Insect exoskeletons react to hypergravity.

A typical feature of biological materials is their ability to adapt to mechanical load. However, it is not known whether the cuticle exoskeleton, one of the most common biological structures, also shares this trait. Here, we show direct experimental evidence that prolonged exposure to hypergravity conditions affects the morphology and biomechanics of an insect exoskeleton. Locusts were raised for several weeks in a custom-designed centrifuge at various levels of hypergravity. Biomechanical measurements and X-ray microtomography show that up to 3 g load Young's modulus of the tibiae increased by about 67%. Higher gravitational loads however decreased the survival rate, body mass and endocuticle thickness. These results directly show that cuticle exoskeletons can react to hypergravity. This ability has so far only been known for bone endoskeletons and plants. Our findings thus add important context to the discussion on general ultimate factors in the evolution of adaptive biological materials and skeletal systems.

Immunomodulation with romiplostim as a second-line strategy in primary immune thrombocytopenia: The iROM study.

Thrombopoietin receptor agonists (TPO-RAs) stimulate platelet production, which might restore immunological tolerance in primary immune thrombocytopenia (ITP). The iROM study investigated romiplostim's immunomodulatory effects. Thirteen patients (median age, 31 years) who previously received first-line treatment received romiplostim for 22 weeks, followed by monitoring until week 52. In addition to immunological data, secondary end-points included the sustained remission off-treatment (SROT) rate at 1 year, romiplostim dose, platelet count and bleedings. Scheduled discontinuation of romiplostim and SROT were achieved in six patients with newly diagnosed ITP, whereas the remaining seven patients relapsed. Romiplostim dose titration was lower and platelet count response was stronger in patients with SROT than in relapsed patients. In all patients, regulatory T lymphocyte (Treg) counts increased until study completion and the counts were higher in patients with SROT. Interleukin (IL)-4, IL-9 and IL-17F levels decreased significantly in all patients. FOXP3 (Treg), GATA3 (Th2) mRNA expression and transforming growth factor-ß levels increased in patients with SROT. Treatment with romiplostim modulates the immune system and possibly influences ITP prognosis. A rapid increase in platelet counts is likely important for inducing immune tolerance. Better outcomes might be achieved at an early stage of autoimmunity, but clinical studies are needed for confirmation.

3D escape: an alternative paradigm for spatial orientation studies in insects.

Arthropods and in particular insects show a great variety of different exoskeletal sensors. For most arthropods, spatial orientation and gravity perception is not fully understood. In particular, the interaction of the different sensors is still a subject of ongoing research. A disadvantage of most of the experimental methods used to date to study the spatial orientation of arthropods in behavioral experiments is that the body or individual body parts are fixed partly in a non-natural manner. Therefore, often only the movement of individual body segments can be used to evaluate the experiments. We here present a novel experimental method to easily study 3D-escape movements in insects and analyze whole-body reaction. The animals are placed in a transparent container, filled with a lightweight substrate and rotating around two axes. To verify our setup, house crickets (Acheta domesticus) with selectively manipulated gravity-perceiving structures were analyzed. The spatial orientation behavior was quantified by measuring the time individuals took to escape toward the surface and the angular deviation toward the gravitational vector. These experiments confirm earlier results and therefore validated our experimental setup. Our new approach thus allows to investigate several comprehensive questions regarding the spatial orientation of insects and other animals.

Dasatinib for treatment of CAR T-cell therapy-related complications.

Cytokine release syndrome (CRS) and immune effector cell-associated neurotoxicity syndrome (ICANS) are severe, potentially life-threatening side effects of chimeric antigen receptor T-cell (CAR T) therapy caused by the release of cytokines by proliferating and activated CAR T-cells. Current mainstay treatment includes interleukin-1 and interleukin-6 (IL-6) blockade and steroids. The use of steroids is still controversial, since they may have the potential to irreversibly damage CAR T-cells and thus increase the risk of relapse. Therefore, additional treatment options need to be explored. We report the successful treatment of a patient with a grade 3 CRS and grade 4 ICANS refractory to IL-6 blockade and steroids with the tyrosine kinase inhibitor dasatinib. The use of dasatinib for treatment of CAR T-cell therapy-related severe complications warrants further studies.

Semi-automated differentiation of insect exo- and endocuticle in X-ray microtomography.

One of the most versatile and complex biological composite materials, the insect exoskeleton shows a huge range in biomechanical properties. The cuticle exoskeleton can be differentiated into two main histologically different layers with distinct properties: the outer, more sclerotized exocuticle and inner, softer endocuticle. For most biomechanical research questions, it is of great importance to be able to selectively characterize geometrical features of these layers. However, most conventional preparation methods (cross-sections, histological staining, SEM) require complex and destructive sample preparation, which provides only two-dimensional information. Here, we present a novel, simple staining method using X-ray microtomography to distinguish between exo- and endocuticle in a 3D environment without sample destruction. We illustrate the power of our method using locust (Locusta migratoria) hindleg tibia, a well characterized biomechanical sample. Our method allows an easy and direct measurement of exo- and endocuticle and their respective geometric features. Applying our method will help to understand the biomechanical role of exo- and endocuticle within an insect exoskeleton and will allow us to understand its composition and morphological features in more detail.

Blast counts are lower in the aspirate as compared to trephine biopsy in acute myeloid leukemia and myelodysplastic syndrome expressing CD56.

INTRODUCTION: CD56 is aberrantly expressed in myeloid neoplasms including myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML). Considering the adhesion effects of CD56, blast quantification in bone marrow might depend on the technique used to obtain respective diagnostic specimens. Therefore, the objective of our study was to investigate the impact of CD56-expression on blast counts in myeloid neoplasms comparing bone marrow aspirates to biopsies. METHODS: We retrospectively analyzed 75 patients diagnosed with MDS and AML. We compared patients with (n = 36) and without (n = 39) CD56-expression by flow cytometry with respect to their blast quantities assessed on bone marrow aspirates versus biopsies. RESULTS: The frequency of CD56-expression on blasts correlated with higher blast counts on biopsies vs. aspirate smears (rs  = 0.52; P = .001). This difference in blast counts was only significant in the CD56 high expressing subgroup (median 68%, 5.5%-95% in biopsy compared to median 32.5%, 1.5%-90% in aspirate; P < .01). The percentage of CD56-positive blasts among the total blast population was lower in the peripheral blood compared to bone marrow (median 31%, 6%-88% vs. 55%, 14%-98%; P = .016). The discrepancy in the blast count between the aspirate and trephine biopsy would have led to misclassification of four cases as MDS instead of AML, if diagnosis had based on the bone marrow aspirate blast count alone. CONCLUSION: Counting blasts in bone marrow aspirates of CD56-positive AML and MDS may be linked to underestimation, potentially leading to misclassification of these myeloid neoplasms, and should therefore be adjusted considering the results obtained on trephine biopsies for reliable diagnosis.

Insect wing damage: causes, consequences and compensatory mechanisms.

The evolution of wings has played a key role in the success of insect species, allowing them to diversify to fill many niches. Insect wings are complex multifunctional structures, which not only have to withstand aerodynamic forces but also need to resist excessive stresses caused by accidental collisions. This Commentary provides a summary of the literature on damage-reducing morphological adaptations in wings, covering natural causes of wing collisions, their impact on the structural integrity of wings and associated consequences for both insect flight performance and life expectancy. Data from the literature and our own observations suggest that insects have evolved strategies that (i) reduce the likelihood of wing damage and (ii) allow them to cope with damage when it occurs: damage-related fractures are minimized because wings evolved to be damage tolerant and, in the case of wing damage, insects compensate for the reduced aerodynamic efficiency with dedicated changes in flight kinematics.

Epidermal Cell Surface Structure and Chitin-Protein Co-assembly Determine Fiber Architecture in the Locust Cuticle.

The geometrical similarity of helicoidal fiber arrangement in many biological fibrous extracellular matrices, such as bone, plant cell wall, or arthropod cuticle, to that of cholesteric liquid mesophases has led to the hypothesis that they may form passively through a mesophase precursor rather than by direct cellular control. In search of direct evidence to support or refute this hypothesis, here, we studied the process of cuticle formation in the tibia of the migratory locust, Locusta migratoria, where daily growth layers arise by the deposition of fiber arrangements alternating between unidirectional and helicoidal structures. Using focused ion beam/scanning electron microscopy (FIB/SEM) volume imaging and scanning X-ray scattering, we show that the epidermal cells determine an initial fiber orientation, from which the final architecture emerges by the self-organized co-assembly of chitin and proteins. Fiber orientation in the locust cuticle is therefore determined by both active and passive processes.

SAUV-A Bio-Inspired Soft-Robotic Autonomous Underwater Vehicle.

Autonomous and remotely operated underwater vehicles allow us to reach places which have previously been inaccessible and perform complex repair, exploration and analysis tasks. As their navigation is not infallible, they may cause severe damage to themselves and their often fragile surroundings, such as flooded caves, coral reefs, or even accompanying divers in case of a collision. In this study, we used a shallow neural network, consisting of interlinking PID controllers, and trained by a genetic algorithm, to control a biologically inspired AUV with a soft and compliant exoskeleton. Such a compliant structure is a versatile and passive solution which reduces the accelerations induced by collisions to 56% of the original mean value acting upon the system, thus, notably reducing the stress on its components and resulting reaction forces on its surroundings. The segmented structure of this spherical exoskeleton protects the encased system without limiting the use of cameras, sensors or manipulators.

What goes up must come down: biomechanical impact analysis of falling locusts.

Many insects are able to precisely control their jumping movements. Once in the air, the properties of the actual landing site, however, are almost impossible to predict. Falling insects thus have to cope with the situation at impact. In particular, for insects jumping to escape predators, a controlled landing movement appears to be a major evolutionary advantage. A quick recovery into an upright and stable body posture minimizes the time to prepare for the next escape jump. In this study, we used high-speed recordings to investigate the falling and in particular the impact behavior of Schistocerca gregaria locusts, a common model organism for studies on the biomechanics of jumping. Detailed impact analyses of free-falling locusts show that most insects typically crashed onto the substrate. Although free-falling locusts tended to spread their legs, they mostly fell onto the head and thorax first. The presence of wings did not significantly reduce impact speed; however, it did affect the orientation of the body at impact and significantly reduced the time to recover. Our results also show that alive warm locusts fell significantly faster than inactive or dead locusts. This indicates a possible tradeoff between active control versus reduced speed. Interestingly, alive insects also tended to perform a characteristic bending movement of the body at impact. This biomechanical adaptation might reduce the rebound and shorten the time to recover. The adhesive pads also play an important role in reducing the time to recover by allowing the insect to anchor itself to the substrate.

Morphology, shape variation and movement of skeletal elements in starfish (Asterias rubens).

Starfish (order: Asteroidea) possess a complex endoskeleton composed of thousands of calcareous ossicles. These ossicles are embedded in a body wall mostly consisting of a complex collagen fiber array. The combination of soft and hard tissue provides a challenge for detailed morphological and histological studies. As a consequence, very little is known about the general biomechanics of echinoderm endoskeletons and the possible role of ossicle shape in enabling or limiting skeletal movements. In this study, we used high-resolution X-ray microscopy to investigate individual ossicle shape in unprecedented detail. Our results show the variation of ossicle shape within ossicles of marginal, reticular and carinal type. Based on these results we propose an additional classification to categorize ossicles not only by shape but also by function into 'connecting' and 'node' ossicles. We also used soft tissue staining with phosphotungstic acid successfully and were able to visualize the ossicle ultrastructure at 2-µm resolution. We also identified two new joint types in the aboral skeleton (groove-on-groove joint) and between adambulacral ossicles (ball-and-socket joint). To demonstrate the possibilities of micro-computed tomographic methods in analyzing the biomechanics of echinoderm skeletons we exemplarily quantified changes in ossicle orientation for a bent ray for ambulacral ossicles. This study provides a first step for future biomechanical studies focusing on the interaction of ossicles and soft tissues during ray movements.

Standardization of 8-color flow cytometry across different flow cytometer instruments: A feasibility study in clinical laboratories in Switzerland.

The EuroFlow Consortium developed a fully standardized flow cytometric approach from instrument settings, through antibody panel, reagents and sample preparation protocols, to data acquisition and analysis. The Swiss Cytometry Society (SCS) promoted a study to evaluate the feasibility of using such standardized measurements of 8-color data across two different flow cytometry platforms - Becton Dickinson (BD) FACSCanto II and Beckman Coulter (BC) Navios, aiming at increasing reproducibility and inter-laboratory comparability of immunophenotypic data in clinical laboratories in Switzerland. The study was performed in two phases, i.e. a learning phase (round 1) and an analytical phase (rounds 2 and 3) consisting of a total of three rounds. Overall, 10 laboratories using BD FACSCanto II (n=6) or BC Navios (n=4) flow cytometers participated. Each laboratory measured peripheral blood samples from healthy donors stained with a uniform antibody panel of reagents - EuroFlow Lymphoid Screening Tube (LST) - applying the EuroFlow standardized protocols for instrument setup and sample preparation (www.EuroFlow.org). All data files were analyzed centrally and median fluorescence intensity (MedFI) values for individual markers on defined lymphocyte subsets were recorded; variability from reference MedFI values was assessed using performance scores. Data troubleshooting and discussion of the results with the participants followed after each round at SCS meetings. The results of the learning phase demonstrated that standardized instrument setup and data acquisition are feasible in routine clinical laboratories without previous experience with EuroFlow. During the analytical phase, highly comparable data were obtained at the different laboratories using either BD FACSCanto II or BC Navios. The coefficient of variation of MedFI for 7 of 11 markers performed repeatedly below 30%. In the last study round, 89% of participants scored over 90% MedFI values within the acceptance criteria (P-score), in line with the results of the EuroFlow quality assessment rounds performed by the EuroFlow expert laboratories(Kalina et al., 2015). Central analysis of data allowed identification of deviations from the standardized procedures and technical issues (e.g. failure to perform correct instrument setup and improper compensation). In summary, here we show that inter-laboratory cross-platform standardization of 8-color flow cytometric measurements in clinical laboratories is feasible and allows for fully comparable MedFI results across BD FACSCanto II and BC Navios instruments. However, adherence to standardized protocols is crucial. Thus, training of the laboratory personnel in the EuroFlow standardized procedures is highly recommended to prevent errors in instrument setup and sample preparation.

RUNX1 Mutations Can Lead to Aberrant Expression of CD79a and PAX5 in Acute Myelogenous Leukemias: A Potential Diagnostic Pitfall.

BACKGROUND: RUNX1 is a crucial transcription factor for hematological stem cells and well-known for its association with acute lymphoblastic leukemia (ALL) and acute myelogenous leukemia (AML). Besides the translocation t(8; 21) that leads to the RUNX1-RUNX1T1 fusion, somatic mutations of RUNX1 have been discovered. METHODS: Four bone marrow trephine biopsies of patients with CD79a-positive and/or PAX5-positive acute leukemias were investigated by immunohistochemistry (IHC), karyotyping, and next-generation sequencing-based genetic analysis. Data were then compared to a historical collective of AML (n = 42) and 42 cases of AML newly diagnosed at our institution between June 2017 and May 2018. RESULTS: We report on 4 cases of acute leukemia with an equivocal immunophenotype showing expression of CD79a and/or PAX5, which led to a preliminary histopathologic classification as probable ALL/unclassifiable acute leukemia. All cases were positive for CD34 and TdT but negative for several myeloid markers on IHC. Mutational analysis revealed point mutations and indels of RUNX1 and further mutations typical for AML such as TET2, DNMT3A, and SRSF2, and 2 cases had tetrasomy 13 characteristic of RUNX1 mutant AML. CONCLUSION: Aberrant CD79a and/or PAX5 expression can be found in AML cases with RUNX1 mutations even without the translocation t(8; 21). Our series shows the expression of CD79a and PAX5 to be a potential pitfall in the classification of RUNX1 mutant acute leukemia.

Fucose-Functionalized Precision Glycomacromolecules Targeting Human Norovirus Capsid Protein.

Norovirus infection is the major cause of nonbacterial gastroenteritis in humans and has been the subject of numerous studies investigating the virus's biophysical properties and biochemical function with the aim of deriving novel and highly potent entry inhibitors to prevent infection. Recently, it has been shown that the protruding P domain dimer (P-dimer) of a GII.10 Norovirus strain exhibits two new binding sites for l-fucose in addition to the canonical binding sites. Thus, these sites provide a novel target for the design of multivalent fucose ligands as entry inhibitors of norovirus infections. In this current study, a first generation of multivalent fucose-functionalized glycomacromolecules was synthesized and applied as model structures to investigate the potential targeting of fucose binding sites in human norovirus P-dimer. Following previously established solid phase polymer synthesis, eight precision glycomacromolecules varying in number and position of fucose ligands along an oligo(amidoamine) backbone were obtained and then used in a series of binding studies applying native MS, NMR, and X-ray crystallography. We observed only one fucose per glycomacromolecule binding to one P-dimer resulting in similar binding affinities for all fucose-functionalized glycomacromolecules, which based on our current findings we attribute to the overall size of macromolecular ligands and possibly to steric hindrance.

Investigating the efficiency of a bio-inspired insect repellent surface structure.

Most insects with smooth or hairy adhesive pads have very little problems in attaching to smooth substrates. A careful selection of surface roughness, however, can effectively limit the contact area of the adhesive organs with the surface. In comparison to conventional toxin-based insect repelling methods, biologically inspired micro- and nanostructured insect repellent surface structures, thus, offer a great potential to effective and environmentally-friendly control insect pests. We here present a simple experimental approach to qualitatively and quantitatively analyse the efficiency of a micro- and nanorough surface structure. Nauphoeta cinerea and Gastrophysa viridula as model organisms for insects with smooth and hairy adhesive pads, respectively, were placed in an enclosed environment. The escape movements of freely running insects over either the repellent or a control surfaces were counted and analyzed in detail. The tested surface structures were able to significantly reduce the escape of cockroaches with smooth adhesive pads by 44.1%. Interestingly, the data indicates that N. cinerea might use mechanical cues from the antenna to discriminate between surfaces before making contact with the adhesive pads. G. viridula with hairy adhesive pads were not significantly affected by the surface structure. By carefully adjusting 'critical' surface topography parameters relevant for insect adhesion, more efficient and selective repellent surfaces might be achieved. Such nanostructure-based insect repellent surfaces could also help to utilize recruitment behavior of certain insect species and might present a novel approach to effectively control insect pests.

Damage, repair and regeneration in insect cuticle: The story so far, and possibilities for the future.

The exoskeleton of an insect can contain countless specializations across an individual, across developmental stages, and across the class Insecta. Hence, the exoskeleton's building material cuticle must perform a vast variety of functions. Cuticle displays a wide range of material properties which are determined by several known factors: the amount and orientation of the chitin fibres, the constituents and degree of cross-linking and hydration of the protein matrix, the relative amounts of exo- and endocuticle, and the shape of the structures themselves. In comparison to other natural materials such as wood and mammal bone, relatively few investigations into the mechanical properties of insect cuticle have been carried out. Of these, very few have focussed on the need for repair and its effectiveness at restoring mechanical stability to the cuticle. Insect body parts are often subject to prolonged repeated cyclic loads when running and flying, as well as more extreme "emergency" behaviours necessary for survival such as jumping, wedging (squeezing through small holes) and righting (when overturned). What effects have these actions on the cuticle itself? How close to the limits of failure does an insect push its body parts? Can an insect recover from minor or major damage to its exoskeleton "bones"? No current research has answered these questions conclusively.

Effect of sample treatment on biomechanical properties of insect cuticle.

Experimental limitations often prevent to perform biomechanical measurements on fresh arthropod cuticle samples. Hence, in many cases short- or long-term storage of samples is required. So far, it is not known whether any of the standard lab-techniques commonly used to fix or store insect cuticle samples in any way affects the biomechanical properties of the respective samples. In this paper we systematically address this question for the first time, with a focus on practical, easily accessible and common lab-methods including storage in water, ethanol, glutaraldehyde, freezing and desiccation. We performed a comprehensive and sensitive non-destructive Dynamic Mechanical Analysis (DMA) on locust hind leg tibiae using a three-point-bending setup. Our results show that from all tested treatments, freezing samples at -20 °C was the best option to maintain the original values for Young's modulus and damping properties of insect cuticle. In addition, our results indicate that the damping properties of locust hind legs might be mechanically optimized in respect to the jumping and kicking direction.

Immune-based guidance of foscarnet treatment duration in a transplant recipient with ganciclovir-resistant cytomegalovirus infection.

A lung and kidney transplant recipient underwent cytomegalovirus (CMV) primary infection with a UL97 mutation. Combined monitoring of viral load and CMV-specific CD4 T-cells allowed reduction of treatment duration with foscarnet, and illustrates how knowledge on the individual immunocompetence towards CMV may be used to individualize duration of antiviral treatment.

Bridging the gap: wound healing in insects restores mechanical strength by targeted cuticle deposition.

If an insect is injured, can it repair its skeleton in a manner which is mechanically strong and viable? Previous work has described the biological processes that occur during repair of insect cuticle, but until now, there has been no biomechanical assessment of the repaired area. We analysed the biomechanics of the injury repair process in the desert locust (Schistocerca gregaria). We show that after an incision, a healing process occurred which almost doubled the mechanical strength of locust tibial cuticle, restoring it to 66% of the original, intact strength. This repair process occurred by targeted cuticle deposition, stimulated by the presence of the injury. The cut surfaces remained unrepaired, but a patch of endocuticle was deposited, reinforcing the area and thus increasing the effective fracture toughness. The deposition rate of endocuticle inside the tibia increased fourfold compared with uninjured controls, but only on the dorsal side, where the incision was placed. The limb is highly loaded during jumping, so this partial restoration of strength will have a profound effect on the fitness of the insect. A finite-element model provided insights into the mechanics of the repair, predicting that the patch material reaches its ultimate strength before the fracture toughness of the existing cuticle is exceeded.