largest diameter in craniocaudal direction. The lesion was slightly hyperdense to the surrounding parenchyma .On the subsequent MRI of the brain, this mass was T1-isointense to grey matter, slightly heterogeneously T2-hyperintense and showed vivid homogeneous contrast enhancement after gadolinium administration . There wDuring surgery, a greyish mass was observed in close relation with and mass effect on the optic chiasm, which was edematous. Due to the firm consistency of the mass, only the lower part could be removed through the subfrontal access and it was decided to perform an additional transcallosal access route. A definite histopathological diagnosis could not be obtained on the frozen section material. The initial working hypothesis was a pituicytoma.Detailed analysis of all available tissue material showed a dominant mucinous component with epithelial cells with eosinophilic matrix and lymphoplasmocytic infiltrate organized in clusters and cords . There wDuring the postoperative hospitalization, the patient developed diabetes insipidus, for which a treatment with desmopressin was started. Later during follow-up, cortisol and thyroid deficiencies and lowered testosterone and growth hormone levels were noted, for which substitution was necessary. Furthermore, intermittent dysregulation of the ionogram with hypernatremia occurred.Eight months later, a transcallosal re-resection of the residual tumour was planned. The patient and his family were informed about the risks and possible aggravation of the hypothalamic insufficiency. During this surgery, further debulking of the very firm tumour was performed, but complete resection was not obtained due to the tight connection with surrounding structures anteriorly and caudally.Postoperative course was complicated with severe apathy and confusion. A pronounced cognitive impairment with mainly short-term memory loss remained, for which years of revalidation were necessary with slow, but steady progression. Follow-up MRI showed no volume increase nor change in morphology during the first two years of follow-up . So, forThe chordoid glioma has been classified by the World Health Organization in 2000 as a grade II tumour. The name of the tumour is derived from the histological appearance, reminiscent of chordoma, but with other characteristics pointing towards its glial origin, such as avidly staining with glial fibrillary acidic protein (GFAP). In the 2007 and 2016 revisions of the WHO brain tumour classification, this tumour has been included as a separate entity. So far, relatively little is known about the genetic underpinnings of this very rare neoplasm.The exact incidence of chordoid gliomas is not yet known, though they appear to be uncommon. They usually occur in middle-aged adults (mean age 46 years old) and have been reported more frequently in female  .Clinical presentation varies from asymptomatic to aggressive and most often symptoms are mild and slowly progressive, with headache, amnesia and visual disturbances as most common features. Specific symptoms like endocrine alterations and visual disturbances are due to the location and its close relation to the hypothalamic and pituitary region and the optic pathways. When obstructive hydrocephalus occurs, this will give rise to more vague symptoms of increased intracranial hypertension.The radiological features are remarkably consistent. A chordoid glioma appears as a third ventricle suprasellar mass, often in close relation to the hypothalamus.On CT, the lesion is a well circumscribed, hyperdense (65%) mass that is homogeneously enhancing after contrast administration (68%). Most often it is oval shaped, with the greatest diameter in craniocaudal direction.On MRI, which is the preferred examination, it appears as a slightly T2-hyperintense mass, T1-isointense to grey matter 63%), with homogeneous enhancement following contrast administration (70%). A minority of the cases (19%) show a peripheral cystic area [%, with hDifferential considerations for a vividly enhancing anterior third ventricle mass are meningioma, ependymoma, pilocytic astrocytoma, chordoma and lymphoma. This entity can sometimes be mistaken for an unruptured aneurysm as well. If there is a cystic component, a craniopharyngioma might also be suspected. Nonfunctioning pituitary tumours may be thought of, but in contrast to pituitary tumours, a chordoid glioma is distinct from the pituitary gland and stalk.The cell of origin is unknown until now, though it has been suggested that they arise from ependymal cells (tanycytes) , 4, 5. TThese tumours are described as well-circumscribed fusiform mucoid lesions. Macroscopically they consist of firm masses with a grey brownish colour . HistoloThyroid transcription factor-1 (TTF-1) expression has also been proposed as a good differential marker for chordoid glioma, but was not performed in our patient. In a multicentric series of 17 cases, all chordoid gliomas expressed TTF-1, as in the developing and adult lamina terminalis. Although TTF-1 is not specific to chordoid glioma, this marker could help in the differentiation of atypical cases , 7. SomeThe chordoid glioma is a slow-growing noninvasive tumour. Complete surgical resection is considered as the treatment of choice for this type of tumour. Nonetheless, the deep location of the tumour, the close relation with the roof of the hypothalamus and the sporadic close relation to cranial nerves and the cavernous sinuses, often prevents complete resection. Intra-operative mortality rates have been reported to be 17–29% of cases, mainly due to bleeding , 8.Postoperative mortality risk is highest during the first month after surgery, caused by pulmonary embolism 42%) and cardiovascular events [2% and caIncomplete resected tumours will continue their slow growth and prompt later re-resection. Stereotactic radiosurgery has been shown to be an effective adjuvant therapy after partial resection and tends to give results superior to external beam radiotherapy.Even though this tumour is low grade, the overall patient outcome tends to be poor, mainly owing to the location, the close proximity of critical neurovascular structures and the difficulty of obtaining a complete surgical resection. Recurrence is very rare after total macroscopic resection and metastasis is not described.The chordoid glioma is a recent but distinct tumour entity. It is a rare low-grade tumour of the anterior wall of the third ventricle with consistent radiological features on MRI. Knowledge and preoperative recognition of this entity is crucial to better estimate and improve patient outcome, by modifying the treatment approach.


A variational principle is derived for two-dimensional incompressible rotational fluid flow with a free surface in a moving vessel when both the vessel and fluid motion are to be determined. The fluid is represented by a stream function and the vessel motion is represented by a path in the planar Euclidean group. Novelties in the formulation include how the pressure boundary condition is treated, the introduction of a stream function into the Euler–Poincaré variations, the derivation of free surface variations and how the equations for the vessel path in the Euclidean group, coupled to the fluid motion, are generated automatically. Moreover, the variational principle captures dynamic coupling with the vessel motion; that is, the variational principle produces both the exact fluid equations in a moving vessel, as well as coupling with the exact equations of motion for the vessel. The vessel motion is a path in the special Euclidean group, SE(2), consisting of rotations and translation in the plane.Variational principles for the coupled motion have been given before, the first by Lukovsky  formulation are the natural continuum versions of Hamilton's principle in classical mechanics and are therefore relatively straightforward. For example, Hsieh  presentsu is studied as a function of position and time. It is the observational approach and the most widely used representation in fluid dynamics. When forecasting the wind in a city, for example, one wants to predict the velocity at a fixed spatial position as a function of time. One is not normally interested in where the parcel of fluid was the day before or the week before. This latter view, where individual fluid parcels are followed for all time, is the LPP description. The position of a parcel of fluid is given byτ = t is the representation of time in the Lagrangian setting. Here n) is a fixed reference space that may be the initial configuration of the fluid, φ = a∈B. The Lagrangian velocity field follows a fluid particle:τ (time derivative with a fixed). The two fields are related byIn the Eulerian representation, the fluid velocity y < h, where y = h represents the free surface for 0 ≤ x ≤ L. Then the natural Lagrangian, based on the kinetic minus potential energy, when the velocity is represented by a stream function, isρ is the constant fluid density and g is the gravitational constant. Taking free variations with respect to h and ψ and setting ψ and h are not the Lagrangian variables of the problem, and hence the classical Hamilton principle with free variations does not apply when such variables are used.Natural variational principles, starting with the kinetic minus the potential energy, fail in the Eulerian setting. For example, take the two-dimensional fluid domain to be in the region 0 < constrained variations induced by the free variations of the Lagrangian variables. For fluids with a fixed boundary, this is known as the Euler–Poincaré framework . However, we will need to introduce two extensions of this theory: firstly, inclusion of a free boundary h and an appropriate variation δh, and, secondly, how to induce a constrained variation, δψ, for the stream function.One thus needs to consider the m et al. , ch. 11)r fields ,14  is scalar valued and a free variation, and { · , · } is the standard -Poisson bracket for scalar valued functions. This variation, and a reduction from the LPP setting to the Eulerian setting, induces a variation at the free surfaceδψ and δh are proved in §The Euler–Poincaré framework was first extended to free boundary flows in Gay-Balmaz et al.  with a cthen use  and a Liet al. )et al.  and its q = 0 and the only vessel degree of freedom is rotation. The irrotational case has been studied in Turner et al. [A canonical problem in the study of dynamic coupling between sloshing and vessel motion is the ‘pendulum-slosh’ problem, illustrated in r et al. . Vorticiα, β, A are arbitrary positive parameters, then the exact steady solution, with a non-trivial vorticity field, ish = h0 with h0 a constant, andm, n are natural numbers. The constant in the pressure is chosen to satisfy the free surface boundary condition givingn cells in the x-direction and m cells in the y-direction. An example of the stream function contours in the case n = 2, m = 1 is shown in There is an exact solution of the sloshing problem with vorticity and a stationary vessel. Suppose the external pressure field evaluated on the free surface has the formSome experiments show evidence of vorticity. In a viscous fluid, vorticity can be generated by boundary layers and vortex shedding. For short times, it is the vorticity in initial data that will dominate. Many sloshing experiments start with a quiescent fluid (e.g. ), genera8.In this paper, a new variational principle has been introduced for two-dimensional inviscid incompressible fluid flow in a moving vessel with coupling to the vessel motion. It is a starting point for new applications and new directions .y = h simplified the analysis, but the variational principle can be generalized to the case where the surface is represented parametrically, x = X and y = Y , for 0 < s < L, thereby allowing for overhanging and a multi-valued free surface. Coupling with elastic vessel motion or elastic pendulum rod is also a potential generalization. Sloshing in microgravity can be modelled by introducing surface tension which fits neatly into the variational framework since surface tension is intrinsically linked to the variational derivative of the free surface mean curvature.The representation of the surface as a graph, et al. [The Lagrangian variational principle here opens the door to generating a Hamiltonian formulation with a Lie–Poisson structure, extending the theory in Lewis et al.  (for incet al.  (for comIn the formulation presented here, there were hints about the structure of the problem in 3D. However there are still technical difficulties in extending the fluid part to 3D. Firstly, a parametrization of divergence-free vector fields is required. The preferred option is a vector stream function, but the pressure boundary condition then presents challenges. The tangential derivative of pressure has two components and the extension of the GKK theory to 3D does not result in a conservation law (see  for the Three-dimensional cylindrical geometry has many applications. Timokha  discusse


A strong variability in cancer incidence is observed between human organs. Recently, it has been suggested that the relative contribution of organs to organism fitness  could explain at least a part of the observed variation. The objective of this study is to investigate theoretically the main factors driving the evolution of tumour resistance mechanisms of organs when their relative contribution to organism fitness is considered. We use a population-scale model where individuals can develop a tumour in a key organ , an auxiliary organ (i.e. in which only a large tumour has a relatively significant impact) or both organs because of metastasis.Our simulations show that natural selection acts in two different ways to prevent cancer in a key and an auxiliary organs. In the key organ, the strategy mostly selected is the highest resistance and only a high cost of resistance mitigates this behavior. Inversely, we observe that a low resistance strategy can be selected in the auxiliary organ when the development of the tumour is slow and the effect of a large tumour on the mortality of the organism is relatively weak. Nevertheless, if the tumour can spread to a key organ, higher resistance strategies are selected in the auxiliary organ.Finally, our study demonstrates that the relative contribution of organs to the organism fitness and the metastatic propensity of the tumour influence the evolution of tumour resistance at organ scale and should be considered by studies aiming to explain the variability in cancer incidence at organ-scale.The online version of this article (10.1186/s12862-018-1298-7) contains supplementary material, which is available to authorized users. In humans, cancer incidence strongly varies among organs Fig. A. For exVarious factors have been proposed to explain the variability of cancer incidence at organ-scale such as the different levels of inherited predispositions, carcinogen exposures or stem cell divisions –5. For Considering that natural selection acts to decrease cancer risk at organ scale, we can expect that the relative significance of organs to reproduction and survival (until reproduction ceases) have played a crucial role on the evolution of organ-specific tumour resistance mechanisms . If a cThe aim of our study is to determine the factors driving the evolution of tumour resistance mechanisms at organ scale when the relative contribution of organs to fitness is considered. We use a population-scale model in which individuals can develop a tumour in a key organ in isolation (scenario 1), in an auxiliary organ in isolation (scenario 2) or in the both organs in interaction through the metastasis (scenario 3). Our simulations show that the mortality rate due to cancer and the mortality rate extrinsic to the cancer are the main drivers of the evolution of tumour resistance mechanisms in the key organ alone and in interaction with the auxiliary organ. Conversely, the evolution of tumour resistance mechanisms in the auxiliary organ strongly depends on its interaction with the key organ. Finally, the relative contribution of organs to the fitness of the organism is likely to have played a crucial role on the current difference of organ vulnerability to cancer and therefore could explain, at least partially, the observed variability of cancer incidence among organs.We model a logistically growing population of individuals that are classified according to their cancer status as follows Fig. A:1\documN and K are the total number of individuals in the population and the carrying capacity respectively. Hrepresents healthy individuals, A and B correspond to individuals with a tumour in a key and an auxiliary organ, respectively. Among healthy individuals, we distinguish those who will never develop a tumour (H0) and individuals who are developing a tumour (H1). The proportion of individuals in each group is shaped by the parameter αwhich represents the rate of tumour emergence in the population and allow to consider both the concomitant influence of genetic susceptibility and environmental exposure to carcinogens. Among individuals with a tumour in a key or an auxiliary organ, we distinguish four tumour stages: tumour initiation , large tumour invading the major part of the organ , tumour that can migrate to other distant organs  and the metastatic stage (stage 4) which corresponds to individuals with a tumour in both the key and the auxiliary organs (M). Healthy individualsH1can develop a tumour of stage 1 in a key organ (A1) or an auxiliary organ (B1) through the tumour initiation rate λa and λb, respectively. Individuals with a tumour of stage 1 (A1 and B1) or stage 2 (A2 and B2) can develop a tumour of stage 2 or stage 3 through the rate τ12 and τ23, respectively. Finally, individuals with a tumour of stage 3 (A3 and B3) can develop a metastatic cancer (M) through the rate τ33. We assume that tumour resistance mechanisms impact only the initiation rates in the key and auxiliary organs , where low values of tumour initiation rates represent high resistance mechanisms., where ff). In other words, we consider a resistance-fecundity trade-off for which high tumour resistance mechanisms have a cost on organism fitness through a reduction in fecundity rate  corresponding to all sources of death that are not directly linked to cancer  and the related-cancer mortality rate (dxy) which depends on the organ affectedx and the tumour stagey   increases by a factor of (1 + c) the mortality of individuals:c is a constant to manage the strength of cancer impact on mortality and varies from 1 to 19  increases significantly and immediately the mortality of the organism whereas only a large tumour (stage 3) in an auxiliary organ (B) has a relatively high impact on the mortality of the organism  in a key organ . The initial population contains 104 healthy individuals (H0 and H1) and the different strategies are equally represented among these individuals. We let the population evolve until reaching a stable equilibrium . Finally, we consider as the selected resistance mechanism in the key organ (λa) or in the auxiliary organ (λb), the resistance mechanism which is the most frequent in those organs within the population at the end of the simulation.We use numerical simulations to determine the level of tumour resistance selected in a key organ and in an auxiliary organ for the different scenarios studied. For each organ, we explore 10 different levels of tumour resistance, corresponding to 100 possible strategies when the organs are in interaction (100 different pairs of λff) in the Additional file We test the effect of all parameters and functions on our results. We evaluate the effects of tumour emergence variability (α) and the effect of the shape of the fecundity-resistance trade-off (m), the cancer-related organism mortality rate (c) and the extrinsic organism mortality rate (dH) are population properties describing the stability of evolutionary strategies in the population and its vulnerability to cancer and all other sources of death. Because the mutation rate never influences the evolution of resistance mechanisms in organs, we no longer discuss on it in this paper. Second, the lethality of tumour stages (δ) is an organ property allowing the distinction between a key organ (δ < 1) and an auxiliary organ (δ > 1)  is a tumour property.The parameters used in the sensitivity analysis describe either the population, the organ or the tumour properties. First, the mutation rate ( 1) Fig. B. Moreov 1) Fig. B. Third,To evaluate the uncertainty inherent to the variability of parameters, we perform a sensitivity analysis using a Latin Hypercube Sampling (LHS) approach provided by the ‘lhs’ package in R 3.4.0. We generate 200 sets of the parameter values through the LHS algorithm, where each parameter varies uniformly from 67 to 100% around its average value, depending on the parameter (Table a and λb) by calculating the Partial Rank Correlation Coefficient (PRCC). The PRCC provides a measure of the strength of the monotonic non parametric correlation between a parameter and the selected resistance strategy, after the linear effects of the remaining parameters have been considered. The strength of the correlation is represented by the absolute value of the PRCC and the direction of the correlation by the sign of the PRCC. By combining the LHS with PRCC, we are able to reasonably assess the sensitivity of our model outcomes to the variability of initial parameters –1917–1933) leads to higher resistance strategies in the auxiliary organ to prevent the spread of tumour to the key organ  may be simultaneously involved with higher susceptibility to cancer and also with increased reproductive potentials , 22. In Second, our results reveal that the efficiency of tumour resistance selected in auxiliary organs may strongly depend on the ability of cancer cells to spread and establish in key organs. In this study, we use a model with two organs which represents the simplest organ network allowing us to study the potential effect of metastases on organ resistance. Considering that the human body is constituted by numerous organs and tissues connected by a complex network of interactions, it appears as an interesting perspective for our work to take into account a larger number of organs, although it would significantly complexify the model (one organ is represented by three equations in the equation system 1). Moreover, the spreading of cancer in our organ network occurs only after the primary tumour has reached the last stage of development (i.e. the third stage). This is a necessary assumption because the modalities of cancer cell spreading in the body are still unknown. Especially it is unclear if cancer cells start spreading with the formation of premalignant lesions, or directly after the primary tumour appears or after the time necessary for the tumour growth to be constraint by the initial environment , 30. HowdH) on the evolution tumour resistance at organ-scale. In our model, the more the extrinsic mortality rate increases and the less the cancer is a likely cause of deaths in the population. Therefore, the variation of the extrinsic mortality rate may represent the variation of the harshness of environmental conditions for our population: a high dH corresponding to a wild population dealing with harsh conditions  and a low dH corresponding to a population enjoying more comfortable conditions in a safer environment . Interestingly, we observe a positive correlation between dH and the tumour initiation rate of the key and the auxiliary organs  which suggests that populations dealing with harsh conditions invest less in tumour resistance mechanisms at organ scale than populations in safer conditions. However, this correlation can also be shaped by the allometric constraint linking fecundity and tumour resistance in our model. The increase of the extrinsic mortality may result in an increase of the selective advantage of individuals with a high fecundity rate which leads to the spread of weak resistance. Even if our model doesn’t allow disentangling the causes-consequences of the co-variations between mortality-fecundity and mortality-resistance, we show that tumour resistance mechanisms at organ-scale are strongly influenced by extrinsic mortality of the population when those mechanisms have a reproductive cost for the organism. This finding suggests that, within the same species, resistance mechanisms selected at organ-scale could differ between wild populations and populations having evolved in safe conditions (such as zoo or protected area). Moreover, our model doesn’t consider that cancer is likely to be an important indirect cause of deaths in wild populations where even a small decrease of body conditions could result in a strong reduction of abilities to find resources or to escape predators, parasites or infections [dH) and could be more fairly estimated by adding a positive interaction between the extrinsic mortality rate (dH) and the cancer-related mortality rate (c).An important aspect of our model is that the evolution of tumour resistance mechanisms is modeled at the population level, allowing us to study the effect of different levels of extrinsic mortality rate  the extended model for different tumour resistance strategies, 2) the mutation function, 3) the fecundity function, 4) the mortality function and a summary of all the functions used in the article (Table S1). (ODT 198 kb)Additional file 2:Supplementary results. This file contains a description of the effect of tumour emergence variability on the tumour resistance mechanisms selected at organ-scale. It also contains a test to determine the influence of using Gompertz function to model the trade-off between tumour initiation rate and fecundity rate in the outcomes. (ODT 1764 kb)


A low voltage 3D parallel electroosmotic flow (EOF) pump composed of two electrode layers and a fluid layer is proposed in this work. The fluid layer contains twenty parallel fluid channels and is set at the middle of the two electrode layers. The distance between fluid and electrode channels was controlled to be under 45 μm, to reduce the driving voltage. Room temperature liquid metal was directly injected into the electrode channels by syringe to form non-contact electrodes. Deionized (DI) water with fluorescent particles was used to test the pumping performance of this EOF pump. According to the experimental results, a flow rate of 5.69 nL/min was reached at a driving voltage of 2 V. The size of this pump is small, and it shows a great potential for implanted applications. This structure could be easily expanded for more parallel fluid channels and larger flow rate. Micropumps are one of the essential components in microfluidics systems . The appAccording to fluid channel type, EOF pumps can be divided into two categories: porous channel pumps and direct channel pumps. Porous channel EOF pumps could generate high flow rates at a relatively low voltage ,9,10. InTo improve the bubble and joule heat problems in direct channel EOF pumps, and to reduce electrophoresis, several solutions are proposed: alternative current (AC) EOF pumps ,16, bubbIn this work, a low voltage three-dimensional non-contact EOF pump with new 3D structure is proposed. The pump is made of PDMS, and gallium-based alloy is used to form electrodes. This type of alloy is in liquid form in room temperature, and can be injected into channels directly by syringe. This alloy is also widely used as micro-electrode , biomateAll three channel layers were designed separately. Each of the two electrode layers contains two symmetric electrode channels. For the convenience of injecting liquid metal, electrode channels were designed in a U shape, with one injection inlet and outlet. The width of those channels is 400 μm, consistently. The distance between electrodes on the same layer is 8000 μm. Fluid channels are set perpendicular to the electrodes. In this work, we use 20 parallel channels as fluid channels. All 20 channels share the same inlet and outlet. The width of these fluid channels is 116 μm each, and the gap between the adjacent channels is 50 μm. The length of fluid channel is 10500 μm. Moreover, in order to reduce the driving voltage of the pump, the membrane layers are made as thin as possible. Due to different fabrication process, the bottom thin membrane is 16μm thick, and top membrane is 43 μm thick. The dimension schematic of this pump is shown in While working, a voltage is applied between two electrodes on both top and bottom electrode layers, as shown in With the design including parallel fluid channels that are not in the same layer as the electrode channels, the number of fluid channels could be easily expanded if needed. Flow rate can be increased by making more parallel fluid channels. The distance between fluid channels and electrode channels is controlled to be under 45 μm, and this lowers the driving voltage applied to the pump. Non-contact electrode design would prevent the bubble generation due to the electrode corrosion, and increase the stability and lifetime for this pump. Also, sample contamination would be prevented for biological applications.66In20.5Sn13.5). The first step of making this pump is to prepare patterned or empty PDMS. All the channels are made by standard soft photolithography. Both top and bottom electrode layers are made by directly pouring PDMS on patterned silicon wafers. In the real fabrication process we found it difficult to make the PDMS membrane exactly as thin as the channel height. So, we combined the top thin membrane layer with fluid channel layer in fabrication process. This combined layer is made by spinning PDMS on a silicon wafer with fluid channel pattern at 1000 RPM, to form a 74 μm thick membrane. With 31 μm high channels, the membrane above fluid channel is 43 μm thick. The bottom thin membrane layer is made in the same way, except an empty silicon wafer is used and the speed is 3000 RPM, resulting in a 16 μm thick membrane. The four layers are then baked on the hot plate to solidify PDMS. The material of the chip is PDMS, and the electrode material is gallium-based alloy  was used to do the alignment. There are two aligning platforms in the machine. Two PDMS layers were stuck on these two platforms respectively and made the alignment under a microscope. Then these two platforms were taken off from the machine together with the PDMS layers and put into the plasma cleaner  to make the plasma treatment. Finally, the platforms were taken out quickly and put back to the align machine to make the final bonding of the two PDMS layers.After finishing the four-layer structure, gallium-based alloy is injected into the electrode channels to form electrodes. Copper lines are used to connect electrode with power source. Package adhesive sealant is used to seal the copper wire and liquid metal, to fasten the connection. TM . The excitation maxima of this particle is 542 nm, and the emission maxima is 612 nm. Zeiss Observer.Z1 microscope  and X-cite Series 120Q  laser source were used to observe particle movement. In the experiment, we used a large droplet to cover both the inlet and the outlet of the fluid channel. After putting a large droplet on the inlet and the outlet, the pressure-driven flow was balanced quickly in just several seconds. In each experiment, before applying voltage we made sure all the fluorescent particles were stagnant in the microchannel. Thus, pressure-driven flow can be neglected, and the fluid flow we observed was only EOF.The performance of this high flow rate 3-D EOF pump was tested in experiments. The working fluid was deionized (DI) water. Fluorescent particles with 0.52 μm diameter were added into the DI water by 1:10000 to test the flow rate. The fluorescent particles were 1% solid Red Fluorescent Polymer Microspheres from Fluoro-Maxr = 0.8 cm, h = 0.5 cm). Two platinum electrodes were put respectively at each end of the diameter of the reservoir. The diameter of the Pt electrode was 0.5 mm, so the distance between two electrodes was 0.7 cm. A 10 V voltage was added between the two electrodes. Particles moved towards and finally gathered at the positive electrode, proving that the particles are negatively charged. During the experiments, 10 particles moving directly from cathode to anode were chosen to estimate While measuring electroosmotic flow (EOF) velocity with fluorescent particles, electrophoresis (EPH) velocity needs to be considered. According to Ref. , measurehttps://consumer.dow.com/) with a value of 2.72. The dielectric constant of water and liquid metal are set according to well-known values. Thus, the electric field strength in the fluid channel is simulated. For the driving voltage of 10 V, the simulated electric field strength was In order to get the electric field strength in the experiment, an electric field simulation of this pump was performed with commercial software Comsol 5.2 . One fluid channel, two pairs of electrodes and a PDMS block are set in the model. All geometric parameters used in the simulation model are the parameters we measured in the real pump, as shown in Flow rate was then calculated with fluid channel width 116 μm, height 31 μm and channel number 20. Results are shown in The results show a linear trend between the flow rate and voltage. The error source might be the choice of particles during calculation. In electroosmotic flow, fluid velocity is higher near the solid edge, and lower in the middle of the channel. We tried to choose these 3 fluorescent particles in every channel as evenly as possible, however errors might exist.During the early stage of this experiment, a 30 μm-thick top membrane was tested, and the membrane was too easy to tear while removing the pattern from its base, because of the existence of the fluid channel. So, we chose a 43 μm-thick top membrane instead. Thinner top and bottom membranes might make a higher electric field in the fluid channel with the same voltage, but are hard to make into a successful pump. Also, even two membranes are with different thickness, the homogeneity of the electric field in fluid channels would stay the same.Placing electrodes on both sides of the fluid channels can increase the electric field strength and uniformity. To verify the merit of this structure, another experiment was performed with only one electrode layer, with the same design parameters and the same data calculation process. Results show that at 10 V, one electrode structure has a fluid velocity of 0.053 mm/min, which is only 1/8 of the velocity of the two electrode-layer structures. Thus, two-layer design makes the pumping more efficient.3 (1.5 cm × 3 cm × 2 mm). To get a similar flow rate by parallel connection 100 of that pump, the volume would be 90 cm3. The size of pump in this work is only 2 cm3 (2 cm × 2 cm × 0.5 cm), about 1/50 of that size.Compared with 2-dimentional single channel EOF pumps , the floThe largest advantages of this pump are its high flow rate and low pumping voltage. By making a thin membrane between fluid channels and electrodes, the pumping voltage can be as low as 2 V, pretty low in noncontact electrode EOF pumps. By using parallel fluid channel design, flow rate is significantly increased. Also, this is an expandable structure. For some former EOF pumps ,13,18, tThere are still limitations to this pump. Even though the flow rate has been increased over 100 times, it is still not enough. To get a 5 mL/min flow rate, 2000 V voltage is needed. To increase the flow rate, a shorter distance between electrodes on the same layer could be used.In this study, we proposed and tested a new 3-dimentioanl low voltage EOF pump. A five-layer structure is fabricated in this pump, including one fluid channel layer, two electrode channel layers and two thin membranes between them. The fluid channel layer contains 20 parallel fluid channels, lying in the middle of the two electrode layers. The distance between fluid channels and both electrode channels is under 45 μm (43 μm and 16 μm). Room temperature liquid gallium-based allay was used as the electrodes. A fluid flow rate of 248.18 nL/min was achieved at 80 V, and the lowest driving voltage is 2 V. In the future, more work would be done to lower the driving voltage and apply it in biological analysis and drug delivery systems.


An osteon consists of a multi-layered bone matrix and interstitial fluid flow in the lacunar–canalicular system. Loading-induced interstitial fluid flow in the lacunar–canalicular system is critical for osteocyte mechanotransduction and bone remodelling.To investigate the effects of the lamellar structure and heterogeneous material properties of the osteon on the distributions of interstitial fluid flow and seepage velocity, an osteon is idealized as a hollow two-dimensional poroelastic multi-layered slab model subjected to cyclic loading. Based on poroelastic theory, the analytical solutions of interstitial fluid pressure and seepage velocity in lacunar–canalicular pores were obtained.The results show that strain magnitude has a greater influence on interstitial fluid pressure than loading frequency. Interestingly, the heterogeneous distribution of permeability produces remarkable variations in interstitial fluid pressure and seepage velocity in the cross-section of cortical bone. In addition, interstitial fluid flow stimuli to osteocytes are mostly controlled by the value of permeability at the surface of the osteon rather than at the inner wall of the osteon.Interstitial fluid flow induced by cycling loading stimuli to an osteocyte housed in a lacunar–canalicular pore is not only correlated with strain amplitude and loading frequency, but also closely correlated with the spatial gradient distribution of permeability. This model can help us better understand the fluid flow stimuli to osteocytes during bone remodelling. Bone is a poroelastic material often subject to physiological cyclic loadings that arise from walking, running, or other daily activities , 2. ThesCortical bone makes up of osteons in interstitial bone tissue. The osteon is a fundamental building unit of the cortical bone at the microscopic scale Fig. a, which Since experimental work at the osteon scale does not seem feasible at the moment, various theoretical models based on Biot’s poroelastic theory have been used to quantitatively evaluate the strain-induced interstitial fluid in cortical bone. Some of the analytical models mentioned above consider the bone tissue as a transverse isotropic poroelastic material , 7, 9. OThe purpose of this study was thus twofold: firstly, to develop a multi-layered poroelastic slab model composed of multiple layers to quantify parameter values for the osteon structures. Secondly, to apply the model with calculated parameter values to investigate the specific influence of spatial gradient distribution of lacunar–canalicular permeability on fluid pressure and velocity in the lacunar–canalicular system.Biot’s poroelasticity theory is used to account for the fluid–solid interactions in this multilayer model of an osteon. After giving the description of the geometry is given and the fundamental equations of poroelasticity are stated, the boundary and initial conditions corresponding to the osteon model are specified.L in the r-direction, where each layer is assumed to be a transverse isotropic material and has a different value of permeability. The model represents the longitudinal cross section of a single cylindrical osteon. Here, we neglected the Haversian canal at the centre of the osteon. The matrix material and fluid are assumed to be compressible. We assumed that fluid can flow freely from the outer and the inner boundaries of the osteon. The model is supposed to be axisymmetric and the symmetry axis of the material is defined as z, so that the interstitial fluid flow can flow only in the radial direction. Physical quantities associated with the i-th layer are recognized by the subscript i. Additionally, all lamellae are assumed to be perfectly bonded between them. To apply axial strain loading on the osteon, two rigid and impermeable plates are placed at the top and bottom of this model.As illustrated in Fig. r-Considering the problem symmetry, the stress components are itten as , 12:1\doContinuity equation is also known as liquid mass conservation equation. The model in this paper based on cylindrical geometry of the osteon and its poroelastic properties. Material is modeled as transverse isotropic. Both fluid and solid phases are supposed to be compressible. Therefore, the flow of interstitial fluid in osteon to satisfy the continuity equation as follows :4\documez-direction.The fluid volumetric variation relation for a transversely isotropic material may be written as:Usually, bone is subjected to low-frequency cyclic loading from the activities of daily life. The equilibrium equations and the strain displacement relation can be written as follows:Substituting Eqs.  and 2)  2) into Then, substituting Eqs.  and 8)  8) into p depend only on r and t. We assume that the axial cyclic loading is in the form of an applied strain [Considering the problem symmetry and fluid flow only in the radial direction, the radial displacement entclass1pt{minimapi,0eiωt , 9, 13, hus, Eq.  can be eSubstituting Eqs.  and 8)  8) into Substituting Eq.  into theApplying ting Eq.  into Eq.ting Eq.  leads top can be described by:The osteon is in a state of balance before cyclic loading is applied, and the fluid pressure in each layer of the osteon is null:At both end of the osteon  int int19\doBy applying Cramer’s rule to Eq. , we can According the governing Eqs. –7), we , we 7), The fundamental solution for Eq.  can be wUsing the boundary condition , the folFrom the above linear equations, the unknown coefficient into Eq. , we get ith layer is derived as follows:To understand the mechanism of mechanical stimuli given to interstitial fluid flow, we investigated the seepage velocity in the lacunar–canalicular pores According to Darcy’s law , the seea = 50 μm and b = 150 μm [For the purpose of investigating the effect of spatial gradients of permeability on interstitial fluid flow and seepage velocity, Eqs.  and 25)25) are u= 150 μm , respect−17 to 10−25 m2 [−18 m2 for the lacunar–canalicular permeability (PLC) as a reference case to investigate lacunar–canalicular permeability effects on the behaviour of interstitial fluid flow. Here, we considered three cases listed in Table Among the material properties listed in Table 10−25 m2 . In our r = 100 μm, owing to the asymmetrical spatial gradient distribution of permeability. For case 2, the spatial gradient of the interstitial fluid pressure in the vicinity of the Haversian canal wall is larger than that around the cement line surface, corresponding to the spatial distribution of permeability. The variation in interstitial fluid pressure p with respect to location r is depicted in Figs. Figures r-direction in cases 1–3 at the interstitial fluid pressure peak. Figure In bone materials, the seepage velocity is closely associated with the mechanical stimuli to osteocytes during the bone remodelling process. We studied the seepage velocity distribution on strain amplitude Considering a poroelastic hollow multi-layered slab model subjected to cyclic loading in the longitudinal direction allowed us to study the effect of lamellar structure characteristics and material properties on interstitial fluid pressure and seepage velocity distribution stimuli to osteocytes embedded in an osteon.−21 m2, and Gatti et al. [−20 m2. The above two references assumed that osteon is homogeneous and listed the permeability span from several orders of magnitude (from 10−23 to 10−21 m2). But in our study, the permeability in different layer was set to be on the same order of magnitude, 10−18 m2 to suit the continuum mechanics as much as possible. Therefore, fluid velocity is index of interstitial fluid flow stimuli given to osteocytes. Setting an appropriate value of permeability according to the experimental findings can help us better understand the load induced fluid stimuli on osteocytes buried in the lacunar–canalicular system.The results in this paper have shown that spatial permeability can cause a remarkable variation in interstitial fluid and seepage velocity distributions in the osteon. To understand how bone permeability affects cellular activities in a remodelling bone, Rémond et al.  built a i et al.  found thSeepage velocity is considered one of the characteristics of mechanical stimuli to osteocytes embedded in lacunar–canalicular poro. Strain amplitude and loading frequency influence the behaviour of the interstitial fluid. Figure −18 m2 for the PLC as a representative permeability to suit the continuum mechanics [There are several limitations related to the assumptions of this model. First, our attention in this paper was only focused on the contribution of lacunar–canalicular porosity (PLC) on interstitial fluid transport and the mechanotransduction phenomenon in osteons and neglected vascular porosity (PV) and collagen–apatite porosity (PCA). However, the results of Cowin’s  study suIn summary, this analysis provides the distributions of interstitial fluid pressure and seepage velocity in cortical bone and emphasizes the importance of lamellar structure characteristics and material properties, such as permeability. Our results suggest that the amplitude strain of cyclic loading affects pore pressure and fluid velocity remarkably more than loading frequency and interstitial fluid pressure is greatly influenced by permeability variations.


Recent advances in cancer immunotherapy have mainly focused on re-activating T-cell responses against cancer cells. However, both priming and activation of effector T-cell responses against cancer-specific antigens require cross-talk with dendritic cells (DCs), which are responsible for the capturing, processing and presentation of tumour-(neo)antigens to T cells. DCs consequently constitute an essential target in efforts to generate therapeutic immunity against cancer. This review will discuss recent research that is unlocking the cancer-fighting potential of tumour-infiltrating DCs. First, the complexity of DCs in the tumour microenvironment regarding the different subsets and the difficulty of translating mouse data into equivalent human data will be briefly touched upon. Mainly, possible solutions to problems currently faced in DC-based cancer treatments will be discussed, including their infiltration into tumours, activation strategies, and antigen delivery methods. In this way, we hope to put together a broad picture of potential synergistic therapies that could be implemented to harness the full capacity of tumour-infiltrating DCs to stimulate anti-tumour immune responses in patients. It is becoming increasingly apparent that our immune system is capable of fighting cancer. Understanding the interplay between our immune system and cancer has led to the development of new treatments that can prolong survival in once-thought terminal patients. The success that immune checkpoint inhibitors (ICIs) have had in the clinic has sparked renewed interest and investment in the tumour immunology field . HoweverOn the other hand, patients who do present a high neoantigen load may not respond to ICIs due to an immunosuppressive tumour microenvironment (TME) . In thesIt is well established that DCs have the ability to link both the innate and adaptive immune systems and to initiate immune responses . In the low, MHC-II+, B220+, Siglec-H+ in mice, and represent the front line of anti-viral immune responses due to their specialised type-I interferon secretion . D. D81]. Ddication . The revte IL-12 . When CDclitaxel F 84]..+ cells 3) ,8685,86]3) . DC matu3) . Combine3) . The tum+ DCs displaying an enhanced maturation and an IL-12 secretory phenotype [Saponin-based adjuvants have also been shown to mediate strong immune responses through the activation of both Th1 and CTL subsets . These ahenotype . When a henotype . After chenotype .The aim of DC-based therapies is to create anti-tumour CTL responses that can clear the tumour mass. Therefore, besides increasing the abundance of TADCs as well as their activation state, the durability of the anti-tumour response should be ensured. There are numerous DC-vaccination strategies described in literature, some of which are currently in clinical trials. However, based on a meta-analysis of cancer vaccine trials between 1991 and 2014, it appears that the number of clinical trials peaked in 2008, and has been declining ever since. Of these trials, very few observed results that were promising enough to warrant advancing from phase II to phase III . CurrentThe success of DC vaccination largely depends on the target antigen. Currently, tumour-lysate based DC vaccines are believed to outperform tumour-associated antigen (TAA)/TAA-peptide pulsed DCs . The modIn contrast to the currently used blood-derived DC-based preparations, a strategy using tumour-derived cDC-based vaccines would not require ex vivo tumour-antigen loading, because these cells have naturally taken up tumour antigens. Indeed, tumour-derived cDC1s and cDC2s could confer protection in B16-OVA melanoma and LLC-OVA lung carcinoma tumours, respectively, when used as a prophylactic vaccination . Tumour-+CD8+ T cell-population in non-responding patients. This same population that was increased in mice allowed them to become newly responsive to PD-1 blockade, which has resulted in a follow-up trial in humans [Tumour-associated antigens can also be directed towards DCs in vivo to induce immune responses. The use of antigens coupled to mAbs or to nanoparticles specific for DC receptors can be a way of directing antigen delivery specifically to DCs. Using the artificial tumour antigen ovalbumin, a group recently showed that it was possible to specifically deliver the ovalbumin antigen when bound to a nanoparticle specific for the complement C3 receptor that is expressed by APCs. The treatment triggered T-cell responses that were able to control a murine lymphoma model that expressed the ovalbumin tumour antigen . Anothern humans . Put togDC-based treatments have great potential in tackling cancer progression. However, DC-based treatments still require a lot of tinkering. The problems of low DC number, immature DC infiltration and antigen availability all need to be addressed. Advances in the understanding of DC biology and how DCs interact with cells infiltrating the TME, open new avenues to explore potential therapeutic interventions. Caution must be taken when new treatments are being translated from mouse to human, as what is true in mice is absolutely not guaranteed to work in humans. Treatments developed to increase DC abundance within tumours can be combined with adjuvants polarising these newly recruited TADCs away from a tolerogenic phenotype and towards a robust, CTL-stimulatory state. Most importantly, any potential DC-based cancer therapy also requires combination with other treatments. Therapies targeting the TME as well as other therapies such as chemotherapy, radiotherapy, ICIs, and tyrosine kinase inhibitors all have important roles to play in cancer treatment. Tumours are capable of exerting their immune evasion mechanisms by exploiting multiple different pathways, and any future treatment should mirror this and fight cancer through the concerted actions of multiple pathways. It is with this in mind that improvements can also be made in the way treatments are evaluated within clinical trials. While survival is still the most important factor to measure, perhaps monitoring the changes to levels of important immune biomarkers can be used as another definition of success. Treatment of terminal patients with DC-based treatments may not be enough to change the course of their disease, sheerly due to the level of progression. However, that same treatment may be incredibly effective in early-stage patients.


Fluid challenge (FC) is one of the most common practices in Intensive Care Unit (ICU). The present study aimed to evaluate whether echocardiographic assessment of the response to FC at the end of the infusion or 20 min later could affect the results of the FC.T0), at the end of FC (T10), then 10 (T20) and 20 min (T30) after the end of FC.This is a prospective, observational, multicenter study including all ICU patients in septic shock requiring a FC of 500 mL crystalloids over 10 min. Fluid responsiveness was defined as a > 15% increase in stroke volume (SV) assessed by velocity-time integral (VTI) measurements at baseline , i.e., became non-responders (NR) at T30 , and 39  patients were persistent responders (PR), i.e., remained responders at T30. Among the 67 NR at T10, 4 became responders at T30, . In the subgroup analysis, no statistical difference in hemodynamic and echocardiographic parameters was found between groups.From May 20, 2014, to January 7, 2016, a total of 143 patients were enrolled in 11 French ICUs . Among the 76/143 (53%) patient responders to FC at This study shows that 51.3% of initial responders have a persistent response to fluid 30 min after the beginning of fluid infusion and only 41.3% have a transient response highlighting that fluid responsiveness is time dependent.ClinicalTrials.gov, NCT02116413. Registered on April 16, 2014The online version of this article (10.1186/s13054-019-2448-z) contains supplementary material, which is available to authorized users. Fluid therapy is the primary resuscitation maneuver of acute circulatory failure management in critically ill patients . AdequatBeyond immediate response to fluid infusion, the efficacy of a fluid bolus over time is affected by various parameters such as blood volume status, cardiac function, type of infused fluid, and capillary leak severity . Little The primary aim of the present study was to determine the proportion of patients changing from a responder status immediately after a 10-min fluid infusion to a non-responder status 20 min after the end of infusion.ClinicalTrials.gov (NCT02116413). Written consent prior to enrolment or in permitted instances, delayed participant or legal surrogate written consent following enrolment was obtained.We conducted a prospective multicenter study involving 11 intensive care units (ICUs) in 8 University Hospitals . The protocol was approved by the Nimes University Hospital local ethics committee  and registered in Adult ICU patients meeting criteria for septic shock and requiring a fluid challenge according to the treating physician were eligible for inclusion.Patient ≥ 18 year old who gave his (her) informed consent (or his (her) relative) or delayedPatient with septic shock according to the Surviving Sepsis Campaign criteria Patient on mechanical ventilation without spontaneous breathingPersistent hypotension despite fluid resuscitation of 20 to 40 mL/kg and requiring vasopressors as indicated in 2012 Surviving Sepsis Campaign criteria 2 < 70% or SvO2 < 65%.ScVO−1 during two consecutive hoursUrine output < 0.5 mL hSkin mottles−1Arterial lactate concentration > 2 mMol LPatients in whom a fluid challenge was required for the following reasons, according to physician decision:The inclusion criteria were:Informed consent not obtainedPrisonerPatient under legal guardianshipParturientSevere mitral or aortic regurgitationPatients with cardiac arrhythmiaPoor echogenicityNon-inclusion criteria were:Exclusion criteria were patients in whom a new fluid challenge was required or requiring an increase in vasopressor infusion rate before the end of the protocol.−1). Fluid responsiveness was defined as a > 15% increase in SV [Included patients were monitored with invasive arterial pressure. The fluid challenge was performed with 500-ml crystalloids (0.9% NaCl or lactated Ringer’s) over 10 min at a constant infusion rate (50 ml minT0) just before the initiation of fluid challenge until 20 min (T30) after the end of fluid challenge. This timing was chosen as fluid challenge is classically performed in 10 to 30 min [T30 was considered as non-ethical as it potentially delayed a new fluid challenge.The different studied parameters were collected from baseline , respiratory rate (RR), FiO2, positive-end expiratory pressure (PEEP), and plateau pressure (Pplat)) and the Score of Organ Failure Assessment (SOFA) were collected at patient inclusion and were not modified during study intervention.Invasive mean arterial pressure (MAP) and heart rate (HR) were continuously monitored.−1), A wave velocity (cm s−1), E/A velocity ratio) coupled to tissue Doppler imaging at the lateral mitral annulus (Ea wave velocity (cm s−1)). For right ventricular function assessment, the right to left surface telediastolic ventricular ratio and the tissue Doppler imaging at the lateral tricuspid annulus (tricuspid S wave velocity (cm s−1)) were measured [T0), at the end of fluid challenge (T10), and 10 (T20) and 20 min (T30) after the end of fluid challenge.Transthoracic echocardiography (TTE) assessment was performed by experienced operator. Patients were preferentially assessed in semi-recumbent position that was not altered during the experiment. The cardiac function was assessed on a five chamber apical view. Stroke volume and its variations during the experiment were assessed by maximal VTI and its variations over time . The maxmeasured , 18. MAPT0, T10, and T30 for measuring PaO2, PaCO2, pH, SaO2, central venous oxygen saturation (ScvO2), arterial lactate (m mol−1) and hemoglobin (g dl−1) concentrations.Blood samples were withdrawn at The primary and secondary analyses were performed according to the intention-to-treat principle. A per-protocol analysis was also performed for the primary criteria by excluding patients for whom the fluid challenge protocol was not respected.T30 among the responders at T10 . This rate was estimated with its 95% confidence interval and compared to a fixed proportion of 10% with a unilateral risk alpha.The primary endpoint was the rate of NR to fluid challenge at T10 were necessary to have a power of 80% to detect a rate of non-responders at T30 greater than 10%, with a unilateral alpha risk of 5%. Under the hypothesis that 50% of patients would be responders at T10, the inclusion of 138 patients was judged necessary. For taking a 5% rate of unusable data into account, 145 patients were included.We calculated that 69 responders to fluid challenge at Quantitative data are expressed as mean and standard deviation (SD) or median and interquartile range (IQR), according to their distribution. Qualitative data are expressed as absolute number and frequency (%).T, Wilcoxon, chi-square, or Fisher’s tests for two group comparisons. When multiple comparisons were performed, a Bonferroni correction was applied. A p value < 0.05 was considered as statistically significant. Statistical analysis was performed using R software version 3.0.2 .Comparison between groups used, when appropriate, ANOVA or Kruskal-Wallis test for three group comparisons and student’s From May 20, 2014, to January 7, 2016, 143 out of 145 patients eligible with septic shock were included . These patients were defined as transient responders (TR). Among the 76 responders, 39  remained responders (R) at T30. These patients were defined as persistent responders (PR) . The per-protocol analysis was achieved on 141 patients, and results are comparable to the results of the intention-to-treat analysis presented above (percentage estimations do not differ more than 0.6%). Hemodynamic and echocardiographic time profile of NR, TR, and PR are shown in Tables Fluid challenge induced a > 15% increase in VTI in 76/143 patients (53%) at p < 0.001)  Fig.  as well The time course of volume expansion after a fluid challenge has been poorly described. In experimental model, Guyton et al. demonstrated in normovolemic anesthetized dogs that fluid infusion increased by two to three times CO and mean circulatory pressure . In thisMoreover, in the present study, we used crystalloids for FC according to international guidelines , 15. SevConversely to previous studies focusing on the time course of fluid effects, we observed that some patients exhibited a sustained response to fluid therapy that we called “persistent response to fluid therapy.” In these patients, the hemodynamic values did not return to baseline 30 min after the start of the infusion. In PR, we could hypothesize that the efficacy of fluid is probably increased because of a significant drop in blood volume leading to very low values of hydrostatic pressure. Hahn and coworkers have nicely demonstrated that the volume efficacy of crystalloids was directly affected by blood volume status , 24. In It is unlikely that vasopressors can influence the type of fluid response, transient or persistent, in the present study as no difference was observed between TR and PR in terms of vasopressor doses.Finally, in the present population with a majority of patients with normal LVEF Table , the absOur results highlight that, among fluid responders, a few proportion of them have a sustained response to fluid. These findings enhance to closely test fluid responsiveness before administering large amounts of fluid (500 ml) and to follow this response over time while looking at the efficacy of fluid infusion on organ dysfunction. Besides, fluid responsiveness should be assessed both at the end of fluid bolus and 30 min after the start in order to identify PR and TR. Further studies are necessary to identify if different fluid strategy should be applied in TR and PR.First, the present study does not report any outcome endpoints.2 were in normal ranges at baseline, suggesting adequate blood volume and cardiac output, and ruling out indication of fluid infusion. However, during sepsis, SvO2 can be elevated due to microcirculatory alterations and may not reflect adequate tissue oxygenation explaining that fluid challenge has been performed despite normal SvO2 value in the present study. Similarly, fluid challenge has also been performed despite a mean 74 mmHg MAP value at baseline as normal MAP does not rule out potential excess of norepinephrine associated with occult hypovolemia.Second, one could argue that MAP and ScvOWe have some limitations to declare:Third, it could be objected that performing fluid responsiveness tests (such as dynamic indices or PLR test) before administering a 500-mL fluid challenge would have been more suitable in order to limit undue fluid infusion. Despite such maneuvers are very informative, they are used in less than 15% of patients in large recent cohort studies , 9. In tWe decided to use TTE to assess the response to fluid challenge even though some authors do not consider TTE as a valuable tool to assess fluid responsiveness . HoweverThis study shows that 51.3% of initial responders have a persistent response to fluid 30 min after the beginning of fluid infusion and only 41.3% have a transient response. These findings highlight that fluid responsiveness is time-dependent and that the issue of optimal timing needs to be addressed in future studies.Additional file 1:Sub-aortic velocity time integral (VTI) individual values. a In persistent responders. b In non-responders. c In transient responders. (PDF 190 kb)


Cellular Dissociation Grade (CDG) composed of tumour budding and cell nest size has been shown to independently predict prognosis in pre-therapeutic biopsies and primary resections of oesophageal squamous cell carcinoma (ESCC). Here, we aimed to evaluate the prognostic impact of CDG in ESCC after neoadjuvant therapy.We evaluated cell nest size and tumour budding activity in 122 post-neoadjuvant ESCC resections, correlated the results with tumour regression groups and patient survival and compared the results with data from primary resected cases as well as pre-therapeutic biopsies.p = 0.016).CDG remained stable when results from pre-therapeutic biopsies and post-therapeutic resections from the same patient were compared. CDG was associated with therapy response and a strong predictor of overall, disease-specific (DSS) and disease-free (DFS) survival in univariate analysis and—besides metastasis—remained the only significant survival predictor for DSS and DFS in multivariate analysis. Multivariate DFS hazard ratios reached 3.3 for CDG-G2 and 4.9 for CDG-G3 neoplasms compared with CDG-G1 carcinomas (CDG is the only morphology-based grading algorithm published to date, which in concert with regression grading, is able to contribute relevant prognostic information in the post-neoadjuvant setting of ESCC. ESCC is often associated with an aggressive disease course, and a substantial percentage of patients are in locally advanced clinical stages at the time of initial diagnosis.14 Classical WHO-based histopathologic grading of ESCC, which is based on the morphological factors keratinization, nuclear pleomorphism and mitotic activity,19 is not applied after neoadjuvant therapy, not only because chemoradiation is known to profoundly influence critical factors of this grading system such as nuclear size21 but also because its prognostic significance is highly controversial even for primary resected ESCC and de facto non-existent after neoadjuvant therapy.18In the neoadjuvant therapy setting, patient prognosis and consequently the selection of postoperative treatment modalities are mainly determined by postoperative histopathologic staging and (to a lesser extent) histopathologic tumour regression grading.24 In ESCC, Cellular Dissociation Grading has been shown to be a stage-independent predictor of the disease course when measured in pre-therapeutic biopsies25 as well as in primary resection specimens.26 However, the feasibility and the prognostic significance of the application of the Cellular Dissociation Grade in ESCC after neoadjuvant treatment has not been investigated so far.Recently, a novel grading approach termed Cellular Dissociation Grade (CDG) based on the evaluation of tumour budding and cell nest size, which are histologic factors that measure the extent of cellular dissociation either from a quantitative (tumour budding) or a qualitative (cell nest size) angle, was proposed for squamous cell carcinomas from various anatomic sites.25 and with data from a cohort of primary resected ESCC.26In order to probe for the transferability of the Cellular Dissociation Grade to the post-neoadjuvant treatment setting in ESCC, we investigated tumour budding and cell nest size in a cohort of 122 ESCC resection specimens after neoadjuvant chemoradiation and correlated the results with postoperative pathological staging , histopathologic tumour regression scoring and overall-, disease-free- as well as disease-specific survival. Furthermore, we compared the Cellular Dissociation Grade derived from resection specimen after neoadjuvant treatment with data from pre-therapeutic biopsies of the same patientsn = 53), the remaining patients  received 5-fluorouracil (5-FU) monotherapy. Of the patients with Platin-based chemotherapy, Cisplatin monotherapy  was the most common therapy approach followed by Oxaliplatin combined with 5-FU  and Carboplatin combined with Paclitaxel . The time span from the date of the initial preoperative diagnosis (as a surrogate time point for the date of initiation of chemoradiation) to surgical resection was available for 113 of the 122 patients (93%) and ranged from 51 to 168 days (mean: 95.7 days). The general surgical procedure applied for all patients with ESCC was abdomino-thoracal resection after Ivor-Lewis,27 which is the standard surgical procedure for oesophageal cancer at our hospital.Our cohort comprised resection specimens from 122 patients suffering from ESCC, who underwent resection after neoadjuvant chemoradiation. Patients were surgically treated between 1991 and 2016 at the University Hospital Rechts der Isar of the Technical University Munich. Mean patient age was 57 years (range: 30–80); male patients were more common than female ones . Neoadjuvant chemotherapy combined with radiation therapy was administered in a synchronous fashion according to the guidelines in place during the time of treatment. The exact chemoradiation scheme was available for 66 of the 122 patients (54%). Radiation dose ranged from 28.8 to 60 Grey . Sixty-three of the 66 patients received a dose of 40 Gy or more. The chemotherapy scheme was Platin-based in 80% of these patients  patients, additional data on the Cellular Dissociation Grade from pre-therapeutic biopsy specimen of the same case were available.n = 6, cT3: n = 107 and cT4: n = 7; cN0: n = 23, cN1: n = 97). Post-neoadjuvant ypTNM data were available for all patients. Histopathological assessment after chemoradiation of the respective resection specimen resulted in 47 patients with carcinomas in a rather early local stage  and 75 patients with locally advanced tumours . Vital lymph node metastases were detected in 47 patients . In total, 95 patients (77.9%) died during follow-up, 86 deaths were tumour specific. A relapse of the disease was noted in 92 (75.4%) patients during follow-up , who was blinded for clinicopathological data and the disease course. Interobserver reproducibility was probed by randomly selecting 45 cases from the cohort, that were evaluated by a second experienced pathologist , fully blinded to the results of the initial evaluation by observer 1.14 subclassifying neoadjuvantly treated ESCCs in complete responders , subtotal responders , partial responders  and nonresponders . Tumours showing complete regression could not be included in the cohort for the obvious reason that in these cases no gradeable tumour was present anymore.Furthermore, all carcinomas were divided into keratinizing, non-keratinizing or basaloid neoplasms. Tumour regression was scored in analogy to the regression score proposed by Becker et al.,The dissociation of small tumour clusters consisting of <5 neoplastic cells into the peritumoural stroma in an invasive fashion was defined as “tumour budding”. Tumour budding activity was investigated throughout the whole slide and scored within the high- power field showing the highest budding activity. The visible area in ×40 magnification was defined as a high-power field (HPF). The field diameter for one HPF (0.55 mm) was determined through division of the field number (22 ×) by the objective magnification (× 40). Low tumour budding activity was stated if 1–4 and high tumour budding activity was stated if ≥5 budding foci were identified , all carcinomas were evaluated for tumour budding within one HPF, reporting the HPF that showed the highest budding activity. In addition, the whole tumour was evaluated for the smallest cell nest size. This approach is in complete analogy to the evaluation algorithm applied in the ESCC biopsy setting.25 A score of 1 received tumours without budding activity, tumours with low budding activity (<5 buds per HPF) were assigned a score of 2 and tumours with high budding frequency (≥5 buds per HPF) received a score of 3. Regarding cell nest size, tumours with large cell nests (>15 cells) received 1 point, while tumours with intermediate (5–15 cells) and small (2–4 cells) nests were scored with 2 and 3 points, respectively. ESCCs with single-cell invasion were scored with 4 points. The sum of both variables resulted in a final grading score ranging from 2 to 7. By using this score, we composed a Cellular Dissociation Grade algorithm for ESCC after neoadjuvant treatment, which is generally identical to the grading algorithm proposed for primary resected ESCC and pre-therapeutic biopsy specimens.26 CDG-G1 (well-differentiated) ESCC had a score ranging from 2 to 3, CDG-G2 (moderately differentiated) neoplasms had a score ranging from 4 to 5 and CDG-G3 (poorly differentiated) carcinomas had a score from 6 to 7. The algorithm for Cellular Dissociation Grading of ESCC after neoadjuvant treatment is summarised in Table We established a grading system that we termed “Cellular Dissociation Grade” (CDG) by using a combined score incorporating both parameters, tumour budding and cell nest size. The modified grade is in analogy to our previous work of primary resected ESCC, that evaluated the extent of both factors in 10 HPFs.2 (chi-square) test as well as χ2 test for trends. Survival analyses were performed by using the Kaplan–Meier method; a log-rank test was used to investigate the significance of differences in survival probabilities. Multivariate survival analysis was performed by using the Cox proportional hazard model. P-values ≤ 0.05 were considered significant. As only planned hypothesis testing was performed, no corrections for multiple testing were necessary in this study.29Statistical analysis was performed by using the Statistic Package for Social Sciences 25.0 . Comparisons of tumour budding, cell nest size and Cellular Dissociation Grade between primary resections, resections after neoadjuvant therapy and pre-therapeutic biopsies were calculated with Mann–Whitney U and Wilcoxon tests. Associations of histopathological characteristics with clinicopathological parameters were calculated with χ14 were grade 3  in 49 out of 122 cases (40.2%), grade 2  in 22/122 cases (18%) and grade 1B  in 51/122 cases (41.8%).In our neoadjuvantly treated cohort, keratinizing tumours  were more frequent than non-keratinizing cancers , 19 ESCCs showed a basaloid morphology . Regression grades according to Beckerr = 0.717, p < 0.001), as seen previously in a primary resected26 and a biopsy cohort.25Tumour budding activity per 1 HPF) was distributed as follows: 15 cases (12.3%) had no budding activity, 49 tumours showed low budding (2–4 buds per one HPF) activity (40.2%) and 58 cancers had high budding (5 or more buds per one HPF) activity (47.5%). The analysis of the minimal cell nest size revealed large cell nests (>15 tumour cells) in 11 cases (9%), intermediate nests (5–15 tumour cells) were present in 4 tumours (3.2%) and 19 cases (15.6%) showed small nests (2–4 tumour cells). In the vast majority of cases , infiltrative clusters of singular tumour cells were observed and these cases were therefore classified as having single-cell invasion  and cell nest size (1–4 points) for each specimen resulted in 15 (12.3%) CDG-G1 (2–3 points) neoplasms, 19 (15.6%) CDG-G2 (4–5 points) tumours and 88 (72.1%) CDG-G3 (6–7 points) carcinomas. Figure –3 pointsn = 135) of almost equal size.26 Tumour budding was significantly higher in the neoadjuvantly treated cohort than in the primary resected population (p = 0.004). The same was true for cell nest size (p < 0.001); specifically the number of cases that showed single-cell invasion was substantially increased. Consequently, overall the group of neoadjuvantly treated cases showed significantly higher Cellular Dissociation Grades than the group of primary resected cancers (p < 0.001). This difference was likely due to the fact that full responders (ypT0)—who are not present in our neoadjuvantly treated cohort since no evaluable tumour is left—often fall into the CDG-G1/G2 category.We compared the frequency of tumour budding, cell nest size and Cellular Dissociation Grade in our neoadjuvantly treated cohort with our own previously published distribution data in a primary resected cohort . The time span between the initial diagnosis and the surgical resection was also not correlated with post-neoadjuvant Cellular Dissociation Grade (p > 0.5).In the subset of patients where the exact chemoradiation scheme was available, neither the type of chemotherapy nor the radiation dose were statistically associated with the post-neoadjuvant Cellular Dissociation Grades . As a consequence, composite Cellular Dissociation Grade was also associated with regression, minimally regressive tumours were significantly less differentiated than tumours with strong regression .Both tumour budding as well as cell nest size were strongly associated with the extent of tumour regression induced by neoadjuvant therapy. The smallest cell nest sizes and the highest budding activity was observed in those tumours that showed minimal to no regression (more than 50% viable tumour) as response to neoadjuvant treatment. In contrast, those cases with strong regression (<10% viable tumour) usually did not show single-cell invasion and strong budding activity (p < 0.05) but not with age, sex, ypT or ypM (Supplementary Table p = 0.09), but not perineural invasion (Pn) or venous invasion (V) was associated with a poorer differentiation according to CDG.When we correlated cell nest size, tumour budding and Cellular Dissociation Grade with staging data, a higher budding activity, smaller cell nest size and poorer differentiation was significantly associated with higher ypN stage  than for cell nest size , disease-specific (DSS) and disease-free (DFS) survival Table . The assp = 0.016, Table p = 0.029, Supplementary Table p = 0.150, Supplementary Table When we performed a multivariate analysis under inclusion of ypT, ypN, ypM, regression grade and Cellular Dissociation Grade, the only prognostically relevant factors that emerged were ypM and Cellular Dissociation Grade. Patients whose tumours were poorly (moderately) differentiated had a hazard ratio of 4.99 3.35) for DFS, when compared with their well-differentiated counterparts (.35 for Dp = 0.006) and DSS (p = 0.016).Although there is a certain overlap with nodal positivity, we additionally performed an exploratory multivariate analysis under inclusion of lymphatic vessel invasion (L0/L1), this resulted in an even stronger p-value for CDG for DFS  and DFS (p = 0.047) compared with well/moderately differentiated tumours (grouped because of low numbers). A clear impact on OS, however, was not observed. Regression grade 3 tumours were almost exclusively poorly differentiated .Interestingly, when we subdivided our cohort according to regression grade and stratified the subgroups for Cellular Dissociation Grade,26 as well as in pre-therapeutic biopsies of oesophageal squamous cell carcinoma,25 to the neoadjuvant setting by evaluating these factors in a large cohort of 122 ESCC specimens resected following neoadjuvant treatment. We were not only able to confirm the prognostic power of the Cellular Dissociation Grade in post-neoadjuvant resection specimens, but were also able to demonstrate that the Cellular Dissociation Grade remained the only prognostic factor for DSS/DFS besides the presence of distant metastases in a multivariate analysis incorporating postoperative pathologic staging , and tumour regression.14In this study, we investigated the transferability of Cellular Dissociation Grading based on tumour budding and cell nest size, which has already been established in primary resection specimens25The proposed grading is reproducible in this setting, which is in line with previous observations not only in biopsy specimens of ESCC but also in other tumour entities with squamous morphology.9 with consecutive surgical resection reflects the common therapy approach for locally advanced but still operable ESCC in the absence of distant metastases.8 In the postoperative setting of neoadjuvantly treated ESCC, histopathologic stage and tumour regression are considered to be among the strongest factors that determine the further course of the disease.14 Nevertheless, a considerable amount of ESCC patients with a partial or even subtotal response to neoadjuvant treatment suffer from disease relapse and consecutively die of the disease,12 underlining the need for additional biomarkers in addition to staging and tumour regression grading that allow for an improved prognostic patient stratification in the postoperative setting.Combined platinum-based chemotherapy and radiation15 and relies on traditional histomorphologic parameters such as nuclear pleomorphism, mitotic activity and degree of keratinization. In neoadjuvantly treated ESCC, WHO-based grading is usually omitted, not only because chemoradiation is known to considerably disarray relevant factors to this algorithm such as nuclear size/pleomorphism21 but also because it is almost irrelevant for clinical decision-making even in primary resected ESCC, due to its at best-limited prognostic significance.19Classical histopathologic grading of ESCC is currently performed in accordance with the algorithm of the WHO classification of tumours of the digestive system36 and other squamous cell carcinoma entities.38 Our proposed grading algorithm that has been shown to nicely discriminate prognostic subgroups in primary resected ESCCs26 as well as in pre-therapeutic ESCC biopsies25 and several other squamous cell carcinoma entities,39 pairs up tumour budding with cell nest size, a second parameter measuring cellular dissociation from a more qualitative angle. Therefore, our novel grading approach has been termed “Cellular Dissociation Grade”. While we already demonstrated preoperative Cellular Dissociation Grading performed on biopsies to be a strong pre-treatment predictor for ESCC survival independent from clinical staging in a previous study,25 the prognostic implications of Cellular Dissociation Grade in the post-neoadjuvant setting of ESCC have not been investigated until now. In this study of 122 post-neoadjuvant ESCC resection specimens, the Cellular Dissociation Grade retained its prognostic power in all survival comparisons, making the Cellular Dissociation Grade—to our knowledge—the first fully and solely morphology-based prognostic system working on tissue after neoadjuvant therapy in ESCC. The prognostic relevance of the Cellular Dissociation Grade is further underlined by our multivariate analysis incorporating the only currently known prognostic factors in the postoperative setting  of ESCC after chemoradiation, where Cellular Dissociation Grade remained the only multivariate prognosticator for DSS/DFS besides distant metastases. Even in the subset of ESCC with high tumour regression , tumour remnants showing a poor differentiation according to the Cellular Dissociation Grade were predictive of shortened DSS/DFS, which means that the grading system is able to further prognostically stratify patients even when they showed a good initial response to neoadjuvant treatment.Tumour budding, a histologic parameter that measures the quantitative extent of cellular dissociation of a neoplasia, has been identified as a highly prognostic factor in ESCCBesides the retained impact of Cellular Dissociation Grade on survival even after neoadjuvant therapy, specifically the association of the grading system with different therapy responder groups is very interesting. When we correlated tumour differentiation according to the Cellular Dissociation Grade with tumour regression grades, almost all of the nonresponders showed a poor differentiation, while well-differentiated and moderately differentiated tumours were much more common in subtotal responders, indicating that poorly differentiated tumours according to the novel grading system likely do not respond well to current chemotherapeutic approaches. This finding is supported by the association of a poor differentiation according to the Cellular Dissociation Grade in pre-therapeutic biopsies with a poor response to the neoadjuvant treatment in the respective resection specimen, although this finding needs further validation in light of the rather small subset of patients where pre- and post-neoadjuvant grading data were available. Another supporting argument is the fact that high Cellular Dissociation Grade cases enrich in the population of neoadjuvantly treated when compared with primary resected tumours, which is likely due to the fact that a certain number of cases with optimal response (ypT0) are for obvious reasons “deselected” from the neoadjuvant resection cohort evaluated here.The Cellular Dissociation Grade itself and thus the capacity of Cellular Dissociation of a given carcinoma does not seem to be strongly influenced by neoadjuvant chemoradiation, which is nicely demonstrated by the largely stable post-therapy grade in the subgroup of patients where pre-therapeutic grading data were available. This finding strongly argues against a mere selection of more poorly differentiated tumour areas of a given neoplasm by neoadjuvant therapy.The lack of predicting factors allowing us to choose the best adjuvant therapy option not only in those patients with but also in those without neoadjuvant therapy is a continuing nuisance for us and dismal for our patients. Although it has not been prospectively tested (and thus proven) regression grade—as an indirect measure of chemotherapeutic sensitivity—might have some potential in this regard. Our data suggest that CDG might be a second factor that could add some information here, since obviously CDG high-grade cases do not respond well, thus arguing that in those cases post-surgery adjuvant treatment might also be less beneficial.In conclusion, our study demonstrates that Cellular Dissociation Grading for oesophageal squamous cell carcinoma provides relevant independent prognostic information in the post-neoadjuvant setting. This makes Cellular Dissociation Grading the only morphology-based grading algorithm for ESCC published to date, which is able to independently contribute prognostic information in the post-neoadjuvant setting in concert with regression grade and pathological/clinical staging. In addition, evidence is mounting that Cellular Dissociation Grade holds potential as a predictor of response to conventional neoadjuvant chemotherapy.Supplemental Material


Recently, we were questioned  about thNew England Journal of Medicine, Cell, and Nature [http://www.partek.com/publications). Also, it is recommended by the NIH (https://ostr.cancer.gov/btep/software/partek).Partek and RLE plot were provided by Yang's group. There is no unqualified sample in GSE32537  and GSE3d Nature , 5. Topihttp://www.chem.agilent.com/cag/bsp/products/1color/OnecolorPerformanceposter_finaldraft.pdf), and we found no unqualified samples.According to the samples from GSE10667  and GSE5


The encapsulation of small molecule drugs in nanomaterials has become an increasingly popular approach to the delivery of therapeutics. The use of emulsions as templates for the synthesis of drug impregnated nanomaterials is an exciting area of research, and a great deal of progress has been made in understanding the interfacial chemistry that is critical to controlling the physicochemical properties of both the encapsulated material and the templated material. For example, control of the interfacial tension between an oil and aqueous phase is a fundamental concern when designing drug delivery vehicles that are stabilized by particulate surfactants at the fluid interface. Particles in general are capable of self-assembly at a fluid interface, with a preference for one or the other of the phases, and much work has focussed on modification of the particle properties to optimize formation and stability of the emulsion. An issue arises however when a model, single oil system is translated into more complex, real-world scenarios, which are often multi-component, with the incorporation of charged active ingredients and other excipients. The result is potentially a huge change in the properties of the dispersed phase which can lead to a failure in the capability of particles to continue to stabilize the interface. In this mini-review, we will focus on two encapsulation strategies based on the selective deposition of particles or proteins on a fluid-fluid interface: virus-like particles and polymer microcapsules formed from particle-stabilized emulsion templates. The similarity between these colloidal systems lies in the fact that particulate entities are used to stabilize fluid cores. We will focus on those studies that have described the effect of subtle changes in core composition on the self-assembly of particles at the fluid-fluid interface and how this influences the resulting capsule structure. Encapsulation in nanomaterials is a powerful approach to the delivery of active components that require protection from harsh external environments. Encapsulation strategies range from mimicking natural delivery vehicles using virus coat protein self-assembly, through to the stabilization of hydrophobic cores by inorganic nanoparticles or polymeric surfactants to form microcapsules. In the case of delivery vehicles based on a capsule structure, significant effort is directed to optimizing the interface used to template the capsule material, be it polymeric or protein in nature. Based on observations in our laboratory and others  can be forced into assemblies of 120 or 180 subunits depending on the molecular weight of the polymer  particles assembled as a monolayer at the interface of perfluorodecalin emulsions Figure ). ChemicThe use of a fluid-fluid interface as a template for the synthesis of polymer microcapsules encapsulating an active component is an exciting area and a great deal of progress has been made in understanding the interfacial chemistry that is critical to controlling the physicochemical properties of both the encapsulated material and the templated material. Control of the interfacial tension between the three phases in an emulsion system  is a fundamental concern when designing a template emulsion and one approach is to use solid nanoparticles as the stabilizer to form a Pickering emulsion. Particles in general are capable of self-assembly at a fluid interface, for example in an oil-water (o/w) emulsion, with a preference for one or the other of the phases and much work has focussed on tailoring the surface chemistry of the particle to control properties such as surface activity and to optimize emulsion properties such as stability and cargo release  sheets have been successfully used to stabilize emulsions with either a water-oil-water (w/o/w) or oil-water (o/w) morphology, depending on the oil used as the dispersed phase, via a Pickering emulsion formation , an endocrine-disrupting chemical found in a wide range of food and drink products (Wang et al., The increasing use of particle-stabilized fluid-fluid interfaces to template the formation of micro/nanostructured capsules that encapsulate an active component has ultimately led to increasing complexity of the components that make up the template emulsion. This is particularly relevant in the field of drug delivery where the drive to incorporate higher concentrations of drug increases the influence of the physicochemical properties of the drug on the emulsion core. The formation and resulting structure of nature's highly defined self-assembling nanocapsules, viruses, is highly sensitive to the composition of the core template and serves as a guiding example to other microcapsule fields. Indeed, computational simulation of assembly has shown that the strength of the interactions between viral subunits and the core, relative to inter-subunit interactions, is a governing principle on assembly around cores that do not match the preferred empty particle geometry (Elrad and Hagan, AT and FS jointly led the initial literature search and drafting. All authors contributed equally to the final writing 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.