Failed Orthodontic PEEK Retainer: A Scanning Electron Microscopy Analysis and a Possible Failure Mechanism in a Case Report.

This study presents a scanning electron microscopy analysis of a failed PEEK retainer in an orthodontic patient. After 15 months of use, the patient reported a gap opening between teeth 41 and 42. The PEEK retainer was removed and sent for electron microscope analysis. To investigate the failure, scanning electron microscopy was employed to assess the microstructure and composition of the retainer at various magnifications. These findings suggest that the failure of the PEEK retainer was multifaceted, implicating factors such as material defects, manufacturing flaws, inadequate design, environmental factors, and patient-related factors. In conclusion, this scanning electron microscopy analysis offers valuable insights into the failure mechanisms of PEEK retainers in orthodontic applications. Further research is necessary to explore preventive strategies and optimize the design and fabrication of PEEK retainers, minimizing the occurrence of failures in orthodontic practice.

Production and characterization of Trichoderma asperellum chitinases and their use in synergy with Bacillus thuringiensis for lepidopteran control.

BACKGROUND: Despite their known negative effects on ecosystems and human health, synthetic pesticides are still largely used to control crop insect pests. Currently, the biopesticide market for insect biocontrol mainly relies on the entomopathogenic bacterium Bacillus thuringiensis (Bt). New biocontrol tools for crop protection might derive from fungi, in particular from Trichoderma spp., which are known producers of chitinases and other bioactive compounds able to negatively affect insect survival. RESULTS: In this study, we first developed an environmentally sustainable production process for obtaining chitinases from Trichoderma asperellum ICC012. Then, we investigated the biological effects of this chitinase preparation - alone or in combination with a Bt-based product - when orally administered to two lepidopteran species. Our results demonstrate that T. asperellum efficiently produces a multi-enzymatic cocktail able to alter the chitin microfibril network of the insect peritrophic matrix, resulting in delayed development and larval death. The co-administration of T. asperellum chitinases and sublethal concentrations of Bt toxins increased larval mortality. This synergistic effect was likely due to the higher amount of Bt toxins that passed the damaged peritrophic matrix and reached the target receptors on the midgut cells of chitinase-treated insects. CONCLUSION: Our findings may contribute to the development of an integrated pest management technology based on fungal chitinases that increase the efficacy of Bt-based products, mitigating the risk of Bt-resistance development. © 2024 Society of Chemical Industry.

Enhancing SEM positioning precision with a LEGO®-based sample fitting system.

Scanning electron microscopy (SEM) is a precious tool in materials science and morphology sciences, enabling detailed examination of materials at the nanoscale. However, precise and accurate sample repositioning during different observation sessions remains a significant challenge, impacting the quality and repeatability of SEM analyses. This study aimed to develop and evaluate a LEGO®-based sample positioning system for SEM analysis. The system was designed to consistently identify and align features across multiple repositioning cycles, maintain accurate positioning along the z-axis, minimize distortion, and provide repeatable and reliable results. The results indicated a high degree of precision and accuracy in the repositioning process, as evidenced by the minimal displacements, deviations in scaling and shearing, and the highly significant results (p < 0.001) obtained from the analysis of absolute translations and rotations. Moreover, the analyses were consistently replicated across six repetitions, underscoring the reliability of the observed results. While the findings suggest that the LEGO-based sample positioning system is promising for enhancing SEM analyses' quality and repeatability, further studies are needed to optimize the system's design and evaluate its performance in different SEM applications. Ultimately, this study contributes to the ongoing efforts to develop cost-effective, customizable, and accurate solutions for sample positioning in SEM, contributing to the advancement of materials science research and all SEM analysis requiring overtime observations of the same sample. RESEARCH HIGHLIGHTS: This study focused on the development and evaluation of a novel LEGO-based sample positioning system specifically designed for SEM analysis. One of the standout features of this system is its ability to consistently identify and align features across multiple repositioning cycles, showcasing its precision and reliability. To further understand the mechanical aspects of the SEM stage, we employed the Rambold Kontroll comparator, which provided a baseline understanding of its mechanical tolerance. The registration process results were particularly noteworthy, as they revealed high accuracy with minimal displacements. Furthermore, the consistent outcomes observed across multiple repetitions emphasize the reliability and robustness of the methods we employed in this research.

Plastics shape the black soldier fly larvae gut microbiome and select for biodegrading functions.

BACKGROUND: In the last few years, considerable attention has been focused on the plastic-degrading capability of insects and their gut microbiota in order to develop novel, effective, and green strategies for plastic waste management. Although many analyses based on 16S rRNA gene sequencing are available, an in-depth analysis of the insect gut microbiome to identify genes with plastic-degrading potential is still lacking. RESULTS: In the present work, we aim to fill this gap using Black Soldier Fly (BSF) as insect model. BSF larvae have proven capability to efficiently bioconvert a wide variety of organic wastes but, surprisingly, have never been considered for plastic degradation. BSF larvae were reared on two widely used plastic polymers and shotgun metagenomics was exploited to evaluate if and how plastic-containing diets affect composition and functions of the gut microbial community. The high-definition picture of the BSF gut microbiome gave access for the first time to the genomes of culturable and unculturable microorganisms in the gut of insects reared on plastics and revealed that (i) plastics significantly shaped bacterial composition at species and strain level, and (ii) functions that trigger the degradation of the polymer chains, i.e., DyP-type peroxidases, multicopper oxidases, and alkane monooxygenases, were highly enriched in the metagenomes upon exposure to plastics, consistently with the evidences obtained by scanning electron microscopy and 1H nuclear magnetic resonance analyses on plastics. CONCLUSIONS: In addition to highlighting that the astonishing plasticity of the microbiota composition of BSF larvae is associated with functional shifts in the insect microbiome, the present work sets the stage for exploiting BSF larvae as "bioincubators" to isolate microbial strains and enzymes for the development of innovative plastic biodegradation strategies. However, most importantly, the larvae constitute a source of enzymes to be evolved and valorized by pioneering synthetic biology approaches. Video Abstract.

Investigating the interfaces of the epiphyseal plate: An integrated approach of histochemistry, microtomography and SEM.

We investigated the interfaces of the epiphyseal plate with over- and underlying bone segments using an integrated approach of histochemistry, microtomography and scanning electron microscopy (SEM) to overcome the inherent limitations of sections-based techniques. Microtomography was able to provide an unobstructed, frontal view of large expanses of the two bone surfaces facing the growth plate, while SEM observation after removal of the soft matrix granted an equally unhindered access with a higher resolution. The two interfaces appeared widely dissimilar. On the diaphyseal side the hypertrophic chondrocytes were arranged in tall columns packed in a sort of compact palisade; the interposed extracellular matrix was actively calcifying into a thick mineralized crust growing towards the epiphysis. Behind the mineralization front, histochemical data revealed a number of surviving cartilage islets which were being slowly remodelled into bone. In contrast, the epiphyseal side of the cartilage consisted of a relatively quiescent reserve zone whose mineralization was marginal in amount and discontinuous in extension; the epiphyseal bone consisted of a loose trabecular meshwork, with ample vascular spaces opening directly into the non-mineralized cartilage. On both sides the calcification process took place through the formation of spheroidal bodies 1-2 µm wide which gradually grew by apposition and coalesced into a solid mass, in a way distinctly different from that of bone and other calcified tissues.

Skeletal calcification patterns of batoid, teleost, and mammalian models: Calcified cartilage versus bone matrix.

This study compares the skeletal calcification pattern of batoid Raja asterias with the endochondral ossification model of mammalians Homo sapiens and teleost Xiphias gladius. Skeletal mineralization serves to stiffen the mobile elements for locomotion. Histology, histochemistry, heat deproteination, scanning electron microscopy (SEM)/EDAX analysis, thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), and Fourier transform infrared spectrometry (FTIR) have been applied in the study. H. sapiens and X. gladius bone specimens showed similar profiles, R. asterias calcified cartilage diverges for higher water release and more amorphous bioapatite. In endochondral ossification, fetal calcified cartilage is progressively replaced by bone matrix, while R. asterias calcified cartilage remains un-remodeled throughout the life span. Ca2+ and PO4 3- concentration in extracellular matrix is suggested to reach the critical salts precipitation point through H2 O recall from extracellular matrix into both chondroblasts or osteoblasts. Cartilage organic phase layout and incomplete mineralization allow interstitial fluids diffusion, chondrocytes survival, and growth in a calcified tissue lacking of a vascular and canalicular system. HIGHLIGHTS: Comparative physico-chemical characterization (TGA, DTG and DSC) testifies the mass loss due to water release, collagen and carbonate decomposition of the three tested matrices. R. asterias calcified cartilage water content is higher than that of H. sapiens and X. gladius, as shown by the respectively highest dehydration enthalpy values. Lower crystallinity degree of R. asterias calcified cartilage can be related to the higher amount of collagen in amorphous form than in bone matrix. These data can be discussed in terms of the mechanostat theory (Frost, 1966) or by organic/inorganic phase transformation in the course evolution from fin to limbs. Mineral analysis documented different charactersof R. asterias vs H. sapiens and X. gladius calcified matrix.

Fin systems comparative anatomy in model Batoidea Raja asterias and Torpedo marmorata: Insights and relatioships between musculo-skeletal layout, locomotion and morphology.

The macroscopic and microscopic morphology of the appendicular skeleton was studied in the two species Raja asterias (order Rajiformes) and Torpedo marmorata (Order Torpediniformes), comparing the organization and structural layout of pectoral, pelvic, and tail fin systems. The shape, surface area and portance of the T. marmorata pectoral fin system (hydrodynamic lift) were conditioned by the presence of the two electric organs in the disk central part, which reduced the pectoral fin surface area, suggesting a lower efficiency of the "flapping effectors" than those of R. asterias. Otherwise, radials' rays alignment, morphology and calcification pattern showed in both species the same structural layout characterized in the fin medial zone by stiffly paired columns of calcified tiles in the perpendicular plane to the flat batoid body, then revolving and in the horizontal plane to continue as separate mono-columnar rays in the fin lateral zone with a morphology suggesting fin stiffness variance between medial/lateral zone. Pelvic fins morphology was alike in the two species, however with different calcified tiles patterns of the 1st compound radial and pterygia in respect to the fin-rays articulating perpendicularly to the latter, whose tile rows lay-out was also different from that of the pectoral fins radials. The T. marmorata tail-caudal fin showed a muscular and connective scaffold capable of a significant oscillatory forward thrust. On the contrary, the R. asterias dorsal tail fins were stiffened by a scaffold of radials-like calcified segments. Histomorphology, heat-deproteination technique and morphometry provided new data on the wing-fins structural layout which can be correlated to the mechanics of the Batoid swimming behavior and suggested a cartilage-calcification process combining interstitial cartilage growth (as that of all vertebrates anlagen) and a mineral deposition with accretion of individual centers (the tiles). The resulting layout showed scattered zones of un-mineralized matrix within the calcified mass and a less compact texture of the matrix calcified fibers suggesting a possible way of fluid diffusion throughout the mineralized tissue. These observations could explain the survival of the embedded chondrocytes in absence of a canalicular system as that of the cortical bone.

Light spectra of biophilic LED-sourced system modify essential oils composition and plant morphology of Mentha piperita L. and Ocimum basilicum L.

Investigating morphological and molecular mechanisms that plants adopt in response to artificial biophilic lighting is crucial for implementing biophilic approaches in indoor environments. Also, studying the essential oils (EOs) composition in aromatic plants can help unveil the light influence on plant metabolism and open new investigative routes devoted to producing valuable molecules for human health and commercial applications. We assessed the growth performance and the EOs composition of Mentha x piperita and Ocimum basilicum grown under an innovative artificial biophilic lighting system (CoeLux®), that enables the simulation of natural sunlight with a realistic sun perception, and compared it to high-pressure sodium lamps (control) We found that plants grown under the CoeLux® light type experienced a general suppression of both above and belowground biomass, a high leaf area, and a lower leaf thickness, which might be related to the shade avoidance syndrome. The secondary metabolites composition in the plants' essential oils was scarcely affected by both light intensity and spectral composition of the CoeLux® light type, as similarities above 80% were observed with respect to the control light treatments and within both plant species. The major differences were detected with respect to the EOs extracted from plants grown under natural sunlight (52% similarity in M. piperita and 75% in O. basilicum). Overall, it can be speculated that the growth of these two aromatic plants under the CoeLux® lighting systems is a feasible strategy to improve biophilic approaches in closed environments that include both plants and artificial sunlight. Among the two plant species analyzed, O. basilicum showed an overall better performance in terms of both morphological traits and essential oil composition. To increase biomass production and enhance the EOs quality (e.g., higher menthol concentrations), further studies should focus on technical solutions to raise the light intensity irradiating plants during their growth under the CoeLux® lighting systems.

The chondro-osseous junction of articular cartilage.

In the synovial joints the transition between the soft articular cartilage and the subchondral bone is mediated by a layer of calcified cartilage of structural and mechanical characteristics closer to those of bone. This layer, buried in the depth of articular cartilage, is not directly accessible and is mostly visualized in histological sections of decalcified tissue, where it appears as a darker strip in contact with the subchondral bone. In this study conventional histology and scanning electron microscopy (SEM) with secondary electron imaging (SE) or backscattered electron imaging (BSE) were used to discriminate the calcified and the uncalcified cartilage in high resolution on native, untreated tissue as well as in deproteinated or demineralized tissue. This approach evidenced a high heterogeneity of the calcified layer of articular cartilage. High resolution pictures revealed that the mineralization process originates by progressive accretion and confluence of individual, small mineral clusters, in a very different way from other hard tissues such as bone, dentin and mineralized tendons. Finally, selective removal of the soft matrix by thermal treatment allowed for the first time a face-on, unrestricted 3D view of the mineralization front.

The combined cartilage growth - calcification patterns in the wing-fins of Rajidae (Chondrichthyes): A divergent model from endochondral ossification of tetrapods.

The relationship between cartilage growth - mineralization patterns were studied in adult Rajidae with X-ray morphology/morphometry, undecalcified resin-embedded, heat-deproteinated histology and scanning electron microscopy. Morphometry of the wing-fins, nine central rays of the youngest and oldest specimens documented a significant decrement of radials mean length between inner, middle and outer zones, but without a regular progression along the ray. This suggests that single radial length growth is regulated in such a way to align inter-radial joints parallel to the wing metapterygia curvature. Trans-illumination and heat-deproteination techniques showed polygonal and cylindrical morphotypes of tesserae, whose aligned pattern ranged from mono-columnar, bi-columnar, and multi-columnar up to the crustal-like layout. Histology of tessellated cartilage allowed to identify of zones of the incoming mineral deposition characterized by enhanced duplication rate of chondrocytes with the formation of isogenic groups, whose morphology and topography suggested a relationship with the impending formation of the radials calcified column. The morphotype and layout of radial tesserae were related to mechanical demands (stiffening) and the size/mass of the radial cartilage body. The cartilage calcification pattern of the batoids model shares several morphological features with tetrapods' endochondral ossification, that is, (chondrocytes' high duplication rate, alignment in rows, increased volume of chondrocyte lacunae), but without the typical geometry of the metaphyseal growth plates. RESEARCH HIGHLIGHTS: 1. The wing-fins system consists of stiff radials, mobile inter-radial joints and a flat inter-radial membrane adapted to the mechanical demand of wing wave movement. 2. Growth occurs by forming a mixed calcified-uncalcified cartilage texture, developing intrinsic tensional stresses documented by morphoanatomical data.

The complex rostral morphology and the endoskeleton ossification process of two adult samples of Xiphias gladius (Xiphiidae).

The authors studied the morphology of the upper and lower jaws, vertebrae and dorsal-fin rays of the teleost fish Xiphias gladius to analyse the skeletal architecture and ossification pattern. The analogies and differences among these segments were investigated to identify a common morphogenetic denominator of the bone tissue osteogenesis and modeling. The large fat glands in the proximal upper jaw and their relationship to the underlying cartilage (absent in the lower jaw) suggested that there is a mechanism that explains rostral overgrowth in the Xiphiidae and Istiophoriidae families. Thus far, the compact structure of the distal rostrum has been interpreted as being the result of remodeling. Nonetheless, no evidence of cutting cones, scalloped outer border of osteons and sequence of bright-dark bands in polarized light was observed in this study, suggesting a primary osteon texture formed by compacting of collagen matrix and mineral deposition in the fat stroma lacunae of the bone, but without being oriented in layers of the collagen fibrils. A similar histology also characterizes the circular structures present in the other examined segments of the skeleton. The early phases of fibrillogenesis carried out by fibroblast-like cells occurred farther from the already-calcified bone surface inside the fat stroma lacunae. The fibrillar matrix was compacted and underwent mineral deposition near the previously calcified bone surface. This pattern of collagen matrix synthesis and calcification was different from that of mammalian osteoblasts, especially concerning the ability to build a lacuno-canalicular system among cells. Necrosis or apoptosis of the latter and refilling of the empty lacunae by mineral deposits might explain the anosteocytic bone formation.

Morphology of joints and patterns of cartilage calcification in the endoskeleton of the batoid Raja cf. polystigma.

The skeleton of the batoid fish consists of a mixture of calcified and uncalcified cartilage with a typical layout of mineral deposition toward the outer border, leaving an uncalcified central core in most of the skeleton segments. An exception is observed in the radials, where mineral deposition is central. Joints and endoskeleton segments were studied in two adult samples of Raja cf. polystigma. Histomorphology, mineral deposition pattern, and zonal chondrocyte duplication activity were compared among several endoskeleton segments, but with particular attention to the fin rays; in the first, the uncalcified cartilage is central with an outer layer ranging from mineralized tesserae to a continuous calcified coating, whereas in the second, the uncalcified cartilage surrounds one or more central calcified columns. The diarthroses have a joint cavity closed by a fibrous capsule and the sliding surfaces rest on the base of mineralized tesserae, whereas the interradial amphiarthroses show a layer of densely packed chondrocytes between the flat, calcified discs forming the base of neighboring radials. In the endoskeleton segments, three types of tesserae are distinguished, characterizing the phases of skeletal growth and mineralization which present differences in each endoskeleton segment. The chondrocyte density between central core, subtesseral layer, and radial external cartilage did not show significant differences, while there was a significant difference in chondrocyte density between the latter zones and the type c tesserae of the pelvic girdle. The histomorphology and morphometry observed in Raja cf. polystigma suggest a model of cartilage growth associated with structural stiffening without remodeling. A key point of this model is suggested to be the incomplete mineralization of the tesseral layer and the continuous growth of cartilage, both enabling fluid diffusion through the matrix fibril network of scattered, uncalcified cartilage zones inside and between the tesserae.

New insights on fibrodysplasia ossificans progressiva: discussion of an autoptic case report and brief literature review.

Fibrodysplasia ossificans progressiva (FOP) is a rare genetic condition with soft tissue progressive ossification, leading to severe disability. We describe a 27-years-old female affected by FOP who died after a fall. An autopsy was performed. Upper and lower extremities resulted in fixed flexion, with kyphoscoliosis of the spine and chest wall deformity. Moreover, a cranial fracture was pointed out. At histology, atypical abundance of corpora amylacea in gray matter was observed. In a sample of macroscopically non-affected muscular tissue, small areas with necrosis of myocytes and hyperplasia of fibroblasts were seen in light microscopy, with intracellular inorganic dystrophic inclusions in transmission electron microscopy. Thyroid gland histology showed diffuse lymphocytic infiltration. Postmortem examination of FOP patients provided precious information about involvement of other tissues, suggesting an initial and widespread inflammatory/dystrophic phase, to be further investigated, because it might reveal new insights about a FOP mutation cascade.

Slippery when wet: The free surface of the articular cartilage.

The free surface of the articular cartilage must withstand compressive and shearing forces, maintain a low friction coefficient and allow oxygen and metabolites to reach the underlying matrix. In many ways it is critical to the physiology of the whole tissue and its disruption always involves deep pathological alterations and loss of the joint integrity. Being very difficult to image with section-based conventional techniques, it was often described by previous research in conflicting terms or entirely overlooked. High-magnification face-on observations with high resolution scanning electron microscopy and with scanning probe microscopy revealed a very thin, delicate superficial layer rich in glycoconjugates, which may explain the very low friction coefficient of the tissue but which was very easily altered and/or dissolved in the preparation. Beneath this superficial sheet lies a thicker coat of thin, highly uniform, slightly wavy collagen fibrils lying parallel to the surface and mutually interconnected by a huge number of interfibrillar glycosaminoglycan bridges. These bridges and the collagen fibrils form an extended reticular structure able to redistribute tensile and compressive stress across a larger area of the surface and hence a greater volume of tissue.

Exploring the role of nasal cytology in chronic rhinosinusitis.

OBJECTIVE: Characterising the eosinophilic profile represents the main step in chronic rhinosinusitis (CRS) endotyping. The aim of the study is to verify the correlation between different methods for tissue eosinophilia quantification. METHODS: 33 CRS patients undergoing endoscopic sinus surgery and 30 controls undergoing non-CRS surgeries were enrolled. Blood venous sampling, nasal biopsy on uncinate process (UP), nasal cytology on inferior turbinate (IT) and middle meatus (MM) were performed. RESULTS: Differences in eosinophil count in blood (P=0.0001), UP (P#x003C;0.0001), IT (P = 0.01) and MM (P = 0.0006) were significant between CRS cases and controls. A weak correlation was found between UP and blood eosinophil count (r = 0.34, P = 0.006) and between UP and IT eosinophil count (r = 0.30, P = 0.017). Moderate correlation between UP and MM (r = 0.51, P #x003C; 0.0001) was shown. ROC analysis predicted eosinophilic CRS with an overall low sensitivity. Once allergic patients were excluded from the analysis, the sensitivity decreased for sampling on IT and increased for MM sampling. CONCLUSIONS: This study suggests that MM cytology gives more accurate information on the degree of tissue eosinophilia. Replication in wide and unbiased cohorts is necessary to verify these results and define accurate thresholds.

3D Reconstruction of HvRNASET2 Molecule to Understand Its Antibacterial Role.

Recent studies performed on the invertebrate model Hirudo verbana (medicinal leech) suggest that the T2 ribonucleic enzyme HvRNASET2 modulates the leech's innate immune response, promoting microbial agglutination and supporting phagocytic cells recruitment in challenged tissues. Indeed, following injection of both lipoteichoic acid (LTA) and Staphylococcus aureus in the leech body wall, HvRNASET2 is expressed by leech type I granulocytes and induces bacterial aggregation to aid macrophage phagocytosis. Here, we investigate the HvRNASET2 antimicrobial role, in particular assessing the effects on the Gram-negative bacteria Escherichia coli. For this purpose, starting from the three-dimensional molecule reconstruction and in silico analyses, the antibacterial activity was evaluated both in vitro and in vivo. The changes induced in treated bacteria, such as agglutination and alteration in wall integrity, were observed by means of light, transmission and scanning electron microscopy. Moreover, immunogold, AMPs (antimicrobial peptides) and lipopolysaccharide (LPS) binding assays were carried out to evaluate HvRNASET2 interaction with the microbial envelopes and the ensuing ability to affect microbial viability. Finally, in vivo experiments confirmed that HvRNASET2 promotes a more rapid phagocytosis of bacterial aggregates by macrophages, representing a novel molecule for counteracting pathogen infections and developing alternative solutions to improve human health.

An in-depth description of head morphology and mouthparts in larvae of the black soldier fly Hermetia illucens.

The larvae of the black soldier fly (BSF) Hermetia illucens are increasingly being used for waste management purposes given their ability to grow on a wide range of organic decaying materials. Although significant efforts have been spent to improve the mass rearing of BSF larvae on specific substrates and their bioconversion capability, little is known about the biology of this insect, especially with regards to the digestive system. In this study, we analyzed the morphology of the head and buccal apparatus of H. illucens larvae by using optical and scanning electron microscopy, evaluating the different mouthparts and their modifications during larval development. Our analysis showed that the larval head of H. illucens presents similarities to those of campodeiform insect larvae, whereas the mandibular-maxillary complex represents a food intake solution typical of Stratiomyidae that enables BSF larvae to ingest semiliquid food. The mouthparts resemble a "tunnel boring machine", where the hypopharynx separates finer organic particles from coarser and inorganic ones.

The synovial surface of the articular cartilage.

The articular cartilage has been the subject of a huge amount of research carried out with a wide array of different techniques. Most of the existing morphological and ultrastructural data on the this tissue, however, were obtained either by light microscopy or by transmission electron microscopy. Both techniques rely on thin sections and neither allows a direct, face-on visualization of the free cartilage surface (synovial surface), which is the only portion subject to frictional as well as compressive forces. In the present research, high resolution visualization by scanning electron microscopy and by atomic force microscopy revealed that the collagen fibrils of the articular surface are exclusively represented by thin, uniform, parallel fibrils evocative of the heterotypic type IX-type II fibrils reported by other authors, immersed in an abundant matrix of glycoconjugates, in part regularly arranged in phase with the D-period of collagen. Electrophoresis of fluorophore-labeled saccharides confirmed that the superficial and the deeper layers are quite different in their glycoconjugate content as well, the deeper ones containing more sulfated, more acidic small proteoglycans bound to thicker, more heterogenous collagen fibrils. The differences found between the synovial surface and the deeper layers are consistent with the different mechanical stresses they must withstand.

Biomarkers of Browning in Cold Exposed Siberian Adults.

Cold-exposure promotes energy expenditure by inducing brown adipose tissue (BAT) thermogenesis, which over time, is also sustained by browning, the appearance, or increase, of brown-like cells into white fat depots. Identification of circulating markers reflecting BAT activity and browning is crucial to study this phenomenon and its triggers, also holding possible implications for the therapy of obesity and metabolic diseases. Using RT-qPCR, we evaluated the peripheral blood mononuclear cells (PBMC) expression profile of regulators of BAT activity (CIDEA, PRDM16), white adipocytes browning (HOXC9 and SLC27A1), and fatty acid ß-oxidation (CPT1A) in 150 Siberian healthy miners living at extremely cold temperatures compared to 29 healthy subjects living in thermoneutral conditions. Anthropometric parameters, glucose, and lipid profiles were also assessed. The cold-exposed group showed significantly lower weight, BMI, hip circumference, and PBMC expression of CIDEA, but higher expression of HOXC9 and higher circulating glucose compared to controls. Within the cold-exposed group, BMI, total cholesterol, and the atherogenic coefficient were lower in individuals exposed to low temperatures for a longer time. In conclusion, human PBMC expresses the brown adipocytes marker CIDEA and the browning marker HOXC9, which, varying according to cold-exposure, possibly reflect changes in BAT activation and white fat browning.

Normal human macula densa morphology and cell turnover: A histological, ultrastructural, and immunohistochemical investigation.

The aim was to analyze the morphology of normal human macula densa (MD), evaluate the cells that may be responsible for its turnover, and collect quantitative data. Of four samples of normal human renal tissue, two were embedded in resin to measure the longitudinal extension and examine the ultrastructure of the MD, the other two were embedded in paraffin to study apoptosis and cell proliferation. The MD is composed of a monolayer tissue about 40 µm long, which includes 35-40 cells arranged in overlapping rows. Ultrastructurally, MD cells show two polarized portions: an apical end, with sensory features, and a basolateral aspect, with paracrine function. MD cells are connected apically by tight junctions, with/without adherens junctions, which form a barrier between the distal tubule lumen and the interstitium. Cells in degeneration, often associated with macrophages, and undifferentiated cells were found in the MD and adjacent distal tubule. A filamentous mat previously described in proximal tubule scattered tubular cells (STCs) was detected in the basal cytoplasm in undifferentiated cells. The tissue was consistently negative for the proliferation marker Ki67 and for the apoptotic markers caspase-3 and caspase-9. This work confirms our earlier morphological findings and provides new data: (a) MD cells display both apical adherens and tight junctions, the latter forming a tubulo-mesangial barrier; (b) the MD is a monolayer made up of about 40 cells arranged in rows; (c) the simultaneous presence of degenerating (8-13%) and undifferentiated (4-13%) cells reminiscent of STCs suggests a non-negligible cell turnover.