Higher Fiber Density Research Articles

Three-dimensional visualization of uterine nerve fiber distribution using fluorescence micro-optical sectioning tomography (fMOST): A pilot study.

This study aimed to observe and quantitatively analyze the morphology and distribution of uterine nerve fibers using fluorescence micro-optical sectioning tomography (fMOST). The goal was to provide an accurate morphological reference for pathological evaluations of uterine nerves. Using fMOST technique, we observed and analyzed the distribution of nerve fibers within the uterus. Our findings revealed a radial dispersion of nerve fibers radiating from myometrium to endometrium. The cervix uteri region exhibited a high density of nerve fibers, displaying terminations in a flower spray pattern. In contrast, nerve fibers in corpus uteri were comparatively sparse. However, we identified a unique "vine-like" pattern of a single nerve fiber extending from myometrium to endometrial layer in areas with concentrated nerves. The fMOST technique is able to effectively elucidate the morphology and distribution of uterine nerve fibers. This method enables three-dimensional visualization of nerves in myometrium and offers a novel approach to observe the pathological changes in uterine nerves.

Robotic Ultrasound and Novel Collagen Analyses for Polycystic Kidney Disease Research Using Mice.

3D imaging and histology are critical tools for assessing polycystic kidney disease ( PKD ) in patients and animal models. Magnetic resonance ( MR ) imaging provides micron resolution, but is time consuming, expensive, and access to equipment and expertise is limiting. Robotic ultrasound ( US ) imaging has lower spatial resolution but is faster, more cost effective, and accessible. Similarly, Picrosirius red ( PSR ) staining and brightfield microscopy is commonly used to assess fibrosis; however, alternative methods have been shown in non-kidney tissues to provide greater sensitivity and more detailed structural characterization. In this study, we evaluated the utility of robotic US and alternative methods of quantifying PSR staining for PKD research. We compared longitudinal total kidney volume ( TKV ) measurements using US and MR. We additionally compared PSR imaging and quantification using standard brightfield with that by circularly polarized light with hue analysis, and fluorescence imaging analyzed using CT-FIRE software for automatic detection of individual collagen fibers. Increased TKV was detected by US in Pkd1RC/RC vs wild type ( WT ) at timepoints spanning early to established disease. US inter-observer variability was greater but allowed scanning in 2-5 minutes/mouse while MR required 20-30 minutes/mouse. While no change in fibrotic index was detected in this cohort of relatively mild disease using brightfield, polarized light showed fibers skewed thinner in Pkd1RC/RC vs WT. Fluorescence imaging showed a higher density of collagen fibers in Pkd1RC/RC vs WT, and fibers were thinner and curvier with no change in length. Additionally, fiber density was higher in both glomeruli and tubules in Pkd1RC/RC , and glomeruli had a higher fiber density than tubules in Pkd1RC/RC , and trended higher in WT. These studies show robotic ultrasound is a rigorous imaging tool for pre-clinical PKD research. Additionally, they demonstrate the increased sensitivity of polarized and fluorescence analysis of PSR-stained collagen.

Fabrication of 3D chitosan/polyvinyl alcohol/brushite nanofibrous scaffold for bone tissue engineering by electrospinning using a novel falling film collector

Despite its advantages, electrospinning has limited effectiveness in 3D scaffolding due to the high density of fibers it produces. In this research, a novel electrospinning collector was developed to overcome this constraint. An aqueous suspension containing chitosan/polyvinyl alcohol nanofibers was prepared employing a unique falling film collector. Suspension molding by freeze-drying resulted in a 3D nanofibrous scaffold (3D-NF). The mineralized scaffold was obtained by brushite deposition on 3D-NF using wet chemical mineralization by new sodium tripolyphosphate and calcium chloride dihydrate precursors. The 3D-NF was optimized and compared with the conventional electrospun 2D nanofibrous scaffold (2D-NF) and the 3D freeze-dried scaffold (3D-FD). Both minor fibrous and major freeze-dried pore shapes were present in 3D-NFs with sizes of 16.11–24.32 μm and 97.64–234.41 μm, respectively. The scaffolds' porosity increased by 53 % to 73 % compared to 2D-NFs. Besides thermal stability, mineralization improved the 3D-NF's ultimate strength and elastic modulus by 2.2 and 4.7 times, respectively. In vitro cell studies using rat bone marrow mesenchymal cells confirmed cell infiltration up to 290 μm and scaffold biocompatibility. The 3D-NFs given nanofibers and brushite inclusion exhibited considerable osteoinductivity. Therefore, falling film collectors can potentially be applied to prepare 3D-NFs from electrospinning without post-processing.

Utilizing Upcycled Graphene Nanoplatelet-Coated Glass Fibers as a Performance Booster and Compatibilizer for Enhanced Mechanical Performance of Polypropylene/Glass Fiber/Graphene Nanoplatelet Composites.

The high usage ratio and elevated density of glass fibers (GFs), often surpassing twice that of polymers, can contribute to undesired increases in the overall density of polymeric materials. One potential solution is the incorporation of graphene as a secondary additive, offering a lower specific gravity and exceptional mechanical properties. In light of this, waste tire-derived graphene nanoplates (GNPs) were optimized for coating onto GFs by considering factors such as surface treatment of the fiber, the dispersion quality of GNP, and the coating technique. The resulting GNP-coated GF (GNP-c-GF) was initially incorporated into pure polypropylene (PP) at low weight percentages (0.1-1 wt %), and 31% increase in the tensile modulus was achieved compared to neat PP. Subsequently, 1 wt % GNP-c-GF was utilized as a compatibilizer in PP/GF/GNP composites to enhance the compatibility between GNP and GF. By strategically incorporating GNP and GNP-c-GF at a lower GF ratio, the detrimental impact on the tensile modulus of 30% GF-filled PP was effectively mitigated, leading to a remarkable enhancement to an impressive value of 5658 MPa. The successful integration of GNP and GNP-c-GF exemplifies their promising potential as additives for achieving superior mechanical properties in composite materials, while concurrently promoting the utilization of recycled content.

Convergent evolution in high-altitude and marine mammals: Molecular adaptations to pulmonary fibrosis and hypoxia.

High-altitude and marine mammals inhabit distinct ecosystems but share a common challenge: hypoxia. To survive in low-oxygen environments, these species have evolved similar phenotypic pulmonary adaptations, characterized by a high density of elastic fibers. In this study, we explored the molecular mechanisms underlying these adaptations, focusing on pulmonary fibrosis and hypoxia tolerance through comparative genomics and convergent evolution analyses. We observed significant expansions and contractions in certain gene families across both high-altitude and marine mammals, closely associated with processes involved in pulmonary fibrosis. Notably, members of the keratin gene family, such as KRT17 and KRT14, appear to be associated with the development of the dense elastic fiber phenotype observed in the lungs of hypoxia-tolerant mammals. Through selection pressure and amino acid substitution analyses, we identified multiple genes exhibiting convergent accelerated evolution, positive selection, and amino acid substitution in these species, associated with adaptation to hypoxic environments. Specifically, the convergent evolution of ZFP36L1, FN1, and NEDD9 was found to contribute to the high density of elastic fibers in the lungs of both high-altitude and marine mammals, facilitating their hypoxia tolerance. Additionally, we identified convergent amino acid substitutions and gene loss events associated with sperm development, differentiation, and spermatogenesis, such as amino acid substitutions in SLC26A3 and pseudogenization of CFAP47, as confirmed by PCR. These genetic alterations may be linked to changes in the reproductive capabilities of these animals. Overall, this study offers novel perspectives on the genetic and molecular adaptations of high-altitude and marine mammals to hypoxic environments, with a particular emphasis on pulmonary fibrosis.

Enhanced properties of TEMPO-oxidized bacterial cellulose films via eco-friendly non-pressurized hot water vapor treatment for sustainable and smart food packaging.

Developing a simple and environmentally friendly method to vary the physical, mechanical, and thermal properties of cellulose films is of great importance. This study aimed to characterize 2,2,6,6-tetramethylpiperidinyl-1-oxyl (TEMPO)-oxidized bacterial cellulose (BC) films prepared using non-pressurized hot water vapor (NPHWV) method. A wet BC-pellicle that had been oxidized with TEMPO was treated with NPHWV for 60, 120, and 240 minutes, respectively. As a control, a TEMPO-oxidized BC (TOBC) film without NPHWV was prepared. The results show that the longer NPHWV duration of the TOBC film increased the tensile and thermal properties. This film became more hydrophobic and showed lower moisture absorption, thermal conductivity and organic solvent uptake, more crystalline structure, and higher fiber density after NPHWV treatment. The acquired results provide a simple, inexpensive, and ecologically friendly method for varying TOBC film properties.

Preparation and characterization of bacterial cellulose synthesized by kombucha from vinegar residue

Bacterial cellulose (BC) has been widely applied in various fields due to its excellent physicochemical properties, but its high production cost remains a challenge. Herein, the present study aimed to utilize the hydrolysate of vinegar residue (VR) as the only medium to realize the cost-effective production of BC. The BC production was optimized by the single-factor test. The treatment of 6 % VR concentration with 3 % acid concentration at 100 °C for 1.5 h and 96 U/mL of cellulase for 4 h at 50 °C obtained a maximum reducing sugar concentration of about 32 g/L. Additionally, the VR hydrolysate treated with 3 % active carbon (AC) at 40 °C for 0.5 h achieved a total phenol removal ratio of 86 %. The yield of BC reached 2.1 g/L under the optimum conditions, which was twice compared to the standard medium. The produced BC was characterized by SEM, FT-IR, XRD, and TGA analyses, and the results indicated that the BC prepared by AC-treated VR hydrolysate had higher fiber density, higher crystallinity, and good thermal stability. Furthermore, the regenerated BC (RBC) fibers with a tensile stress of 400 MPa were prepared successfully using AmimCl solution as a solvent by dry-wet-spinning method. Overall, the VR waste could be used as an alternative carbon source for the sustainable production of BC, which could be further applied to RBC fibers preparation.

Microplastics in the maximum chlorophyll layer along a north-south transect in the Mediterranean Sea in comparison with zooplankton concentrations

The aim of this study was to characterize and quantify microplastics (MPs) at the chlorophyll maximum layer (CML), around 30 to 60 m depth, during a cruise dedicated to the study of contaminants in plankton, the MERITE-HIPPOCAMPE project, along a north-south transect in the western Mediterranean Sea (Tedetti et al., 2023). Plankton were collected by horizontal net tows in this layer using a multinet Hydrobios Midi equipped with 60 μm mesh-size nets. The collected plankton were fractionated through a sieve column for various later contaminant measurements and for zooplankton analysis (Fierro-González et al., 2023). For all stations, samples were also fully examined for microplastics (MPs) for fractions >300 μm. MPs were found at all stations in the CML layer (mean: 42.9 ± 45.4 MPs m−3), of which 96 ± 4 % were fibers. The ratios of mesozooplankton/MPs and detritus/MPs in this CML were respectively 223 ± 315 and 2544 ± 2268. These data are analyzed together with MPs concentrations from sea- surface sampled with a 300 μm net-size Manta net at the same stations. Overall, our observations highlight the very high density of fibers at the CML, mainly associated with aggregates, raising the hypothesis of their interactions with marine snow. Therefore, the importance of marine snow and vertical layering will have to be considered in future MP distribution modelling efforts.

An experimental platform for stochastic analyses of single serotonergic fibers in the mouse brain.

The self-organization of the serotonergic matrix, a massive axon meshwork in all vertebrate brains, is driven by the structural and dynamical properties of its constitutive elements. Each of these elements, a single serotonergic axon (fiber), has a unique trajectory and can be supported by a soma that executes one of the many available transcriptional programs. This "individuality" of serotonergic neurons necessitates the development of specialized methods for single-fiber analyses, both at the experimental and theoretical levels. We developed an integrated platform that facilitates experimental isolation of single serotonergic fibers in brain tissue, including regions with high fiber densities, and demonstrated the potential of their quantitative analyses based on stochastic modeling. Single fibers were visualized using two transgenic mouse models, one of which is the first implementation of the Brainbow toolbox in this system. The trajectories of serotonergic fibers were automatically traced in the three spatial dimensions with a novel algorithm, and their properties were captured with a single parameter associated with the directional von Mises-Fisher probability distribution. The system represents an end-to-end workflow that can be imported into various studies, including those investigating serotonergic dysfunction in brain disorders. It also supports new research directions inspired by single-fiber analyses in the serotonergic matrix, including supercomputing simulations and modeling in physics.

A novel batch-scale fabrication method for hundred-micron YAG transparent ceramic fibers

YAG ceramic fiber is the preferred material for the gain medium of the new generation fiber laser. However, the fabrication of hundred-micron and defect-free YAG ceramic fibers is still the primary challenge. In this paper, YAG ceramic fibers with the diameter of 147 μm and length of 113 mm were successfully batch fabricated by the combination of gelcasting and mold design, and the diameter fluctuation rate was only 1.4%. The solid loading of the slurries (46~54 vol.%) were systematically investigated to achieve high density and net-shape forming of green fibers. It was found that the YAG ceramic fiber was highly transparent with the optimized solid loading of 52 vol.%, and the transmittance of the YAG ceramic (2 mm thick) at 800 nm was 82.6%. Its bending strength was as high as 1124 ± 86 MPa. The ideas and methods of this study will promote the development of gelcasting in the field of hundred-micron ceramic fibers.

In vitro-in vivo wound healing efficacy of Tridax Procumbens extract loaded Carboxymethylcellulose film

In this work, Tridax Procumbens Extracts (TPE) were blended with Carboxymethylcellulose (CMC), and film was developed through the casting method. The phytochemical screening of the TPE/CMC film was carried out and found the presence of carbohydrates, tannins, saponins, and cardiac glycosides. The presence of elements such as C, O, Na, P, Cl, K, Ca, Mn, and Nb in TP/CMC film was confirmed through EDX. The melting point of the film was found around 291 ± 0.5 °C which was determined through the DSC curve. The maximum tensile strength of the TPE/CMC was found as 14 ± 0.5 MPa. The film showed antibacterial activity against Escherichia coli, Streptococcus pyogenes, Staphylococcus aureus, and Vibrio cholera compared to the control. Cell viability study exhibited 95 % and 98 % cell proliferation for the test film after interacting with the L929 cell line for 18 h and 24 h. The optical contact angle of the TPE/CMC film was also determined. The in-vivo, wound healing studies on adult mice showed healing within 10 days only and the histopathological results revealed the maximum number of fibroblasts with a high density of collagen fibers in the test group indicating that the prepared film can be an effective wound dressing material.

(090) Immunohistochemical Staining of Elastin in the Vestibule

Abstract Introduction The vestibule is an external genital structure lying 90° to the vaginal canal opening. During the thrusting movements associated with penetrative sexual activity, the vestibule undergoes significant stretching during both invagination and evagination. The vestibule also undergoes extreme stretching during childbirth. The rete pegs in the non-keratinized squamous epithelium facilitate the ability of the vestibule to mechanically stretch. More importantly, however, the vestibular lamina propria, with nerves, blood vessels, glands, needs to contain a high density of extracellular matrix (ECM) proteins with stretching capabilities. In particular, the ECM protein elastin endows skin structures with the ability to stretch, recoil, and have elasticity. Elastin has previously been shown to influence mechanical stretching properties of tendons. Objective The objective of this study was to examine the density of elastin proteins in the vestibule in reproductive age patients as there are currently limited data. Methods A total of 7 reproductive age patients with normal sex steroid hormone values and complaints of entrance dyspareunia were diagnosed with neuroproliferative vestibulodynia and subsequently underwent complete vestibulectomy. Tissue blocks were created from the specimen, and 5 μm sections studied with immunohistochemical staining with primary antibody. Sections were then counterstained with Gill-modified hematoxylin. Results In all vestibular specimen, there was very high density of elastin underneath the epithelium in rete pegs and throughout the entire lamina propria (Fig 1). There is a density gradient with the highest density immediately deep to the basement membrane of the epithelium. Elastin fibers were arranged in all orientations: coronal (frontal), sagittal (longitudinal), and transverse (axial) planes. Conclusions The high density of elastin fibers in rete pegs and throughout the lamina propria in a multi-planar orientation is suggestive of extreme stretching capability in eugonadal vestibular tissues. The highest density of elastin staining was seen immediately below the basement membrane of the epithelium, which may provide tissue integrity and shear resilience to the epithelial layer. In sex steroid hormone insufficiency states such as menopause or with use of combined hormonal contraceptives, individuals may develop loss of introital elasticity and fissures due to a deficiency of the ECM protein elastin. More research is needed on elastic fiber production, organization, and integration with other cutaneous extracellular matrix proteins, and how they may be affected by sex steroid hormones. Disclosure Any of the authors act as a consultant, employee or shareholder of an industry for: Endoceutics, Inc.

MP27-14 ELASTIN STAINING OF THE VESTIBULE

You have accessJournal of UrologyCME1 Apr 2023MP27-14 ELASTIN STAINING OF THE VESTIBULE Alyssa Yee, Alain Dury, Johanne Ouellet, Barry Komisaruk, Sue W Goldstein, and Irwin Goldstein Alyssa YeeAlyssa Yee More articles by this author , Alain DuryAlain Dury More articles by this author , Johanne OuelletJohanne Ouellet More articles by this author , Barry KomisarukBarry Komisaruk More articles by this author , Sue W GoldsteinSue W Goldstein More articles by this author , and Irwin GoldsteinIrwin Goldstein More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003255.14AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: The vestibule is an external genital structure lying 90° to the vaginal canal opening. The vestibule consists of non-keratinized squamous epithelium with numerous rete pegs, underlying lamina propria with nerves, blood vessels, glands, and extracellular matrix (ECM) proteins. During thrusting movements associated with penetrative sexual activity, the vestibule undergoes significant stretching during invagination/evagination. The vestibule also undergoes extreme stretching during childbirth. Elastin is an ECM protein in microfibrils that endows skin structures with ability to stretch, recoil, and have elasticity. Elastin has previously been shown to influence mechanical properties of tendons; degraded or abnormal elastin structures caused by aging or disease may contribute to pain and increased risk of injury. There are limited data on elastin in the vestibule in reproductive age patients and its role in vestibular elasticity/integrity. METHODS: 7 reproductive age patients with entrance dyspareunia from neuroproliferative vestibulodynia, with normal values of testosterone, estradiol, and progesterone, underwent complete vestibulectomy. Tissue blocks were created from the specimen, and 5 υm sections studied with IHC primary antibody. Finally, sections were counterstained with Gill-modified hematoxylin. RESULTS: In all vestibular specimen, there was very high density of elastin underneath the epithelium in rete pegs and throughout the entire lamina propria. There is a density gradient with the highest density immediately deep to the basement membrane of the epithelium. Elastin fibers were arranged in all orientations: coronal (frontal), sagittal (longitudinal), and transverse (axial) planes. CONCLUSIONS: The high density of elastin fibers in rete pegs and through the lamina propria in a multi-planar orientation is suggestive of extreme stretching capability in eugonadal vestibular tissues. The highest density of elastin staining was seen immediately below the basement membrane of the epithelium, which may provide tissue integrity and shear resilience to the epithelial layer. More research is needed on elastic fiber production, organization, and integration with other cutaneous extracellular matrix proteins, and how this may change during androgen insufficiency states such as menopause or with use of combined hormonal contraceptives. Source of Funding: None © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e367 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Alyssa Yee More articles by this author Alain Dury More articles by this author Johanne Ouellet More articles by this author Barry Komisaruk More articles by this author Sue W Goldstein More articles by this author Irwin Goldstein More articles by this author Expand All Advertisement PDF downloadLoading ...

Quantitative Image Analysis of Fibrillar Collagens Reveals Novel Diagnostic and Prognostic Biomarkers and Histotype-Dependent Aberrant Mechanobiology in Lung Cancer

Fibrillar collagens are the most abundant extracellular matrix components in non–small cell lung cancer (NSCLC). However, the potential of collagen fiber descriptors as a source of clinically relevant biomarkers in NSCLC is largely unknown. Similarly, our understanding of the aberrant collagen organization and associated tumor-promoting effects is very scarce. To address these limitations, we identified a digital pathology approach that can be easily implemented in pathology units based on CT-FIRE software (version 2; https://loci.wisc.edu/software/ctfire) analysis of Picrosirius red (PSR) stains of fibrillar collagens imaged with polarized light (PL). CT-FIRE settings were pre-optimized to assess a panel of collagen fiber descriptors in PSR-PL images of tissue microarrays from surgical NSCLC patients (106 adenocarcinomas [ADC] and 89 squamous cell carcinomas [SCC]). Using this approach, we identified straightness as the single high-accuracy diagnostic collagen fiber descriptor (average area under the curve = 0.92) and fiber density as the single descriptor consistently associated with a poor prognosis in both ADC and SCC independently of the gold standard based on the TNM staging (hazard ratio, 2.69; 95% CI, 1.55-4.66; P < .001). Moreover, we found that collagen fibers were markedly straighter, longer, and more aligned in tumor samples compared to paired samples from uninvolved pulmonary tissue, particularly in ADC, which is indicative of increased tumor stiffening. Consistently, we observed an increase in a panel of stiffness-associated processes in the high collagen fiber density patient group selectively in ADC, including venous/lymphatic invasion, fibroblast activation (α-smooth muscle actin), and immune evasion (programmed death-ligand 1). Similarly, a transcriptional correlation analysis supported the potential involvement of the major YAP/TAZ pathway in ADC. Our results provide a proof-of-principle to use CT-FIRE analysis of PSR-PL images to assess new collagen fiber–based diagnostic and prognostic biomarkers in pathology units, which may improve the clinical management of patients with surgical NSCLC. Our findings also unveil an aberrant stiff microenvironment in lung ADC that may foster immune evasion and dissemination, encouraging future work to identify therapeutic opportunities.

In Situ Detection of the Flexural Fracture Behaviors of Inner and Outer Bamboo-Based Composites

This paper investigated the fracture toughness and enhancement mechanism for each component in bamboo-based composites at the cellular level. In situ characterization techniques identified the fracture behaviors of bamboo-based composites in three-point bending tests, and scanning electron microscope (SEM) further visualized the crack propagation of the fracture surface. In addition, the improvement mechanism of bamboo-based composites was illustrated by mechanical properties at the cellular level assisted with nanoindentation tests. Our in situ test results showed that the bamboo-based composites exhibited a longer deformation and higher bending load compared with bamboo. The fracture was non-catastrophic, and crack propagated in a tortuous manner in bamboo-based composites. Microstructural analysis revealed that phenol-formaldehyde (PF) resin pulled out and middle lamella (ML) breaking rather than transverse transwall fracturing occurred in parenchymal cells. The higher density of fibers in the bamboo-based composites triggered massive interfacial delamination in the middle lamella (ML), which was a weak mechanical interface. Furthermore, indented modulus and hardness illustrated that phenol-formaldehyde (PF) resin improved the mechanical strength of cell walls, especially parenchymal cells. The crosslinks of PF resin with the cell walls and massive fibers were the primary mechanisms responsible for the fracture toughness of bamboo-based composites, which could be helpful for advanced composites.

Remodelling of the fibre-aggregate structure of collagen gels by cancer-associated fibroblasts: A time-resolved grey-tone image analysis based on stochastic modelling.

Solid tumors consist of tumor cells associated with stromal and immune cells, secreted factors and extracellular matrix (ECM), which together constitute the tumor microenvironment. Among stromal cells, activated fibroblasts, known as cancer-associated fibroblasts (CAFs) are of particular interest. CAFs secrete a plethora of ECM components including collagen and modulate the architecture of the ECM, thereby influencing cancer cell migration. The characterization of the collagen fibre network and its space and time-dependent microstructural modifications is key to investigating the interactions between cells and the ECM. Developing image analysis tools for that purpose is still a challenge because the structural complexity of the collagen network calls for specific statistical descriptors. Moreover, the low signal-to-noise ratio of imaging techniques available for time-resolved studies rules out standard methods based on image segmentation. In this work, we develop a novel approach based on the stochastic modelling of the gel structure and on grey-tone image analysis. The method is then used to study the remodelling of a collagen matrix by migrating breast cancer-derived CAFs in a three-dimensional spheroid model of cellular invasion imaged by time-lapse confocal microscopy. The structure of the collagen at the scale of a few microns consists in regions with high fibre density separated by depleted regions, which can be thought of as aggregates and pores. The approach developped captures this two-scale structure with a clipped Gaussian field model to describe the aggregates-and-pores large-scale structure, and a homogeneous Boolean model to describe the small-scale fibre network within the aggregates. The model parameters are identified by fitting the grey-tone histograms and correlation functions of the images. The method applies to unprocessed grey-tone images, and it can therefore be used with low magnification, noisy time-lapse reflectance images. When applied to the CAF spheroid time-resolved images, the method reveals different matrix densification mechanisms for the matrix in direct contact or far from the cells. We developed a novel and multidisciplinary image analysis approach to investigate the remodelling of fibrillar collagen in a 3D spheroid model of cellular invasion. The specificity of the method is that it applies to the unprocessed grey-tone images, and it can therefore be used with noisy time-lapse reflectance images of non-fluorescent collagen. When applied to the CAF spheroid time-resolved images, the method reveals different matrix densification mechanisms for the matrix in direct contact or far from the cells.

White matter microstructure predicts measures of clinical symptoms in chronic back pain patients

Chronic back pain (CBP) has extensive clinical and social implications for its sufferers and is a major source of disability. Chronic pain has previously been shown to have central neural factors underpinning it, including the loss of white matter (WM), however traditional methods of analyzing WM microstructure have produced mixed and unclear results. To better understand these factors, we assessed the WM microstructure of 50 patients and 40 healthy controls (HC) using diffusion-weighted imaging. The data were analyzed using fixel-based analysis (FBA), a higher-order diffusion modelling technique applied to CBP for the first time here. Subjects also answered questionnaires relating to pain, disability, catastrophizing, and mood disorders, to establish the relationship between fixelwise metrics and clinical symptoms. FBA determined that, compared to HC, CBP patients had: 1) lower fibre density (FD) in several tracts, specifically the right anterior and bilateral superior thalamic radiations, right spinothalamic tract, right middle cerebellar peduncle, and the body and splenium of corpus callosum; 2) higher FD in the genu of corpus callosum; and 3) lower FDC – a combined fibre density and cross-section measure – in the bilateral spinothalamic tracts and right anterior thalamic radiation. Exploratory correlations showed strong negative relationships between fixelwise metrics and clinical questionnaire scores, especially pain catastrophizing. These results have important implications for the intake and processing of sensory data in CBP that warrant further investigation.

Plant‐based diets fed to juvenile meagre Argyrosomus regius with low methionine and taurine supplementation led to an overall reduction in fish performance and to an increase in muscle fibre recruitment

Methionine and taurine are amino acids (AA) that are usually deficient when fish meal is replaced by plant proteins. In this study, three diets were tested in juvenile meagre (initial weight: 13.4 g) for 8 weeks. The D1 diet had 0.2% methionine and 1% taurine supplementation; the D2 and D3 diets had 0.6% methionine and 1% and 2% taurine supplementation, respectively. The results showed that meagre fed the D1 diet had lower specific growth rate (2.2 to 2.5), lower feed efficiency (0.9 to 1.2) and higher food conversion rate (FCR, 1.1 to 0.8) as well as a lower activity of the alanine aminotransferase (ALAT) enzyme. Furthermore, a higher recruitment of muscle fibres (46% compared to 36%) as well as a higher fibre density was observed (1019 compared to 870 fibres mm-2 ). This study shows that meagre requires a sufficient quantity of methionine in plant-based diets to avoid a reduction in fish performance. Furthermore, taurine supplementation in the D1 diet was not able to mitigate the effects of methionine deficiency. A higher taurine supplementation did not improve meagre performance.

5-HT7 receptor-dependent intestinal neurite outgrowth contributes to visceral hypersensitivity in irritable bowel syndrome

AbstractIrritable bowel syndrome (IBS) is characterized by visceral hypersensitivity (VH) associated with abnormal serotonin/5-hydroxytryptamine (5-HT) metabolism and neurotrophin-dependent mucosal neurite outgrowth. The underlying mechanisms of VH remain poorly understood. We investigated the role of 5-HT7 receptor in mucosal innervation and intestinal hyperalgesia. A high density of mucosal nerve fibres stained for 5-HT7 was observed in colonoscopic biopsy specimens from IBS patients compared with those from healthy controls. Staining of 5-HT3 and 5-HT4 receptors was observed mainly in colonic epithelia with comparable levels between IBS and controls. Visceromotor responses to colorectal distension were evaluated in two mouse models, one postinfectious with Giardia and subjected to water avoidance stress (GW) and the other postinflammatory with trinitrobenzene sulfonic acid-induced colitis (PT). Increased VH was associated with higher mucosal density of 5-HT7-expressing nerve fibres and elevated neurotrophin and neurotrophin receptor levels in the GW and PT mice. The increased VH was inhibited by intraperitoneal injection of SB-269970 (a selective 5-HT7 antagonist). Peroral multiple doses of CYY1005 (a novel 5-HT7 ligand) decreased VH and reduced mucosal density of 5-HT7-expressing nerve fibres in mouse colon. Human neuroblastoma SH-SY5Y cells incubated with bacteria-free mouse colonic supernatant, 5-HT, nerve growth factor, or brain-derived neurotrophic factor exhibited nerve fibre elongation, which was inhibited by 5-HT7 antagonists. Gene silencing of HTR7 also reduced the nerve fibre length. Activation of 5-HT7 upregulated NGF and BDNF gene expression, while stimulation with neurotrophins increased the levels of tryptophan hydroxylase 2 and 5-HT7 in neurons. A positive-feedback loop was observed between serotonin and neurotrophin pathways via 5-HT7 activation to aggravate fibre elongation, whereby 5-HT3 and 5-HT4 had no roles. In conclusion, 5-HT7-dependent mucosal neurite outgrowth contributed to VH. A novel 5-HT7 antagonist could be used as peroral analgesics for IBS-related pain.

Mirror peripheral neuropathy and unilateral chronic neuropathic pain: insights from asymmetric neurological patterns in leprosy.

Leprosy-related multiple mononeuropathy offers a pattern of impairment where neuropathy with and without neuropathic pain (NeP) are present in the same individual, thus allowing to investigate peripheral sensory and innervation in both conditions. This cross-sectional study collected data on clinical and neurological examination, pain assessment questionnaires, quantitative sensory test, and intraepidermal nerve fiber density of patients with leprosy and divided the cohort into 2 groups: with NeP (P+) and without NeP (P-). Furthermore, we assessed mirror body areas in the same NeP individuals with bilateral neuropathy also presenting unilateral NeP. Pain-free patients having unilateral neuropathy were controls. A total of 37 P+ and 22 P- patients were evaluated. Limb areas with NeP had signs of C-fiber dysfunction and hyperesthesia on quantitative sensory testing compared with limb areas having neuropathy without NeP. Skin denervation was found in all patients with leprosy. Comparisons of limbs with and without neuropathy and with and without NeP revealed that higher heat pain thresholds (HPTs) were associated with neuropathic pain areas, whereas less altered HPT was correlated with higher fiber density. Furthermore, a relationship was found between time of leprosy treatment termination and more intense neuropathy, expressed by HPT increasing 0.03°C each month. As expected, interindividual comparisons failed to show differences in intraepidermal nerve fiber density and subepidermal plexus areas between P+ and P- patients ( P = 0.2980, P = 0.9044; respectively). Higher HPT and lower mechanical detection threshold were related to NeP. This study pointed out the relevance of intraindividual comparisons including mirror areas when assessing local changes in peripheral NeP.