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Biology subjects

Alam, P.

Publications and source records attributed to Alam, P..

At least 19 recordsLinked to original sources

Deep-learning based 3D segmentation of heterogeneous lizard claw tissue from CT data

The accurate segmentation of lizard claws is important as they are materially heterogeneous, comprising both bone and keratinous tissue. This study presents a deep learning framework for the automated segmentation of lizard claw tissues, specifically bone and keratin, from CT imaging data. A dataset comprising 14 lizard claws was used in this work, with annotations generated through a superpixel based labeling approach to provide ground truth reference segmentations. To evaluate the effect of spatial context on segmentation performance, both 2D and 2.5D CNN architectures using DeepLabV3 with ResNet-50, ResNet-101, and Inception-ResNet-v2 backbones were investigated, with predictions subsequently reconstructed into three-dimensional volumes for analysis. Performance was assessed using a leave one out cross validation (LOOCV) strategy and evaluated with 3D Dice Similarity Coefficient (DSC), Intersection over Union (IoU), Sensitivity (Recall), 95th Percentile Hausdorff Distance (HD95), and Relative Volume Error (RVE). Experimental results demonstrate that 2.5D CNN architectures consistently outperform their 2D counterparts across all evaluation metrics, highlighting the importance of incorporating inter-slice contextual information for volumetric tissue segmentation. From amongst the models, the 2.5D Inception-ResNet-v2 achieved the best overall performance, reaching a validation accuracy of 97.5% and producing segmentation results that closely align with ground-truth tissue structures. Our findings demonstrate the effectiveness of 2.5D deep learning approaches for the high accuracy segmentation of heterogeneous lizard claw tissues from CT data, whilst providing a robust framework for automated morphological analysis in comparative anatomical studies.

zoology↗

AI-BioMech: Deep Learning Prediction of Mechanical Behavior in Aperiodic Biological Cellular Materials

We introduce AI-BioMech, a deep learning based framework that directly predicts the mechanical response of cellular structures from 2D images, eliminating the need for manual geometry definition and traditional finite element simulations. The framework is trained on synthetic datasets representing biological cellular structures and benchmarked against real experimental data. Finite element analysis (FEA) based labeling is used to generate pixel level annotations for semantic segmentation, enabling accurate identification of stress and strain distributions. By learning spatial and hierarchical patterns from these annotations, the model automatically extracts complex features to predict cellular material responses under compressive loading conditions. Transfer learning with fine tuning by using the DeepLabv3 architecture with ResNet50, ResNet101, and Inception ResNetV2 backbones enhances prediction accuracy and generalization from limited datasets. Model predictions are validated against experimental results and Digital Image Correlation (DIC) measurements, demonstrating strong agreement with physical observations. The results show that AI-BioMech achieves up to 99% prediction accuracy while significantly outperforming traditional methods in computational speed and scalability.

biophysics↗

Grip and grasp: lizard claw inspired robotic manipulators

This paper considers the selection of a lizard claw to apply as an end effector on a bioinspired robotic manipulator, to improve grasping efficiency. Our work draws on morphological parameters that influence mechanical grip. By considering geometrical ratios that are built on parameters such as the arm-to-tip angle, the claw inner radius, and the slip distance, our research identifies optimal parameters that influence grip strength and stability. We demonstrate that these parameters, when combined to form two specific indices (the geometric mean, IGM, and the arithmetic mean of deviations, IAMD), provide a basis for more extensive comparisons between the different claw morphologies researched. The claw of Crotaphytus collaris is deduced as being the most suitable candidate for application as a bioinspired end effector in a robotic manipulator used for grasping objects, as it closely aligns with our optimal design criteria. To practically demonstrate gripping and grasping, we use a lightweight ping pong ball as well as larger and heavier objects such as an abalone shell, boxing mitts and a printer spool. Our robotic manipulator integrates dual grasping strategies. These include pinching for delicate objects and scooping for larger, irregular objects, thus enhancing its versatility across different environments and for different applications. Overall, this paper offers valuable new insights into biomimetic manipulator design using claws as end effectors, highlighting the importance of lizard claw selection based on morphology.

bioengineering↗

Kinetics of elastic recoil in the wings of the cicada, Dundubia rufivena

In this paper, we quantitatively characterise the kinetics of elastic recoil in the wings attached to the cicada Dundubia rufivena. We use high speed videography to deduce the velocities of recoil motion in both bending (B) and torsion (T), in downstroke and upstroke modes of flapping. Elastic recoil is faster in downstroke (DS) than it is in upstroke (US), reaching average velocities, [Formula], 244 {degrees}.s-1 in DS-B vs 6434 {degrees}.s-1 in US-B, and 5887 {degrees}.s-1 in DS-T vs 4545 {degrees}.s-1 in US-T. Our results also therefore evidence that bending velocities during elastic recoil, are higher to the velocities in torsion, and conclude that this is a result of the necessary geometrical distances that need to be covered over a stroke. The wings do not act alone as biological springs, but rather, we find that the stiffnesses of the wings (1.7-3.6 GPa) are higher when attached to the cicada body, than they are when detached. This evidences thoracical involvement as part of the biological spring enabling elastic recoil, indicating that elastic recoil of flapping wings should be approached from a systems perspective, rather than solely through a localised understanding of wing mechanics.

zoology↗

Gait analysis of Pak Biawak: a necrobot lizard built using the skeleton of an Asian water monitor (Varanus salvator )

In this paper,we consider the feasibility of mimicking the sprawling gait of a live varanid (Varanus salvator) using a necrobot (named: Pak Biawak), a robot constructed using the skeletal parts of a deceased varanid of the same species. Pak Biawak is manufactured using simple joints and components, and limb motion is coupled to passive spine bending to enable the sprawling gait. Here, we assess both the lateral and dorsal kinematics of Pak Biawak at different speeds, and compare the metrics from each to those of a similarly sized live varanid. When assessing lateral view shape metrics (stride aspect ratio, stride circularity, normalised stride swept area, normalised stride swept area perimeter), we find that Pak Biawaks gait is consistent across all speeds and the majority of Pak Biawaks lateral shape metrics are kinematically aligned with those of the live varanid. This also proves true when comparing Pak Biawaks lateral trajectory metrics (radial distance of swept area, normallised root mean squared error) against those of the live varanid, and at different speeds of sprawling. Pak Biawaks dorsal metrics include the spine bending amplitude and period, and these are not found to be significantly different to those of the live varanid, however, Pak Biawaks amplitude is affected by sprawling speed. We use three metrics to compare forward and reverse limb sweeps including, angular curvature, differential curvature, and a normalised arc length. Of these, a preponderance of highly significant differences (p[≤] 0.001) are observed on comparing the forward sweep arc length of Pak Biawak at every sprawling speed against the forward sweep arc length of the live lizard. All other kinematic metrics in the necrobot are nevertheless very close to those of the live lizard. Finally, when comparing the trackway width of Pak Biawak against the live lizard, we again find there is very close kinematic compatibility between the two, and conclude that our necrobot can be designed and manufactured to mimic the sprawling gait of a real varanid, even when using simple kinematic linkages in unison with a passive spine bending differential applied at only one central location in the necrobot spine.

bioengineering↗

Rational Generation of Monoclonal Antibodies and Intrabodies Selective for Pathogenic TDP-43

TAR DNA-binding protein 43 (TDP-43), encoded by the TARDBP gene, is a ribonucleoprotein associated with the pathogenesis of amyotrophic lateral sclerosis (ALS), frontotemporal dementia (FTD), and Alzheimers disease (AD). Under physiological conditions, TDP-43 is predominantly localized in the nucleus, where it participates in a variety of cellular functions related to RNA splicing, transport, and stability, as well as miRNA biogenesis. In disease, it is disproportionately mislocalized to the cytoplasm where it forms aggregates, which contribute to neurotoxicity and prion-like cell-to-cell propagation of pathogenic TDP-43. Targeting of misfolded aggregates of TDP-43 represents an attractive therapeutic strategy. However, development of effective immunotherapeutic agents remains a challenge, as they require stringent selectivity for misfolded TDP-43 in order to maintain the essential functions of physiologically native TDP-43. To address this issue, monoclonal antibodies (mAbs) and intrabodies were generated against an epitope in the N-terminal domain of TDP-43 that is only exposed when the protein is misfolded, but not in its properly folded form. We show that mouse and rabbit mAbs against this epitope displayed high binding affinities by surface plasmon resonance analysis and selectively reacted with pathological TDP-43 in post-mortem tissues from ALS, FTD, and AD patients. In a cell line system, human embryonic kidney (HEK) 293T cells, mAbs and corresponding intrabodies specifically reacted with cytoplasmic aggregates of transfected misfolded TDP-43 lacking the nuclear localization signal, TDP-43{Delta}NLS. Functionally, mAbs inhibited cell-to-cell transmission of misfolded TDP-43 and the seeding activity of misfolded TDP-43 from FTLD brain homogenates by a novel RT-QuIC assay. Intrabodies promoted the degradation of intracellular aggregates of TDP-43 in HEK293T cells and in induced pluripotent stem cell-derived motor neurons (iPSC-MNs) from ALS patients. The results provide proof-of-concept evidence that supports selective targeting of misfolded toxic aggregates of TDP-43 as a potentially safe and effective avenue to treat neurodegenerative diseases associated with TDP-43 proteinopathy.

immunology↗

A Standardized Method for Insect Color Analyses using Open Source Software: AInsectID Version 1.1 Color Merge

The accurate representation of color is important in applications involving species identification. Environmental variations introduce inconsistencies in color perception, affecting the reliability of automated image processing algorithms. In previous work, we developed a hybrid algorithm, AInsectID Version 1.1 Color Merge, to overcome challenges posed by over-segmentation and under-segmentation in insect wing color clustering. We achieved this by using color differences between superpixels to measure homogeneity during the superpixels segmentation process. Nevertheless, our algorithm remains sensitive to environmental effects, affecting its performance and accuracy in color analyses. Here, we introduce a standard imaging method for insect species, as a pre-requisite to analysis in AInsectID Version 1.1 Color Merge. We systematically examine the effects of varying lighting conditions, angle of observation, and working distance in a controlled environment to assess their impact on the performance of the algorithm. We find that by meticulously controlling lighting, working distance, and lighting angle, we develop an evidence-based standard approach to imaging colors that is robust and repeatable. By following our standardized procedure, consistent color analyses are possible under varying environmental conditions. The method was tested using the Delta E2000 ({Delta}E) color difference metric with a threshold of 1, demonstrating that our standard approach maintains perceptual accuracy within the Just Noticeable Difference (JND) range, while improving the reliability of color analyses of insect wings in diverse environments. Finally, to validate the robustness of our standardization method, we evaluated the certainty of our results at different levels of confidence.

zoology↗

Sundoli: A necro(w)bot with multi-stiffness joints built using geared mechanical metastructures

In this communication, we design and analyse Sundoli, a necrobot (a bionically engineered robot using decreased animal parts). It is manufactured using a crow endoskeleton, supported and rearticulated by a geared mechanical metastructure to enable controllable passive deformation. The metastructures and bone braces are designed to affix the femur bone to the tibiotarsus, whilst still permitting kinematic movement between the tibiotarsus and the tarsometatarsus of the crow skeleton. The rearticulated hips function as a fulcrum between the upper and lower body parts, whilst concurrently enabling sagittal rotation of the crow skeleton about the hips. Static compression tests, finite element analyses, and in-situ tests conducted using Sundoli shows that the deformation behaviours of metastructures with and without supports are acutely sensitive to the angle of the tarsometatarsus relative to both the ground and the loading direction, highlighting the importance of designing the metastructure holistically and with consideration to the entire skeletal structure. At different loads and angles, the metastructures exhibit variable stiffnesses over their full deformational ranges, demonstrating their effectiveness in protecting the brittle biological bones. Using a metastructure as a mechanism for passive joint rearticulation enables Sundoli an ability to support a payload 8.7 times its body weight without lateral support (an 870% payload ratio) and 14 times its body weight with lateral support (a 1400% payload ratio). This payload capacity is achievable through the full range of its upper body movement in the sagittal plane throughout the full range of its upper body movement in the sagittal plane.

bioengineering↗

MCM2 mediates post-MI cardioprotection by promoting the pro-angiogenic cardiosome signaling

AbstractO_ST_ABSBackgroundC_ST_ABSIn the past decade, induced cardiac rejuvenation has emerged as a leading approach to repair cardiac injury. Recent studies demonstrate that promoting cell cycle reentry in adult cardiomyocytes (CM) enhances cardiac rejuvenation by influencing paracrine signaling. We previously demonstrated that the inhibition of two cell cycle inhibitors, Retinoblastoma 1 (Rb1) and Meis homeobox 2 (Meis2), in the adult CM enhances angiogenesis and cardiac function following ischemic injury, but the underlying mechanisms have yet to be elucidated. The goal of this study is to determine the mechanisms by which inhibition of Rb1 and Meis2 promotes cardiac rejuvenation in a mouse model of myocardial infarction. MethodsMyocardial infarction was induced in adult C57/Bl6 mice via permanent LAD occlusion followed by direct injection of either control or Rb1+Meis2 siRNA cocktail to the ischemic myocardium. MCM2 overexpression done via direct myocardial injection of an MCM2-plasmid DNA expression cassette. Cardiac function and LV wall motion was assessed via echocardiography, and fibrosis and CM hypertrophy were assessed via histology. RNA-sequencing was performed on isolated adult murine CMs with siRNA-mediated Rb1+Meis2 knockdown to delineate the downstream mechanisms. Further identification of Rb1, Meis2, and MCM2-dependent mechanisms were done using in vitro techniques in isolated CMs and HUVEC cells. ResultsWe show that siRNA-mediated knockdown of Rb1 and Meis2 in vivo reduces pathological LV remodeling and preserves cardiac structure and function in adult mouse hearts post-MI. RNA-seq analyses revealed MCM2 as a potential downstream target of Rb1/Meis2 to enhance protective paracrine signaling in primary adult CMs. Indeed, re-expression of MCM2, which is developmentally lost from neonatal to adult CM in the heart, improves cardiac function and LV wall motion while reducing myocyte hypertrophy and fibrotic scar size post-MI. Mechanistically, re-expression of MCM2 promotes the secretion of pro- angiogenic factors from adult CM, and transfer of conditioned media from MCM2 expressing CM induced vasculogenesis in HUVEC cells. Proteomic analysis of the MCM2 interactome confirmed a significant enrichment of angiogenic mediators and suggests an MCM2-dependent protein packaging of pro- angiogenic factors in CM-derived small extracellular vesicles (cardiosomes). ConclusionRe-expression of MCM2 in adult CM promotes the secretion of pro-angiogenic cardiosomes that induce paracrine revascularization of endothelial cells and mitigates cardiac injury post-MI. SUMMARY DIAGRAM O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=193 SRC="FIGDIR/small/628232v1_ufig1.gif" ALT="Figure 1"> View larger version (60K): org.highwire.dtl.DTLVardef@17eaeforg.highwire.dtl.DTLVardef@40f0f5org.highwire.dtl.DTLVardef@18b7664org.highwire.dtl.DTLVardef@109cb1c_HPS_FORMAT_FIGEXP M_FIG C_FIG

physiology↗

Genetic silencing of KCa3.1 inhibits atherosclerosis in ApoE null mice

Increased expression of KCa3.1 has been found in vascular smooth muscle (SMC), macrophages, and T cells in atherosclerotic lesions from humans and mice. Proliferating SMC cells increase the expression of KCa3.1, such that it becomes a dominant K+ channel and contributes to SMC cell migration. The efficacy of pharmacological inhibition of KCa3.1 in limiting atherosclerosis progression has been demonstrated in mice and pigs, however direct, loss-of-function, i.e. gene silencing, studies are absent. To investigate the role of KCa3.1, we used CRISPR/Cas9 to generate KCa3.1-/-Apoe-/- (DKO) mice and assessed lesion development in the brachiocephalic artery (BCA) of DKO versus Apoe-/- mice on a Western diet for 3 months. Notably, the loss of KCa3.1 did not affect serum total cholesterol or body weight. In BCAs of DKO mice, lesion size (0.036 mm{superscript 2} vs. 0.118 mm{superscript 2}, p<0.05) and relative stenosis (13.9% vs. 43.0%, p<0.05) were reduced by 70% compared to Apoe-/- mice, with no effect on medial or lumen area. Additionally, DKO mice exhibited a significant reduction in macrophage content within atherosclerotic plaques compared to Apoe-/- mice, independent of sex. In vitro migration assays further showed a significant reduction in migration of bone marrow-derived macrophages (BMDMs) from DKO mice compared to those from Apoe-/- mice. Furthermore, in vitro experiments using rat aortic smooth muscle cells (RAOSMCs) revealed significant inhibition of PDGF-BB-induced MCP1/Ccl21 expression upon KCa3.1 inhibition, while activation of KCa3.1 further enhanced MCP1/Ccl21 expression. Both in vivo and in vitro analyses showed that silencing KCa3.1 and sex had no significant effect on the collagen content of plaque. RNAseq analysis of BCA samples from DKO and Apoe-/- mice revealed PPAR-dependent signaling as a potential key mediator of the reduction in atherosclerosis due to KCa3.1 silencing. Overall, this study provides the first genetic evidence that KCa3.1 is a critical regulator of atherosclerotic lesion development and composition and provides novel mechanistic insight into the link between KCa3.1 and atherosclerosis.

physiology↗

Poka: a necro-robot beetle with a measured payload ratio of 6847%

This paper is concerned with the design, manufacture and validation of Poka, a novel millimetre-scale necro-robot aimed at bridging the performance gap between miniature robots and insects. To create Poka, we use the exoskeleton of a deceased five-horned rhinoceros beetle (Eupatorus gracilicornis) as a mechanical chassis, which is mechatronically functionalised to enable ambulation. When comparing the payload ratio, PR, of Poka against reported values of the rhinoceros beetle Xyloryctes thestalus, we find that Pokas PR is more than 2-fold higher, reaching a measured maximum of 6847% (i.e. 68.47 times its own body weight). The specific power at maximum payload, Ps,t, is nevertheless of the same order of magnitude in both Xyloryctes thestalus (0.21 W/kg) and Poka (0.28 W/kg). Pokas highest average speed, [Formula] is achieved at a PR = 2739%, after which it progressively decreases with increasing payload ratio, reaching its minimum [Formula] at maximum payload ratio. When comparing Pokas maximum measured PR of 6847% against those of sixteen other ambulating robots, we find that Pokas PR far exceeds that of any other robot to date, the highest being otherwise from SuperBot who has a PR = 530%. Pokas payload ratio is therefore the highest robot payload ratio recorded to date and we attribute this to (a) the use of the beetle body as a natural composite chassis with high specific properties, and (b) the additive manufacture of bionic beetle parts using low density but stiff polylactic acid, designed with structurally stable geometries.

bioengineering↗

The stab resistance of Bombyx mori silk cocoons

This paper considers the mechanical response of Bombyx mori silk cocoons to knife stabbing, a simple but controlled way of simulating predaceous penetration. Here, we stab test both entire cocoons (EC) and cocoon wall segments (CWS) statically and dynamically, and note that the process can be broken down in three stages. The first stage involves material deflection, the second is knife penetration, and the third is knife perforation. We find that ca. 95 % of the kinetic energy is lost during the penetration stage. There are noticeable differences in strain between the equatorial [Formula] and meridional [Formula] directions before and after the stabbing of EC specimens (p < 0.001). The apparent area of the cocoon is noted to be on average 7 % lower after stabbing than it is prior to being stabbed (p < 0.01). We find that while compression of the cocoon from stabbing results in equatorial expansion (with a Poissons ratio,{nu} = 0.25), in the meridional direction the cocoon contracts ({nu} = -0.05) thus showing auxetic behaviour. Force-deflection curves are different in CWS specimens as compared to EC specimens, and this is attributable to natural curvatures in CWS specimens remaining even after a being flattened for mounting and testing. Differences between EC and CWS specimens are also noticeable in the sizes of the stab footprints, with EC samples exhibiting 33 % smaller footprints than CWS samples (p < 0.001). We conclude that testing whole cocoon structures provides a more accurate understanding of their properties as compared to cut and flattened structures. This is because flattening cocoon wall specimens induces delamination and multiple failure zones, reducing the natural stab resistance of the material.

zoology↗

AInsectID Version 1.1: an Insect Species Identification Software Based on the Transfer Learning of Deep Convolutional Neural Networks

AInsectID Version 1.11, is a GUI operable open-source insect species identification, color processing2 and image analysis software. The software has a current database of 150 insects and integrates Artificial Intelligence (AI) approaches to streamline the process of species identification, with a focus on addressing the prediction challenges posed by insect mimics. This paper presents the methods of algorithmic development, coupled to rigorous machine training used to enable high levels of validation accuracy. Our work integrates the transfer learning of prominent convolutional neural network (CNN) architectures, including VGG16, GoogLeNet, InceptionV3, MobileNetV2, ResNet50, and ResNet101. Here, we employ both fine tuning and hyperparameter optimization approaches to improve prediction performance. After extensive computational experimentation, ResNet101 is evidenced as being the most effective CNN model, achieving a validation accuracy of 99.65%. The dataset utilized for training AInsectID is sourced from the National Museum of Scotland (NMS), the Natural History Museum (NHM) London and open source insect species datasets from Zenodo (CERNs Data Center), ensuring a diverse and comprehensive collection of insect species.

zoology↗

Fast skeletal myosin binding protein-C expression exacerbates dysfunction in heart failure

During heart failure, gene and protein expression profiles undergo extensive compensatory and pathological remodeling. We previously observed that fast skeletal myosin binding protein-C (fMyBP-C) is upregulated in diseased mouse hearts. While fMyBP-C shares significant homology with its cardiac paralog, cardiac myosin binding protein-C (cMyBP-C), there are key differences that may affect cardiac function. However, it is unknown if the expression of fMyBP-C expression in the heart is a pathological or compensatory response. We aim to elucidate the cardiac consequence of either increased or knockout of fMyBP-C expression. To determine the sufficiency of fMyBP-C to cause cardiac dysfunction, we generated cardiac-specific fMyBP-C over-expression mice. These mice were further crossed into a cMyBP-C null model to assess the effect of fMyBP-C in the heart in the complete absence of cMyBP-C. Finally, fMyBP-C null mice underwent transverse aortic constriction (TAC) to define the requirement of fMyBP-C during heart failure development. We confirmed the upregulation of fMyBP-C in several models of cardiac disease, including the use of lineage tracing. Low levels of fMyBP-C caused mild cardiac remodeling and sarcomere dysfunction. Exclusive expression of fMyBP-C in a heart failure model further exacerbated cardiac pathology. Following 8 weeks of TAC, fMyBP-C null mice demonstrated greater protection against heart failure development. Mechanistically, this may be due to the differential regulation of the myosin super-relaxed state. These findings suggest that the elevated expression of fMyBP-C in diseased hearts is a pathological response. Targeted therapies to prevent upregulation of fMyBP-C may prove beneficial in the treatment of heart failure. Significance StatementRecently, the sarcomere - the machinery that controls heart and muscle contraction - has emerged as a central target for development of cardiac therapeutics. However, there remains much to understand about how the sarcomere is modified in response to disease. We recently discovered that a protein normally expressed in skeletal muscle, is present in the heart in certain settings of heart disease. How this skeletal muscle protein affects the function of the heart remained unknown. Using genetically engineered mouse models to modulate expression of this skeletal muscle protein, we determined that expression of this skeletal muscle protein in the heart negatively affects cardiac performance. Importantly, deletion of this protein from the heart could improve heart function suggesting a possible therapeutic avenue.

physiology↗

In-silico evaluation of Mirror repeats in some selected genes ofCandida albicans

All cellular processes in a living cell are controlled by its genetic material. DNA in majority of the domains acts as a regulatory molecule by controlling various vital functions. The genetic makeup of DNA or RNA (in viruses) is unique in all the domains. Their uniqueness is determined by the presence of various types of repetitive patterns of bases. These includes inverted repeats, tandem repeats, VNTRs, palindromes etc. Among many repetitive pattern types, Mirror repeats (MR) found to be dispersed throughout the genes or genomes. These sequences are associated with various functional features like their involvement in H-DNA formation, in replication & transcription, nervous system related diseases development etc. The major focus of this investigation is to identify MR sequences from some selected genes of Candida albicans using a bioinformatics based pipeline. The approach refers to as FPCB which utilized some manual steps to extract out MR sequence from any targeted gene or genome. The current study find out that the identified Mirror repeats found to be dispersed throughout the selected genes along with variable length. Among them the maximum & minimum MR sequences were reported in the gene FAS2 (108) & HIS1 (15) respectively. The present study will be helpful to provide a new insight for molecular as well as computational based studies of Candida albicans as well as other related fungal groups.

bioinformatics↗

Mode III tear resistance of Bombyx mori silk cocoons

This paper concerns the tear properties and behaviour of Bombyx mori silk cocoons. The tear resistance of cocoon layers is found to increase progressively from the innermost layer to the outermost layer. Importantly, the increase in tear strength correlates with increased porosity, which itself affects fibre mobility. We propose a microstructural mechanism for tear failure, which begins with fibre stretching and sliding, leading to fibre piling, and eventuating in fibre fracture. The direction of fracture is then deemed to be a function of the orientation of piled fibres, which is influenced by the presence of junctions where fibres cross at different angles and which may then acts as nucleating sites for fibre piling. The interfaces between cocoon wall layers in Bombyx mori cocoon walls account for 38% of the total wall tear strength. When comparing the tear energies and densities of Bombyx mori cocoon walls against other materials, we find that the Bombyx mori cocoon walls exhibit a balanced trade-off between tear resistance and lightweightness.

zoology↗

HuR inhibition reduces post-ischemic cardiac remodeling by dampening acute inflammatory gene expression and the innate immune response

Myocardial ischemia/reperfusion (I/R) injury and the resulting cardiac remodeling is a common cause of heart failure. The RNA binding protein Human Antigen R (HuR) has been previously shown to reduce cardiac remodeling following both I/R and cardiac pressure overload, but the full extent of the HuR-dependent mechanisms within cells of the myocardium have yet to be elucidated. In this study, we applied a novel small molecule inhibitor of HuR to define the functional role of HuR in the acute response to I/R injury and gain a better understanding of the HuR-dependent mechanisms during post-ischemic myocardial remodeling. Our results show an early (two hours post-I/R) increase in HuR activity that is necessary for early inflammatory gene expression by cardiomyocytes in response to I/R. Surprisingly, despite the reductions in early inflammatory gene expression at two hours post-I/R, HuR inhibition has no effect on initial infarct size at 24-hours post-I/R. However, in agreement with previously published work, we do see a reduction in pathological remodeling and preserved cardiac function at two weeks post-I/R upon HuR inhibition. RNA-sequencing analysis of neonatal rat ventricular myocytes (NRVMs) at two hours post-LPS treatment to model damage associated molecular pattern (DAMP)-mediated activation of toll like receptors (TLRs) demonstrates a broad HuR-dependent regulation of pro-inflammatory chemokine and cytokine gene expression in cardiomyocytes. We show that conditioned media from NRVMs pre-treated with HuR inhibitor loses the ability to induce inflammatory gene expression in bone marrow derived macrophages (BMDMs) compared to NRVMs treated with LPS alone. Functionally, HuR inhibition in NRVMs also reduces their ability to induce endocrine migration of peripheral blood monocytes in vitro and reduces post-ischemic macrophage infiltration to the heart in vivo. In summary, these results suggest a HuR-dependent expression of pro-inflammatory gene expression by cardiomyocytes that leads to subsequent monocyte recruitment and macrophage activation in the post-ischemic myocardium.

molecular biology↗

Drug repurposing screens identifies compounds that inhibit α-synuclein oligomers' membrane disruption and block antibody interactions

Small soluble oligomers of the protein -synuclein (SO) have been linked to disruptions in neuronal homeostasis, contributing to the development of Parkinsons Disease (PD). While this makes SO an obvious drug target, the development of effective therapeutics against SO are challenged by its low abundance and structural and morphological complexity. Here we employ two different approaches to neutralize toxic interactions made by SOs with different cellular components. Firstly, we use available data to identify four neuronal proteins as likely candidates for SO interactions, namely Cfl1, Uchl1, Sirt2 and SerRS. However, despite promising results when immobilized, all 4 proteins only bind weakly to SO in solution in microfluidic assays, making them inappropriate for screening. In contrast, the formation of stable contacts formed between SO and vesicles consisting of anionic lipids not only mimics a likely biological role of SO but also provided a platform to screen two small molecule libraries for disruptors of these contacts. Of the 11 leads obtained in this way, 2 significantly impaired SO contacts with other proteins in a sandwich ELISA assay using SO-binding monoclonal antibodies and nanobodies. In addition, 5 of these leads suppressed -synuclein amyloid formation. Thus a repurposing screening that directly targets a key culprit in PD pathogenesis shows therapeutic potential. HighlightsO_LIThe toxic oligomer formed by -synuclein (SO) is an important drug target. C_LIO_LINeuronal proteins found by pull-down assays do not bind SOs in solution. C_LIO_LILiposome assay identifies 7 approved drugs reducing SO membrane disruption. C_LIO_LIWe identify different inhibitory mechanisms used by different compounds. C_LIO_LITwo top drug hits disrupt SO binding to oligomer-specific antibodies. C_LI Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=123 SRC="FIGDIR/small/511078v1_ufig1.gif" ALT="Figure 1"> View larger version (37K): org.highwire.dtl.DTLVardef@184611aorg.highwire.dtl.DTLVardef@33049org.highwire.dtl.DTLVardef@1573ae4org.highwire.dtl.DTLVardef@1db6f38_HPS_FORMAT_FIGEXP M_FIG C_FIG

biophysics↗