Astragalus root, significantly alleviates IPF by inhibiting transforming growth factor β1 (TGF-β1) induced epithelial-mesenchymal transition (EMT). Further investigations into the influence of ASV on lncRNAs expression will be helpful to delineate the complex regulatory networks underlying the biological function of ASV. Here, we found sirt1 AS expression was significantly decreased in BLM-induced pulmonary fibrosis. We further found that sirt1 AS effectively inhibited TGF-β1-meidated EMT in vitro and alleviated the progression of IPF in vivo. Mechanistically, sirt1 AS was validate to enhance the stability of sirt1 and increased sirt1 expression, thereby to inhibit EMT in IPF. Furthermore, we demonstrated that ASV treatment increased sirt1 AS expression and silencing of sirt1 AS impaired anti-fibrosis effects of ASV on IPF. Collectively, sirt1 AS was critical for ASV-mediated inhibition of IPF progression and targeting of sirt1 AS by ASV could be a potential therapeutic approach for IPF.Long noncoding RNAs sirt1 antisense (sirt1 AS) was reported to play crucial roles in the progression of organ fibrosis. However, the roles of sirt1 AS in idiopathic pulmonary fibrosis (IPF) are still unknown. In addition, we have previously demonstrated that astragaloside IV (ASV), a bioactive saponin extract of the  Idiopathic pulmonary fibrosis (IPF) a progressive interstitial pneumonia with poor prognosis . The patLong noncoding RNAs (lncRNAs) are a large class of noncoding transcripts that are>200 nt long and that lack protein-coding capacity . EmerginAstragalus root, could attenuate IPF by inhibiting TGF-β1-dependent EMT [Sirt1, a member of sirtuins family, is suggested to regulate the balance between myoblast proliferation and differentiation . Overexpdent EMT  and ASV dent EMT . We thusFirstly, we examined the expression levels of sirt1 AS in TGF-β1 treated RLE-6TN cells, with the finding that 10 ng/ml TGF-β1 treatment down-regulated sirt1 AS expression in a time-dependent manner . We therTo validate the effect of sirt1 AS in IPF, mice were administrated with bleomycin (BLM) to induce IPF. The expression of sirt1 AS was also found to be down-regulated in lung tissues from IPF mice, and its expression was significantly up-regulated in IPF mice transfected with Ad-sirt1 AS . We founWe then further explore the mechanism how sirt1 AS regulates the IPF progress. A previously studies found sirt1 could control EMT progress of lung fibrosis , and a rOn the base of above findings, we therefore investigated whether sirt1 AS influenced the EMT progress in a sirt1 dependent way. The results of western blot analysis in We previously reported that ASV has inhibitory effects on TGF-β1-induced EMT in IPF, we then investigate whether sirt1 AS is involved in the function of ASV on IPF. We examined the effects of ASV on sirt1 AS expression and found that 40 μg/ml ASV started to up-regulated sirt1 AS expression and 100 μg/ml ASV influenced sirt1 AS expression at most . ThereafTo validate the implication of sirt1 AS in the anti-fibrosis role of ASV on IPF, a loss-of-function approach was employed. Mice treated with sh-sirt1 AS showed significant decrease in sirt1 AS expression when compared with those treated with sh-Scram . KnockdoIn the present study, we characterized that sirt1 AS inhibited TGF-β1-induecd EMT of alveolar epithelial cells in IPF. sirt1 AS was a negative regulator of EMT and has anti-fibrosis function to alleviate the progression of IPF. The results further revealed that sirt1 AS directly bound sirt1 mRNA to increase the stability of its mRNA, therefore to upregulate sirt1 expression; Sirt1 AS suppressed EMT of alveolar epithelial cells in a sirt1-dependent way. Above data suggested that manipulating sirt1 AS represents promising therapeutic strategies for IPF. Moreover, our results indicated the changes in sirt1 AS expression induced by ASV and sirt1 AS silencing could abolish the protective effects of ASV on IPF. These findings demonstrated a new pathway from ASV to sirt1 AS in regulating EMT for the treatment of IPF.Many studies have demonstrated that antisense lncRNAs participates in the process of organ fibrosis –24. HoweNATs alter the recruitment of members of the spliceosome to sense transcript by forming RNA-RNA duplexes with the sense transcript . It has Astragalus membranaceus and reported to have protective effects on IPF [Astragaloside IV (ASV) is one of the active ingredients of s on IPF , 33. Reps on IPF . ASV alss on IPF . These fs on IPF . And we s on IPF . This stOur study has certain limitations. First, we manipulated the expression of sirt1 AS in mice by intratracheal injection of virus vectors. Gene editing, such as CRISP system. may also be the good approaches. Second, although we identified that sirt1 AS alleviated IPF progression by upregulating sirt1, do other regulators, such as miRNAs, transcription factors and splicing factors, participate in the binding process between sirt1 AS and sirt1 mRNA remain unknown. These questions need to be addressed in future studies. In sum, the present study demonstrated that sirt1 AS attenuated IPF through transcriptionally activating sirt1. In addition, our results showed that ASV/lncRNA sirt1 AS/sirt1/Akt/foxo3 signaling pathway participates in modulating TGE-β1 induced EMT and thus alleviated BLM-induced pulmonary fibrosis, providing a novel molecular basis for the application of ASV in the therapy of IPF.th and pulmonary tissue sections were collected for further studies.C57Bl/6J mice were obtained from Shanghai SLAC Laboratory Animal Co., Ltd. and maintained in a 12-hour light-dark cycle and all experimental process were approved by the Ethics Committee of Affiliated Hospital of Shandong University of Traditional Chinese Medicine. The mice of IPF model was achieved by intratracheally injecting with bleomycin  dissolved in a total of 50 microliter sterile saline; the control group were instead with equal amounts of sterile saline. After the administration of BLM for three days, adenovirus vectors named adenovirus-control (ad-NC), ad-sirt1 AS, specific short hairpin RNA for sirt1 AS (sh-sirt1 AS) or sh-NC were intratracheally injected into the mouse pulmonary tissues, as previously described . ASV  were cultured in F12 medium  with 10% Fetal Bovine Serum , penicillin (100 U/mL) and streptomycin (100 μg/mL) in a humidified incubator at 37 °C with 5 % COescribed , treatme5 RLE-6TN cells were plated in six-well culture dishes prior to small RNA transfection. On the following day, cells were washed with serum-free medium and starved for 6 h. Subsequently, sh-RNA/si-RNA and Lipofectamine 2000 (Invitrogen) were separately mixed with Opti-MEM I Reduced Serum Medium for 5 min and the two mixtures were incubated at room temperature for 20 min. The Lipofectamine sh-RNA/si-RNA mixture was added to the cells and incubated at 37 °C for 6 h and fresh medium containing 10% FBS was added. The cells were maintained in culture until subsequent experiments. For the transfection in vivo, mice were intratracheally delivered with adenovirus control, Ad-sirt1 AS + Ad-si-NC, or Ad- sirt1 AS + Ad-si-sirt1 on the third day after BLM treatment.The adenovirus overexpression sirt1 AS (ad-sirt1 AS) were synthesized by Shanghai Genechem Co.,Ltd. Ad-sirt1 AS was used to infect with RLE-6TN cells as previously described . LentiviThe isolated lung tissues were fixed in 4% paraformaldehyde, decalcified, dehydrated, cleared with dimethylbenzene and embedded in paraffin. The 5 μm sections were stained with either Hematoxylin and Eosin or Masson's trichrome staining using standard procedures. Immunohistochemistry was performed to measure the expression of collagen1 or fibronectin1. Sections were incubated with 3% hydrogen peroxide to block endogenous peroxidase activity. Following incubation with the primary antibody at 4°C overnight, the slides were treated with DAB  to visualize the immunoreactivity. The images were obtained using the Leica DM3000 microscope .After treatment, cells were fixed with 4% formaldehyde for 5 mins. Following incubation with anti-α-SMA  or E-cadherin  antibodies and fluorescent-labeled secondary antibodies (Alexa Fluor® 488 or Alexa Fluor® 594), DAPI (Life Technologies Corporation) was used to stain nuclear counter. Images were captured by a Nikon Eclipse 800 epifluorescence microscope with the appropriate filters.RLE-6TN cells were fixed with 4% paraformaldehyde and then incubated with 0.2mol/L HCl followed by proteinase K treatment. After pre-hybridization, cells were incubated with pre-hybrid buffer with 0.1 μg FITC-labeled sirt1 AS probe at 37 °C overnight and washed with 2 × SSC  and 1 × SSC  in order. Cells were then incubated with FITC-labeled streptavidin (1:400) at room temperature, washed with PBS and DAPI (sigma) was used for nuclear staining.−ΔΔCt method.Real-time quantitative PCR (qPCR) was performed to detect the expression of sirt1 AS, and fibrogenesis-related genes. GAPDH was included as an endogenous control. In briefly, total RNA was extracted using TRIzol (Invitrogen). Concentration of the RNA was assessed with NanoDrop-2000 Spectrophotometer , then reverse transcription using a PrimeScript reagent Kit With gDNA Eraser (Takara). The primers of target genes were synthesized by Shanghai bioengineering Co.Ltd. . QPCR waRLE-6TN cells were treated according to the manufacturer’s instructions (Thermo Fisher Scientific) to obtain separate nuclear and cytoplasm parts. RNA extraction and qPCR were then conducted for nuclear and cytoplasm parts, respectively.For mRNA stability measurements, RLE-6TN cells were treated with 2 μg/ml actinomycin D , which suppresses transcription. Total RNA was extracted at indicated time point  to determine the residual mRNAs by using qPCR analysis. β-actin was served as the internal control.Cultured cells and tissues were lysed with RIPA buffer  with phosphatase inhibitor cocktail  and then protein for further western blot following the protocol as describe previously . The ant5/ml. The lower chamber contained 500 μl of F12 medium containing 10% FBS. After 24 h’ culture, the cells that migrated to the other side of the upper chamber were fixed with 4% paraformaldehyde and subjected to crystal violet staining. The number of transmembrane cells was artificially counted.RLE-6TN cells with a cell density of 5,000 cells/well were seeded in 96-well plates. After incubating for 24 h, the cells were treated with varying interventions and then the cells were incubated with CCK-8 solution  for 2 h. A Microplate Reader  was used to measure the absorbance at 450 nm. After transfection, cells were trypsinized, harvested, and resuspended in serum-free F12 medium and seeded in the upper chamber of a Transwell chamber at a cell density of 1 × 10Data are expressed as the mean ± SD. Student’s t-test or ANOVA was used to compare the means of two or three groups. All statistical analyses were conducted using GraphPad Prism v8 . The significance level was set at P<0.05.Supplementary Tables


One-step microwave heating can simplify the reduction step of graphene oxide and provide a convenient, economical, and effective method of preparing graphene composites. The structural analysis shows that LiFePO4/graphene has high phase purity and crystallinity. The morphological analysis shows that LiFePO4/graphene microspheres and micron blocks are composed of densely aggregated nanoparticles; the nanoparticle size can shorten the diffusion path of lithium ions and thus increase the lithium-ion diffusion rate. Additionally, the graphene sheets can provide a rapid transport path for electrons, thus increasing the electronic conductivity of the material. Furthermore, the nanoparticles being packed into the micron graphene sheets can ensure stability in the electrolyte during charging and discharging. Raman analysis reveals that the graphene has a high degree of graphitization. Electrochemical analysis shows that the LiFePO4/graphene has an excellent capacity, high rate performance, and cycle stability. The discharge capacities are 166.3, 156.1, 143.0, 132.4, and 120.9 mAh g−1 at rates of 0.1, 1, 3, 5, and 10 C, respectively. The superior electrochemical performance can be ascribed to the synergy of the shorter lithium-ion diffusion path achieved by LiFePO4 nanoparticles and the conductive networks of graphene.In this study, micro/nanoscale LiFePO Li/Li+). However, LiFePO4 materials have some drawbacks, such as inferior electronic conductivity (ca.10−9–10−10 Scm−1) as well as slow one-dimensional lithium ion diffusion, which is a formidable obstacle to the high performance of lithium-ion batteries  suspension. The above experimental methods are very complicated, and most of them require long-term high-temperature treatment and atmosphere protection, which lead to high energy consumption and cost. Additionally, the graphene and active materials agglomerate easily and distribute unevenly. Therefore, simplifying the preparation technology and obtaining a product with a small and homogeneous distribution remain great challenges for preparing LiFePO4/graphene composites. Microwave heating is a convenient, economical, and environmentally friendly route for the preparation of graphene composites in a way that addresses the deficiency of graphene modification. Microwave heating can simplify the reduction step of graphene oxide, as, due to the microwave-absorbing properties of graphene oxide, microwave irradiation can restore it into graphene directly without any reductive agent and atmosphere.Small particle size can decrease the migration distance of lithium ions from the interior to the surface and increase the diffusion rate , LiOH·H2O (95%), H3PO4 (85%), ethylene glycol, and sucrose were purchased from Sinopharm Chemical Reagent Co. Ltd. Graphene oxide was synthesized from natural graphite powder (325 mesh) using a modified Hummers' method  with Cu Kα radiation operated at 40 kV and 40 mA. The contents of graphene and carbon in the LiFePO4/graphene and LiFePO4/C composites were calculated from TG-DSC , which was carried out from room temperature to 700°C under an air atmosphere at a rate of 10°C min−1. The morphologies of LiFePO4/graphene composites were observed using a scanning electron microscope  and transmission electron microscopy . The Raman spectra of LiFePO4/graphene and LiFePO4/C composites were recorded from 100 to 3,200 cm−1 on a Renishaw Raman microprobe  using a 514.5 nm argon-ion laser at room temperature.The structures of LiFePO4/graphene and LiFePO4/C composites were evaluated with 2,025 coin-type batteries. The cathode electrodes were prepared by mixing 80 wt% active materials (LiFePO4/graphene or LiFePO4/C) and 10 wt% carbon black (TIMCAL) with 10 wt% polytetrafluoroethylene  in isopropyl alcohol solution . A uniform slurry was formed and pasted onto Al foils, dried at 120°C for 12 h, and then cut into circular electrodes with a diameter of 10 mm. Lithium metal  was used as the anode, Celgard polypropylene (Celgard 2400) as the separator, and 1M LiPF6 dissolved in ethylene carbonate and dimethyl carbonate (with a 1:1 volume ratio) as the electrolyte . The cells were assembled in an argon-filled glove box . Cells were charged and discharged at room temperature using a LAND-CT2001A battery cycler  within the voltage range of 2.7–4.2 V (vs. Li+/Li). Cyclic voltammetry (CV) was performed with an Auto Potentiostat 30 system at a scan rate of 0.1 mVs−1 between 2.5 and 4.2 V. Electrochemical impedance spectroscopy (EIS) profiles were obtained at the same open-circuit voltage by applying a 5-mV amplitude of the AC voltage with the frequency ranging from 100 kHz to 0.01 Hz.The electrochemical behaviors of the LiFePO4/graphene and LiFePO4/C composites are here investigated. XRD patterns of the composites are shown in 4/graphene and LiFePO4/C composites. All the sharp diffraction peaks corresponding to the (200), (101), (210), (011), (111), (211), (301), (311), (121), (410), (221), (401), (112), (222), and (123) planes can be indexed to the orthorhombic olivine-type structure LiFePO4 with the Pnma space group (JCPDS card No. 83-2092)  is observed, proving that the graphene oxide has already been reduced into graphene directly without any reducing agent or atmosphere. This is mainly because the graphene oxide with a large amount of oxygen functional groups on the surface that can absorb microwaves easily, and electromagnetic energy is converted into heat and molecular kinetic energy; the reactive oxygen groups are then exfoliated, and, finally, the graphene oxide is restored into graphene. Also, the introduction of graphene has no effect on the structure of LiFePO4. Moreover, the diffraction pattern of LiFePO4/C shows no diffraction peaks corresponding to residual carbon, indicating that the carbon decomposed from sucrose in the sample exists in an amorphous state.The phase constitution and crystal structure of the synthetic LiFePO4/graphene and LiFePO4/carbon composites, as shown in 4 can be completely oxidized to Li3Fe2(PO4)3 and Fe2O3 under air flow, and the total weight gain is about 5.07% in theory  and symmetric (ν1) stretching of the P–O bonds. The modes at 626 and 587 cm−1 correspond to the symmetric (ν2) and anti-symmetric (ν4) bending of the O-P-O angles. The mode at 395 cm−1 corresponds to the lithium cage and oxygen ion breathing cage. The modes in the 100–300 cm−1 range are induced by translation of Fe and coupled translation and vibration of Fe and −1 and a G band at around 1,590 cm−1  is inversely proportional to the degree of graphitization of carbon materials. The ID/IG in LiFePO4/graphene composites is 1.18, while the ID/IG in LiFePO4/C composites is 1.43. This implies that the graphene has a higher degree of graphitization than the carbon decomposed from sucrose. The higher the degree of graphitization, the better the conductivity of the carbon. A high degree of graphitization is favorable for electron transfer and improves the electrochemical performance of the cathode. Additionally, the strong signals of the graphene (D band and G band) weaken and override the bands of LiFePO4 in the high-frequency region.Raman scattering spectroscopy was employed to recognize the chemical structure of the LiFePO4/graphene are shown in 4/graphene composites are composed of micron-scale spheres and blocks with average dimensions of ~2 μm. In 4/graphene microspheres and micron blocks are composed of densely aggregated nanoparticles. This structure forms because the self-heating effect induced by the microwave heating can greatly shorten the reaction time, and the graphene wrapping the surface of LiFePO4 particles can inhibit the growth of grains. Under the action of graphene, the nanoparticles assembled into microspheres and micron blocks. When the highly conductive electrolyte penetrates into the cathode material, the nanoparticles have a high specific surface area, which increases the contact area with the electrolyte. The nanoparticle size can shorten the diffusion paths of electrons and lithium ions and improve the conductivity of the cathode material significantly. Moreover, the micron structure formed by the aggregation of nanoparticles does not collapse during the process of lithium-ion intercalation and deintercalation, which ensures the stability of cathode material in the electrolyte. TEM and HRTEM images of the micron/nanoscale LiFePO4/graphene composite are shown in 4 particles. 4 are highly graphitic. The highly efficient and stable conducting network can give the material desirable electrochemical properties. In the energy spectrum, elements of P, O, Fe, and C are found, as shown in SEM images of the graphene oxide and LiFePO4/graphene composites is illustrated in The formation process of the LiFePO4/graphene electrode dynamics is shown in − and Li+) from their “reservoirs” toward the LiFePO4 particles  is shown as step C. It can be seen that graphene can provide a high-speed channel for the rapid diffusion of electrons and cause the electrons to reach the reactive site quickly, thus increasing the electronic conductivity of the materials. Meanwhile, the nanoparticles can shorten the transport path of Li+ from the surface to the interior of solid active particles and improve the diffusion coefficient of lithium ions. Moreover, the nanoparticles are surrounded by the micron graphene sheets, and the micron structure guards the stability of the material. Therefore, LiFePO4/graphene composites are expected to have excellent electrochemical performance.A schematic diagram of LiFePO4/graphene and LiFePO4/C cathode materials. 4/graphene and LiFePO4/C composites. In the first scan, there is a pair of redox peaks corresponding to the Fe2+/Fe3+ couple (Ding et al., 4/graphene, the potential difference between the oxidation and reduction peaks decreases from 0.27 to 0.26 V, which means that the phase is stabilized in subsequent cycles. 4/C composites. During the second scan, the potential difference increases from 0.31 to 0.33 V, which proves that detrimental polarization becomes more and more serious. The results show that LiFePO4/graphene composites have very high reversibility and better electrochemical activity.Cyclic voltammetry was performed to investigate the electrochemical kinetics of LiFePO4/C electrode is explained by the electron transfer pathway, as shown in + ions intercalate. In contrast, for LiFePO4/graphene, due to the one-dimensional Li+ ion mobility in the framework, the graphene can ensure that electrons reach particles from all directions and alleviate the polarization, as shown in 4/graphene composites, with well-defined peaks and smaller potential difference, have higher electrochemical reactivity.The polarization of the LiFePO4/graphene and LiFePO4/C at progressively increasing C rates from 0.1 to 10 C are shown in 4/graphene composites, the initial discharge capacity is 166.3 mAhg−1 at 0.1 C, and the discharge capacity decreases to 156.1 mAhg−1 with an increase in the discharge rate to 1 C. At a higher discharge rate of 5 C, the cell delivers a capacity of 132.4 mAhg−1. Even at a 10 C rate, the capacities can reach 120.9 mAhg−1, and a good voltage plateau remains above 3 V. For LiFePO4/C, the discharge capacity is 154.8, 133.8, 121.6, 105.9, and 86.4 mAh g−1 at 0.1, 1, 3, 5, and 10 C rates, respectively. The cycling performances of the LiFePO4/graphene and LiFePO4/C from 0.1 to 10 C are shown in 4/graphene, the specific capacity decreases with increasing current rate, the capacity retention remains very good for all of the different rates; the discharge capacity retentions are, respectively, 99.5, 99.2, 99.4, 99.1, and 97.1% at 0.1, 1, 3, 5, and 10 C current rates after being cycled 10 times. While for LiFePO4/C, the discharge capacity retentions are, respectively, 97.7, 96.9, 93.0, 87.1, and 79.5% at 0.1, 1, 3, 5, and 10 C current rates. All of the results demonstrate that LiFePO4/graphene composites have better rate performance and cycling stability. This can be attributed to the excellent electrical conductivity of graphene, which can improve the conductivity and stability of materials.The charging and discharging capacity profiles of the LiFePO4/graphene and LiFePO4/C cathodes. 4/graphene and LiFePO4/C cathodes. The experimental EIS data is simulated by Zview2.1 software according to the equivalent circuit as shown in + ions at the electrode/electrolyte interface and the charge transfer process. Meanwhile, a low-frequency sloping line is related to the Warburg impedance of the lithium-ion diffusion in the electrode (Zhang et al., RS is the internal resistance of the cell and corresponds to the electrodes, electrolyte, and the separator resistance, Rct is associated with the charge-transfer resistance, CPE is associated with the capacitance contributed by the surface of the active material (Guo et al., Rct value of the LiFePO4/graphene cathode is 79 Ω, which is smaller than the 129 Ω value of the LiFePO4/C cathode. The result shows that graphene can reduce the charge transfer resistance of Li-ion insertion and extraction between the electrode/electrolyte and increase the conductivity of the LiFePO4/graphene cathode.Electrochemical impedance spectroscopy was used to investigate the electrochemical behaviors of LiFePO4/graphene composite was successfully prepared as cathode material through one-step microwave heating. The graphene oxide, which has excellent microwave-absorbing properties, can react with microwaves quickly and be restored into high-quality graphene directly without any reducing agent or atmosphere. The introduction of graphene does not impact the structure of LiFePO4, and LiFePO4 nanoparticles are packed into micron graphene sheets. The graphene network, which has a high degree of graphitization, can provide a high-speed channel for the rapid transfer of electrons and thus increase the electronic conductivity of materials. Meanwhile, the nanoparticles can improve the diffusion coefficient of lithium ions. Moreover, because the nanoparticles are surrounded by the graphene sheets, the micron structure guards the stability of the material. The electrochemical analyses reveal that the LiFePO4/graphene composites have excellent high-rate performance and cycling life. The outstanding electrochemical performance, as well as the fast and efficient method, make this technology commercially viable.A LiFePOThe datasets generated for this study are available on request to the corresponding author.SL, PY, and WS performed the experiments. SL, PY, HL, and XZ performed the data analysis. SL and PY wrote the paper. All authors contributed to the theoretical analysis and the general discussion.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Imaging sequences for detection of meningeal and parenchymal lesions are critical in intracranial pathology. Our study analysed FLAIR MRI sequence for evaluating postcontrast enhancement.  FLAIR imaging sequences have been used in evaluation of enhancement in the brain. We conducted a study of FLAIR imaging sequences to better delineate postcontrast enhancement.  In this prospective hospital-based observational study, postcontrast T1 MTC and delayed postcontrast T2 FLAIR and T1 FLAIR images of 66 patients with intracranial pathology were assessed by experienced radiologists from November 2017 to November 2019.  28 cases of meningeal enhancement were identified in delayed postcontrast T2 FLAIR images. Low-grade gliomas included in the study showed postcontrast enhancement on postcontrast T1 MTC images. Multiple sclerosis lesions were better seen on postcontrast T1 FLAIR. In extraaxial lesions of 11 cases of meningioma, brighter enhancement was seen on delayed postcontrast T2 FLAIR images.  We found that delayed postcontrast T2 FLAIR was better in detection of meningeal enhancement in infectious meningitis and in meningitis carcinomatosis than T1 MTC images. In delayed postcontrast T2 FLAIR images, intra-axial parenchyma lesions appeared more conspicuous or similar to T1 MTC images. Delayed postcontrast T1 FLAIR images provided better anatomic delineation of intra-axial lesions. Some., 5. We cIn this prospective hospital-based observational study, postcontrast T1 MTC and delayed postcontrast T2 FLAIR and T1 FLAIR images of 66 patients—32 patients with meningeal enhancement , 23 parenchymal enhancing lesions (7 patients with infection (5 cases tuberculomas and 2 toxoplasmosis), 2 patients with demyelination (multiple sclerosis), 6 patients with primary tumors (4 low-grade gliomas and 2 high-grade gliomas), and 8 patients with metastatic lesions), and 11 patients with extraaxial lesions(meningiomas)—were assessed by experienced radiologists during a study period from November 2017 to November 2019.The study was approved by the institutional ethics committee. Data were collected from all age groups of patients who were referred to the department of radio diagnosis for MRI brain study with contrast, to rule out meningeal or parenchymal lesions.Data were collected from the patients referred to the department of radio diagnosis, for MR imaging of the brain, in the teaching hospitals attached to Kasturba Medical College, Mangalore. MR examinations were performed on 1.5 Tesla (Magnetom Siemens Avanto).Gadolinium dimeglumine (0.5 mmol/ml) was administered.Sequential order of postcontrast sequences was as follows: T1W postcontrast sequence was run prior to delayed postcontrast FLAIR (fluid attenuation inversion recovery) sequences.  TR:9000  TE:92  TI:2500  Slice thickness: 5 mmImaging parameters for delayed (10 min) postcontrast T2 FLAIR imaging:  TR:2000  TI:860  TE:59  Slice thickness: 5 mmImaging parameters for postcontrast T1W FLAIR:  TR:741  TE:17  Slice thickness: 5 mmImaging parameters for postcontrast T1W MTC images:Patients referred for gadolinium-enhanced brain imaging.Patients with intracranial aneurysm clips.Patients with intraorbital metal fragments, any electrically, magnetically, and mechanically activated implants .Patients having renal failure or raised creatinine (1.6 mg/dl) levels.Out of the 28 cases with clinical diagnosis of leptomeningitis, in all of the 28 cases, meningeal enhancement was easily identified in delayed postcontrast T2 FLAIR images. In comparison, in postcontrast T1 MTC images, differentiation of meningeal enhancement from vascular enhancement was difficult and could have been missed easily. CSF studies were positive in all 28 cases. On the contrary, delayed postcontrast T1 FLAIR images demonstrated minimal meningeal enhancement in 26 cases out of the 28 cases of leptomeningitis, and the enhancement was inferior to T1 MTC images in all of these 28 cases .Out of the 23 cases of intra-axial parenchymal lesions included in the study, in 2 cases, postcontrast T1 FLAIR was not done. In rest of 21 cases, delayed postcontrast T1 FLAIR was done, out of which 13 cases showed brighter and thicker enhancement of solid components and peripheral walls of the lesions. It provided better delineation of lesion. These 13 cases included 4 cases of solid cystic parenchymal mass lesions (2 primary and 2 metastatic), 2 cases of peripherally enhancing primary cystic tumors, 2 cases of cystic metastasis, 4 cases of infection (3 tuberculomas and 1 toxoplasmosis), and 1 case of demyelination (multiple sclerosis). In such cases, characteristics of the wall of peripherally enhancing tumors/infectious lesions were better defined by delayed postcontrast T1 FLAIR images as compared to postcontrast T1 MTC.In three out of four low-grade glioma included in the study, subtle postcontrast enhancement seen on postcontrast T1 MTC images could not be well appreciated on delayed postcontrast T2 FLAIR images due to the intrinsic FLAIR hyperintense nature of the lesions . Subtle Postcontrast enhancement could not be well appreciated on delayed postcontrast T2 FLAIR images in two cases of multiple sclerosis as compared to postcontrast T1 MTC images. In patients with multiple lesions delayed T1 postcontrast FLAIR images picked up almost the same number of lesions as compared to postcontrast T1 MTC images.Out of the 11 cases of meningioma, brighter enhancement was seen on delayed postcontrast T2 FLAIR images as compared to postcontrast T1 MTC images. Dural tail was easier to identify on delayed postcontrast T2 FLAIR images in 8 out of 11 cases due to brighter enhancement as compared to postcontrast T1 MTC images. In one case of meningioma, dural tail was less well appreciated than in postcontrast T1 MTC images. CSF cleft was better appreciated on postcontrast T2 FLAIR images than on postcontrast T1 MTC images in 8 out of 11 cases .Dural tail and extraaxial nature of the lesion was better appreciated on postcontrast T2 FLAIR images than postcontrast T1 MTC images.Our results are similar to a report published by Kremer et al.  which coParmar et al.  in an inStudies done by Splendiani et al. and Artzi et al.  show simWe have found that, in delayed postcontrast T1 and T2 FLAIR images, most of the intra-axial parenchyma lesions appeared more conspicuous or similar to T1 MTC images. Flair images show increased signal intensity because the T1 and T2 effects of various tissues supplement each other.In a study done by Hakyemez et al.  in patieHowever, in our study, we found that delayed postcontrast FLAIR images can be helpful in differentiating between metastatic disease and high-grade glioma as a greater degree of perilesional signal change representing edema is more indicative of metastatic disease, as described in another study .In our study, we found that delayed postcontrast T1 FLAIR images were not sensitive for detection of leptomeningeal enhancement. Any leak of gadolinium into the CSF space may shorten the TI of CSF enough so that the longitudinal magnetization recovers before the null point of CSF on T2 FLAIR images; however, this leak of gadolinium is not enough for the recovery of CSF signal intensity on T1 FLAIR images due to the shorter TI time and more T1 weightage of the images as compared to T2 FLAIR images.Postcontrast T1 FLAIR images provided better anatomic delineation, with a better contrast between the gray and white matter and also between gray matter and the lesion, and T2 FLAIR showed delineation between lesion and CSF due to the darker CSF on T2 FLAIR images.We found that delayed postcontrast T1 FLAIR images provided better anatomic delineation with intense enhancement of high-grade tumors, large metastatic, and infectious ring-enhancing lesions .Thickened walls of chronic lesions have increased vascularity which leads to accumulation of gadolinium within the lesions, and high-grade tumors with increased perfusion show increased gadolinium retention in blood pool or within the tumor. Retention of gadolinium results in decrease of TI in CSF so that it is no longer nulled by the inversion pulse and appears bright. The cystic nonenhancing components within the lesion appear even more hypointense (than on T1 MTC) due to the inversion pulse.Increased contrast between enhancing hyperintense and nonenhancing cystic component of the lesions along with the intrinsic high CNR of lesions on T1 FLAIR images provides excellent delineation of lesions on delayed postcontrast T1 FLAIR images. Our findings were consistent with those of Al-Saeed et al. .The overall image contrast was judged to be superior on T1-weighted FLAIR images compared with T1-weighted FSE images by all neuroradiologists  Figure .Delayed postcontrast T2 and T1 FLAIR images showed better enhancement of meningeal and parenchymal lesions and detected more number of lesions as compared to postcontrast T1 MTC. Delayed postcontrast T2 FLAIR images provided better delineation of extraaxial mass lesions with more conspicuous enhancement and better demarcation of dural tail.Hence, we concluded that delayed postcost contrast T2 FLAIR images should be included in the routine protocol for contrast MRI brain study.On the contrary, postcontrast T1 FLAIR had no added advantage over T1 MTC images in detection of meningeal lesions. Although T1 FLAIR images provided better delineation of the extraaxial mass lesions, the degree of enhancement was either inferior or similar to T1 MTC images.Postcontrast T1 FLAIR provides better enhancement and delineation of intraparenchymal high-grade vascular tumors and large tuberculomas than postcontrast T1 MTC.


The processing of the powders has been made using a colloidal route, and after that, a spark plasma sintering process was performed in order to densify the samples. Creep tests were performed at temperatures between 1200–1250 °C in an argon atmosphere. The microstructure obtained by SEM of the sintered and tested specimens was characterized quantitatively to elucidate the deformation mechanism. Raman spectroscopy was carried out to check the integrity of the graphene. The average grain size was in the order of 1 µm and the shape factor was 0.7 for all the studied materials. The integrity of the graphene was checked before and after the creep experiments. The careful analysis of the creep tests shows that graphene oxide or its reduced version are not efficient phases for creep resistance improvement in general, contrary to what is reported elsewhere. However, the results permit the suggestion of a creep improvement in nanocomposites at a very high temperature regime due to an enhanced reactivity of oxygen between carbon and alumina interfaces. In the case of zirconia, the results give us the conclusion that the oxidation of graphene is a highly detrimental issue regarding the improvement of high-temperature plasticity. This paper aims to give an answer to the following question: is the oxidation of graphene a critical issue for high-temperature plasticity in graphene-reinforced ceramics? To give a convincing reply, we will focus on two very different graphene-based ceramic composites: reduced graphene oxide (rGO)-reinforced alumina (α-Al Traditionally, a secondary phase is used in the world of materials to reinforce them and to try and change its mechanical properties. The result is a composite with enhanced properties ,2. In th2O3) there are a lot of studies trying to improve the creep behavior by adding a secondary phase. For example, Lessing et al. [2O3 doped with a 10 vol.% of carbon nanotubes had a better creep behavior than the pure material with a similar microstructure. In another study, Zapata-Solvas et al. [2O3/single-wall carbon nanotube composites. An extensive variety of alumina exists, with a large scatter in the diffusion kinetics processes which can also affect the mechanical properties [2O3/ carbon nanofiber composite decreased with respect to pure alumina, but at 1250 °C they found a similar behavior of the composite and the pure material. The same authors reported that the Al2O3/graphene oxide composite has a better creep resistance than the pure material. Zapata-Solvas et al. [Concerning alumina,  is the most important mechanism of deformation at high temperatures of yttria-stabilized-tetragonal zirconia (3YTZP) ,22. Somemposites . Cano-Crmposites  found thThe scientific problem is the comprehension of the importance of the reactivity of oxygen between the graphene and the alumina or zirconia grains, and if this reaction is detrimental in the mechanical properties at high temperatures. In order to give an answer, several creep experiments at very elevated temperatures using graphene as the secondary phase were performed for the first time of our knowledge. Consequently, the main goal of this study is to prepare a homogeneous dispersion of reduced graphene oxide (rGO) into alumina and zirconia matrices and to obtain nearly full density compacts by spark plasma sintering with enhanced creep resistance at high temperatures compared to pure materials. To achieve this aim, several experimental techniques were employed: scanning electron microscope, Raman spectroscopy, and creep at high temperatures. The creep experiments were analyzed according to the Equations (1)–(3) presented in the 2O3  has an average particle size of 0.35 µm and a specific surface area of 9.5 m2/g. The powder of tetragonal-ZrO2 stabilized with 3% mol of Y2O3  has an average particle size of 90 nm and a specific surface area of 6.7 m2/g. Both kinds of powders have a high purity and higher than 99.9%.The starting powders of α-alumina and tetragonal zirconia used were the same as in ,29. The 2/g was used as the secondary phase [Reduced graphene oxide  with a length of 1–4 μm, a thickness of 0.7–1.2 nm and a surface area of 103 mry phase ,31. PartN-methyl-2-pyrrolidone (NMP) with magnetic stirring, as this is considered elsewhere an excellent non-aqueous vehicle to achieve the highly efficient dispersion of reduced graphene oxide. Alumina was dispersed in water using an ammonium salt of poly(acrylic) acid  in a concentration of 0.5 wt.% on a dry solids basis and sonicated with different sonication times. Concentrated suspensions of alumina-reduced graphene oxide were prepared to solid loadings of 30 vol.% and homogenized with 1 min of ultrasonication.First of all, reduced graphene oxide was dispersed in To obtain a homogeneous dispersion of reduced graphene oxide in zirconia, both materials were dispersed independently and furtherly mixed. Zirconia was dispersed in water to a concentration of 30 vol.% using an ammonium salt of poly(acrylic) acid  in a concentration of 0.5 wt.% on a dry solid basis. Reduced graphene oxide was dispersed in 1-methyl-2-pyrrolidone at the same solids content (30 vol.% reduced graphene oxide) and both slurries were subjected to mechanical homogenization and sonication for 1 min . Once dispersed, proper fractions of both suspensions were mixed altogether to obtain two relative contents of reduced graphene oxide in the zirconia matrix of 2 and 6.7 vol.%, applying 1 min of additional sonication. The rheological behavior of the different suspensions was analyzed using a rotational rheometer.The optimized suspensions of the mixture were frozen in a rotatory chamber using liquid nitrogen as refrigerant (−196 °C). The frozen suspensions were introduced in a freeze-dryer for 24 h. The condensator temperature was −50 °C, and the conditions of the storage camera were 20 °C and 0.050 mbar .A2rGO: 98 vol.% α-alumina and 2 vol.% reduced graphene oxide.A6.7rGO: 93.3 vol.% α-alumina and 6.7 vol.% reduced graphene oxide.Z2rGO: 98 vol.% t-zirconia and 2 vol.% reduced graphene oxide.Z6.7rGO: 93.3 vol.% t-zirconia and 6.7 vol.% reduced graphene oxide.The following resultant mixtures of powders were obtained:As reference materials, pure alumina and pure zirconia commercial powders were used for the sake of comparison. These powders are named A and Z, respectively.The sintering process employed was the same as in ,29. The Raman spectroscopy was performed in order to note the integrity of the reduced graphene oxide. Measurements were performed in the same way as described in ,29.The microstructural characterization was performed by scanning electron microscopy. The surfaces of the samples were polished with diamond paste. The microstructure was described by measuring the grain size and the shape factor. Fracture surfaces were also observed to check the presence and distribution of reduced graphene oxide; to this end a high resolution scanning electron microscope  was employed in the mode secondary electrons.A is a dimensionless constant, G is the shear modulus, b is the magnitude of the Burgers vector, k is the Boltzmann constant, T is the absolute temperature, d is the grain size, p is the inverse of the exponent of the grain, σ is the stress, n is the stress exponent and D a diffusion coefficient [The creep at high temperatures of these new materials was investigated in the same way as explained in ,29. It mfficient .n) and the activation energy (Q). A value of stress exponent was obtained by changing the applied load as it can be seen in the Equation (2) and a value of the activation energy was obtained by modifying the temperature (3).The characteristics parameters of the creep experiments were the stress exponent  demonstrating their good dispersibility. Zirconia was dispersed in water, whereas reduced graphene oxide was dispersed in a non-aqueous medium (NMP) in order to improve the dispersion. The final suspensions had a solid loading of 30 vol.%. −1 observed in 2O3 stabilized ZrO2 and in Al2O3 ceramics in which there is grain boundary sliding with diffusion as an accommodation mechanism [T and σ data. Therefore, additional values of the stress exponents and activation energies were obtained. These facts and the same microstructure before and after creep permit us to affirm that the most important deformation mechanism is grain boundary sliding accommodated by diffusion [All these experimental facts are in good concordance with a general model of the creep deformation in Yechanism ,39. The iffusion ,39. The iffusion . The strOne aspect which that is very important is the analysis of the creep resistance due to the introduction of this carbon phase. To this end, and assuming that the grain size exponent is three because the accommodation process is the diffusion along the grain boundary, we are using this value to normalize strain rate versus stress plots in log–log scales in The reactivity of oxygen, present in the lattice of graphene, could be a crucial issue for the prediction of high-temperature creep resistance. Further work must be conducted to elucidate the local chemistry bonds involved in the ceramic composite at high temperatures.The microstructure is constant during the creep at high temperatures and the deformation mechanism is grain boundary sliding.The analysis of the high temperature mechanical properties allows for the conclusion that reduced graphene oxide does not provide an improvement of more than an order of magnitude in comparison to pure alumina, but it is potentially attractive in the range of nanocomposites or very high temperatures (from 1250 °C).The reduced graphene oxide reinforced zirconia composites are, inside the experimental uncertainty, similar to pure zirconia regarding high-temperature creep properties. The ability of the carbon phase to “wet” the grain boundaries and distribute homogeneously  during deformation is a critical element to predict the high-temperature plasticity.


User-interaction event logs provide rich and large data sets that can provide valuable insights into how people engage with technology. Approaches such as ecological momentary assessment (EMA) can be used to gather accurate real-time data in an individual’s natural environment by asking questions at any given instant.The purpose of this study was to evaluate user engagement and responses to EMA questions using InspireD, an app used for reminiscence by persons with dementia and their caregivers. Research findings can be used to inform EMA use within digital health interventions.A feasibility trial was conducted in which participants (n=56) used the InspireD app over a 12-week period. Participants were a mean age of 73 (SD 13) and were either persons with dementia (n=28) or their caregivers (n=28). Questions, which they could either answer or choose to dismiss, were presented to participants at various instants after reminiscence with personal or generic photos, videos, and music. Presentation and dismissal rates for questions were compared by hour of the day and by trial week to investigate user engagement.21=21.4, P<.001). Questions asked following reminiscence with personal media, especially those asked after personal photos, were less likely to be answered compared to those asked after other media. In contrast, questions asked after the user had listened to generic media, in particular those asked after generic music, were much more likely to be answered.Overall engagement was high, with 69.1% of questions answered when presented. Questions that were presented in the evening had the lowest dismissal rate; the dismissal rate for questions presented at 9 PM was significantly lower than the dismissal rate for questions presented at 11 AM (9 PM: 10%; 11 AM: 50%; χThe main limitation of our study was the lack of generalizability of results to a larger population given the quasi-experimental design and older demographic where half of participants were persons with dementia; however, this study shows that older people are willing to participate and engage in EMA. Based on this study, we propose a series of recommendations for app design to increase user engagement with EMA. These include presenting questions no more than once per day, after 8 PM in the evening, and only if the user is not trying to complete a task within the app. Digital health and well-being products such as health apps are becoming increasingly popular given that mobile technology is ubiquitous in daily life. In addition to the user data that are recorded during use of these platforms, all user interactions and events can be logged to represent usage. Such user interaction and event logs provide rich and large data sets that can reveal valuable insights into how people engage with technology. This case study explored user engagement with ecological momentary assessment (EMA) in an app designed for persons with dementia and their caregivers.Dementia includes a group of symptoms associated with ongoing cognitive decline and is highly prevalent with approximately 50 million cases worldwide . Within Many different techniques (such as EMA or experience sampling methodology) can be used to gather accurate data on daily living. These methodologies are used to capture real-time data in an individual’s natural environment through repeated sampling . This caIn our previous work, we developed an app for reminiscence which incorporated EMA for the Individual Specific Reminiscence in Dementia (InspireD) feasibility study. The study used a quasi-experimental design and investigated the use of an iPad app to allow people living with mild to moderate dementia and their family caregivers to reminisce . In phasReminiscing made up the largest proportion of total app interactions for persons with dementia (71%) and their caregivers (47%) . Across Participants living with dementia attained statistically significant increases in mutuality, quality of caregiver and patient relationship, and subjective well-being from the beginning to end of the study . AdditioIn this study, we sought to address four research questions: (1) What is the temporal engagement with EMA questions over hours of the day and across the trial period? (2) How differently do persons with dementia engage in responding to questions compared to how caregivers engage? (3) How do persons with dementia and their caregivers engage in responding to questions after reminiscing with video, audio, and photos? (4) How do persons with dementia and their caregivers engage in responding to questions after reminiscing with personal media compared to how they respond after reminiscing with generic media? The aim of this study was to evaluate engagement with EMA questions while using an app with the overall goal to inform EMA use within digital health interventions.A user engagement study was conducted in which participants used a digital health intervention, an app for reminiscence, at home for a trial period of 12 weeks. EMA questions on mutuality were presented to users at various points during the trial period while they were reminiscing using the app.The study received ethical approval in March 2016 (16/NI/0035) in line with regional and national Health Service Trust research governance. Ethical considerations were ensuring voluntary participation by supporting separate informed consent for the persons with dementia and their caregivers, secure handling and storage of data in line with University policy, and reminding participants of their right to withdraw from the study at any time. In this study, for each person with dementia who participated, a family caregiver also participated. Each dyad (person with dementia and their caregiver) was given an iPad and asked to use the app at home for a 12-week period. Participants were encouraged verbally and in writing to use the app at least 3 times per week during the trial period. Information technology training was provided on 3 occasions; 2 sessions were provided before the trial period began, and 1 session was provided in week 6 of the trial period. The purpose of the training was to provide guidance on how to use the app, how to upload media, and to provide general app support. The app recorded user event logs locally on each iPad using an SQLite database (a public domain structured query language database). User event log data were later collected in person from the iPads using a portable storage device. Specific activities and activity types were logged: entry (logging in), administrative activities , reminiscing , queries at a given instant (EMA questions), and exiting (logging out). The app allowed individuals to upload their own photos, videos, and music or to access media online. For the purposes of this study, generic media was defined as photos, videos, and music that were accessed online through the app, for example, a photograph of street where the individual lived as a child. Personal media was defined as media that was uploaded, such as an old family photograph.The EMA questions were a subset of items derived from the Mutuality Scale . Over thR studio (version 3.6.0) and the R programming language were used for data wrangling and statistical programming. For the first part of the analysis, the user log data were filtered to only contain data related to queries at a given instant in order to look at overall engagement with questions. Over the course of the 12-week trial period, a total of 832 questions were presented. Of these questions, 77 were asked during training days and so were excluded from this analysis, leaving 755 questions asked during the trial period.Data were aggregated by time of day (hourly), and by week (weeks 1 to 12) to obtain question presentation counts and dismissal rates for each user type (persons with dementia or caregivers). Data were aggregated over the 12-week trial period to obtain total question presentation counts and dismissal rates for each user type , media format , media source , and the last logged activity . Some media could not be categorized into generic or personal media and were, therefore, only included in the aggregate media category . For last logged activity, 124 out of 628 (16%) EMA questions were filtered out of the data since the previous recorded event had been an EMA question and not an action that could trigger a question .P<.05 was considered statistically significant. Chi-square tests were used to compare the proportion of questions answered versus dismissed across hours of the day, across trial weeks and for other logged activities—log-in, add or delete photo, exit app. Pairwise chi-square tests (n=36) adjusted for multiple testing using Bonferroni correction were used to compare dismissal rates for each question following reminiscence with all photos, videos and music, and then segregated into personal and generic photos, videos and music .Spearman rank correlation coefficients were used for association analysis between the number of questions presented and the number of interactions with the app, number of app interactions and the number of questions presented across trial weeks, dismissal rate and time of day, dismissal rate and trial week, number of questions presented and the number of questions dismissed per hour, where A total of 30 dyads (person with dementia and their caregiver) were recruited to the study; however, usage data was found to be corrupted in the iPad software used by 2 participants with dementia and their caregivers. Therefore, only 28 sets of tracking data (n=56 participants) were analyzed in this study. The characteristics of the participants are shown in P<.001), with roughly 1 question asked for every 10 app interactions. The overall dismissal rate for questions asked during the trial period excluding training days was 30.9%. Hence, 522 out of 755 (69.1%) questions were answered. Persons with dementia used the app more in the trial period than caregivers did [Five different EMA questions from the Mutuality Scale  were prevers did , but desA breakdown of presentation and dismissal for each question is shown in P=.001).The number of questions presented per hour and per week are shown in 21=21.4, P<.001); questions asked at 9 PM  were 5 times more likely to be answered than questions asked at 11 AM . The dismissal rate for questions was high at the start and end of the trial and was lowest in the middle of the trial .The hourly and weekly dismissal rates are shown in he trial . For exaP<.001, ρ=0.81). Weekly dismissal rates differed between user types .Dismissal rates by time of day and by week for each user type are shown in er types . There wP<.001, ρ=0.84) and persons with dementia . From investigating the residuals, there were significantly less questions dismissed at the hours of 2 PM , 9 PM, and 10 PM for both caregivers and persons with dementia. For both user types, questions in the evening were more likely to be answered than dismissed.There was a strong correlation between the number of questions presented and the number of questions dismissed per hour for caregivers  and generic music versus personal music . There was no significant difference between proportion of questions of dismissed and answered between generic videos and personal videos .As shown in 21=10.9, P<.001). The dismissal rate for questions asked after viewing personal photos  was significantly higher than the rates of those asked after generic music , personal music , and generic photos . The dismissal rate for questions asked after interacting with generic music  was significantly lower than those of personal music , personal videos , or personal photos . These results show that questions asked following reminiscence with personal media, especially those asked after personal photos, were less likely to be answered compared to those asked after other media. In contrast, questions asked after the user had listened to generic media, in particular those asked after generic music, were much more likely to be answered.Significantly more questions were dismissed after viewing any type of photo than after listening to any type of music . There was no significant difference between questions answered and questions dismissed when caregivers logged in , when caregivers added or deleted photos , or when caregivers or persons with dementia exited the app .Questions were also presented to users as they were completing other logged activities. There were significantly more questions answered than were dismissed when a person with dementia logged into the app  was deemed sufficient to meet the objectives of a feasibility study.Another limitation was the recording of log data. User events were logged in real time and were later collected in person from the iPads using portable storage devices. As a result, we could not control for lost local data due to operating system failures, app crashes, bugs, and updates. This was the case with some of the log data which was recorded. For 16% of all EMA questions asked, the event which was logged previously was also an EMA question instead of one of the actions  that shoThis study explored engagement with EMA questions using an app for reminiscence where the users were caregivers and persons with dementia. Our results show that older people are willing to participate and engage in EMA; however, more can be done to increase engagement. Notwithstanding the limitations, based on this study we propose a set of recommendations for the use of EMA to optimize user engagement within a digital intervention. EMA questions should add value and help to validate the use of the digital health app in line with the study objectives. It is important that EMA questions do not distract or interfere with the overall purpose of the app which was to allow persons with dementia and their caregivers to reminisce. To avoid overprompting the user, no more than one question should be presented per day and ideally in the evening after 8 PM as this is when people were most likely to engage. If the user is trying to complete a task within the app such as adding content then a question should not be presented. These recommendations can be broadly applied to EMA use in similar settings. Future work will be carried out to study the engagement on a larger scale with more participants, which will further support these recommendations. Future work will also involve studying app usage patterns, for example, if a user engages in an EMA question, when will they next engage, and can we predict engagement based on user log analysis.


Model organisms like C. elegans and Drosophila are powerful systems to help dissect molecular genetic aspects of complex biological processes. Here we will present a number of biological questions and approaches that have been used to study questions of neural integrity and healthful aging using these systems.


Dendropanax morbifera, which is endemic to southern regions of Asia, have long been used as a folk medicine to treat variety of diseases. However, the sap of this plant has not been widely studied and its bioactive properties have yet to be clearly elucidated. Here, we isolated extracellular vesicles from D. morbifera sap with the goal of improving the intracellular delivery efficiency and clinical effectiveness of bioactive compounds in D. morbifera sap. We further investigated the anti-metastatic effects of D. morbifera sap-derived extracellular vesicles (DMS-EVs) using a cancer metastasis model based on 3D microfluidic system that closely mimics the in vivo tumor environment. We found that DMS-EVs exerted a concentration-dependent suppressive effect on cancer-associated fibroblasts (CAFs), which are important mediators of cancer metastasis. DMS-EVs also altered expression level of genes, especially growth factor and extracellular matrix (ECM)-related genes, including integrin and collagen. Our findings suggest that DMS-EVs can act as anti-CAF agents to reduce CAFs in the tumor microenvironment. They further indicate the utility of our 3D microfluidic model for various drug-screening assays as a potential alternative to animal testing for use in validating therapeutic effects on cancer metastasis.Natural medicinal plants have attracted considerable research attention for their potential as effective drugs. The roots, leaves and stems of the plant,  Extracellular vesicles (EVs), which contain signal proteins, lipids, micro RNAs (miRNAs) and functional mRNAs, play an important role in intercellular communication in the extracellular space ,2. In adDendropanax morbifera is a plant endemic to southern parts of Asia, a region with a rich folk medicine tradition and a history of commercial production of D. morbifera golden varnish. Recent studies have reported that extracts from roots, leaves and stems of D. morbifera exhibit antioxidant, anticancer and anti-diabetic effects, as well as anti-complement, anti-inflammatory and antiplasmodial activities  fetal bovine serum  and 1% [v/v] penicillin-streptomycin . Before detachment of cells from the culture dish, the culture medium was replaced with Dulbecco’s Modified Eagle Medium (DMEM), supplemented with 10% exosome-depleted FBS and 1% [v/v] penicillin-streptomycin to remove interference from FBS exosomes in media. The culture medium was then collected and concentrated by tangential flow filtration using a membrane with a 100-kDa molecular weight cutoff . The resulting exosomes were filtered with a 0.22-μm filter and stored in a −80 °C freezer until use.B16BL6 murine melanoma cells  were cultured in minimum essential media alpha , supplemented with 10% [5.0 mg/mL solution of collagen gel (pH 7.4) was prepared by add 10× phosphate buffered saline , 0.5N NaOH, distilled deionized water and collagen solution  in the given order. The hydrogel solution was then injected into the gel-filling region of microfluidic devices. Hydrogel-filled devices were place in a humidified, 37 °C incubator for 30 min. After incubation, all channels of the devices were filled with endothelial basal medium-2 , and the devices were again placed in the incubator.2 environment and used at fewer than 5 passages in all experiments. Approximately 50–60 μL of HUVEC suspensions (5 × 106 cells/mL) was seeded into the central channel of microfluidic devices and allowed to attach. After attachment of HUVECs, all reservoirs of devices were filled with EGM-2 medium, and HUVECs were incubated for 2 days to allow formation of an endothelial monolayer.HUVECs  were cultured in endothelial growth medium  at 37 °C in a 5% COHUVEC monolayers were reconstituted within 3D microfluidic devices supported by the collagen gel, which recapitulates a 3D tumor microenvironment. Cancer (melanoma)-derived exosomes (C-EVs) were delivered at a concentration of 50 µg/mL, after which interstitial fluid flow was provided through channels on one side through the collagen by virtue of height differences between side reservoirs. Our previous studies showed that the predicted interstitial flow rates in the device were range from 0.1–0.7 µm/s, which was pathophysiological level of interstitial flow ,16. CAFsCellular uptake of DMS-EVs was analyzed by fluorescence microscopy. DMS-EVs extracted as described above were tagged by incubating with red Di-I stain (1 µg/mL), a lipophilic dye. After DMS-EV treatment and uptake by HUVECs and CAFs in the 3D microfluidic device, the medium was removed and cells were fixed by incubating with 4% paraformaldehyde. The cells were prepared for staining by adding 0.15% Triton X-100 and incubating for 30 min at room temperature, after which all solutions were removed and wells were filled with 3% bovine serum albumin (BSA). After incubating for 1 h at room temperature, the solutions were again removed and Actin green 488 was added to stain the cytoplasm of the cells. Cells were then incubated for 30 min at room temperature, washed three times with 1% BSA, and incubated at room temperature for 15 min with Hoechst33342, which stains nuclei. The wells were then washed with 1% BSA and filled with PBS, after which cells were imaged under a fluorescence microscope .The viability of mammalian cells was tested using a LIVE/DEAD viability/cytotoxicity kit  as described by the manufacturer. Briefly, after culturing as described in the text, the medium was removed and cells were washed three times with PBS. Staining solution, prepared by mixing calcein AM and ethidium homodimer-1, was then added and cells were incubated for about 30 min at room temperature. The percentage of live cells stained by calcein with green fluorescence converted from calcein-AM by intracellular esterase activity and dead cells stained with orange/red by ethidium homodimer-1 (EthD-1) was assessed by imaging cells under a fluorescence microscope .v/v) for measurement of zeta potential. Diluted samples were inserted into the folded capillary cell  and detected using the same equipment indicated above.Hydrodynamic size and zeta potential of DMS-EVs were determined using dynamic light scattering (DLS) and a Zetasizer . Isolated EVs were placed in a thermostatic cell at 20 °C. The size distribution was determined by measuring hydrodynamic size. Thereafter, samples were mixed with distilled water at a ratio of 95:3 , which assesses the combined properties of light scattering and Brownian motion. Samples suspended in liquid were diluted and visualized with a Nanosight instrument  at a temperature of 25 °C using a 488 nm laser.Nanostring nCounter assays  were conducted according to the procedure outlined by the manufacturer. All tags were removed except for the DNA tags ligated onto the 3′-end of each miRNA in the samples. After each miRNA was uniquely represented, the samples were hybridized with tag-specific nCounter capture and barcoded reporter probes by reacting with 5 mL of 5×-diluted sample preparation reaction solution. Total RNA samples (100 ng) were incubated at 64 °C for at least 18 h. All excess capture and reported probes were removed. Target RNA molecules in the sample were quantified and classified by counting each fluorescent barcode with the nCounter digital analyzer.p-value < 0.05 was considered statistically significant.All data were obtained through three replicate experiments. Statistical analyses were performed using two-way analysis of variance (ANOVA) followed by Dunnett’s multiple comparisons test with GraphPad Prism . 9 particles/mL, and particles displayed a negative zeta potential value of −25 mV. Zeta potential provides a measure of dispersion stability, a characteristic property of the population of exosomes that reflects interactions among charged particles. A higher zeta potential results in greater electrostatic mobility between particles and a greater tendency to resist aggregation. Vesicles with minimum zeta potentials of ±20 mV are moderately stable [DMS-EVs were isolated using a sequential centrifugation step and filtration approach based on mammalian exosome isolation techniques, and their size distribution was evaluated by nanoparticle tracking analysis (NTA) and dynamic light scattering (DLS) analysis a,b. BothFluorescent DMS-EVs, labeled with red Di-I and incubated with CAFs as described in Materials and Methods, were widely distributed within cells, including in the cytoplasm and around nuclei e, indicaA 3D microfluidic model was used to monitor differentiation of CAFs from HUVECs a,b. To tAs a control, normal fibroblasts (NFs) were seeded into the medium channel, and inhibition rates for CAFs and NFs were determined in real time by counting the number of CAFs or NFs per field of view after single treatment with 1 or 10 µg/mL of DMS-EVs c,d. As sReprograming of fibroblasts into CAFs by cancer cells is mediated by miRNA. However, CAFs derived from tumor stroma have been reported to differ from normal stromal fibroblasts in terms of the expression of ECM proteins, pro-tumor factors, and cytokines ,21. ThusTo further assess the effect of concentration and treatment procedure on the suppression of CAFs by DMS-EVs, we investigated three different conditions: repetitive treatment, pre-treatment before adding cancer-derived EVs (C-EVs) and co-treatment with C-EVs and DMS-EVs together .For repetitive treatment, CAFs were treated with DMS-EVs every 24 h. With this experimental paradigm, after the initial DM-EV treatment, the inhibition rate and number of CAFs were similar regardless of the concentration of DM-EVs, and 4 days were required to completely suppress CAFs a. Thus, For pre-treatment, CAFs were treated with 1 or 10 µg/mL of DMS-EVs followed by treatment with C-EVs. In these experiments pre-treatment with 1 or 10 µg/mL of DMS-EVs decreased the survival of CAFs by 37% and 47%, respectively, on day 4 compared with DMS-EV–untreated CAFs. To date, there is limited evidence of the effectiveness of natural products in preventing cancer or cancer recurrence . Thus, oIn the co-treatment paradigm, DMS-EVs and C-EVs were applied together on a human endothelial monolayer our in vitro model. Co-treatment with 1 or 10 µg/mL of DMS-EVs together with C-EVs decreased CAF survival by 55% and 76%, respectively, on day 4 compared with DMS-EV–untreated CAFs. There is increasing evidence that natural products from dietary sources can make an important contribution to cancer development, and that diet may affect outcomes after cancer diagnosis ,27. Our To identify the major CAF-associated genes that are affected by DMS-EV treatment, we profiled mRNA expression using the Nanostring nCounter platform, which includes a panel of 770 genes known to be involved in metastasis. To this end, CAFs were treated with 1 or 10 μg/mL of DMS-EVs and differences in gene expression compared with untreated CAFs were assessed. Of the 770 genes in the Nanostring nCounter analysis system panel, approximately 20% are CAF-related genes. Heat maps used to compare differentially expressed gene levels between untreated CAFs and CAFs treated with 1 or 10 µg/mL of DMS-EVs showed clusters based on similarities and dissimilarities among groups a. IntereWe then performed a Gene Ontology (GO) enrichment analysis of differently expressed mRNAs with critical functions in CAFs, comparing untreated CAFs with CAFs treated with 1 or 10 µg/mL of DMS-EVs, as shown in To further investigate genes whose expression was changed in CAFs by DMS-EVs in a concentration-dependent manner, we focused on 10 genes that showed different degrees of differential expression between treatment with 1 and 10 µg/mL of DMS-EVs a: CD44, TFG-β2, which was significantly upregulated in CAFs in the absence of DMS-EVs treatment, was decreased by 5.8 and 6.2-fold at 1 and 10 µg/mL of DMS-EVs, respectively, indicating a dominant effect of DMS-EVs on CAFs b. The TFDuring cancer progression, CAFs are directly involved in dysregulated collagen turnover, which leads to increased stiffening of the tissue ,39. The Although these findings reveal changes in CAF-associated genes with DMS-EV treatment, whether DMS-EVs contribute directly to the inhibition of genes associated with the reduction in the number CAFs will require further investigation, because these genes exhibit interactions among the various ECM-related pathways. The traditional therapeutic modalities against cancer are chemotherapy, radiation therapy, and antiangiogenic therapy . HoweverWe investigated the inhibitory effects of DMS-EVs against CAFs in a 3D microfluidic model. We quantitatively analyzed the concentration-dependent effects of DMS-EVs on the phenotypic properties and viability of CAFs in real time, and performed a genotypic analysis to identify and determine the function of the dominant genes associated with CAFs that were affected by DMS-EVs treatment. Results obtained using our 3D in vitro model suggest that naturally derived nano-vesicles from plants have the potential to exert concentration- and time-dependent preventive and suppressive effects on the tumor microenvironment. Genes that showed differential expression in response to treatment with DMS-EVs preferentially included growth factors and ECM-related proteins, including integrins and collagens. On the basis of our results, we propose that an integrated nano-medicine approach using natural plant-based nanotechnology may provide a new direction for improving targeted stromal therapy. Further in vivo pre-clinical research and clinical studies are needed to develop safer and more effective anticancer metastasis treatments in the future.


Despite the availability of multi-“omics” strategies, insights into the etiology and pathogenesis of sarcoidosis have been elusive. This is partly due to the lack of reliable preclinical models and a paucity of validated biomarkers. As granulomas are a key feature of sarcoidosis, we speculate that direct genomic interrogation of sarcoid tissues, may lead to identification of dysregulated gene pathways or biomarker signatures.To facilitate the development sarcoidosis genomic biomarkers by gene expression profiling of sarcoidosis granulomas in lung and lymph node tissues (most commonly affected organs) and comparison to infectious granulomas (coccidiodomycosis and tuberculosis).Transcriptomic profiles of immune-related gene from micro-dissected sarcoidosis granulomas within lung and mediastinal lymph node tissues and compared to infectious granulomas from paraffin-embedded blocks. Differentially-expressed genes (DEGs) were profiled, compared among the three granulomatous diseases and analyzed for functional enrichment pathways.STAB1, HBEGF, and NOTCH4), excluded others and identified new genomic markers present in lung and lymph nodes, ADAMTS1, NPR1 and CXCL2. Comparisons between sarcoidosis and pathogen granulomas identified pathway divergences and commonalities at gene expression level.Despite histologic similarities, DEGs and pathway enrichment markedly differed in sarcoidosis granulomas from lymph nodes and lung. Lymph nodes showed a clear immunological response, whereas a structural regenerative response was observed in lung. Sarcoidosis granuloma gene expression data corroborated previously reported genomic biomarkers (These findings suggest the importance of tissue and disease-specificity evaluation when exploring sarcoidosis genomic markers. This relevant translational information in sarcoidosis and other two histopathological similar infections provides meaningful specific genomic-derived biomarkers for sarcoidosis diagnosis and prognosis. Sarcoidosis is a multisystemic disease of unknown etiology characterized by the formation of granulomatous lesions, especially in lung tissues and thoracic lymph nodes , 2. The Coccidioides immitis and C. posadasii, a soil-dwelling fungi disease, are endemic in the arid southwestern USA and northern Mexico.The etiology of sarcoidosis is unknown, and the diagnosis remains one of exclusion . SpecifiGranulomas are conglomerates of epithelioid and multinucleated giant cells surrounded by CD4+ and CD8+ T lymphocytes that result from the complex immunopathogenesis between host genetic factors, environmental or infectious triggers . SarcoidAs expression profiling of sarcoidosis granuloma using next generation sequencing expression has not been reported, one goal of the present study was to compare gene expression profiles in two sarcoidosis granulomatous tissues: lung and lymph node. The second goal of this study was to assess the ability of granuloma gene expression profiles to discriminate sarcoidosis from TB and CM. Our bioinformatic analyses included determination of unique pathways associated with sarcoidosis and overlapping pathways with CM and TB. Our results indicate that despite histologic similarities, sarcoidosis lung and lymph node granulomas exhibit distinct expression profiles. In addition, sarcoidosis pathway expression was significantly divergent compared to TB and CM. These findings suggest the importance of tissue-specific pathobiology considerations to be employed when exploring potential sarcoidosis genomic markers for potential diagnostic use and discriminate from other granulomatous diseases.We used microdissected granulomatous tissue from lung  and medi2 thickness of a 5 μm FFPE tissue were obtained, 5–10 sections per subject were lysed followed by RNA extraction-free chemistry method using the HTG®’s assay kit. Library preparation for the HTG Edge processor for nuclease protection steps was used, followed by PCR tagging, library amplification, quantitation and normalization. Library was then sequenced on an Illumina MiSeq using 150-cycle V3 kits [Formalin-fixed, paraffin-embedded (FFPE) microdissections from lung and lymph node granulomatous tissue and healthy tissue (controls) were assayed using Next Generation Sequencing-based gene expression by HTG EdgeSeq Oncology-biomarker . This system uses targeted capture sequencing to quantitate RNA expression levels of gene targets in FFPE tissues. The panel included 2535 probes and 15 housekeeper genes for quantitative analysis of targeted mRNAs. A list of the complete biomarker panel genes and the biological pathways assayed are in Additional file  V3 kits .We generated the transcriptome index using the salmon package  and a faWe validated our transcriptome results in three independent cohorts of microarray datasets publicly available in the Gene Expression Omnibus (GEO), GSE16538 and GSE63548  and an uMicrodissected lung and lymph node granulomas from 18 sarcoidosis subjects, four TB subjects, three CM subjects and six healthy controls were assessed. There were no significant differences in age (p < 0.1) in the three granulomatous disease groups  was obtained as the ratio between the averages. We identified 250 significantly dysregulated genes between all granulomatous-diseased tissues and healthy control tissues. Figure NR1H3 and CXCL9 were upregulated whereas the remainder  showed downregulation in both lung and lymph nodes. Interrogation of the expression levels of these 10 sarcoidosis genes to TB and CM DEGs revealed CCL14, CXCL9, FABP4, NR1H3 were also significantly dysregulated in TB. Only six genes  were exclusively dysregulated in both lung and lymph node in sarcoidosis granulomas but not in CM or TB . Only three DEGs, SLAMF7, DC27, and CXCL13, were dysregulated in all three diseases . Expression of CXCL13, a chemokine and B-lymphocyte chemoattractant associated to calcium influx was downregulated in both sarcoidosis (lymph node) and TB granulomas but upregulated CM granulomas. A list of the top dysregulated genes by granuloma tissue origin is available in Additional file Gene expression profiles from CM lung granulomas exhibited the fewest number of dysregulated genes, 34 DEGs with a balanced pattern of dysregulation, 18 DEGs downregulated, 16 DEGs upregulated. A total of 15 DEGs were exclusively dysregulated in CM granulomas. Eight upregulated transcripts were common to sarcoidosis lung granulomas , cytokine-cytokine receptor interaction, chemokine signaling, VEGFA-VEGFR2 and focal adhesion accounted for the most significant pathways with the most number of proportionally dysregulated genes involved, we then mapped the pathways associated to the genes that were exclusively present in sarcoidosis. Sarcoidosis pathways showed similar altered pathways to the rest of the granulomatous diseases with the exception of nuclear receptors meta-pathway . However, CXCL9 was also upregulated in the TB granulomas. Downregulated genes included FABP4, a fatty acid-binding protein associated with activation of proinflammatory macrophages in breast cancer, obesity and leukemia progression [CCL14, a chemokine ligand associated with changes in intracellular calcium concentration in monocytes. Hypercalcemia associated to granulomatous diseases is commonly reported [CXCL13 and CXCL14, both DEG in sarcoidosis and TB needs further investigation.Only ten DEGs were common to both sarcoidosis lung granulomas and lymph nodes granulomas. gression , 40. FABgression , 41. We reported –44, the CXCL13, CD27 and SLAMF7, a finding that points to diverse pathogenic mechanisms in granulomatous diseases that result in a similar histopathological conglomeration of CD4+ T cells surrounded by CD8+ T cells, fibroblasts and B cells. CXCL13 is a chemokine with key role in B cell migration by regulation of Ca++ influx [SLAMF7 DE but only in sarcoidosis lymph node granulomas.Despite similarities at the histological level, our analysis revealed sarcoidosis granulomas to be significantly divergent at the genomic level when compared to CM granulomas. TB and sarcoidosis granuloma profiles share stronger similarity at the transcriptional and pathway level. Interestingly, the number of DE genes in each comparison using the same thresholds varied significantly, indicating that the number of dysregulated genes does not correlate with the number of samples per comparison, or the size of the microdissection, suggesting that the granulomas are qualitatively distinct depending on the disease and tissue of origin. We identified only three shared dysregulated transcripts in CM, TB and sarcoidosis (lymph nodes); + influx , while S+ influx . FurtherThe activation of Th1 cells capable of producing interferon (IFN)-γ is important in the immunological pathogenesis of the granuloma formation . InterfeCXCL9, a chemoattractant for lymphocytes previously proposed as a marker of sarcoidosis severity [NOTCH4 expression to be exclusively present in lung sarcoidosis. NOTCH signaling is initiated after activation of toll-like receptors in macrophages and it has been previously reported that Notch receptor ligand Dll4 is expressed in cutaneous sarcoidosis granulomas [NOTCH4 have been reported as a sarcoidosis-associated locus in genome-wide associated study in African Americans [This study also served to demonstrate the lack of specificity of prior suggested potential sarcoidosis genomic markers due to common dysregulation in fungal or mycobacterium granulomas. For example, severity , was alsseverity . On the anulomas . Supportmericans .SERTAD1, HBEGF, KLRB1) as DEGs by oncopanel profiling. KLRB1 expression was also downregulated in TB and SERTAD1 was significantly downregulated in CM. Therefore, these two genes appear to be part of common process in granulomatous diseases. In contrast, HBEGF, a heparin-binding epidermal growth factor, was down-regulated in sarcoidosis lung granulomas  and was not dysregulated in CM or TB. HBEGF is involved in epithelialization, wound contraction and angiogenesis with an active pro-inflammatory role in skin and lung alveolar regeneration [HBEGF expression was up-regulated in contrast to the observed down-regulation in granuloma tissues. This variation in gene expression across tissues may be explained by either heritability at cis loci, by trans regulation across tissues [STAB1 as part of a miRNA-derived gene signature for complicated sarcoidosis [IL6ST was significantly dysregulated only in TB granulomas (FC − 1.42 and p 0.0008), while STAB1 gene was significantly downregulated in lymph nodes in sarcoidosis . STAB1 codes for a transmembrane receptor that is expressed in endothelial cells and lymph nodes, with functions in angiogenesis, lymphocyte homing and cell adhesion which are key aspects in the chronicity of the granuloma.We have previously generated a 20-sarcoidosis gene signature by microarray in peripheral blood from PBMCs . We now neration , pneumonneration . Importa tissues , 54 or bADAMST1, CXCL2 and FABP4, that were identified DEGs by the oncopanel. These three genomic markers were present exclusively in sarcoidosis in both lung and lymph node. NPR1, Natriuretic Peptide Receptor 1, is associated with cardiac hypertrophy and fibrosis [ADAMTS1, gene encodes a disintegrin and metalloproteinase with thrombospondin motif associated with various inflammatory processes and a potentially important regulator of pathogenic granuloma formation in sarcoidosis in both compartments.Leveraging the power derived from publicly available microarray data sets from independent cohorts, we successfully validated fibrosis  and has There are some limitations of the current study. One is related to RNA from FFPE tissues, despite the use of quantitative nuclease protection chemistry that enabled extraction-free quantitation of mRNA, RNA quantity and quality are limited in FFPE compared to cryopreserved tissues. Other limitations include the use of a sequencing panel and not whole genome sequencing, that limited number of gene probes and the expression levels of some genes was undetected. Our comparisons were against tissues and not immune cells, that and the difference in platforms (RNAseq vs microarrays) can explain the limited overlap to already published data sets. Last, the only gene expression data available in CM was microarray data in mice strains limiting the validation of our results in CM. Despite the observed differences at the pathway level in sarcoidosis granulomas at lung and lymph node that indicate disease progression, further investigations are needed to corroborate the expression of these genes in sarcoidosis heterogeneous subphenotypes since limited clinical data was available to establish these correlations.Sarcoidosis is a complex systemic disease of unknown cause with clinical care hampered by the paucity of available tools for diagnosis and prognosis. Our study is the first to identify RNAseq-derived transcriptome differences in granulomas from lung and lymph nodes of sarcoidosis subjects compared to CM and TB. Our data suggest that immunological signaling related-pathways distinguishes sarcoidosis from other granulomatous diseases and fills a crucial gap in knowledge of gene expression within granulomas. This study corroborates previous genomic markers suggested for sarcoidosis such as STAB1, HBEGF, FABP4 and NOTCH4, with HBEGF and NOTCH4 only expressed in lung granulomas. We have identified new genomic markers for sarcoidosis, ADAMTS1, CXCL2, and NPR1. This relevant translational information in sarcoidosis and other two histopathological similar infections provides important information for the future development of meaningful specific genomic-derived biomarkers for sarcoidosis diagnosis and prognosis.Additional file 1: Table S1. Oncopanel gene list. List of the complete biomarker panel genes and the biological pathways assayed.Additional file 2: Table S2. Sarcoidosis clinical phenotype. Scadding stage based on radiographic imaging which consist of Stage I: hilar enlargement only; Stage II: hilar enlargement plus interstitial lung disease; Stage III: interstitial lung disease and Stage IV: lung fibrosis. Lung involvement and other organs affected based on clinical data. All subjects have at least one reported clinical symptom related to lung involvement . NA. Information not available.Additional file 3: Table S3. Microarrays data sets used for validation of the DEG identified by the oncopanel.Additional file 4: Table S4. Sarcoidosis, TB and CM top common dysregulated genes. Common DEGs in granulomas derived this oncopanel in lung sarcoidosis and CM and in granulomas derived from lymph nodes from TB and Sarcoidosis.Additional file 5: Fig. S1. Microarray validation. Comparison of granuloma DEGs identified by the oncopanel to microarray datasets. Heat maps representing expression profiles of validated gene sets in A) Sarcoidosis Lung 14 transcripts validated, B) Sarcoidosis Lymph nodes, 30 transcripts validated and C). TB lymph nodes, 46 transcripts validated. Transcripts differentially regulated showed a fold change ±2 & FDR<0.01.