Minimum Feret Diameter Research Articles

A User-Friendly Approach for Routine Histopathological and Morphometric Analysis of Skeletal Muscle Using CellProfiler Software

Adult skeletal muscle is capable of active and efficient differentiation in the event of injury in both physiological and pathological conditions, such as in Duchenne muscular dystrophy (DMD). DMD is characterized by different features, such as continuous cycles of degeneration/regeneration, fiber heterogeneity, chronic inflammation and fibrosis. A well-defined and standardized approach for histological and morphometric analysis of muscle samples is necessary in order to measure and quantify specific regenerative parameters in myopathies. Indeed, non-automatic methods are time-consuming and prone to error. Here, we describe a simple automatized computational approach to quantify muscle parameters with specific pipelines to be run by CellProfiler software in an open-source and well-defined fashion. Our pipelines consist of running image-processing modules in CellProfiler with the aim of quantifying different histopathological muscle hallmarks in mdx mice compared to their wild-type littermates. Specifically, we quantified the minimum Feret diameter, centrally nucleated fibers and the number of macrophages, starting from multiple images. Finally, for extracellular matrix quantification, we used Sirius red staining. Collectively, we developed reliable and easy-to-use pipelines that automatically measure parameters of muscle histology, useful for research in myobiology. These findings should simplify and shorten the time needed for the quantification of muscle histological properties, avoiding challenging manual procedures.

A comparison of phenotypic variation in Triticum durum Desf. genotypes deposited in gene banks based on the shape and color descriptors of kernels in a digital image analysis.

Durum wheat is the tenth most valuable crop on a global scale. The aim of this study was to compare the phenotypic variation of T. durum accessions of different origin with contemporary spring cultivars of this cereal species. One hundred and two accessions and 12 contemporary cultivars of Triticum durum Desf. as well as Kamut® wheat (T. turanicum), a Triticum species closely related to T. durum, were analyzed. The aim of this study was to describe the degree of variation in the phenotypic traits of grain and selected traits associated with technological quality. The examined genotypes were characterized by considerable phenotypic variation, and they can be a valuable source of material for genetic recombination in durum wheat breeding. The analyzed accessions were characterized by a higher average content of protein (16.48 vs. 14.56%) and wet gluten (38.04 vs. 32.07%), higher Zeleny sedimentation values (69.7 vs. 60.4ml), and higher flour strength (W index values of 404.64 vs. 353.47) than the reference cultivars. The kernels of the evaluated accessions and cultivars did not differ significantly in average crease depth, but significant differences were observed in the values of descriptors directly linked with kernel size, especially kernel image area and minimal Feret diameter. The traits responsible for the processing suitability of grain were more strongly correlated with color descriptors than shape descriptors, which suggests that color parameters can be used to select high-quality breeding material. The analyzed accessions have two major weaknesses, namely relatively low yields (22.6 dt ha-1 on average) and undesirable grain color, indicative of low carotenoid concentration. The accessions deposited in gene banks do not meet the relevant agronomic requirements. However, both grain yield and carotenoid concentration are polygenic traits which can be improved if desirable combinations of QTLs are assembled in breeding lines and cultivars.

Towards a generic protocol for measuring the constituent particle size distribution of E171 in food by electron microscopy

Food additive E171 (titanium dioxide, TiO 2 ) is authorized to be used as food colorant in the European Union. E171 is a particulate material, containing a fraction of nanoparticles. A major challenge for food control laboratories is the lack of harmonized, validated analytical methods for measuring the physicochemical properties of nanoparticles in food. This study proposes a generic method for the determination of the number-based size distribution of the constituent particles of E171 in food products based on transmission electron microscopy (TEM). As proof of concept, the protocol was evaluated on 20 food products containing E171 and one personal care product containing titanium dioxide. The total titanium concentration in the products measured by inductively coupled plasma - mass spectrometry (ICP-MS) ranged from 0.1 to 4.0 g/kg, corresponding to 0.2–6.7 g TiO 2 /kg. The required initial mass of starting product needed for TEM analysis was estimated from the ICP-MS concentration. Sample preparation included matrix digestion, a first centrifugation step to remove the remaining food matrix, sonication to reduce agglomeration, and a second centrifugation step to concentrate the sample. Descriptive TEM analysis showed that five food products, labelled to contain E171, contained smaller rutile TiO 2 particles (20–40 nm) coated with mica, which are potassium aluminium silicate-based pearlescent pigments. For 14 other food products and the personal care product, the protocol resulted in a homogenous distribution of the TiO 2 particles over the grid surface. In general, variation between products in the number of particles on the grids was limited; on average, 672 ± 173 particles were measured in 10 images. The median of the minimum Feret diameter distribution varied from 72 ± 7 nm to 135 ± 14 nm. The percentage of nanoparticles (minimum Feret diameter smaller than 100 nm) ranged from 23% to 82%. The corresponding mass percentages of these fractions ranged from 3% to 41%. The percentages of E171 particles smaller than 30 nm was very low (3% or less) while corresponding mass fractions were negligible. For only one food product, the TiO 2 concentration was too low to apply the protocol without adaptations. • A generic protocol for particle size measurement of E 171 in food is proposed. • The protocol is designed for transmission electron microscopy. • The protocol was evaluated on 20 food products and one personal care product. • The median Fmin varied from 72 ± 7 nm to 135 ± 14 nm. • The percentage of nanoparticles (<100 nm) ranged from 23% to 82%.

Particle size distributions following chewing: Transformation of two-dimensional outcome from optical scanning to volume outcome from sieving

Following chewing, the particle size distribution (PSD) by cumulative volume is characterized by X 50 , and b . Volume data are lacking when using less time-consuming optical scanning instead of sieving for characterizing PSDs. The aim was to transform 2D scanning data to 3D ones. Optosil® and almond were used as artificial and real solid foods, and minimal Feret-diameter (min-FD) as a measure of particle size. Border min-FDs were calibrated as a function of sieve aperture to determine size classes. Flakes produced by chewing were considered as ellipsoids. The ratio between the largest and the intermediate ellipsoid's radius was known by scanning. The ratio between the smallest and the intermediate radius was adjusted with Solver of Excel® to minimize the difference between the estimated total volume of the particles, and the known total volume. Estimated values of X 50 and b from scanning closely approached those from sieving, the ‘gold standard’. • Analysis of particle size distributions (PSDs) of a chewed solid artificial food. • PSDs by weight (volume) are usually obtained by labour and time consuming sieving. • Transformation of 2D optical scanning data to 3D volumes can replace sieving data. • Mathematical basis of 2D-3D transformation, assuming an ellipsoid particle shape. • The solution for 2D-3D transformation is also effective for real solid food (almond).

Comparison of Shifts in Skeletal Muscle Plasticity Parameters in Horses in Three Different Muscles, in Answer to 8 Weeks of Harness Training.

Training-induced follow-up of multiple muscle plasticity parameters in postural stability vs. locomotion muscles provides an integrative physiological view on shifts in the muscular metabolic machinery. It can be expected that not all muscle plasticity parameters show the same expression time profile across muscles. This knowledge is important to underpin results of metabolomic studies. Twelve non-competing Standardbred mares were subjected to standardized harness training. Muscle biopsies were taken on a non-training day before and after 8 weeks. Shifts in muscle fiber type composition and muscle fiber cross-sectional area (CSA) were compared in the m. pectoralis, the m. vastus lateralis, and the m. semitendinosus. In the m. vastus lateralis, which showed most pronounced training-induced plasticity, two additional muscle plasticity parameters (capillarization and mitochondrial density) were assessed. In the m. semitendinosus, additionally the mean minimum Feret's diameter was assessed. There was a significant difference in baseline profiles. The m. semitendinosus contained less type I and more type IIX fibers compatible with the most pronounced anaerobic profile. Though no baseline fiber type-specific and overall mean CSA differences could be detected, there was a clear post-training decrease in fiber type specific CSA, most pronounced for the m. vastus lateralis, and this was accompanied by a clear increase in capillary supply. No shifts in mitochondrial density were detected. The m. semitendinosus showed a decrease in fiber type specific CSA of type IIAX fibers and a decrease of type I fiber Feret's diameter as well as mean minimum Feret's diameter. The training-induced increased capillary supply in conjunction with a significant decrease in muscle fiber CSA suggests that the muscular machinery models itself toward an optimal smaller individual muscle fiber structure to receive and process fuels that can be swiftly delivered by the circulatory system. These results are interesting in view of the recently identified important fuel candidates such as branched-chain amino acids, aromatic amino acids, and gut microbiome-related xenobiotics, which need a rapid gut–muscle gateway to reach these fibers and are less challenging for the mitochondrial system. More research is needed with that respect. Results also show important differences between muscle groups with respect to baseline and training-specific modulation.

Concurrent influences of tool offset and ultrasonic vibration on the joint quality and performance of dissimilar Al/Cu friction stir welds

Dissimilar joints made of aluminium alloy (Al) and copper (Cu) have great prospective applications in the electrical and electronic industries. Still, it is a challenging task to join them and obtain good performance. In this study, dissimilar AA6061-T6 and C10100 joints were produced by friction stir welding (FSW) and ultrasonic vibration-assisted friction stir welding (UVaFSW) processes with three different tool offset degrees from the Al side to the Cu side. Tool offset towards Al base material and in the Al/Cu interface produced defect-free joints with sound strengths in both joints made with and without ultrasonic vibration. On the other hand, tool offset towards Cu base material produced unsound joints with defective features such as wormholes and a lack of surface fill in both joints made with and without ultrasonic vibration. Ultrasonic vibration addition favoured a drastic reduction in the intermetallic compound (IMC) layer thickness, particularly amongst the welds made with tool offset in the Al/Cu interface, than those made with tool offset towards Al base material. In addition, the influence of tool offsets and the ultrasonic vibration addition remarkably affects the sizes (particle areas and the minimum Feret diameters) and the concentration of the Cu particles distributed in the weld nugget. In the joints produced with tool offset towards Al base material, the particle areas and the minimum Feret diameters were decreased by ~73.3% and ~42.6%, respectively, with the ultrasonic vibration addition. On the other hand, ultrasonic vibration in the joints produced with tool offset in the Al/Cu interface resulted in decreased particle areas and the minimum Feret diameters by ~60.6% and ~24.6%, respectively. The joints made with ultrasonic vibration have higher tensile strength than those made by conventional FSW.

Comparison between normal and reverse orientation of graft in functional and histomorphological outcomes after autologous nerve grafting: An experimental study in the mouse model.

Autologous nerve grafting has been considered the gold standard for the treatment of irreparable nerve gaps. However, the choice of effective proximodistal orientation of autografts (normal or reversed) is controversial. Therefore, we compared functional and histological outcomes between normal and reversed orientations of autografts in a mouse sciatic nerve model. Thirty C57BL/6J mice weighing 20-25 g were assigned to the donor, normally oriented autograft, and reverse-oriented autograft groups (n= 10 per group). A 10-mm section of the sciatic nerve was harvested from a donor mouse. Half the harvested nerve was grafted onto an irreparable gap in a recipient mouse using either a normal or reversed orientation. The sciatic functional index (SFI) was measured biweekly for up to 12 weeks postoperatively. Morphological analysis was performed using immunofluorescence staining for neurofilament (NF) and myelin protein zero (P0) in cross-sectional and whole-mount nerve preparations in 12 weeks postoperatively. Additionally, morphological analysis of the tibialis anterior muscle was performed using hematoxylin and eosin staining. NF or P0-expressing axons were counted and cross-sectional area (CSA) and minimum Feret's diameter of myofibers were measured. The SFI recovered gradually up to 12 weeks after autografting, but there were no significant differences in the SFI between the normal and reversed orientations. The number of NF-expressing axons in center of graft was significantly higher in the normal orientation than in the reversed orientation (P < .05). However, there were no significant differences in the number and mean intensity of P0-expressing axons between the orientations. The CSA of myofibers was significantly larger in the normal orientation than in the reversed orientation (P < .05). Normally oriented autografts promote axonal regrowth and prevent neurogenic muscular atrophy compared with reverse-oriented autografts. However, despite these positive histomorphometric effects, the proximodistal orientation of the autograft does not affect functional outcomes.

Local (4‐Aminopyridine)‐PLGA‐PEG Treatment Improves Functional Recovery and Muscle Morphology after Traumatic Peripheral Nerve Injury in Mice

Traumatic peripheral nerve injury (TPNI) represents a major medical problem resulting in loss of both motor and sensory function, and in severe cases, permanent disability. Inadequate axonal regeneration and delayed muscle reinnervation contribute to muscle atrophy and poor functional recovery. To date, there is no effective pharmacologic treatment which can promote neuromuscular outcomes after TPNI. In repurposing studies, we have shown that daily systemic 4-aminopyridine (4-AP), an FDA-approved potassium channel blocker for multiple sclerosis, enhances functional recovery, improves myelination, and attenuates muscle atrophy after TPNI. However, 4-AP at higher doses cause adverse effects such as seizures. Thus, we sought to develop a novel locally injectable formulation of 4-AP which could be administered to a nerve injury site and deliver 4-AP at a sustained rate for several weeks. We developed (4-AP)-Poly(lactide-co-glycolide)-b-Poly(ethylene glycol)-b-Poly(lactide-co-glycolide) (PLGA-PEG-PLGA), consisting of two FDA-approved, biodegradable, and biocompatible hydrogels. It is unknown whether this locally injectable formulation of 4-AP would have an impact on TPNI-induced functional recovery, as well as on muscle atrophy. In this study, an established mouse model of right sciatic nerve crush injury was used and mice received one of 4 treatments: (4-AP)-PLGA-PEG, PLGA-PEG vehicle, systemic 4-AP, or saline control (n = 5/group). Functional recovery was assessed using sciatic function index, grip strength, and von Frey filament testing. At post-injury day 28, gastrocnemius (GN) and tibialis anterior (TA) muscles were harvested from both injured (innervated by crushed nerve) and uninjured (contralateral healthy) hind limbs for histomorphological analysis. Our results show that a single local administration of (4-AP)-PLGA-PEG remarkably enhanced motor and sensory functional recovery on post-sciatic nerve crush injury days 1, 3, 7, 14, and 21. Histomorphometric analysis of the muscles revealed that (4-AP)-PLGA-PEG treatment significantly increased right TA cross-sectional area (CSA) (1488.2 µm2 vs. 438.4 µm2, p<0.01) and minimum Feret's diameter (MFD) (53.9 µm vs. 30.3 µm, p<0.01) compared with the other groups. Similarly, (4-AP)-PLGA-PEG treatment significantly increased right GN CSA (1213.8 µm2 vs. 471.9 µm2, p<0.001) and MFD (48.5 µm vs. 32.6 µm, p<0.01) compared with the other groups. These preliminary findings on global functional and muscle fiber morphological parameters demonstrate that (4-AP)-PLGA-PEG can be used locally for neuromuscular outcomes after TPNI. Further cellular, molecular, and biochemical studies will define the mechanistic insights for this promising long-acting local therapeutic agent in TPNI.

Comparative study of histological and histo-chemical image processing in muscle fiber sections of broiler chicken

The image processing of muscular fibers has seen significant improvements over the last decades, from light microscopy acquisition to virtual microscopy and from manual segmentation and fiber typing to automatic. The present study aims to discuss the main tools employed in different histo-enzymological image processing phases applied to muscle fibers, which are respectively: the acquisition, segmentation and fiber typing as well as their efficiency in the determination of morphometric parameters. First, the acquisition: to count the cell numbers within the muscle fiber sections, optical microscopic images with different magnifications (x100, x200, x250, and x400) were compared with virtual slides digitized by Slide Scanner. Second, the segmentation: Three softwares (Fiji “Digitizing pen, Mouse”, Image Pro Plus 10 [semi-automatic], and Cytoinformatics LLC [automatic]) were compared for image segmentation quality and the determining of fiber numbers. Third, manual fiber typing: employing Fiji of segmented images using the 3 previously mentioned softwares were performed to calculate the accuracy of the morphometric parameter (Cross sectional Area [ CSA ], perimeter and Minimal Feret's Diameter [ MFD ]). The results of the acquisition showed that the scanner slides have a better resolution and detected a higher number of cells than when optical microscopes with different magnifications were used; with the latter, the processing can be performed only with an acceptable resolution, where the number of cells is lesser, which unfortunately requires several repetitions and exhausting work. Our findings regarding segmentation indicate that the Cytoinformatics LLC showed the best processing time and the highest quality followed by IP and Fiji. Finally and for the ultimate step, the morphometric parameter calculation: showed that the best accuracy was attained by using Fiji followed by Cytoinformatics and finally by IP. The findings of this study suggest that Fiji (semi-automatic) showed the best quality/price ratio (open access software) for segmentation and fiber typing, but it proved to be time consuming when compared to Image Pro Plus 10 (semi-automatic) and Cytoinformatics LLC (automatic).

Characterisation of TiO2-containing pearlescent pigments with regard to the European Union labelling obligation of engineered nanomaterials in food

ABSTRACT A wide range of trendy food colourants and ready-to-eat foods containing pearlescent pigments providing glitter effects is currently on the market. These pearlescent pigments consist of mica (potassium aluminium silicate) platelets generally coated with titanium dioxide and/or iron oxides. All single components are approved food additives in the European Union (EU) (E 555, E 171 and E 172). However, the European Food Safety Authority (EFSA) has stated recently, that pearlescent pigments should be evaluated as new food additives. Food grade titanium dioxide was already shown to contain a considerable fraction of nanoparticles. Thus, the question about ‘nano’-labelling of TiO2-containing pearlescent pigments according to the ‘Novel Food’ and ‘Food Information to Consumers’ regulations arose. In order to provide data for dealing with these issues, in this study four commercially available products of different food categories containing pearlescent pigments were characterised with focus on the structure, size and chemical composition of these pigments. The measurement methods used were flow particle image analysis (FPIA), static light scattering (SLS) and scanning electron microscopy (SEM) combined with energy-dispersive x-ray spectroscopy (EDX). After isolation from various food matrices, the glitter pigments could be easily identified and differentiated by fast FPIA screening from any remaining organic food matrix particles due to their typical platelet-like shape and transparency. The particle size distribution of the platelets was determined by means of SLS and found to be in the range of 8–167 µm. SEM was identified as the most suitable technique for the analysis of the nano-structured coating. For all constituent metal oxide particles (TiO2 and/or Fe2O3) a median minimum Feret diameter (Fmin) of 29.9–46.8 nm was obtained by quantitative SEM image analysis.

Characterization and toxicity evaluation of air-borne particles released by grinding from two dental resin composites in vitro

ObjectiveThe project aims to evaluate whether inhalation of particles released upon grinding of dental composites may pose a health hazard to dentists. The main objective of the study was to characterize the dust from polymer-based dental composites ground with different grain sized burs and investigate particle uptake and the potential cytotoxic effects in human bronchial cells. MethodsPolymerized blocks of two dental composites, Filtek™ Z250 and Filtek™ Z500 from 3M™ ESPE, were ground with super coarse (black) and fine (red) burs inside a glass chamber.Ultrafine airborne dust concentration and particle size distribution was measured real-time during grinding with a scanning mobility particle sizer (SMPS). Filter-collected airborne particles were characterized with dynamic light scattering (DLS) and scanning electron microscopy (SEM).Human bronchial epithelial cells (HBEC-3KT) were exposed to the dusts in dose-effect experiments. Toxicity was measured with lactate dehydrogenase (LDH) assay and cell counting kit-8 (CCK8). Cellular uptake was observed with transmission electron microscopy (TEM). ResultsAirborne ultrafine particles showed that most particles were in the size range 15−35 nm (SMPS). SEM analysis proved that more than 80% of the particles have a minimum Feret diameter less than 1 μm. In solution (DLS), the particles have larger diameters and tend to agglomerate. Cell toxicity (LDH, CCK8) is shown after 48 h and 72 h exposure times and at the highest doses. TEM showed presence of the particles within the cell cytoplasm. SignificanceProlonged and frequent exposure through inhalation may have negative health implications for dentists.

Nanoparticle size and 3D shape measurement by electron tomography: An Inter-Laboratory Comparison

Electron tomography (ET) has been used for quantitative measurement of shape and size of objects in three dimensions (3D) for many years. However, systematic investigation of repeatability and reproducibility of ET has not been evaluated in detail. To assess the reproducibility and repeatability of a protocol for measuring size and three-dimensional (3D) shape parameters for nanoparticles (NPs) by ET, an inter-laboratory comparison (ILC) has been performed. The ILC included six laboratories and six instruments models from three instrument manufacturers following a standard measurement protocol.A technical specification describing the normative steps of the protocol is published by the International Standards Organization (ISO). Gold NPs with 30 nm nominal diameter contained within a rod-shaped carbon support were measured. The use of a rod-shaped sample support eliminated the missing wedge effect in the experimental tilt series of projected images for improved quantification. A total of 443 NPs were initially measured by NRC-NANO and then 115 out of the 443 NPs were measured by five other labs to compare measurands such as the Volume (V), maximum Feret diameter (Fmax), minimum Feret diameter (Fmin), volume-equivalent diameter (Deq) and aspect ratio (Frat) of the NPs. The results of the five labs were compared with the results obtained at NRC-NANO. The maximum disagreement in measurements of Fmin and Fmax obtained by the participating labs did not exceed 7 %. The measured Deq was between 27.5 nm and 30.3 nm in agreement with the NP manufacturer’s specification (28 nm–32 nm). In addition to the above, the influence of the missing wedge effect and beam-induced NP movement was quantified based on the differences of the results between labs.

Physico-chemical characterisation of the fraction of silver (nano)particles in pristine food additive E174 and in E174-containing confectionery

ABSTRACT Silver (E174) is authorised as a food additive in the EU. The unknown particle size distribution of E174 is a specific concern for the E174 risk assessment. This study characterised the fraction of silver (nano)particles in 10 commercially available pristine E174 food additives and 10 E174-containing products by transmission electron microscopy (TEM) and single-particle inductively coupled plasma-mass spectrometry (spICP-MS). TEM analysis showed that all samples contained micrometre-sized flakes and also a fraction of (nano)particles. Energy-dispersive X-ray spectroscopy (EDX) and electron diffraction confirmed that the (nano)particles and micrometre-sized flakes consisted of silver. A higher amount of (nano)particles was observed in the products than in the food additives. In addition, the surface of the micrometre-sized flakes was rougher in products. The median of the minimum external dimension, assessed as minimal Feret diameter, of the fraction of (nano)particles determined by quantitative TEM analysis was 11 ± 4 nm and 18 ± 7 nm (overall mean ± standard deviation), for food additives and products, respectively. Similar size distributions were obtained by spICP-MS and TEM, considering the limit of detection of spICP-MS. The median of the equivalent spherical diameter of the fraction of (nano)particles determined by spICP-MS was 19 ± 4 nm and 21 ± 2 nm (overall mean ± standard deviation), for food additives and products, respectively. In all samples, independent of the choice of technique, the nano-sized particles represented more than 97% (by number) of the silver particles, even though the largest mass of silver was present as flakes.

Functional recovery with histomorphometric analysis of nerves and muscles after combination treatment with erythropoietin and dexamethasone in acute peripheral nerve injury.

IntroductionPeripheral nerve injury (PNI) often leads to significant functional loss in patients and poses a challenge to physicians since treatment options for improving functional outcomes are limited. Recent studies suggest that erythropoietin and glucocoticoids have beneficial effects as mediators of neuro-regenerative processes. We hypothesized that combination treatment with erythropoietin and glucocoticoids would have a synergistic effect on functional outcome after PNI.Materials and methodsSciatic nerve crush injury was simulated in ten-week-old male C57BL/6 mice. The mice were divided into four groups according to the type of drugs administered (control, erythropoietin, dexamethasone, and erythropoietin with dexamethasone). Motor functional recovery was monitored by walking track analysis at serial time points up to 28 days after injury. Morphological analysis of the nerve was performed by immunofluorescent staining for neurofilament (NF) heavy chain and myelin protein zero (P0) in cross-sectional and whole-mount nerve preparations. Additionally, morphological analysis of the muscle was performed by Hematoxylin and eosin staining.ResultsCombination treatment with erythropoietin and dexamethasone significantly improved the sciatic functional index at 3, 7, 14, and 28 days after injury. Fluorescence microscopy of cross sectional nerve revealed that the combination treatment increased the ratio of P0/NF-expressing axons. Furthermore, confocal microscopy of the whole-mount nerve revealed that the combination treatment increased the fluorescence intensity of P0 expression. The cross-sectional area and minimum Feret’s diameter of the muscle fibers were significantly larger in the mice which received combination treatment than those in the controls.ConclusionOur results demonstrated that combination treatment with erythropoietin and dexamethasone accelerates functional recovery and reduces neurogenic muscle atrophy caused by PNI in mice, which may be attributed to the preservation of myelin and Schwann cell re-myelination. These findings may provide practical therapeutic options for patients with acute PNI.

The wasting-associated metabolite succinate disrupts myogenesis and impairs skeletal muscle regeneration.

Muscle wasting is a debilitating co-morbidity affecting most advanced cancer patients. Alongside enhanced muscle catabolism, defects in muscle repair/regeneration contribute to cancer-associated wasting. Among the factors implicated in suppression of muscle regeneration are cytokines that interfere with myogenic signal transduction pathways. Less understood is how other cancer/wasting-associated cues, such as metabolites, contribute to muscle dysfunction. This study investigates how the metabolite succinate affects myogenesis and muscle regeneration. We leveraged an established ectopic metabolite treatment (cell permeable dimethyl-succinate) strategy to evaluate the ability of intracellular succinate elevation to 1) affect myoblast homeostasis (proliferation, apoptosis), 2) disrupt protein dynamics and induce wasting-associated atrophy, and 3) modulate in vitro myogenesis. In vivo succinate supplementation experiments (2% succinate, 1% sucrose vehicle) were used to corroborate and extend in vitro observations. Metabolic profiling and functional metabolic studies were then performed to investigate the impact of succinate elevation on mitochondria function. We found that in vitro succinate supplementation elevated intracellular succinate about 2-fold, and did not have an impact on proliferation or apoptosis of C2C12 myoblasts. Elevated succinate had minor effects on protein homeostasis (~25% decrease in protein synthesis assessed by OPP staining), and no significant effect on myotube atrophy. Succinate elevation interfered with in vitro myoblast differentiation, characterized by significant decreases in late markers of myogenesis and fewer nuclei per myosin heavy chain positive structure (assessed by immunofluorescence staining). While mice orally administered succinate did not exhibit changes in overall body composition or whole muscle weights, these mice displayed smaller muscle myofiber diameters (~6% decrease in the mean of non-linear regression curves fit to the histograms of minimum feret diameter distribution), which was exacerbated when muscle regeneration was induced with barium chloride injury. Significant decreases in the mean of non-linear regression curves fit to the histograms of minimum feret diameter distributions were observed 7 days and 28 days post injury. Elevated numbers of myogenin positive cells (3-fold increase) supportive of the differentiation defects observed in vitro were observed 28 days post injury. Metabolic profiling and functional metabolic assessment of myoblasts revealed that succinate elevation caused both widespread metabolic changes and significantly lowered maximal cellular respiration (~35% decrease). This study broadens the repertoire of wasting-associated factors that can directly modulate muscle progenitor cell function and strengthens the hypothesis that metabolic derangements are significant contributors to impaired muscle regeneration, an important aspect of cancer-associated muscle wasting.

What role would the pores related to brittle minerals play in the process of oil migration and oil & water two-phase imbibition?

This study aimed to investigate the geometry distinction of the pores formed by brittle minerals in shale oil reservoir and dig out its significance in hydrocarbon migration and exploitation.14050 pores related to brittle minerals in typical shale oil reservoir samples are selected as the research objects, high-resolution field emission scanning electron microscopy (FE-SEM), imbibition test, Nuclear Magnetic Resonance(NMR), high-resolution image technique, construction of new parameters, statistics and analysis of large numbers and other means are used to discover this issue.Results firstly show that the development of pores in the two directions (Angle between two directions is 90 degrees or approximately 90 degrees) are complementary and promote each other. Reversely, the data points begin to be scattered, which means that the development of the pore in the above two directions begins to “lose stability”, rather than promote each other all the time. This could prove that the rectangle (width, height), Legendre ellipse (major axis and major axis) and maximum and minimum Feret diameter of pores can all be used as the effective criteria for the development of pore shape related to brittle minerals. Sufficient evidences could prove that the pores related to brittle minerals contribute to oil and gas migration and oil & water imbibition.The conclusion of this study will provide an important theoretical basis for clarifying the oil occurrence mechanism of shale oil reservoir and looking for the geological and engineering dessert from the microscopic viewpoint.

Measurement of the Pediatric and Adult Osseous External Auditory Canal: Implications for Transcanal Endoscopic Ear Surgery.

To develop a method for measuring the diameter of the osseous external auditory canal (OEAC) in pediatric and adult patients. Retrospective analysis of imaging data from patients with a cholesteatoma who underwent transcanal endoscopic ear surgery (TEES) to remove the cholesteatoma by analyzing preoperative sagittal cone beam computed tomography (CBCT) images using ImageJ. Pediatric and adult patients were included who underwent TEES for a cholesteatoma between December 2011 and March 2015 and had available preoperative CBCT scans. Sagittal CT imaging data were analyzed by ImageJ to measure the maximum and minimum Feret diameters along the OEAC. The output data were then analyzed to determine the smallest maximum and smallest minimum Feret diameters for each patient. Fifty-one patients ranging in age from 3 to 83 years old were included in this study. The smallest maximum Feret diameter ranged from 6.5 to 10.1 mm (mean: 8.0 mm) for pediatric patients (<16 years of age) and 6.9 to 15.0 mm (mean: 8.9 mm) for adult patients, while the smallest minimum Feret diameter ranged from 3.6 to 5.9 mm (mean: 5.1 mm) for pediatric patients and 3.4 to 6.4 mm (mean: 5.4 mm) for adult patients. Moreover, the smallest maximum Feret diameters of children were significantly smaller than those of adults. We have developed a novel method for measuring the diameter along the OEAC and determining its narrowest site. This method can be incorporated into the preoperative evaluation of patients who are candidates for TEES.

Double Diffusion Encoding for Probing Radiation-Induced Microstructural Changes in a Tumor Model: A Proof-of-Concept Study With Comparison to the Apparent Diffusion Coefficient and Histology.

Microstructure analyses are gaining interest in cancer MRI as an alternative to the conventional apparent diffusion coefficient (ADC), of which the determinants remain unclear. To assess the sensitivity of parameters calculated from a double diffusion encoding (DDE) sequence to changes in a tumor's microstructure early after radiotherapy and to compare them with ADC and histology. Cohort study on experimental tumors. Sixteen WAG/Rij rats grafted with one rhabdomyosarcoma fragment in each thigh. Thirty-one were imaged at days 1 and 4, of which 17 tumors received a 20 Gy radiation dose after the first imagery. 3T. Diffusion-weighted imaging, DDE with flow compensated, and noncompensated measurements. 1) To compare, after irradiation, DDE-derived parameters (intracellular fraction, cell size, and cell density) to their histological counterparts (fraction of stained area, minimal Feret diameter, and nuclei count, respectively). 2) To compare percentage changes in DDE-derived parameters and ADC. 3) To evaluate the evolution of DDE-derived parameters describing perfusion. Wilcoxon rank sum test. 1) Intracellular fraction, cell size, and cell density were respectively lower (-24%, P < 0.001), higher (+7.5%, P < 0.001) and lower (-38%, P < 0.001) in treated tumors as compared to controls. Fraction of stained area, minimal Feret diameter, and nuclei count were respectively lower (-20%, P < 0.001), higher (+28%, P < 0.001), and lower (-34%, P < 0.001) in treated tumors. 2) The magnitude of ADC's percentage change due to irradiation (16.4%) was superior to the one of cell size (8.4%, P < 0.01) but inferior to those of intracellular fraction (35.5%, P < 0.001) and cell density (42%, P < 0.001). 3) After treatment, the magnitude of the vascular fraction's decrease was higher than the increase of flow velocity (33.3%, vs. 13.3%, P < 0.001). The DDE sequence allows quantitatively monitoring the effects of radiotherapy on a tumor's microstructure, whereas ADC only reveals global changes. 2. Stage 4. J. Magn. Reson. Imaging 2020;52:941-951.

Direction-specific Disruption of Paretic Arm Movement in Post-stroke Patients.

ABSTRACTObjective:This study aimed to characterize reaching movements of the paretic arm in different directions within the reachable workspace in post-stroke patients. Methods:A total of 12 post-stroke patients participated in this study. Each held a ball with a tracking marker and performed back-and-forth reaching movements from near the middle of the body to one of two targets in front of them located on the ipsilateral and contralateral sides of the arm performing the movement. We recorded and analyzed the trajectories of the tracking marker. The stability of arm movements was evaluated using areas and minimum Feret diameters to assess the trajectories of both the paretic and non-paretic arms. The speed of the arm movement was also calculated. Results:For the paretic arm, contralateral movement was more impaired than ipsilateral movement, whereas for the non-paretic arm, no difference was observed between the directions. The maximum speed of the contralateral movement was significantly slower than that of the ipsilateral movement in both the paretic and non-paretic arms. Conclusion:The paretic arm shows direction-specific instability in movement toward the contralateral side of the arm.

Rice in vitro digestion: application of INFOGEST harmonized protocol for glycemic index determination and starch morphological study.

Starch is the main sugar source present in staple foods. Understanding starch hydrolysis during digestion and the resulting glucose release can be important to strategically modulate starch digestion and glucose absorption. In vitro digestion methodologies are fundamental to evaluate starch hydrolysis length and rate, but the lack of uniformity between protocols prevent the comparison of results. In this context, three different Carolino rice varieties (i.e., Carolino white-Cw, Carolino brown-Cb and Carolino Ariete brown-CAb) were submitted to the INFOGEST harmonized in vitro digestion protocol for the evaluation of starch hydrolysis and subsequent glycemic index (GI) determination, and starch granules morphological study. Samples of Carolino rice presented total starch percentages between 64.52 (for Cb) to 71.52% (for Cw) with low amylose content (16.19-19.95%, varying in the following order Cb < Cab ≈ Cw). During digestion, between 39.43 (for CAb) to 44.48% (for Cb) of starch was hydrolyzed, classifying samples as medium GI foods (61.73-69.17). Starch hydrolysis was accompanied by a decrease of starch granules dimensions. For all samples, area decrease was higher than 59%, perimeter decrease was higher than 37%, feret diameter decrease was higher than 39% and minimum feret diameter decrease was higher than 32%. This work provides new insights to describe, both qualitatively and quantitatively, the fate of rice during digestion, and allowed establishing a comparative basis for the development of rice-based recipes with a lower GI.