Tissue engineering is a multidisciplinary field that relies on the development of customized biomaterial to support cell growth, differentiation and matrix production. Toward that goal, we designed the grafting of silane groups onto the chitosan backbone (Si-chito) for the preparation of in situ setting hydrogels in association with silanized hydroxypropyl methylcellulose (Si-HPMC). Once functionalized, the chitosan was characterized, and the presence of silane groups and its ability to gel were demonstrated by rheology that strongly suggests the presence of silane groups. Throughout physicochemical investigations, the Si-HPMC hydrogels containing Si-chito were found to be stiffer with an injection force unmodified. The presence of chitosan within the hydrogel has demonstrated a higher adhesion of the hydrogel onto the surface of tissues. The results of cell viability assays indicated that there was no cytotoxicity of Si-chito hydrogels in 2D and 3D culture of human SW1353 cells and human adipose stromal cells, respectively. Moreover, Si-chito allows the transplantation of human nasal chondrocytes in the subcutis of nude mice while maintaining their viability and extracellular matrix secretory activity. To conclude, Si-chito mixed with Si-HPMC is an injectable, self-setting and cytocompatible hydrogel able to support the in vitro and in vivo viability and activity of hASC. Tissue engineering is a multidisciplinary approach that incorporates biology, medicine and engineering . As a fiThroughout the literature, several strategies have been developed to prepare the ideal scaffold ,7,8. HydNumerous biomaterials have been used for the fabrication of hydrogels, including natural materials derived from animals or plants  and synthetic materials with a wide range of synthetic polymers . ScaffolIn the past few years, the authors have been developing a cellulose-based hydrogel (Si-HPMC). The silanization of HPMC with pendant alkoxysilane or silanolate groups allows reversible, pH-dependent, polycondensation by sol–gel route without any reactants that are often cytotoxic, thus allowing the injected hydrogel to fit and remain at the damaged area. The deficiencies of silated HPMC is that it is a low degradable hydrogel , and it To overcome these problems, one of the candidates of interest as natural polymeric material for scaffold preparation would be chitosan (chitin-derived polymer) . Chitin Chitosan, a deacetylated chitin, is a polymer that has free amine groups within its backbone chain, thus has the characteristics of a polymeric hydrogel owing to a high water absorption capacity . ChitosaThis paper deals with the functionalization of chitosan with silane groups towards the development of an injectable self-setting chitosan-based hydrogel. First of all, we reported the protocol and the efficacy of the functionalization of chitosan. Then, the formulation of the silated chitosan (Si-chito) was described. Following the preparation of the hydrogels, their rheological and mechanical characterizations were performed, and the cell viability in 2D and 3D was evaluated. Finally, the preclinical relevance of our Si-chito hydrogels was investigated in subcutaneous nude mouse cells.The 4-(2-hydroxyethyl)-1-piperazine-ethanesulfonic acid , (3-Isocyanatopropyl)triethoxysilane  and the chitosan  were purchased from Sigma Aldrich, Darmstadt, Germany. The NaCl  and the HCl  were ordered from VWR, Radnor, PA, USA.v/v) to eliminate unreacted GPTMS, and HPMC-Si powder was dried at 37 °C. Silanized chitosan (Si-chito) was synthesized as follows: 100 mL of chitosan  was poured in a round-bottomed flask and from 1 to 2.5 eq (1.55 mL to 3.875 mL) of (3-Isocyanatopropyl)triethoxysilane (IPTS) was added. The mixture was then stirred from 1 h to 5 h at room temperature. The synthesis conditions are listed in Silanized hydroxypropyl methylcellulose (Si-HPMC) and the acidic buffer solution  were prepared as previously described ,29,30. Wwt/wt) was measured as 1.5%.SEM-EDX monitoring was used to determine the efficiency of the grafting, and coupled plasma atomic emission spectroscopy (ICP-ES) was used to determine the amount of grafted silane (for the selected condition), and the percentage  were performed in 0.09 M NaOHaq. The dialyzes eliminate the non-grafted silane derivatives used for siloxane grafting onto the HPMC and Chitosan. The hydrogel precursor solution was then obtained by mixing 1 volume of the above Si-HPMC/Si-chito basic solution contained in one Luer-lock syringe, with 0.5 volume of acidic BS in another Luer-lock syringe, by the interconnection of both syringes; the final pH is 7.4. This precursor mixture was still injectable at room temperature during 30–40 min until the gel point was reached. A 0 wt % Si-chito hydrogel consists of 98 wt % of water and 2 wt % of Si-HPMC polymer, and a 2 wt % Si-chito hydrogel consists of 96% wt % of water and 2 wt % of Si-HPMC and 2 wt % of Si-chito polymers.Si-HPMC polymer (3 wt %) was dissolved in 0.2 M NaOH aqueous solution; then Si-chito (from 0 to 3 wt %) was dissolved in 3 wt % Si-HPMC aqueous solution (0.2 M NaOH). Finally, two dialyzes with a molecular weight cutoff at 6–8 kDa  were autoclaved at 120 °C for 20 min with an Alphaklave 23.®, Dreieich, Germany) with a titanium cone-plate geometry . Steady shear tests were carried out at 25 °C on different Si-HPMC and Si-HPMC/Si-chito solutions (2 and 3 wt %). The operating shear rate ranged from 0.1 to 9000 s−1. Different flow curves were fitted and extrapolated to lower shear rates by the Cross equation [Rheological measurements were performed using the Haake MARS rheometer (ThermoHaakeequation .Injectability properties were investigated using a compression test device (TAHDplus) with a 5 kg load cell utilized for the measurements at an injectability rate of 17 sec per mL through an 18G needle. A syringe containing 1 mL of hydrogel precursor, after mixing Si-HPMC or Si-chito with the acidic buffer, was set, and the injection force was measured.The gel points were obtained on HAAKE RheoStress RS300 rheometer  using cone/plate geometry . Liquid hydrogel precursor solutions were injected onto the plate immediately after mixing, and the measurements started 1–2 min later. Storage modulus (G′) and loss modulus (G″) was monitored as a function of time under oscillation frequency sweep (from 1 to 22 Hz) at constant temperature (23 °C). The gel point was achieved when tan d = G″/G′ became independent on frequency. Each sample was measured in triplicate.The viscoelastic moduli (storage modulus G’ and loss modulus G″) and the breaking strength (σ) were obtained using a HAAKE MARS rheometer (Thermo Scientific) using plate geometry . Liquid hydrogel precursor solutions were injected into the well of 12 well plates immediately after mixing, stored under a humid atmosphere, and the measurements started 14 days later (full crosslinking of the network). Storage modulus (G′), loss modulus (G″) and breaking strength (σ) were monitored as a function of strain (from 0.1 to 3000 Pa) at constant frequency and temperature (1 Hz and 23 °C). Each sample was measured in triplicate.The adhesion of the hydrogels was evaluated via a traction test using a TAHD plus device. First, the hydrogel was molded between two layers of the desired substrate .To do so, a spacing disc was first prepared with 6 mm external diameter, 4 mm internal diameter and 3/10 mm-thickness A. This sThen, the samples were prepared as follows: first, a disc of the desired substrate was put into a well of a 12 well plate. Then the spacing disc was put onto the substrate, and the 4 mm well was filled with the desired hydrogel and covered with a second disc of the substrate B. After −1, and the force was monitored (N).Once the hydrogel was fully crosslinked, the structure was pasted onto the TAHD plus using cyanoacrylate super glue to anchor each faces to the device C. The tr2 incubator at 37 °C in Dulbecco’s modified Eagle’s medium (DMEM) . Culture media was supplemented with 10% fetal bovine serum, 1% penicillin/streptomycin and 1% L-glutamine and changed each 2–3 days. Human adipose stromal cells (hASC) and human chondrosarcoma cell line (SW1353) were cultured in a 5% CO2. After 24 h, the culture medium was removed, and 500 µL/well of precursor solution of pure Si-HPMC or SI-HPMC/Si-chito were added. The hydrogel samples were incubated at 37 °C for 1 h before adding 1 mL of culture medium. As a positive control, cells were cultured in the absence of Si-HPMC hydrogel. As a negative control, cells were cultured in the presence of actinomycin-D (5 µg mL−1), which is a transcription inhibitor. After 1, 2, 3 and 7 days of culture, the hydrogels and culture medium were removed by aspiration. Trypan blue exclusion dye assays were performed to count the cells. In order to evaluate their mitochondrial activity, the methyl tetrazolium salt (MTS)  test was performed by adding the MTS solution to each well for 1 h. The optical density of the formazan dye was measured in a spectrophotometer at 490 nm. Each condition was tested in quadruplicate.SW1353 cells were allowed to attach to 24-well plates at a final density of 10,000 cells per cm−1) was used as an internal control for cell death. In each well, the culture medium was replaced by 200 µL of a solution containing 2.5 µL of DMEM medium supplemented with 0.25 µL of calcein-AM and 5 µL of ethidium homodimer-1. After 45 min the dye mixture was removed, and the hydrogels were intensively rinsed with phosphate-buffered saline solution before being observed with a confocal laser scanning microscope (Nikon D-eclipse C1) equipped with an argon/krypton laser. Each condition was tested in quadruplicate, and for each sample, two random positions  were chosen within the hydrogel, and a series of images were recorded starting from these positions along the z-axis. The 210 images obtained per sample were analyzed with a quantimeter Q550 .3D culture cell viability was assessed by using LIVE/DEAD Assay Kits  along with confocal image analysis. hASC cells were dispersed in the hydrogel precursor solutions within 5 min following their preparation (thus before gelification) at a final concentration of 1,000,000 cells per mL of hydrogel. Five hundred microliter of each mixture was molded into 24-wells plates and incubated at 37 °C for 1 h to allow the gelification of the hydrogels. Afterward, 1 mL of DMEM medium was added per well, and the samples were incubated for 1, 2, 3, and 7 days before the Live/Dead assays. Actinomycin-D treatment  were used for the in vivo study . All animals were treated in accordance with the Nantes University of Medical Animal Care Guidelines. Four experimental conditions were designed. Two control groups were designed: Si-HPMC was injected alone as negative control and Si-HPMC + hNC as a positive control. Then, 2 experimental groups were designed: Si-HPMC/Si-chito alone and Si-HPMC/Si-chito + hNC. Four samples per mice were implanted subcutaneously in the back along each side of the mice dorsum. Animals were under general anesthesia using isoflurane gas , and the implantations were performed under aseptic conditions. After 6 weeks, mice were anesthetized and euthanized. Hydrogels were individually dissected and removed from the subcutaneous site. Histological analyses were performed.All histological samples were fixed in 10% formalin and embedded in paraffin. Embedded samples were sectioned (5 µm-thick). Thereafter paraffin sections were deparaffinized using toluene, rehydrated through a graded series of ethanol and rinsed in distilled water. Tissue sections were stained with hematoxylin–eosin-safran (HES), Alcian blue (AB) and Masson’s trichrome (MT). Sections were then visualized using a light microscope . HES stains the nucleus in purple, the cytoplasm in pink, collagen fibers in orange, and AB reveals the presence of glycosaminoglycans (GAGs) in blue. Finally, MT stains the nucleus in blue/black, the cytoplasm in pink/red, hematins and keratin in bright red and collagen in green.p < 0.05.Results are expressed as mean ± SD of replicate determinations. Comparative studies of means were performed by using two-way ANOVA with minimal statistical significance at The functionalization of the chitosan backbone was developed through the modification of several chemical parameters. As summarized in In a second step, the ability of the functionalized chitosan to form a hydrogel was evaluated. The different steps of the formulation  were monitored and are reported in The selected formulation was then characterized to determine their potential usefulness as hydrogel for tissue engineering. Throughout the physicochemical characterizations, we evaluated and selected the right formulation (in terms of concentration) for biological investigation. First, we measured the viscosity of the solutions. As reported in Usually, the viscosity profile of solutions can be quite informative of the injectability behavior. Nevertheless, the syringeability of the Si-HPMC/Si-chito solutions was monitored. 50 to 70 N is the maximum force that can be applied by hand . As expeThe gel point, the time point at which the solution becomes a solid, was determine by monitoring the Tan d as a function of time. The time points are reported in After evaluating the rheological properties of our hydrogels, we investigated the mechanical behavior to determine either the Si-HPMC/Si-chito could withstand the mechanical force in vivo or not. To do so, the storage modulus (G′) and the breaking strength (σ) at the end of the crosslinking process were measured. The results, plotted in ® as a positive control. The results  compared to Si-HPMC alone, HPMC as negative control and cyanoacrylate super glue results  showed aMoreover, the confocal analysis allowed us to have a look at the shape of the embedded cells. While the hASC usually showed a round spherical shape (data not are shown), specific of the non-adherent cell, after 7 days of culturing within Si-HPMC/Si-chito hydrogels, they displayed a stretching of their shape with filaments , leadingThe aim of the cellular studies was to evaluate the cytotoxicity of our mixed hydrogels in 2D (hydrogel above the cell layer) and in 3D (cell proliferation within the hydrogel).Throughout the physicochemical characterizations, we have selected the best condition to synthesize and formulate the Si-chito: 1 equivalent of IPTS with no pH control for 3H followed by its formulation in association with Si-HPMC as copolymer.The selected Si-chito was formulated with an increasing ratio, and the cell metabolism was monitored using MTS assay on days 1, 2, 3 and 7. The results were plotted in 6 cells/mL) were mixed with Si-HPMC or Si-HPMC/Si-chito hydrogels and injected in subcutis for 6 weeks. Histological staining demonstrated that the conditions without chondrocytes (Si-HPMC and SI-HPMC/Si-chito) showed no cells. However, some colorations were observed within the graft as the chitosan interacted with the histological staining molecules. These staining can suggest that both silated polysaccharides are not fully miscible and areas with more concentrated Silated chitosan seem to be visible by the 3 stainings. Towards the utilization of Si-HPMC/Si-chito hydrogels for tissue engineering purposes, the next step was to investigate if they may be able to support the chondrogenic activity of chondrocytes by detecting the presence of chondrogenic extracellular matrix components. Consequently, we performed an in vivo implantation experiment in nude mice subcutis. After harvest, isolation and culture, hNC  ,34,35,36The strategy of the self-setting formulation of the hydrogel was based on the silane chemistry. This process was selected with regard to previous work of the laboratory dealing with Silated HPMC and in the objective to get functionalized polymers using the same chemistry for the development of tunable hydrogels.+) into siloxanes (Si-O-Si) under physiological conditions [Si-HPMC allows in situ forming hydrogels based on inorganic–organic chemistry, leveraging the self-condensation of grafted silanolate moieties . The adhesion onto the surface of the targeted tissue is essential for tissue engineering. Indeed, the maintenance on the site will give the time required for the cellularized hydrogel to produce the extracellular matrix molecules and potentially heal the damaged tissue.In this paper, we focus on the use of chondrocytes to evaluate the potential use of our hydrogels for skeletal tissue engineering. The ideal biomaterial should not only allow for the maintenance of the chondrocyte differentiation but should also enable their transplantation via a mini-invasive surgical protocol in vivo. Consequently, to decipher whether Si-HPMC/Si-chito hydrogels could represent this ideal scaffold, we investigated its ability to support the transplantation of chondrogenic cells and the formation of cartilage tissue in vivo. To address this issue, we focused our attention on the use of cells exhibiting a reliable and robust chondrogenic potential such as hNC ,18. InteIn conclusion, we have developed the synthesis of silanized chitosan, which can be formulated with Si-HPMC to obtain a stiffer hydrogel showing a stronger adhesion onto the surface of tissues. This self-setting hydrogel showed good cytocompatibility and can support cell viability and extracellular matrix secretion in a subcutaneous pocket in nude mice. The efficiency in a relevant animal model for cartilage or osteochondral defects regeneration will now be conducted as a preclinical investigation.


Phyllostachys nidularia (Bambusoideae: Poaceae), widely distributed in the Yangtze River Basin and various provinces (regions) in southern China, is one of the most important small and medium-sized bamboo species used in both bamboo shoots and timber. In the present study, we assembled a complete chloroplast genome of the economically important bamboo form Phyllostachys nidularia f. farcta H.R.Zhao & A.T.Liu using whole genome sequencing data previously reported. The complete chloroplast (cp) genome is 139,706 bp in length. A total of 129 unique genes were annotated, including 82 protein-coding, 39 tRNA, and eight rRNA genes. Phylogenetic analysis results supported that P. nidularia f. farcta was closely related to Phyllostachys reticulata. This work would help us better understand the evolution of the Phyllostachys cp genome. Phyllostachys nidularia  is one of the most widely distributed bamboo species in China, with high economic value and ecological benefits. Phyllostachys nidularia is closely related to people's life and production, its bamboo shoots are edible, bamboo stalks are important construction, manufacturing, and paper industry materials, and P. nidularia is one of the most important economic bamboo species in China  is a form of P. nidularia. It traditionally identifies the analyzed specimen as Phyllostachys nidularia f. farcta in China. Compared with the original form, its culm is solid or nearly solid  method. The alignment was achieved with MAFFT . Total genomic DNA was extracted from bamboo shoot bud tissue and the specimen voucher is deposited in the college of forestry, Nanjing Forestry University NJFU-2020779). The raw data was uploaded to NCBI with Accession number-SRS6922745. The Hiseq 2500 sequencer was used to obtain 35.4 gb of raw data. After filtering and pruning with fastp , eight tRNA genes , and all rRNA genes  were located in the IR regions. The GC content of complete cp genome was 38.88%. The GC content of IRs (44.22%) was higher than that of LSC (36.97%) and SSC (33.16%).The cp genome was 139,706 bp in size and is quadripartite, consisting of a pair of inverted repeats , a large single copy , and a small single copy . There were 129 unique genes, including 82 protein-coding, 39 tRNA, and eight rRNA genes. Seven protein-coding genes (P. nidularia f. farcta was closely related to P. reticulata (P. nidularia f. farcta will provide useful genetic information for the further study and conservation of bamboo species. Phyllostachys is divided into two groups (Sect. Phyllostachys and Sect. Heteroclada). We have checked the data several times and found that Phyllostachys nidularia and Phyllostachys reticulata (MN537808.1) are more similar based on pairwise genetic distance, and do not show closer Phyllostachys nigra var. henonis of Sect. Heteroclada in genetic distance. Previous studies (Zhang et al. Phyllostachys species may have hybridized during their long evolutionary history, which has deepened our understanding of the phylogeny of Phyllostachys.The phylogenetic analysis indicated that ticulata . The chl


The biological and pathological functions of ADAMTS proteases are determined broadly by their respective substrates and their interactions with proteins in the pericellular and extracellular matrix. For some ADAMTS proteases, substrates have been identified and substrate cleavage has been implicated in tissue development and in disease. For other ADAMTS proteases, substrates were discovered in vitro, but the role of these proteases and the consequences of substrate cleavage in vivo remains to be established. Mutations in ADAMTS10 and ADAMTS17 cause Weill–Marchesani syndrome (WMS), a congenital syndromic disorder that affects the musculoskeletal system , the eyes (lens dislocation), and the heart . WMS can also be caused by mutations in fibrillin-1 (FBN1), which suggests that ADAMTS10 and ADAMTS17 cooperate with fibrillin-1 in a common biological pathway during tissue development and homeostasis. Here, we compare and contrast the biochemical properties of ADAMTS10 and ADAMTS17 and we summarize recent findings indicating potential biological functions in connection with fibrillin microfibrils. We also compare ADAMTS10 and ADAMTS17 with their respective sister proteases, ADAMTS6 and ADAMTS19; both were recently linked to human disorders distinct from WMS. Finally, we propose a model for the interactions and roles of these four ADAMTS proteases in the extracellular matrix.Secreted  In developing lungs, Adamts10 was expressed in the connective tissue between the bronchi and the accompanying blood vessels. Adamts10 was also expressed in the stomach, duodenum, pancreas, dorsal root ganglia, and primary ossification centers of vertebrae, but was absent in the liver. In the developing musculoskeletal system, Adamts10 mRNA was detected between the cartilaginous metacarpals and metatarsals of the hand and feet and in dense connective tissues such as the joint capsule and developing tendons and ligaments. At E17.5 Adamts10 expression was strong in the cartilage of developing bones and in the wall of large arteries. Adamts10 was expressed in several ocular tissues during development, including non-pigmented ciliary epithelium, lens fiber cells, and parts of the retina [Adamts10 continued to be expressed in adult chondrocytes, tendon, and skeletal muscle in mice [ADAMTS10 mRNA was expressed in the heart, brain, lung, pancreas, but ADAMTS10 expression was notably low or absent in skeletal muscle [Adamts10 knockout mice apparently undergo normal development, with limited embryonic lethality (see below) [The biological functions of ADAMTS6, 10, 17, and 19 are not only determined by differential posttranscriptional and posttranslational modifications, which may guide ECM localization and substrate binding and cleavage, but also by the cell types and tissues that express these proteases. ectoderm ,33. RNA e retina . Express in mice . In humae below) .ADAMTS10, ADAMTS17 is widely expressed in many fetal and adult tissues in humans [ADAMTS17, isoform a (22 exons) and isoform b (16 exons), with distinct expression patterns, were identified. Both ADAMTS17 isoforms were expressed in lung, brain, and the eye, including the retina. Immunostaining in human normal and cancer tissues showed widespread expression in multiple tissues to varying degrees [Adamts17 expression in the eye, especially at the lens equator and in the trabecular region [Adamts17 mRNA was detected in the perichondrium, the intervertebral disc, and in the smooth muscle cell layer of blood vessel walls. The most pronounced expression of Adamts17 was observed in lung parenchyma, skin epidermal basal cell layer, and in developing hair follicles. Adamts17 mRNA was notably absent from the growth plate of long bones, the endothelium of blood vessels, and the bronchial epithelium in lung tissue.Similar to n humans . Two iso degrees . However degrees . In-situr region . Apart fADAMTS6 and ADAMTS19 is currently available. ADAMTS6 mRNA was detected in the mouse heart, specifically in the outflow tract, the heart valves, the atria, and the ventricular myocardium [ADAMTS6 mRNA was weakly expressed in placental tissue, but was not detected in mouse embryonic development or in adult human tissues [ADAMTS19 mRNA was detected in fetal lung and osteosarcoma tissue [Adamts19 was shown to be expressed in the heart valve at all stages of heart valve formation and elongation and in bone and cartilage anlagen at E10.5 [Less detailed information about the expression patterns of ocardium . In nort tissues . ADAMTS1a tissue . More reat E10.5 .ADAMTS10 and ADAMTS17 show broad, both overlapping and distinct gene expression patterns in tissues. Therefore, it is possible that ADAMTS10 and ADAMTS17 may compensate for each other in mouse knockout models of the individual proteases at least in some tissues (see below). If these proteases can indeed compensate for each other in murine loss-of-function models, similar to what has been observed for the ADAMTS7/ADAMTS12 and ADAMTS9/ADAMTS20 pairs, needs to be explored [ADAMTS10 with ADAMTS17 or vice versa appears to occur in tissues affected in WMS, specifically the eye and the musculoskeletal system. However, a lack of cardiac involvement and joint contractures in WMS4 due to ADAMTS17 mutations leaves the possibility that compensation with ADAMTS10, but not vice versa, causes the lack of cardiac and joint presentations in WMS4 (see below). ADAMTS6 and ADAMTS19 appear to have more tissue-specific expression patterns, which are reflected in the specific diseases associated with mutations in ADAMTS6 and ADAMTS19, as described in the next section.Taken together, explored ,20. No cADAMTS10 and ADAMTS17 cause WMS [ADAMTS10, but is typically not observed in individuals with WMS4 due to ADAMTS17 mutations. Similarly, joint stiffness is underrepresented in WMS4. Given that there are only a few individuals with WMS reported worldwide, definitive genotype–phenotype correlations are currently not feasible. Therefore, it is unclear if WMS caused by mutations in ADAMTS10 or ADAMTS17 represent different ends of a spectrum of WMS severity or if WMS due to mutations in ADAMTS10 or ADAMTS17 represent distinct disease entities, as suggested by the prior use of “WMS-like syndrome” for WMS4 [ADAMTS10 and ADAMTS17 mutations causing WMS are spread out over the entire protein with no specific mutation hot spots  of more than 12,000 cases versus 390,000 controls as susceptibility loci likely to cause carpal tunnel syndrome [ADAMTS10 and ADAMTS17 were identified in a GWAS for genes influencing differential body fat distribution in women compared to men [ADAMTS10 or ADAMTS17 to traits governed by many genes are expected to be small. However, the possibility of studying double knockout mice for ADAMTS10 and ADAMTS17 may provide an opportunity to gain some insights into the contribution of these protease to multigenic traits, such as carpal tunnel syndrome or body fat distribution. Mutations in ause WMS ,25,26,27ot spots a. Howevein place ,45,46,47syndrome . In addid to men . The indADAMTS6 are associated with altered ventricular conduction in the heart and cause a prolonged QRS interval observed by electrocardiography [ADAMTS19 cause non-syndromic heart valve disease [Mutations in iography . A prolo disease . In functional studies, disease-associated point mutations in recombinant ADAMTS6, 10, or 17 collectively abolished or reduced secretion of these proteases ,26,34. TAdamts6, Adamts10, Adamts17, and Adamts19 [Adamts6 knockout was identified in an ethylnitrosourea (ENU) mutagenesis screen for congenital heart defects [Adamts6-deficient mice show developmental heart defects, including double outlet right ventricles, atrioventricular septal defects, and ventricular hypertrophy. Adamts6-deficient mice die at or around birth.Phenotypes of knockout mice have been reported for Adamts19 ,34,42,50Adamts10 were reported that show overlapping and, somewhat surprising, also distinct phenotypes. In the first strain, 41 nucleotides in exon 5 of Adamts10 were deleted via homologous recombination resulting in a reading frame shift and in presumed nonsense-mediated ADAMTS10 mRNA decay [Adamts10 knockout mice showed some embryonic lethality in a pure C57BL/6 background which was not observed after backcrossing in a mixed C57BL/6 × 129/Sv background. Adamts10 knockout mice showed reduced weight gain, but no abnormalities in the musculoskeletal system that would be characteristic for WMS. ADAMTS10-deficient eyes developed a mild shallowing of the anterior chamber, indicative of a potential abnormal positioning of the lens. However, the ciliary zonule was intact. Despite the presence of an intact ciliary zonule, immunostaining showed that fibrillin-2, which is normally not detected in the adult ciliary zonule, persisted beyond prenatal and early postnatal development, suggesting that ADAMTS10 may play a role in the degradation or masking of fibrillin-2 in the postnatal ciliary zonule [ADAMTS10 mutation (R237X) was introduced at the homologous location in the mouse Adamts10 gene (S236X) using CRISPR-Cas9 genome editing [Adamts10 models, homozygous Adamts10-S236X mice developed hallmark features of WMS, i.e., shorter long bones, thicker skin, and increased muscle mass. The long bones were 6%–10% shorter in ADAMTS10-deficient mice compared to wild type controls and alterations in the growth plate, i.e., a shorter resting zone with increased proliferation and an extended hypertrophic zone, were observed. The increased skeletal muscle mass in Adamts10-S236X knockout mice was attributed to the presence of more, but smaller myofibers compared to controls. The eyes of Adamts10-S236X mice had a smaller ciliary body with altered ciliary processes and showed a similar persistence of fibrillin-2 in the ciliary zonule of adult ADAMTS10-deficient eyes compared to the Adamts10-strain described above. Since both mouse strains show overlapping and distinct features, notably in the WMS-related musculoskeletal features which were present in Adamts10-S236X mice, it is tempting to speculate if the propeptide domain of ADAMTS10, which could be detected in the Adamts10-S236X mice, exerts a dominant negative effect contributing to the development of the musculoskeletal manifestations in Adamts10-S236X mice. This is especially relevant given the fact that the ADAMTS10 propeptide cannot be cleaved by furin and as such may not be released as a distinct peptide entity in wild type mice or humans.Two different mouse strains targeting NA decay . These Ay zonule ,51. HoweAdamts17 knockout mouse was generated by Cre/Lox-mediated deletion of a portion of exon 4, which generates a frameshift mutation and results in ADAMTS17 mRNA depletion [Adamts10 knockout mice, Adamts17 knockout mice showed some neonatal or perinatal lethality. Adamts17 knockout mice showed features characteristic of WMS, including shorter long bones, brachydactyly, and thick skin. Bone shortening in Adamts17 mice was also attributed to alterations in the growth plate. In contrast to Adamts10-S236X mice, a shortened hypertrophic zone in the growth plate of the tibia, with no alterations in chondrocyte proliferation, was observed in Adamts17-deficient mice. Mechanistically, canonical bone morphogenetic protein (BMP) signaling was attenuated in Adamts17-deficient primary chondrocytes. An eye phenotype was not reported for the Adamts17 knockout mice.An epletion . SimilarAdamts19 knockout mice, recapitulating the heart valve phenotype in human non-syndromic heart valve disease caused by ADAMTS19 mutations [Adamts19-deficient mice develop thickened aortic valves that can result in aortic regurgitation and severe aortic valve stenosis. However, the cardiac valve phenotype was present in only ~40% of Adamts19 knockout mice, suggesting partial compensation by other proteases, potentially ADAMTS10 or ADAMTS17. Expression of ADAMTS10 was indeed observed in the leaflets of the aortic valve and WMS1 due to ADAMTS10 mutations can result in mitral valve prolapse [Very recently, a cardiac valve phenotype was described in utations . Adamts1prolapse ,52.ADAMTS10 or ADAMTS17 or dominant mutations in FBN1 can all cause WMS, a role for both of these ADAMTS proteases in fibrillin microfibril formation and homeostasis was predicted and subsequently confirmed. Wild-type ADAMTS10 cleaves recombinant fibrillin-1 inefficiently, but fibrillin-1 proteolysis was significantly enhanced when the consensus furin site in recombinant ADAMTS10 was restored [Given that recessive mutations in restored . More rerestored . This inrestored ,54,55. Irestored . ADAMTS1restored ,56,57,58restored . The perrestored ,60,61.ADAMTSL2 cause geleophysic dysplasia, which, like WMS, belongs to the acromelic dysplasia group of disorders which all share the musculoskeletal and skin phenotypes [Much less is known about the molecular function and substrate spectrum of ADAMTS6, the sister protease of ADAMTS10. It was shown that ADAMTS6 binds to a similar N-terminal region of fibrillin-1 (exons 8–11) and that ADAMTS6 binding was not affected by a WMS mutation introduced in recombinant fibrillin-1 . Overexpenotypes . Mutatioenotypes ,69.ADAMTS17 mutations, we observed a reduction in fibronectin and fibrillin-1 secretion and their deposition in the ECM [ADAMTS10 mutation were analyzed, fibrillin microfibril deposition was similarly reduced [ADAMTS17 mutations, would suggest a potential role for ADAMTS17 as a chaperone to facilitate fibrillin-1 secretion and possibly secretion of other ECM proteins [ADAMTS17 binds to fibrillin-1 microfibrils and recombinant fibrillin-2 but it did not cleave either of the fibrillin isotypes in vitro . In cont the ECM . These f the ECM . When WM reduced . The obsproteins . When thIn contrast to ADAMTS17, its sister protease ADAMTS19, undergoes furin processing, but does not undergo autoproteolysis . So far, no substrates for ADAMTS19 have been reported. Similar to the ADAMTS6 and ADAMTS10 pair, the possibility exists that ADAMTS17 may act more like an ADAMTS-like protein through its bioactive peptides and ADAMTS19 may be a true protease involved in proteolytic ECM remodeling in vivo.So far, the spectrum of substrates for the ADAMTS6/ADAMTS10 and ADAMTS17/ADAMTS19 protease pairs and their involvement in biological and pathogenic mechanisms are only emerging. Some of the proposed interactions of these ADAMTS proteases and the open questions associated with processes in which these ADAMTS proteases might play a role, are summarized in ADAMTS10, ADAMTS17, or FBN1 result in WMS, one possibility is that ADAMTS17 activates ADAMTS10 when both proteases are sequestered to fibrillin microfibrils in the ECM. Fibrillin-1 binding could induce conformational changes in ADAMTS10 that would expose a putative cryptic cleavage site for ADAMTS17. The resulting proteolysis by ADAMTS17 would activate ADAMTS10 and convert the inactive enzyme into a more efficient protease. Alternatively, ADAMTS17 or its autocatalytic peptides could activate ADAMTS10 independent of propeptide processing through allosteric mechanisms modulating ADAMTS10 protein conformation. We are currently exploring the ADAMTS10-ADAMTS17-fibrillin-1 microfibril network experimentally with mouse models and with in-vitro systems. The connection of ADAMTS6 and ADAMTS10 is so far only studied in epithelial cells where ADAMTS10 suppresses the expression of ADAMTS6 and both ADAMTS proteases show opposing effects on fibrillin microfibril formation [ADAMTS19 were linked to a human disorder only very recently [Adamts19 knockout mice. It will be equally exciting to see how the ADAMTS-fibrillin connections will be further disentangled and what the identification of novel substrates will suggest about the biological functions of this evolutionary related subgroup of ADAMTS proteases.Given the fact that mutations in ormation . The funrecently . It will


Image-guided tumour ablation has developed into a well-established treatment paradigm for solid cancers. As reflected by the increasing number of key publications in CVIR, several needle-guided thermal and non-thermal ablation techniques have evolved into reliable and widely adopted therapies in modern clinical oncology. Although complete tumour eradication has always been the primary objective, fundamental changes to this basic concept may be in the offing. Anecdotal reports of spontaneous regression of distant tumours following local tumour ablation  triggered research of the underlying systemic immune mechanisms. This effort is gradually evolving into novel treatment approaches where focal tumour ablation is combined with passive or active immunotherapy.Immunomodulating properties have been identified for all energy-based needle-guided ablative modalities . EvidencLocal tumour progression (LTP) due to an incomplete ablation remains a major drawback of needle-guided tumour ablation. In this issue of CVIR, we are treated to an article by Zhang et al.  who wereHowever, some degree of reservation towards the results is required as the follow-up time was short, side effects of the combination therapy were not reported and, foremost, because animal results can be far-off translation into clinical practice. Furthermore, 2 of the surviving mice developed rapid and massive tumour growth after tumour re-challenging suggestive for latent pro-oncogenic effects.In conclusion, this study supports the rationale to combine thermal ablation with checkpoint inhibition for HCCs. This proof of concept now warrants further investigation of the optimal dosage, timing, and sequencing of both therapies taking into account potential pro-oncogenic effects. If results can be clinically reproduced, the combination regimen may not only provide an alternative strategy to prevent LTP after HCC ablation, even a paradigm shift towards intentional incomplete ablations combined with immune enhancing drugs is conceivable. Interventional oncologists are well positioned in the centre of oncological care and, given the rise of immunotherapy, the integration of immuno-oncology in interventional oncology is inevitable, as stated in the Society of Interventional Oncology White Paper on the emerging role of immunotherapy in interventional oncology . Conside


P = 0.002), while there was no significant difference in comparison of UC patients with CD patients (P = 0.585). Furthermore, there was a negative correlation between adropin and fecal calprotectin , whereas in the total study population, we found a significant negative correlation with fasting glucose levels . A multivariable logistic regression showed that serum adropin was a significant predictor of positive IBD status when enumerated along with baseline characteristics . Our findings imply that adropin could be involved in complex pathophysiology of IBD, but further larger scale studies are needed to address these findings.Adropin is a novel peptide mostly associated with energy homeostasis and vascular protection. To our knowledge, there are no studies that investigated its relationship with inflammatory bowel diseases (IBD). The aim of this study was to compare serum adropin levels between 55 patients with IBD (30 Ulcerative colitis (UC) patients, 25 Crohn’s disease (CD) patients) and 50 age/gender matched controls. Furthermore, we explored adropin correlations with IBD severity scores, hsCRP, fecal calprotectin, fasting glucose and insulin levels. Serum adropin levels were significantly lower in patients with IBD in comparison with the control group (2.89 ± 0.94 vs 3.37 ± 0.60 ng/mL,  Global prevalence of IBD is on the rise, as the number of countries adopting “western lifestyle” increases3. Although the exact etiology of IBD is unknown, it is currently considered to be multifactorial and that is fabricated through complex interactions between genetic predisposition, environmental factors, immune disorders and microbial disturbances4.Inflammatory bowel disease (IBD) is a chronic relapsing inflammatory disorder of unknown etiology and unpredictable course. It is mainly classified into two disorders: ulcerative colitis (UC) and Crohn’s disease (CD). They both have a similar clinical manifestation whereas they differ in the location and depth of the inflammation. They are also associated with extraintestinal manifestations which can affect the skin, joints, eyes, liver and neurocognitive performance5. It is encoded by Energy Homeostasis Associated Gene (ENHO) expressed in the liver and brain, but its presence is also proven in muscle, heart, pancreas, and kidneys6. A study performed on mice pointed to the role of adropin as a physiological regulator of glucose metabolism and oxidation of fatty acids. It was found that mice with diet induced obesity, when treated with adropin, showed increased glucose tolerance, reduced insulin resistance, and the promotion of carbohydrates in oxidative reactions7. A study conducted on patients presented the first evidence pointing to the link between adropin, obesity and risk of metabolic syndrome. It was found that the concentration of adropin were decreased in subjects with obesity and insulin resistance, and that loss of body weight led to an increase in adropin levels8. Regarding cardiovascular effects, several recent studies linked low adropin levels with high blood pressure. Negative correlation of adropin with arterial blood pressure, as with the levels of vasoconstriction factor endothelin-1, speaks in favor of the beneficial effect of adropin as a vascular protector10. Moreover, adropin is involved in neovascularization and vascular protection through the promotion of endothelial nitric oxide (NO) by regulating vascular endothelial growth factor receptor – 2 (VEGF2) and endothelial nitric oxide synthase (eNOS) pathways11.Special interest of this study is adropin, a novel peptide which acts as an energy regulator through lipid and glucose metabolism15. Inflammation is considered to play a major role in these metabolic disorders and some studies are pointing that they are conditioned by the active phase of IBD16. TNF-α is lately regarded as one of the main inflammatory drivers in the exacerbation of IBD17, and amid its numerous proinflammation effects, TNF-α down regulates eNOS which consequently leads to reduced perfusion, higher leukocyte infiltration, impaired wound healing and endothelial dysfunction18. Given that it was reported by several studies conducted on chronic inflammatory diseases that adropin has a significant correlation with TNF-α21 and since a study conducted on nonhuman primates reported expression of ENHO gene in terminal ileum and colon22, its worthy exploring could adropin be somehow involved in complex IBD pathophysiology. The possible metabolic, immunomodulatory and protective role of adropin stresses the importance of further necessary research in this field.It is indicative by several studies that patients with IBD develop hyperglycemia, insulin resistance and they also have a higher prevalence of diabetes mellitus compared to the general populationWith extensive analysis of available literature, there is no published study that investigated adropin association with IBD. Hence, the aim of this study was to determine serum adropin levels in patients with IBD in comparison with healthy controls. Additionally, we wanted to evaluate the relationship between adropin and UC and CD severity scores, glucose metabolism parameters and inflammatory biomarkers.This cross-sectional study was performed at the University Hospital of Split and the University of Split School of Medicine during the period from 1 December 2017 to 1 June 2018.All subjects were informed about the procedures, course and purpose of the study in a timely manner. Before the start of the study every participant individually signed an informed written consent. The study was approved by the Ethics Committee of University Hospital of Split  and University of Split School of Medicine , and was conducted in accordance with all ethical principles of the Seventh Revision of the Helsinki Declaration from 2013.23. The control group consisted of healthy volunteers, matched with the age and gender of the investigated group. Inclusion criteria were: disease duration of at least one year, stable disease activity in the past 3 months and age between 18 and 65 years. Stable disease activity is referring to clinical evaluation of the disease activity. Using the medical database, we checked did any of the patients contacted a physician regarding new disturbances, worsening and/or had a need for therapy revision. Exclusion criteria were: diabetes; cardiovascular disorders; therapy with corticosteroids during 3 months prior to study onset; substance abuse and consumption of alcohol more than 40 grams/day. We checked medical records of the control subjects regarding gastrointestinal conditions and additionally we performed screening for abdominal pain presence, symptoms related to defecation, change in frequency and form of stool according to the Rome IV criteria for irritable bowel syndrome24, as well as any other symptoms which could suggest gluten and lactose intolerance. If any of these conditions were present, we excluded the subject from the control group. Furthermore, all potential control group subjects undergone detailed physical examination along with laboratory analysis of the complete blood count, differential blood count and levels of high sensitivity C-reactive protein. We excluded all participants who showed any sign of inflammation in any of these steps.This study included 55 adult patients with pre-diagnosed IBD (30 patients with UC and 25 patients with CD) and 50 healthy controls. Diagnosis of UC and CD is based on medical history and clinical, radiological, endoscopic and histological features in accordance with the European Consensus on Crohn’s Disease and Ulcerative Colitis (ECCO)Disease activity was evaluated using clinical and endoscopic indices. Assessment was performed by the same experienced gastroenterologist according to the latest guidelines and the colonoscopy needed for the evaluation of the disease activity was performed within 2 weeks of blood sampling.25.Ulcerative colitis endoscopic index of severity (UCEIS) is a quantitative score for grading mucosal inflammation based on the specific endoscopic findings. Three parameters are graded: vascular pattern; bleeding; erosions and ulcers. Depending on the score there are four possible grades for disease activity: remission (0–1); mild (2–4); moderate (5–6); and severe (7–8)26.Mayo endoscopic score (MES) is an instrument used for evaluation of UC through endoscopic findings. There are four possible stages: 0 - remission; 1 - mild disease with erythema, mild friability and decreased vascular patterns; 2 - moderate disease with marked erythema, friability, erosions and absence of vascular patterns; 3 - severe disease with spontaneous bleeding and ulcerations27.Mayo score is a score used for evaluation of UC activity based on four categories . The results correlate with disease severity: <2 remission; 3–5 mild; 6–10 moderate; 10–12 severe28.Simple endoscopic score for Crohn’s disease (SES-CD) is a grading system used for endoscopic evaluation of CD activity. According to the majority of studies the threshold values for interpretation of the results are: ≤2 remission; 2–7 mild activity; 7–16 moderate activity; and ≥16 severe disease29.Crohn’s disease activity index (CDAI) is a quantitative score for the assessment of CD activity based on clinically reported signs, laboratory results and patient reported symptoms in a 7 day period. The three possible grades depending on the result are: clinical remission <150; mild to moderate activity 150–450; and severe disease >45030.Harvey-Bradshaw index (HBI) is a simplified version of the CDAI for assessment of CD activity. Threshold values for the four possible grades are: clinical remission <5; mild activity 5–7; moderate activity 8–16; and severe disease>1631. Hence, patients were classified using only the endoscopic scores, while clinical index scores were only descriptively reported. UC disease activity was assessed with two endoscopic index scores  and CD disease activity evaluation was performed with one endoscopic score (SES-CD). In the case of discordance between UCEIS and MES score, results of UCEIS were followed due to its methodological superiority and validity which is in accordance to the latest ECCO guidelines.According to the latest ECCO guidelines, clinical indices should be cautiously used since they are not well-validated in the clinical practice and inconsistencies could be seenBlood samples were taken after 12-hour fasting from the cubital vein in test tubes with anticoagulant (K3-EDTA) via polyethylene catheter, and after extraction were centrifuged and stored at −80 °C for further analysis. Serum concentrations of adropin were determined using the dual enzyme-linked immunosorbent assay (ELISA) of human adropin , according to the manufacturer’s instructions. The assay detects immunoreactivity of adropin (34–76) in human, rat and mice serum, and there is a 100% homology between peptides. Calibrations were double measured, optical density (OD) values were in accordance with predefined OD values stated in manufacturer instructions and coefficient of variability (CV) of paired calibrations were <15%. Concentration of the analyzed quality control sample which arrived from the manufacturer was within predefined acceptable deviation. The linear range of the assay was 0.3–8.2 ng/mL and sensitivity was 0.3 ng/mL. CV within the probe was less than 10%, and between probes was less than 15%. All blood samples were processed according to international standards, in the same laboratory, by the same, experienced medical biochemist. Moreover, biochemist was blinded to the subjects group in the study. Other biochemical parameters were analyzed according to standard laboratory procedures.Stool samples were received by a trained laboratory technician in sterile containers within 3 days of sampling. Fecal extraction and dilution to the final concentration of 1:500 was performed with Buhlmann Calex Cap device  according to the manufacturer instructions. Fecal calprotectin (FC) concentrations were measured by turbidimetric immunoassay method using Buhlmann fCAL turbo assay  and quantification was performed with the biochemical analyzer Beckman Coulter AU680 .All participants were subjected to detailed anamnesis, physical examination and measurements of anthropometric characteristics - body weight, body height, body mass index (BMI), waist circumference and hip circumference. For measurement of body weight and height, a calibrated medical scale with built-in heights  was used. BMI was calculated according to the formula = [body weight (kg)] / [height per square (m2)]. The waist circumference was measured in the middle distance between the bottom of the rib cage in middle axillary line and the tip of the iliac crest in the standing upright position of the examinee. The circumference of the hips was measured at the level of the largest circumference of the gluteal muscles, above the line connecting the large trochanters of the femur.Sample size analysis has been conducted using data from a pilot study on 10 randomly selected subjects from the patient population (5 patients with UC and 5 patients with CD). Also, data from 10 randomly selected matched control patients was obtained. The value of serum adropin levels, which was the main outcome of the study, was used for the calculation. In IBD patients, the mean adropin levels were 2.99 ± 0.95 ng/mL, and 3.58 ± 0.78 ng/mL in the control group. With type I error of 0.05, and the power of 90%, the required sample size was 47 participants per group.P < 0.05.Collected data was analyzed with statistical software MedCalc . Quantitative data was expressed as mean ± standard deviation or median and interquartile range, while qualitative data was expressed as whole number and percentage. Kolmogorov-Smirnov test was used to estimate the normality of data distribution. Comparison of serum adropin levels and other parameters between patients with IBD and control subjects was done by Student t-test for independent samples. For comparison of qualitative variables, Chi-squared test was used. Pearson’s correlation or Spearman’s correlation was used to estimate the correlation between biochemical, anthropometric and clinical parameters with serum adropin levels. Additionally, multivariable logistic regression analysis of independent predictors for positive IBD status was performed, with reporting OR, 95% CI and p-value. The level of statistical significance is set at P > 0.05; for all analysis) (Table P = 0.001), urea  and albumin levels  in IBD patients compared to the control group  Table . AccordiP = 0.002) and there was no significant difference in comparison of UC patients with CD patients   whereas in the total study population there was a significant negative correlation  (Table P = 0.025), whereas in the total study population, a significant negative correlation with fasting glucose levels was found   after model adjustment for age, BMI and gender, with serum adropin levels as dependent variable.Moreover, multiple linear regression analysis showed that adropin levels retained significant association with fecal calprotectin , and Mayo  scores, whereas in CD patients we found a significant negative correlation between serum adropin levels and SES-CD , HBI , and CDAI  scores   and it is well established that endothelial dysfunction in IBD patients is associated with the decreased production of NO, whereas studies showed that administration of anti TNF-α therapy in IBD leads to significant improvement of endothelial dysfunction36. This could also explain why the risk of coronary heart disease and heart failure is higher in IBD patients38, while the prevalence of traditional cardiovascular risk factors, such as high BMI, diabetes, hyperlipidemia, obesity, hypertension and lack of physical activity is relatively lower in IBD patients than in the general population39. Another possible mechanism through which adropin could be associated with IBD pathophysiology is its antioxidative effect. A recent study conducted on mice with induced nonalcoholic steatohepatosis showed that knockout of adropin significantly exacerbated steatosis, inflammation and fibrosis while on the other hand intraperitoneal administration of adropin led to reduced expression of inflammation genes and upregulated nuclear factor erythroid 2-related factor 2 (Nrf2)40, one of the most prominent regulators of cellular resistance to oxidative stress41. It is possible that adropin in a similar way of action through Nrf2 alleviates oxidative stress which is, proven by several studies, closely connected with inflammation in IBD43. Nevertheless, additional studies are needed to clarify the role of adropin in IBD patients and its part in this complex cascade of pathophysiological pathways.Adropin is a peptide mostly associated with energy homeostasis and vascular protection but through its network of pathways and interactions it could also be linked with inflammation. Recently several studies reported a correlation between serum adropin and different disorders such as diabetes mellitus, atherosclerosis, polycystic ovary syndrome and obstructive sleep apnea44. All of the previously mentioned studies on low-grade inflammatory conditions found the presumed association30. This result could be circumstantial due to inclusion of only those patients who had a stable IBD activity in the past 3 months.Contrary to our expectations, we didn’t find a significant correlation between adropin and hsCRP in IBD patients. It is acknowledged that even so CRP is insufficient to substitute endoscopic or radiographic findings in IBD, it’s well used for evaluation of the disease severity46. Adropin correlation with FC implies that adropin could be associated with IBD activity and severity, especially when enumerated with our finding of adropin significant association with UC and CD severity scores, which are the clinically among most important instruments for evaluation of the disease activity. However, it is also possible that these results are due to adropin down regulation through previously mentioned inflammation mechanism. Higher disease activity amplifies FC levels but also reactively up regulates proinflammatory cytokines such as TNF-α and possibly subsequently reduces adropin levels.Another interesting finding of our study was the negative correlation between adropin and FC. Several studies showed that FC correlates strongly with the laboratory and diagnostic signs of disease activity47. A recent study on mice showed that intraperitoneal administration of adropin results in a significant reduction of blood glucose, serum insulin levels and increase of serum adropin levels48. Another animal study showed that there is an inverse relationship between adropin levels and dysregulation of glucose metabolism22. However, a study conducted on human participants reported an increase of plasma adropin levels with fructose intake which was most prominent in subjects who exhibited hypertriglyceridemia49. In our recent study on patients with obstructive sleep apnea we also found a negative correlation between adropin and fasting glucose levels and furthermore a negative correlation between adropin and HOMA-IR and HbA1c. We discussed this through the possibility of adropin interactions with eNOS and therefor increased systemic bioavailability of NO. Consequently, decreased adropin would lead to lower insulin sensitivity and increased inflammation burden as showed by adropin correlation with HOMA-IR, TNF-α and IL-6 in that study32. Furthermore, a recent study showed that adropin treatment reduced expression of gluconeogenic regulatory enzymes in the liver which subsequently inhibited hepatic glucose production while improving hepatic insulin sensitivity50. This is in alignment with the results of an animal study which proved that adropin levels are higher in fed mice and lower in fasting ones48. It is possible that adropin is down regulated by fasting conditions so gluconeogenesis wouldn’t be inhibited and peripheral fatty acid would be used to support hepatic glucose production, while blood glucose would be spared to support the central nervous system. On the contrary, during re‐feeding adropin levels increase and glucose utilization is activated.Additional significant finding of our study is the negative correlation of serum adropin levels with fasting glucose levels in the whole study population. That is in line with the established fact that adropin is a regulator of glucose and energy homeostasisThe limitation of our study was its cross-sectional design which disables establishment of causal relationship. Moreover, it had single center patient inclusion with a relatively small sample size. Additionally, due to technical reasons we were not able to measure FC in the whole control group.In conclusion, this is the first study that reported decreased serum adropin levels in patients with IBD and demonstrated a negative correlation between adropin levels and IBD severity scores. Altogether our study implies that adropin could be involved in the complex pathophysiology of IBD and even potentially serve as a novel predictor of the disease activity. However, future larger scale studies are necessary to evaluate significance of these findings.


In the article titled “The Threshold of the Severity of Diabetic Retinopathy below Which Intensive Glycemic Control Is Beneficial in Diabetic Patients: Estimation Using Data from Large Randomized Clinical Trials” [1], there was an error in the Acknowledgments section, where Grant No. 0040205301D50 should be corrected to Grant No. 2015SZ0228.


Recent data highlights an imbalance in research grant success among groups underrepresented within the biomedical workforce, including racial/ethnic minorities and women. However, there is no data on grant success for researchers with disabilities.For these analyses, aggregate data on self-reported disability status for National Institute on Health (NIH) research grant applicants and awardees was obtained from 2008 to 2018, including disability category: mobility/orthopedic, hearing, visual disabilities, and other disabilities. The percentage of applications and awards, as well as grant success rates (% of applicants receiving awards), by Principal Investigators (PIs) disability status were calculated. Data was desegregated, and logistic models determined trend of applicants reporting disability over time. The percentage of NIH grant applicants with PIs reporting a disability significantly declined from 1.9% in 2008, to 1.2% in 2018 (p<0.001). Data on grant awardees was similar, 1.9% of awards in 2008, declining to 1.2% in 2018 (p<0.001) had PIs reporting a disability. Across all years, the percentage of applications and awards with PIs reporting visual disabilities was lower than the percentage reporting mobility/orthopedic, or hearing disabilities . Overall grant success rates differed by disability status . The lowest overall grant success rate was among PIs reporting unknown disability status or who withheld this status (18.6%). These results underscore the underrepresentation of researchers with disabilities among grant applicants and awardees, and indicate lower grant success rates among PIs reporting disabilities. Calls for greater diversity in science and medicine have been strengthened by recent data highlighting an imbalance in research grant success among groups underrepresented within the biomedical workforce, including women –4 and raWomen, Minorities, and Persons with Disabilities in Science and Engineering” is among the only sources of data on the representation of persons with disabilities in science. This report indicates that researchers with disabilities are underrepresented in science, as only 10% of employed scientists and engineers report having a disability[Despite the stated commitment to the inclusion of persons with disabilities, this population remains largely omitted from biomedical workforce diversification efforts. This omission is evident by the limited reporting and surveillance of the representation of persons with disabilities in the biomedical workforce . For exaisability, and is isability. HoweverFor persons with disabilities, the barriers to a successful research career are multifactorial and mirror those experienced by other underrepresented groups including social and cultural bias, lack of concordant mentoring, and structural barriers to engaging in the research process . Social Grant funding is an important indicator of research career success . Recent Documented accounts of disability bias during grant review exist , yet theHere, we present data on the number and percentage of National Institutes of Health (NIH) grant applicants and awardees. We hypothesize that researchers reporting disabilities will have decreased research grant success as compared to those not reporting disabilities.The NIH collects data on grant applicants’ disability status via self-report under the demographics section within the eRACommons portal. The Principal Investigator’s (PI) disability status is determined by their response to the question “Do you have a disability?”  No Disability, defined as applications or awards where the PI indicated “no” to the question “Do you have a disability?”; and (3) Unknown/Withheld Disability Status, which is a combined category calculated as the sum of applications or awards where the query “Do you have a disability?” was left blank by the PI (defined as “unknown disability status”) or where the PI selected “Do not wish to provide” in response to the “Do you have a disability?” question (defined as “withheld disability status”). The data for this category was provided from the NIH as the composite of both the unknown and withheld groups  request. Aggregate data was provided, which included the annual number and percentage of NIH grant applications and awards from PIs classified into one of three disability categories within the eRACommons : (1) RepThe data provided included disability information for PIs on the following grant mechanisms: R, P, M, S, K, U, DP1, DP2, DP3, DP4, DP5, D42, G12. The resulting aggregate data included information from Multiple Principal Investigators (MPI) grants, and included only competing applications. American Recovery and Reinvestment Act (ARRA) grants, UC6 grants, and applications withdrawn prior to review were excluded from these data. Data were provided at the level of applications and awards and were provided in aggregate, independent of how many grants each applicant/PI submitted or was awarded in any given year.The percentage of application and awards with PIs reporting disability by category was calculated as the sum of responses for each disability category divided by the total number reporting a disability. Grant success rate was calculated as the percentage of grant applications that received funding.To examine differences and trends over time, data was desegregated and logistic models were used to determine differences in the proportion of applicants reporting disabilities over time. Trends of the grant success rate over time were not linear and yearly data was combined using a Mantel-Haenzel approach to determine differences in funding rates by disability status; odds ratios and 99% confidence intervals are reported.Analyses were performed using SAS  software.In 2008, 1.9% of NIH grant applications included a PI reporting a disability . This peThere was no change in the proportion of NIH grant applications and awards from PIs reporting any type of disability  between 2008 and 2018 (p = 0.54 for applicants and p = 0.58 for awardees). . HoweverGrant success rate was examined using combined data over the entire period (2008 to 2018). Overall funding rates were compared by disability status . These funding rates were significantly lower for PIs reporting disabilities (27.2%) than for those not reporting disabilities (29.7%)  over this period. The overall funding rate, however, was lowest for applications with PIs reporting unknown disability status or where the PI withheld disability status  , Fig 3. These data indicate that less than 2% of NIH applications and awards include PIs that report a disability, and that the percentage of NIH grant applications and awards from PIs reporting disabilities declined significantly between 2008 and 2018. These results may reflect one or more potential scenarios: (1) that the underrepresentation of biomedical researchers with disability may be worsening over time, (2) that despite increased overall applications to NIH, researchers with disabilities are submitting fewer applications, on average, then researchers without disabilities or (3) that more researchers with disabilities are withholding disability status over time. The percentage of applicants and awards with PIs with unknown or withheld disability status, however, declined from 2008 to 2018 (see below for further discussion), suggesting that the observed decline in the percentage of NIH grant applications and awards from PIs reporting disabilities is not driven by a decline in PI’s willingness to disclose disability. These results mirror similar research that suggest the barriers for female researchers begin in the processes leading up to submitting a grant proposal [When examined by disability category, the percentage of grant applications and awards with PIs reporting visual disabilities was lower than other disability categories. This result suggest that researchers with visual disabilities may have unique barriers that limit applying for and obtaining NIH grants. The aggerate data used for these analyses do not allow us to investigate the reasons for this difference, but do highlight the need for and value of collecting and examining data by disability type to examine and identify barriers to inclusion, as each disability and area of research results in unique barriers rendering a “one-size-fits-all” approach to inclusion antiquated.Our results further indicate that the grant success rates of PI’s reporting disabilities were significantly lower than PIs not reporting disability over this same period. This result suggests that researchers with disabilities may be disadvantaged in the grant peer review process and mirrors similar research suggesting barriers to grant funding for female and underrepresented researchers. Recent research by Hoppe et al. suggestsThe percentage of NIH applications and awards from PIs reporting unknown disability status or who withheld reporting this status declined from 2008 to 2018 , suggesti.e. psychiatric, learning, and chronic health). Data on unknown/withheld disability status suggest a reluctance to disclose disability status within eRACommons. While this likely underreporting results in underestimates of the data presented, the underreporting is, itself, a problem that needs to be addressed. The reluctance to disclose disability status is reflective of the negative stereotypes and stigma that surround disability in the biomedical workforce and academia [The limitations of this study include missing information from both PIs who withheld disability responses and those who reported unknown disability status. This potentially influences our reported estimates; however, we cannot determine the direction or magnitude of this impact. Therefore, the results from this group should be interpreted with caution. Additional limitations of these data should also be considered. First, disability status is likely underestimated within this dataset for two reasons: (1) disability is self-reported within eRACommons, and (2) not all categories of disability are queried (academia ,15,24–27Despite the limitations in this data, our analysis is among the first reports of grant application and awardee data by disability status. These data indicate that individuals with disabilities are underrepresented within the biomedical workforce, may be less likely to apply for research funding, and have lower grant success rates than researchers not reporting disabilities.Further, our results underscore the urgency to identify and address the barriers to grant funding success for researchers with disabilities. These efforts should consider personal accounts of disability in research and medicine ,29, and These data also suggest that efforts to realize the NIH’s commitment to the inclusion of biomedical researchers with disabilities will require more specific, focused efforts, programmatic funding and accountability for existing requirements to recruit and retain persons with disabilities. Building on recommendations for other underrepresented groups ,31 increOur results indicate that researchers with disabilities are underrepresented among NIH grant funding applicants and have lower grant success rates as compared to researchers not reporting disabilities. Though efforts are underway to enhance the representation of persons with disabilities in the biomedical workforce,18,19, t


It provides a validation process of the developed prototype by experimental verification. The developed design was assessed in a comparison experiment with a commercially available local school backpack. An experimental study was used which recruited thirty healthy college students (aged 19 to 23 years). Two independent variables evaluated were school backpack type  and load levels as a percentage of body weight. Three load levels were employed 10%, 15%, and 20%. These variables were measured on the responses: bag comfort scale and the percent of maximum voluntary contraction (%MVC) of six muscles . The developed backpack provided astonishing performance at levels of 15% and 20% of body weight in terms of subjective measure and electromyography (EMG) responses. It also showed that increasing the carried weight more than 10% result in reducing activity on the erector spinal muscles, while it increases on abdominal oblique muscles. The developed backpack design confirmed the efficiency of its bases by distributing the carried weight among the trunk through side pockets, attached to the body through two upper and lower straps. It helped the body to distribute the carried weight and avoid concentrating pressure on specific areas.Carrying a heavy school backpack has extensively been reported as a prime cause of children's body strain. It is suggested that the load should not exceed 10 percent of the child's body weight; however, ensuring this requires continuous monitoring. The study explores how ergonomically designed school backpack based on the user anthropometric data ( Carrying school backpack is a common practice observed across all stages of academic life. School students begin to carry their bags from the first year in the school until last year at the University. Several studies report this activity as a risk to the student's health –3. AlthoStudies denote weight as prime and critical elements of backpack carriage –8. Hong Few studies emphasized that ergonomically designed backpack with an adaptation of anthropometric characteristics substantially decrease musculoskeletal disorders (MSDs) in students . This stAnother study conducted in Saudi Arabia showed that pain generally experienced in the shoulder (70%) and back (18%) . EarlierThis study consists of two phases. The first phase was concerned with developing a school backpack design using users' anthropometric data, where the anthropometric measures were collected from a sample of 280 students. Then, the school backpack was developed and manufactured with specified design dimensions based on the collected anthropometric data. Also, the second phase was concerned with testing and validating the developed school backpack by comparing body muscle EMGs and subjective perception of the comfort for 30 participants between the newly designed backpack and commercial one.Distributing weight among other body parts instead of concentrating forces on shoulders and back onlyAttach the weight as close as possible to the body by adding a strap in the lower and upper sides of the bagThe first phase was conducted to figure out the best school backpack design, including its dimensions. Consequently, some studies focused on positioning the carried weights on the back through testing shoulder strap tightness , 45. LifThe initial ergonomics designed backpack was drafted based on ergonomics features and standards, anthropometries and weight distributions, and with consideration of the user's requirements, preferences, and desires. While keeping the back pocket, two side pockets were added to the bag, as shown in Fiberglass tapes Spreading caliper (0-600 mm)Sliding caliper (Martin type with length 0-200 mm and depth of 0-50 mm)Fixed anthropometry (0-2100 mm with straight probes and provided with curved measuring branches).The first phase started with recruited 280 volunteer male college students from the university. The anthropometric measurements were adopted based on its acknowledgment as effective in the previous studies , 30, 38.A calibrated balance scale  was used to measure participants' weights. After collecting information, data was processed through normality testing, uniformity testing, homogeneity testing, and percentage data calculation. The designing of the backpack incepted by specifying the correct dimensions based on the data processing, following the creation of a prototype. Anthropometric measurements were gathered as shown in The second phase started with recruited the minimum number of participants that required in this study according to equation . This eqN is the required sample size numbers. CV is the coefficient of variation, and α is the percentage of the desired relative accuracy.Where α = 5% relative accuracy is required for the 5th and 95th percentiles, whereas values of coefficient of variation CV are adapted from Ramadan and Al-Shayea  and . Results for all muscles are demonstrated in F  = 7.683, p < 0.01, η2 = 0.209] and . The backpack type significantly affected both measured muscular activities, which were substantially lower when students wore the ergonomic backpack than when wearing the commercial one for the right and left of abdominal muscles.For abdominal muscles, ANOVA analysis pointed out that backpack type significantly affected %MVC for both right and left abdominal muscles  and . p < 0.023, and p < 0.05, for the right and left abdominal muscles, respectively.The analysis testified that backpack load significantly affected %MVC for both right and left abdominal  and . For erector spinae muscles, ANOVA analysis exhibits that backpack type substantially affected %MVC for both right and left erector spinae . p < 0.001 and p < 0.045, respectively. Also, the result shows that the developed bag exerts less effort into both erector spinae muscles, as shown in Analysis escorts that load percentage significantly affected %MVC for left erector spinae [The study aimed to achieve two objectives: first, developing a backpack design in the light of studies done primordially. The second one was testing the developed backpack by comparing it with the commercial one. Preceding studies reported that shoulders and back were the main two areas that cause pain when an increased load on the back is carried , 50. SimParticipants who wore developed backpacks felt more comfort than participants who wore the commercial ones regardless of the weight they carry. It was found that wearing the developed backpacks make the participants feeling more comfortable by 24.26% as compared to the commercial ones. The results revealed that as carried load percent increased, comfort feeling decreased by 23.89% at 20% when compared to 15% and by 38.83% when compared to 10%; as well as at 15% when compared to 10%, comfort score decreased by 12.06%.On the other hand, the muscular activity of selected muscles  expressed in %MVC (maximum voluntary contraction) was measured. It caught a sight that the developed backpack reduced the muscular activity of trapezius, erector spinae, and abdominal compared to the commercial bag. Noticeably, the results of this study ushered that load percentage has a major influence on the muscular activity of left and right abdominals and right and left erector spinae. As expected, as the load percentage increased, muscular activity increased. This agrees with a study Al-Khabbaz et al.  saying tNotwithstanding, it was noticed that abdominal muscle activity increased as the load percentage increased. This was observed that abdominal muscles had more EMG activity than the rest of the selected muscles. It could be concluded that the human body adopts extra loads through transferring some forces to the abdominal muscles. This result agrees with Son . Son 5757 investUnfortunately, early and effective treatment should be provided as it can lead to severe and everlasting back issues. Standardizing a limit on the school bag load was repeatedly highlighted as a preventive solution. In contrast, different researches have tried to develop schoolbags designed ergonomically. In this study, a developed backpack design was developed to reduce the adverse effect of carrying a schoolbag on students. The developed backpack was designed and assessed practical. The assessment was based on a comparative approach in which a developed backpack was tested in comparison with a commercial one. Thirty university students aged between 20 and 30 participated in the experiment. Assessment of developed backpack was performed at different three levels of weight. Weight level expressed as a percentage of body weight. Therefore, three weight levels employed: 10%, 15%, and 20% of the participant's body weight. It was demonstrated that developed backpack proved better performance at levels of 15% and 20% of body weight in terms of subjective measure and EMG responses. However, at a level of 10%, it was not performing as desired. In reality, a large number of students carry more than 10% of their body weight, as demonstrated by various studies , 14, 17.The findings of the current study showed that increasing the carried weight of more than 10% while wearing a newly developed schoolbag would result in reducing trunk flexion when compared to the commercial one. This action could cause some relief on the erector spinae muscles. The design of a developed backpack might help the body to achieve this result through the side pockets in better functioning. Also, the contact area between the bag and the student body would be increased to reduce the pressure felt by students. It is also recommended to add more user-friendly delicate materials to be added to eliminate the probable size issue, which might add more stress because of friction. Softer material must be used in the inner sides of the side pocket to eliminate friction stress. These considerations were not possible in this study because of the limited time.In this experiment, schoolbags weights were varying from participant to another, because the weight was expressed as a percentage of body weight. However, in reality, schoolbags mostly are equivalent in weight with minimal differences. Therefore, use three-fixed weight in future research could show very different results. Finally, the developed design confirmed the efficiency of its bases. Distributing weight among the whole body through side pockets, attach weight to the body through two upper and lower straps were confirmed in the new design. Moreover, the developed design confirmed that it helped the body to distribute weight and avoid concentrating pressure on specific areas. This is a vital factor in reducing the probable risk of carrying a schoolbag. Also, it enforced the pocket volume to be in a specific volume to keep the distributed weights as designed for.The ergonomic measurements of the school backpack, including the backpack height and width, were determined for the students in a range of age 19 to 23 years by utilizing anthropometric dimensions. Many studies addressed the physiological and psychological effects and the imposed forces and stresses on the students' bodies caused by wearing backpacks . In addiThe findings of the study are limited, given its inclusion of only male participants and recruitment from a single university and region. To help expand the study scope, the same results are suggested to be replicated for the female participants and across different age groups; thus, the findings may not be generalizable to other genders or age groups. Accordingly, it is suggested that early educational interventions should be improved for minimizing the risk that emerges as a result of carrying the excessive load. Also, standardizing the schoolbag design can help reduce the complaints of back pain as a result of carrying bag load. In addition to the conclusions drawn regarding each previous hypothesis, this investigation did not study individuals who are obese and younger who carry backpacks. Therefore, those have higher instability and great changes in the posture leading to potentially at high-risk populations, and it should be investigated in future work to identify risks of injuries and falls. This study investigated the effects of short-term backpack load carriage; thus, the observations may not be applicable to long-term carriage, because trunk muscle activity may vary with fatigue and have significant effects on lumbar joint loads. Only three backpack loading conditions were evaluated, which might be a concern of generalizing the conclusions of this study. More detailed and larger-scale research is recommended for future study on the effect of backpack loads on the trunk and head inclinations, lumbar spine loading, gait analysis, cardiac effort (heart rate variability), respiration system, or at the joints of the lower limbs during walking biomechanically.