Cerebellar Granule Cells Research Articles

Second Harmonic Generation Microscopy as a Novel Intraoperative Assessment of Rat Median Nerve Injury

Nerves that are functionally injured but appear macroscopically intact pose the biggest clinical dilemma. Second Harmonic Generation (SHG) Microscopy may provide a real-time assessment of nerve damage, with the ultimate goal of allowing surgeons to accurately quantify the degree of nerve damage present. The aim of this study was to demonstrate the utility of SHG microscopy to detect nerve damage invivo in an animal model. Ten Sprague-Dawley rats were anesthetized and prepared for surgery. After surgical exposure and using a custom-made stretch applicator, the right median nerves were stretched by 20%, corresponding to a high strain injury, and held for 5 minutes. The left median nerve served as a sham control (SC), only being placed in the applicator for 5 minutes with no stretch. A nerve stimulator was used to assess the amount of stimulation required to induce a flicker and contraction of the paw. Nerves were then imaged using a multiphoton laser scanning microscope. Immediately after injury (day 0), SHG images of SC median nerves exhibited parallel collagen fibers with linear, organized alignment. In comparison with SC nerves, high strain nerves demonstrated artifacts indicative of nerve damage consisting of wavy, undulating fibers with crossing fibers and tears, as well as a decrease in the linear organization, which correlated with an increase in the mean stimulation required to induce a flicker and contraction of the paw. Second Harmonic Generation microscopy may provide the ability to detect an acute neural stretch injury in the rat median nerve. Epineurial collagen disorganization correlated with the stimulation required for nerve function. In the future, SHG may provide the ability to visualize nerve damage intraoperatively, allowing for better clinical decision-making. However, this is currently a research tool and requires further validation before translating to the clinical setting.

A Bioinspired Orthopedic Biomaterial with Tunable Mechanical Properties Based on Sintered Titanium Fibers.

Inadequate mechanical compliance of orthopedic implants can result in excessive strain of the bone interface, and ultimately, aseptic loosening. It is hypothesized that a fiber-based biometal with adjustable anisotropic mechanical properties can reduce interface strain, facilitatecontinuous remodeling, and improve implant survival under complex loads. The biometal is based on strategically layered sintered titanium fibers. Six different topologies are manufactured. Specimens are tested under compression in three orthogonal axes under 3-point bending and torsion until failure. Biocompatibility testing involves murine osteoblasts. Osseointegration is investigated by micro-computed tomography and histomorphometry after implantation in a metaphyseal trepanation model in sheep. The material demonstrates compressive yield strengths of up to 50MPa and anisotropy correlating closely with fiber layout. Samples with 75% porosity are both stronger and stiffer than those with 85% porosity. The highest bending modulus is found in samples with parallel fiber orientation, while the highest shear modulus is found in cross-ply layouts. Cell metabolism and morphology indicate uncompromised biocompatibility. Implants demonstrate robust circumferential osseointegration in vivo after 8 weeks. The biometal introduced in this study demonstrates anisotropic mechanical properties similar to bone, and excellent osteoconductivity and feasibility as an orthopedic implant material.

Long-term depression–inductive stimulation causes long-term potentiation in mouse Purkinje cells with a mutant thyroid hormone receptor

Thyroid hormones (THs) regulate gene expression by binding to nuclear TH receptors (TRs) in the cell. THs are indispensable for brain development. However, we have little knowledge about how congenital hypothyroidism in neurons affects functions of the central nervous system in adulthood. Here, we report specific TH effects on functional development of the cerebellum by using transgenic mice overexpressing a dominant-negative TR (Mf-1) specifically in cerebellar Purkinje cells (PCs). Adult Mf-1 mice displayed impairments in motor coordination and motor learning. Surprisingly, long-term depression (LTD)-inductive stimulation caused long-term potentiation (LTP) at parallel fiber (PF)-PC synapses in adult Mf-1 mice, although there was no abnormality in morphology or basal properties of PF-PC synapses. The LTP phenotype was turned to LTD in Mf-1 mice when the inductive stimulation was applied in an extracellular high-Ca2+ condition. Confocal calcium imaging revealed that dendritic Ca2+ elevation evoked by LTD-inductive stimulation is significantly reduced in Mf-1 PCs but not by PC depolarization only. Single PC messenger RNA quantitative analysis showed reduced expression of SERCA2 and IP3 receptor type 1 in Mf-1 PCs, which are essential for mGluR1-mediated internal calcium release from endoplasmic reticulum in cerebellar PCs. These abnormal changes were not observed in adult-onset PC-specific TH deficiency mice created by adeno-associated virus vectors. Thus, we propose the importance of TH action during neural development in establishing proper cerebellar function in adulthood, independent of its morphology. The present study gives insight into the cellular and molecular mechanisms underlying congenital hypothyroidism-induced dysfunctions of central nervous system and cerebellum.

Phosphacan acts as a repulsive cue in murine and rat cerebellar granule cells in a TAG-1/GD3 rafts-dependent manner.

Phosphacan, a chondroitin sulfate proteoglycan, is a repulsive cue of cerebellar granule cells. This study aims to explore the molecular mechanism. The glycosylphosphatidylinositol-anchored neural adhesion molecule TAG-1 is a binding partner of phosphacan, suggesting that the repulsive effect of phosphacan is possibly because of its interaction with TAG-1. The repulsive effect was greatly reduced on primary cerebellar granule cells of TAG-1-deficient mice. Surface plasmon resonance analysis confirmed the direct interaction of TAG-1 with chondroitin sulfate C. On postnatal days 1, 4, 7, 11, 15, and 20 and in adulthood, phosphacan was present in the molecular layer and internal granular layer, but not in the external granular layer. In contrast, transient TAG-1 expression was observed exclusively within the premigratory zone of the external granular layer on postnatal days 1, 4, 7, and 11. Boyden chamber cell migration assay demonstrated that phosphacan exerted its repulsive effect on the spontaneous and brain-derived neurotrophic factor (BDNF)-induced migration of cerebellar granule cells. The BDNF-induced migration was inhibited by MK-2206, an Akt inhibitor. The pre-treatment with a raft-disrupting agent, methyl-β-cyclodextrin, also inhibited the BDNF-induced migration, suggesting that lipid rafts are involved in the migration of cerebellar granule cells. In primary cerebellar granule cells obtained on postnatal day 7 and cultured for 7 days, the ganglioside GD3 and TAG-1 preferentially localized in the cell body, whereas the ganglioside GD1b and NB-3 localized in not only the cell body but also neurites. Pre-treatment with the anti-GD3 antibody R24, but not the anti-GD1b antibody GGR12, inhibited the spontaneous and BDNF-induced migration, and attenuated BDNF-induced Akt activation. These findings suggest that phosphacan is responsible for the repulsion of TAG-1-expressing cerebellar granule cells via GD3 rafts to attenuate BDNF-induced migration signaling.

A U-bent fiberoptic absorbance biosensor array (ArFAB) for multiplexed analyte detection

Compact and dip-type U-bent fiber optic sensor (U-FOS) probes exhibit remarkably high evanescent wave absorbance (EWA) sensitivity and are highly suitable for label-free multiplexed detection of proteins, virii and bacteria by exploiting their intrinsic optical absorption property in the UV region. Here, we show the design and development of a novel EWA based eight-channel array fiber optic absorbance biosensor (ArFAB) using U-FOS probes for multi-analyte detection or multi-sample analysis in real-time. A proof-of-concept ArFAB is designed to consist of a UV LED (λmax = 285 nm) and a highly sensitive CMOS linear image sensor coupled to a sensor probe module through fan-out (1-to-8) and parallel (8 nos) fiber bundles respectively. The sensor probe module is designed to hold and efficiently couple the light to/from 8 U-FOS probes with the help of ceramic ferrules and mating sleeves. The CMOS line sensor, with 8 coupling fibers equidistantly placed along its length, distinctly quantifies the intensity response from each of the eight U-FOS probes. The ArFAB was systematically validated for reproducible optical intensity measurements and label-free multiplexed detection of proteins and bacteria was realized. Moreover, a novel sandwich immunoassay with plasmonic labels for immunoglobulin G (IgG) with picomolar analyte detection limits at 280 nm wavelength is demonstrated. With mass-producible U-FOS probes and a low-cost optoelectronic instrumentation that can be easily customized for LSPR or any other sensing phenomenon, the ArFAB is highly promising for cost-effective biomolecular interactions, and cell analyses and clinical diagnostic applications.

Zerumbone, a major sesquiterpene from Syringa pinnatifolia Hemsl., exerts the sedative effect by regulating GABAergic nervous system

Ethnopharmacological relevanceZerumbone (ZER) is a humulane sesquiterpenoid isolated from Syringa pinnatifolia Hemsl. (SP), its content accounts for 64.7% of volatile oil and 0.86% of total ethanol extract (TEE), representing one of characteristic ingredient of SP. As a representative Mongolian medicine with anti-“Khii”, anti-asthma, and clearing-heat effects, SP has been used for the treatment of cardiovascular diseases, upset, insomnia, and other symptoms. Aim of studyPrevious results showed that TEE has sedative effect, but the pharmacological substances and its sedative mechanism remains unclear. This study aims to determine whether ZER, as one of major and characteristic sesquiterpenoids of SP, contributes to the sedative effect of SP and its underlying mechanism. Materials and methodsLocomotor activity and threshold dose of pentobarbital sodium sleep experiments were used to evaluate the sedative effects in mice. ELISA assay was used to examine the level of GABA/Glu ratio in rats hippocampus, cortex and hypothalamus tissue. The binding ability of ZER with glutamic acid decarboxylase 67 (GAD67) and Gephyrin protein were predicted by molecular docking. Western blot and Immunohistochemistry assay were used to determine the expression of GABAergic nerve system related proteins (GAD67, Gephyrin) in rat’s hypothalamus. ZER was co-administrated with flumazenil and bicuculline (GABAA antagonist) to determine whether it acts on GABAA receptor. Furthermore, MQAE assay was used to test the effect of ZER on the chloride ion concentration in cerebellar granule cells. ResultsCurrent data demonstrated that ZER dose-dependently (5–20 mg/kg) reduces the locomotor activity and sleep latency of mice, and extend sleeping time of mice. The results of ELISA showed that ZER increases the level of GABA/Glu in rats brain tissue, in particular in hypothalamus. Molecular docking results revealed that ZER has a strong affinity to GAD67 and Gephyrin protein. The Western blot and Immunohistochemistry data indicated that ZER up-regulates the expression of GAD67 and Gephyrin protein in rat’s hypothalamus. Antagonism test results demonstrated that flumazenil and bicuculline reverse the effect of ZER on threshold dose of pentobarbital sodium sleep experiments. In addition, ZER also could dose-dependently (5–20 μM) increase the chloride ion concentration in cerebellar granule cell, suggesting that ZER induces the opening of chloride channel, exerts central inhibitory effect. ConclusionZER has a significant sedative effect in mice and rat, and the effect is associated with GABAergic nervous system. The present results suggest that ZER, as one of the major bioactive ingredients of SP, contributes to the sedative effect and provide substantial evidence for its traditional use of anti-“Khii” in clinic of Syringa pinnatifolia.

A cerebellar-prepontine circuit for tonic immobility triggered by an inescapable threat.

Sudden changes in the environment are frequently perceived as threats and provoke defensive behavioral states. One such state is tonic immobility, a conserved defensive strategy characterized by powerful suppression of movement and motor reflexes. Tonic immobility has been associated with multiple brainstem regions, but the underlying circuit is unknown. Here, we demonstrate that a strong vibratory stimulus evokes tonic immobility in larval zebrafish defined by suppressed locomotion and sensorimotor responses. Using a circuit-breaking screen and targeted neuron ablations, we show that cerebellar granule cells and a cluster of glutamatergic ventral prepontine neurons (vPPNs) that express key stress-associated neuropeptides are critical components of the circuit that suppresses movement. The complete sensorimotor circuit transmits information from sensory ganglia through the cerebellum to vPPNs to regulate reticulospinal premotor neurons. These results show that cerebellar regulation of a neuropeptide-rich prepontine structure governs a conserved and ancestral defensive behavior that is triggered by an inescapable threat.

Prenatal and postnatal development of the rat cerebellar granule cells following maternal administration of thyme and tramadol

Prenatal and postnatal development of the rat cerebellar granule cells following maternal administration of thyme and tramadol

Selective transduction of cerebellar Purkinje and granule neurons using delivery of AAV-PHP.eB and AAVrh10 vectors at axonal terminal locations.

Adeno-associated virus (AAV)-based brain gene therapies require precision without off-targeting of unaffected neurons to avoid side effects. The cerebellum and its cell populations, including granule and Purkinje cells, are vulnerable to neurodegeneration; hence, conditions to deliver the therapy to specific cell populations selectively remain challenging. We have investigated a system consisting of the AAV serotypes, targeted injections, and transduction modes (direct or retrograde) for targeted delivery of AAV to cerebellar cell populations. We selected the AAV-PHP.eB and AAVrh10 serotypes valued for their retrograde features, and we thoroughly examined their cerebellar transduction pattern when injected into lobules and deep cerebellar nuclei. We found that AAVrh10 is suitable for the transduction of neurons in the mode highly dependent on placing the virus at axonal terminals. The strategy secures selective transduction for granule cells. The AAV-PHP.eB can transduce Purkinje cells and is very selective for the cell type when injected into the DCN at axonal PC terminals. Therefore, both serotypes can be used in a retrograde mode for selective transduction of major neuronal types in the cerebellum. Moreover, our in vivo transduction strategies are suitable for pre-clinical protocol development for gene delivery to granule cells by AAVrh10 and Purkinje cells by AAV-PHPeB.

Morphological Determinants of Cell-to-Cell Variations in Action Potential Dynamics in Substantia Nigra Dopaminergic Neurons.

Action potential (AP) shape is a critical electrophysiological parameter, in particular because it strongly modulates neurotransmitter release. As it greatly varies between neuronal types, AP shape is also used to distinguish neuronal populations. For instance, AP duration ranges from hundreds of microseconds in cerebellar granule cells to 2-3 ms in SNc dopaminergic (DA) neurons. While most of this variation across cell types seems to arise from differences in the voltage- and calcium-gated ion channels expressed, a few studies suggested that dendritic morphology also affects AP shape. AP duration also displays significant variability in a same neuronal type, although the determinants of these variations are poorly known. Using electrophysiological recordings, morphological reconstructions, and realistic Hodgkin-Huxley modeling, we investigated the relationships between dendritic morphology and AP shape in rat SNc DA neurons from both sexes. In this neuronal type where the axon arises from an axon-bearing dendrite (ABD), the duration of the somatic AP could be predicted from a linear combination of the ABD and non-ABDs' complexities. Dendrotomy experiments and simulation showed that these correlations arise from the causal influence of dendritic topology on AP duration, due in particular to a high density of sodium channels in the somatodendritic compartment. Surprisingly, computational modeling suggested that this effect arises from the influence of sodium currents on the decaying phase of the AP. Consistent with previous findings, these results demonstrate that dendritic morphology plays a major role in defining the electrophysiological properties of SNc DA neurons and their cell-to-cell variations.SIGNIFICANCE STATEMENT Action potential (AP) shape is a critical electrophysiological parameter, in particular because it strongly modulates neurotransmitter release. AP shape (e.g., duration) greatly varies between neuronal types but also within a same neuronal type. While differences in ion channel expression seem to explain most of AP shape variation across cell types, the determinants of cell-to-cell variations in a same neuronal type are mostly unknown. We used electrophysiological recordings, neuronal reconstruction, and modeling to show that, because of the presence of sodium channels in the somatodendritic compartment, a large part of cell-to-cell variations in somatic AP duration in substantia nigra pars compacta dopaminergic neurons is explained by variations in dendritic topology.

Point-by-point inscribed sapphire parallel fiber Bragg gratings in a fully multimode system for multiplexed high-temperature sensing.

We study the point-by-point inscription of sapphire parallel fiber Bragg gratings (sapphire pFBGs) in a fully multimode system. A parallel FBG is shown to be critical in enabling detectable and reliable high-order grating signals. The impacts of modal volume, spatial coherence, and grating location on reflectivity are examined. Three cascaded seventh-order pFBGs are fabricated in one sapphire fiber for wavelength multiplexed temperature sensing. Using a low-cost, fully multimode 850-nm interrogator, reliable measurement up to 1500°C is demonstrated.

Adhesive Wear and Frictional Performance of Banana Fibre Reinforced Epoxy (BaFRE) Composite.

This current work is an attempt to investigate the tribological performance of banana fibre-reinforced epoxy (BaFRE) composite in dry contact conditions. The test is conducted on a wear test rig of type Block on Ring (BOR) based on ASTM G77, G137-95 standard. Different applied normal loads (5–30 N) subjected to a stainless steel counterface at different sliding speeds (1.7–3.96 m/s) and fixed sliding distance of 6.72 km were set as the experimental parameters. The test was conducted using neat epoxy (NE) as the control tests’ specimen while BaFRE composite was in anti-parallel (AP) and parallel (P) fibre strips orientation (O). The specific wear rate (Ws), friction coefficient, temperature variation and surface roughness (RA) of the NE and BaFRE composite in AP-O and P-O were investigated. The worn surface morphology of the test samples was examined under a high-resolution metallurgy microscope. The outcome of the work revealed that AP-O exhibited excellent wear performance when compared to P-O and NE. Moreover, the lowest friction coefficient of 0.0121 is achieved when AP-O is subjected to 30 N of applied load and 3.96 m/s of sliding velocity. BaFRE composite in AP-O demonstrated the lowest range of temperature variation when it was subjected to 30 N at 3.96 m/s of counterface sliding velocity. Due to the high shear resistance at the rubbing zone incurred by the AP-O test specimen and counterface, the RA values were remarkably high for the test specimen (i.e., 1.122 µm) and the counterface wear track zone (0.166 µm) as compared to the P-O and NE test samples. The predominant wear mechanism for the AP-O test specimen was plastic deformation, detached fibre, micro crack, minor fibre debonding and micro ploughing. In conclusion, there are improvements in terms of wear and friction performance of the composite when banana fibres are used as a reinforced element in epoxy resin. The improvement in Ws and friction coefficient for the AP test specimen was 29.4% and 48.6%, respectively, as compared to NE at 30 N of applied load, 6.72 km of sliding distance and 2.83 m/s of sliding speed.

Numerical Simulation of Convective Diffusion of Point Particles in a Laminar Flow Past a Row of Profiled Hollow Fibers

The numerical modeling of transverse laminar flow past a new type of hollow-fiber membranes with external profiling has been performed. A model system of parallel fibers with symmetrical parallel protrusion obstacles or grooves is considered. The absorption of point particles (solute or gas molecules) from a laminar transverse flow of a viscous incompressible liquid (gas) is calculated for a row of fibers, and the dependences of the efficiency of retention of particles by fibers on the Peclet (Pe), Reynolds (Re), and Schmidt (Sc) numbers and on the distance between neighbor fibers in a row are determined. The flow velocity and concentration fields are calculated by numerical solution of the Navier–Stokes equations and the convective diffusion equation in a wide range of Peclet numbers Pe = 0.1 − 105 for Sc = 1, 10, 1000 and Re ≤ 100.

Rac-deficient cerebellar granule neurons die before they migrate to the internal granule layer

Granule neurons are the most common cell type in the cerebellum. They are generated in the external granule layer and migrate inwardly, forming the internal granule layer. Small Rho GTPases play various roles during development of the nervous system and may be involved in generation, differentiation and migration of granule neurons. We deleted Rac1, a member of small Rho GTPases, by GFAP-Cre driver in cerebellar granule neurons and Bergmann glial cells. Rac1flox/flox; Cre mice showed impaired migration and slight reduction in the number of granule neurons in the internal granule layer. Deletion of both Rac1 and Rac3 resulted in almost complete absence of granule neurons. Rac-deficient granule neurons differentiated into p27 and NeuN-expressing post mitotic neurons, but died before migration to the internal granule layer. Loss of Rac3 has little effect on granule neuron development. Rac1flox/flox; Rac3+/−; Cre mice showed intermediate phenotype between Rac1flox/flox; Cre and Rac1flox/flox; Rac3−/−; Cre mice in both survival and migration of granule neurons. Rac3 itself seems to be unimportant in the development of the cerebellum, but has some roles in Rac1-deleted granule neurons. Conversely, overall morphology of Rac1+/flox; Rac3−/−; Cre cerebella was normal. One allele of Rac1 is therefore thought to be sufficient to promote development of cerebellar granule neurons.

P16-06 Effect of Imidacloprid on NMDA receptor gene expression and glutamate-induced toxicity in cultured mouse cerebellar granule neurons

P16-06 Effect of Imidacloprid on NMDA receptor gene expression and glutamate-induced toxicity in cultured mouse cerebellar granule neurons

Bacillus subtilis programs the differentiation of intestinal secretory lineages to inhibit Salmonella infection.

The role of intestinal microbiota on fate determination of intestinal epithelial cells has not been extensively examined. In this study, we explore the effect of Bacillus subtilis on programmed intestinal epithelial differentiation. We find that B.subtilis stimulates the differentiation of intestinal secretory cells. Moreover, B.subtilis inhibits the Notch pathway to reduce the expression of hairy and enhancer of split 1, thereby shifting intestinal stem cell differentiation toward a secretory cell fate. Moreover, we demonstrate that the programming effect of B.subtilis on intestinal differentiation is Toll-like receptor 2 pathway dependent. B.subtilis is associated with increased numbers of Paneth and goblet cells in the intestine. This results in the production of antimicrobial peptides to protect the intestinal mucosal barrier against Salmonella typhimurium. This study demonstrates that B.subtilis contributes to the differentiation of secretory cells by affecting Notch pathway signaling to maintain the intestinal barrier.

P75NTR prevents the onset of cerebellar granule cell migration via RhoA activation.

Neuronal migration is one of the fundamental processes during brain development. Several neurodevelopmental disorders can be traced back to dysregulated migration. Although substantial efforts have been placed in identifying molecular signals that stimulate migration, little is known about potential mechanisms that restrict migration. These restrictive mechanisms are essential for proper development since it helps coordinate the timing for each neuronal population to arrive and establish proper connections. Moreover, preventing migration away from a proliferative niche is necessary in maintaining a pool of proliferating cells until the proper number of neuronal progenitors is attained. Here, using mice and rats, we identify an anti-migratory role for the p75 neurotrophin receptor (p75NTR) in cerebellar development. Our results show that granule cell precursors (GCPs) robustly express p75NTR in the external granule layer (EGL) when they are proliferating during postnatal development, however, they do not express p75NTR when they migrate either from the rhombic lip during embryonic development or from the EGL during postnatal development. We show that p75NTR prevented GCP migration by maintaining elevated levels of active RhoA. The expression of p75NTR was sufficient to prevent the migration of the granule cells even in the presence of BDNF (brain-derived neurotrophic factor), a well-established chemotactic signal for this cell population. Our findings suggest that the expression of p75NTR might be a critical signal that stops and maintains the GCPs in the proliferative niche of the EGL, by promoting the clonal expansion of cerebellar granule neurons.

Immune Microenvironment and Lineage Tracing Help to Decipher Rosette-Forming Glioneuronal Tumors: A Multi-Omics Analysis.

Rosette-forming glioneuronal tumors (RGNT) are rare low-grade primary central nervous system (CNS) tumors. The methylation class (MC) RGNT (MC-RGNT) delineates RGNT from other neurocytic CNS tumors with similar histological features. We performed a comprehensive molecular analysis including whole-exome sequencing, RNAseq, and methylome on 9 tumors with similar histology, focusing on the immune microenvironment and cell of origin of RGNT. Three RGNT in this cohort were plotted within the MC-RGNT and characterized by FGFR1 mutation plus PIK3CA or NF1 mutations. RNAseq analysis, validated by immunohistochemistry, identified 2 transcriptomic groups with distinct immune microenvironments. The "cold" group was distinguishable by a low immune infiltration and included the 3 MC-RGNT and 1 MC-pilocytic astrocytoma; the "hot" group included other tumors with a rich immune infiltration. Gene set enrichment analysis showed that the "cold" group had upregulated NOTCH pathway and mainly oligodendrocyte precursor cell and neuronal phenotypes, while the "hot" group exhibited predominantly astrocytic and neural stem cell phenotypes. In silico deconvolution identified the cerebellar granule cell lineage as a putative cell of origin of RGNT. Our study identified distinct tumor biology and immune microenvironments as key features relevant to the pathogenesis and management of RGNT.

A 3D Monte Carlo Simulation of Convective Diffusional Deposition of Ultrafine Particles on Fiber Surfaces

The microscale simulation of ultrafine particle transport and deposition in fibrous filtration media was achieved with a novel particle tracking model using a 3D Monte Carlo model. The particle deposition process is governed by the convection–diffusion field. Simulations were performed by considering the fibrous filtration media as an array of identical parallel fibers, in which the flow field was accurately described by an analytical solution. The model of particle movement was described by the random probability distribution characterized by a dimensionless factor, the Peclet number (Pe), based on a convection–diffusive equation of particle transport in fluid. The influence of the particle Peclet number (Pe) on the particle deposition process and the resulting deposition morphology was investigated. The results were analyzed in terms of dust layer growth, particles’ trajectories and dust layer porosity for a vast range of Peclet numbers. The development of distinct deposition morphologies was found by varying the Peclet number (Pe). With a small Peclet number, diffusion dominated deposition and led to the formation of a more open and looser dust layer structure, while with larger Peclet numbers, convection dominated deposition and was found to form compact deposits. According to the change in the location of the packing densities along the dust layer height direction, the dust layer structure could be divided into three typical parts: the substructure, main profile and surface layer. In addition, the deposit morphologies observed for a high Pe were in good agreement with the experimental results found in the literature.

Applicability of ARFI elastography in detecting elasticity changes of the equine superficial digital flexor tendon with induced injury.

The objectives of this prospective, experimental study were to describe changes in the stiffness of the equine superficial digital flexor tendon (SDFT) after induced injury, deep digital flexor tendon (DDFT), accessory ligament (AL-DDFT), and suspensory ligament (SL) during 90 days of healing using acoustic radiation force impulse (ARFI) elastography. Eight healthy horses were selected. Preinjury B mode and ARFI evaluations were performed bilaterally in the palmar metacarpal region. Injury was induced only on the left forelimb (G2) by a single injection of collagenase in SDFT, 15 cm distal to the accessory carpal bone. The right forelimb was used as a control (G1). Evaluations were performed at eight timepoints: one before injury (T0) and seven (T1-T7) after injury (3, 15, 30, 40, 60, 75, and 90 days post-induction). Tendinopathies were visualized as hypoechoic areas with loss of parallel tendon fiber pattern. Injured SDFTs presented mainly cool colors (soft) from T1 to T3, and from T4, there was an increase in warm colors (hard), close to the appearance of tendons of G1. In the first four timepoints, there was a decrease in stiffness compared to G1 (P < 0.001). On T1 and T2, a cutoff value <6.21 m/s to determine tendinopathy of the SDFT was established (75.8% sensitivity and 92.03% specificity). Stiffness changes in the DDFT, AL-DDFT, and SL of injured limbs occurred at different timepoints. Tendinopathy significantly altered the stiffness of the injured tendon and the adjacent tissues. ARFI made it possible to detect these changes, helping to monitor the reparation of this injury.