SPAG5 has antitumor effects has not been determined clinically.For certain human cancers, sperm associated antigen 5 (SPAG5) exerts important functions for their development and progression. However, whether RNA interference (RNAi) targeting SPAG5 oncogene (siSPAG5) efficiently. The nanomaterials were designed to prolong the half-life of siSPAG5 in blood, increase tumor cell-specific uptake, and maximize the efficiency of SPAG5 silencing. In vitro, FeSiNTs carrying siSPAG5 inhibited the growth, migration, and invasion of bladder cancer cells. In vivo, the FeSiNTs inhibited growth and metastasis in three models of bladder tumors  with no obvious toxicities. Mechanistically, we showed that FeSiNTs/siSPAG5 repressed PI3K/AKT/mTOR signaling, which suppressed the growth and progression of tumor cells.The results indicated that Fe-doped chrysotile nanotubes (FeSiNTs) with a relatively uniform outer diameter (15–25 nm) and inner diameter (7–8 nm), and a length of several hundred nanometers, which delivered an siRNA against the The results highlight that FeSiNTs/siSPAG5 caused no activation of the innate immune response nor any systemic toxicity, indicating the possible therapeutic utility of FeSiNTs/siSPAG5 to deliver siSPAG5 to treat bladder cancer.The online version contains supplementary material available at 10.1186/s12951-021-00935-z. A wip53 , BCL2 (egulator) , and SPAtigen 5) , could rtigen 5) : The adm3Si2O5(OH)4) is a 1:1 hollowed tubular morphology clay mineral consisting of one tetrahedral SiO4 sheet outer surface and one octahedral gibbsite Mg(OH)3 sheet inner surface. Despite asbestos long-term exposure could cause chronic pleural diseases, pulmonary fibrosis and lung cancers, which strongly depending on length-diameter ratio and possibly metal content, the synthetic hydrosilicate chrysotile with a length of several hundred nanometers possess little toxic potential compared to natural chrysotile with a length above 10 μm. According to previous literatures, lung cancer and pulmonary fibrosis caused by asbestos long-term exposure with longer than ~ 15 µm and thicker than 0.1 µm, mesotheliomas and pleural plaques caused by asbestos long-term exposure with longer than ~ 4–5 µm and thinner than ~ 0.1 µm. Geoinspired synthetic chrysotile nanotubes have been prepared using hydrothermal synthesis with the composition 3Si2O5(OH)4, and specific properties, such as optical, electronic, and magnetic properties, have been achieved. Previous studies suggest that natural minerals could be synthesized and tailored for cancer therapeutic applications [Naturally aluminosilicate minerals with nanosized structures  have been constructed to efficiently deliver siRNAs because of their stable chemical composition, micro-morphology structures, and unique physicochemical properties. Precise control over the main physicochemical features, such as the size, shape, and charge, is essential for the natural aluminosilicate minerals used in the field of bio-medical engineering. Chrysotile  to deliver an anti-cancer siRNA targeting the patients –18. Accosurvival . SPAG5 i in vivo –21. Moregnancies –21.In this study, we created Geoinspired synthetic FeSiNTs, consisting of a special nanostructure combining Fe cations occupying octahedral sites and a tubular morphology, which could be used for the encapsulation, sustained release, and intracellular delivery of siRNAs. We used these siRNA-delivering nanoparticles to treat bladder cancer. Moreover, FeSiNTs could efficiently and safely deliver siSPAG5 into bladder cancer cells, where they escaped from endosomes into the cytosol. We showed that siSPAG5 delivered by FeSiNTs nanoparticles could effectively inhibit the migration, invasion, and proliferation of bladder cancer cells.2 adsorption isotherms  assay. Cytotoxicity was assessed using FeSiNTs delivering the NC-siRNA in T24 cells at different concentrations. No obvious toxicity was observed even at the highest concentration of FeSiNTs complexes (200 μg/mL), a level that was 20 times higher than the concentration required for effective transfection -siRNA complexes by T24 cells was determined using inverted fluorescence microscopy. After 6 h of incubation, FAM-siRNA could be observed inside T24 cells, demonstrating FeSiNTs internalization  increased in a FeSiNTs/FAM-siRNA dose-dependent manner  compared with that of the cells treated with PBS, FeSiNTs, and FeSiNTs/siNC  was higher compared with that in cells treated with PBS, FeSiNTs, or FeSiNTs/siNC. The inhibition of SPAG5 on T24 cell behavior, we performed a wound-healing assay in transformed T24 cells. The migration of FeSiNTs/siSPAG5-transfected T24 cells was significantly reduced in a time-dependent manner. By contrast, the controls showed no such reduction  of siRNA. Local administration via direct intratumoral injection of siRNA complexes has been achieved in mouse xenograft models. For example, polyethyleneimine (PEI)-siRNAs could inhibit glioblastoma xenograph tumor growth after intratumoral injection . IntraveWe next determined whether FeSiNTs nanoparticle-coated siSPAG5 could reach the tumor site by passing through the blood stream, and if they were stable in circulation. PBS, free FAM-siSPAG5, FeSiNTs, or FeSiNTs/FAM-siSPAG5 were injected into mice’s tail veins. The biodistribution of the complexes was determined using Xenogen IVIS Lumina system in mice at 0.5, 4, 8, and 16 h after injection -mediated gene silencing [We next evaluated the depletion of the SPAG5 protein level in the tumors induced by the FeSiNTs-siSPAG5 complexes. The FeSiNTs-siSPAG5 showed a greater inhibitory effect on SPAG5 protein levels compared with that of the PBS control . Furthermore, treatment with FeSiNTs-siSPAG5 resulted in a lower tumor stage in most of the bladder cancers examined (stage pTa/T1), which contrasted with that in the other groups, in which most of tumors were at a stage higher than pT2. This suggested that the FeSiNTs-siSPAG5 treatment had a markedly better therapeutic effect than PBS treatment. This conclusion was confirmed by histological examination of excised bladders and H&E staining of tissue slices . The rapid proliferation and growth of tumor cells can promote the occurrence of tumor metastasis and invasion [We demonstrated highly efficient accumulation of intact FeSiNTs-siSPAG5 in tumors; therefore, the tail vein injection lung metastasis model was used to evaluate the therapeutic efficacy of FeSiNTs-siSPAG5. Pulmonary metastatic nodules were enumerated in the excised lungs from each group of mice. In contrast to those in the PBS and other control groups, almost no macroscopic tumor metastases were seen in the lungs of the FeSiNTs-siSPAG5 group Fig. A, B. Theinvasion , thus thWe next investigated the in vivo toxicity of the FeSiNTs-siSPAG5 complexes. H&E staining of kidney, lung, spleen liver, and heart sections revealed no obvious histological differences between the FeSiNTs-siSPAG5-treated and the control-treated mice 3 sheet outer surface. The negatively charged siSPAG5 could be effectively anchored on the positively charged external surface through electrostatic interaction, but it was hard to package into the negatively charged internal surface due to the surface tension and the like charges repel. As other studies in the literature have reported, vacuum impregnation process have been used to increase the encapsulation of Fe-doped chrysotile nanotubes aggregation. Summary, Fe-doped chrysotile nanotubes show that the mixed-charge nanoparticles covered with positively charged external surface and negatively charged internal surface, could passive transport siSPAG5 in bladder cancer cells while no cytotoxicity towards normal cells. In addition, the siRNA and its nanocarriers were cleared by phagocytes initially, and then accumulated in the kidneys and liver before being cleared from the body. Moreover, siSPAG5 induced markedly increased in vivo antitumor activity, as evidenced by the attenuation of PI3K/AKT/mTOR signaling. The PI3K/AKT/mTOR pathway is a major oncogenic driver that exerts vital functions in cancer growth, survival, and progression, and is frequently activated during carcinogenesis. Our findings suggest that the components of FeSiNTs-siSPAG5 each have specific functions, resulting in multiple regulation mechanisms to treat bladder cancer at different stages of delivery.The application of nanoparticles comprising siRNAs has significant advantages over traditional genetic approaches, such as the capacity to alter the expression levels of several genes without recourse to animal breeding, its flexible experimental design, and its translational potential. Although therapies using siRNAs have shown promise, there are significant barriers to their delivery that prevent their clinical translation. The FeSiNTs nanoparticles developed in the present study were designed to incorporate a method of tumor-specific accumulation to solve the challenges presented by siRNA delivery: Protection of siRNAs, their residence time in circulation, cell uptake, site-specific biodistribution, and escape from the endosomal pathway. SPAG5 promotes cell growth and progression in culture; therefore, we hypothesized that silencing of er cells , 19–21. Conventional nontargeted small-RNA therapeutics cause side effects. To minimize these side effects, we tested the hypothesis that selective delivery to specific cells could be affected by the materials’ physiochemical properties alone. The developed FeSiNTs nanoparticles could transfect bladder cancer cells efficiently in vitro and in vivo, but had no effect on matched normal cells. Immune activation induced by siRNAs causes transient increase cytokines, whose levels increase, peak, and decrease at various times from 1 to 24 h after the first siRNA treatment . Other nSPAG5 expression and reduced bladder cancer progression and growth. The developed nanoparticles could potentially be used as an siRNA delivery method for robust bladder-directed therapy. Moreover, FeSiNTs-siSPAG5 was well tolerated, without causing marked dose-dependent or treatment-related toxicity. The significance of the present study lies its demonstration of siSPAG5 selective delivery to bladder tumors. Our previously study also showed that > 50% of cases of bladder express SPAG5 [In summary, we have developed FeSiNTs nanoparticles for siRNA targeted delivery to treat bladder cancer cells that overexpress SPAG5. These nanoparticles can efficiently avoid endosome degradation and deliver the therapeutic siRNA into the cytoplasm rapidly, resulting in significantly inhibited expression of ss SPAG5 , suggest2. Molecular Probes  provided DAPI. Dojindo Laboratories  provided the CCK-8 and BD PharMingen  provided the Annexin V-FITC Apoptosis Detection kit. Genepharma  synthesized all the siRNAs. The siRNA targeting SPAG5 comprised the sense strand 5′-AUCUUAAGGAGAGCCAUGATT-3′ and the antisense strand 5′- UCAUGGCUCUCCUUAAGAUGC-3′. The scrambled negative control (NC)-siRNA comprised the sense strand 5′-UUCUCCGAACGUGUCACGUUU-3′ and the antisense strand 5′-ACGUGACACGUUCGGAGAAUU-3′. The fluorescent -labeled NC-siRNAs had the same sequence, with the dye was attached to the antisense strand. The PCR primers used to detect SPAG5  and β-actin  were synthesized by Sangon Biotech .T24, a human bladder tumor cell line, was obtained from the ATCC . Dulbecco’s modified Eagle’s medium , containing 10% fetal bovine serum , 100 units/mL penicillin and 100 mg/mL streptomycin was used to culture the T24 cells at 37 °C with 5% CO2, 4MgCO3⋅Mg(OH)2⋅5H2O and FeCl2⋅4H2O with a Si/Mg + Fe molar ratio = 1.50% was used to synthesize the Fe-doped chrysotile nanotubes. The SiO2, 4MgCO3⋅Mg(OH)2⋅5H2O and FeCl2⋅4H2O concentration are 10 mM, 14.8 mM to 13.9 mM and 0.148 mM to 1.0 mM, respectively. Aqueous NaOH (0.4 M) was used to increase the pH of the mixture to 12–14. Hydrothermal treatment was applied at 210–220 °C. The reaction was allowed to proceed for 1 to 3 days, after which the reaction products were dried at 150 °C for 2 h. Fourier transform infrared spectroscopy , X-ray diffraction , and Transmission electron microscopy  were used to characterize the microstructure of the composite material.A gel mixture of SiOSiRNA loading: Thirty microliters of a 3 mg/mL FeSiNTs aqueous solution was mixed with the siRNA solution , subjected to vacuum impregnation for 3 h, and then centrifuged at 1000 rpm for 5 min. Gel electrophoresis of the supernatant was used to evaluate the siRNA loading efficiency.The FeSiNTs NPs and siRNA were mixed at mass ratios of 1:1, 10:1, 20:1, 30:1, 50:1, and 100:1, and incubated for 30 min at room temperature. Centrifugation for 5 min at 5000 rpm was used to remove insoluble particles. The free siRNA and the complexes in the supernatant were subjected to electrophoresis through a 3% agarose gel containing ethidium bromide . A UV transilluminator was used to visualize the bands.The FeSiNTs/siRNA and free siRNA were incubated in medium and serum. Centrifugation for 5 min at 5000 rpm was used to remove insoluble particles. At designated time points, samples were stored at − 20 °C until all samples were collected. Samples were then thawed on ice for electrophoresis.4 cells/well and then incubated overnight at 37 °C and 5% CO2 until they reached 60–80% confluence. The cells were then transfected using various predetermined formulations. At 6 h after transfection, FAM-siSPAG5 uptake by the cells was assessed using an inverted fluorescence microscope .In three 24-well plates, T24 cells were seeded at 5 × 104) were placed into a 35 mm glass bottom culture dish  and cultured for at 37 °C in 5% CO2 for 24 h, after which the culture medium was replaced by 500 mL of serum-free DMEM containing a 30:1 mass ratio of FeSiNTs/FAM-siSPAG5 complexes. At 6 h after transfection, cells were washed, fixed, and stained using DAPI. Images were obtained using Olympus FluoView confocal laser scanning microscopy (CLSM) and subjected to analysis using FV10-ASW viewer software (Olympus).T24 cells  was used for cell staining for 1 h. Then, stained cells were cultured with free FAM-siRNA  and FeSiNTs-siRNA for 1 h at 37 °C. The treated cells were rinsed and cultured in fresh medium for a further 1–6 h. At predetermined time points, aliquots of cells were removed, fixed using 4% PFA, and observed under a confocal microscope. The LysoTracker Red and FAM-siRNA excitation wavelengths were 550 and 480 nm, respectively.To determine the lysosomal escape of siRNAs, T24 cells were seeded in a confocal dish at 1 × 10SPAG5-specific primers and the 7900 Fast RT–PCR system  to assess the mRNA expression of SPAG5. The endogenous control gene ACTB (encoding beta actin) was used to normalize the expression data.The TRIzol reagent (Invitrogen) was used to extract total cellular RNA, which was converted to cDNA and then subjected to quantitative real-time PCR (qPCR) using For western blotting analysis, total proteins were prepared from transfected cells. For each treatment, 50 μg of protein was resolved on a 10% SDS gel and the separated proteins were transferred electrophoretically to a polyvinylidene difluoride membrane, which was incubated overnight at 4 °C with rabbit monoclonal antibodies recognizing SPAG5 , phosphorylated (p)-phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K) , p-protein kinase B (AKT) , and p-mechanistic target of rapamycin (mTOR) . Thereafter, the membrane was incubated horseradish peroxidase-linked goat anti-rabbit secondary antibody at 37 °C for 1 h . The internal control was beta-actin . The ChemiDoc XRS System  was used to detect the western blotting signals.Protein expression was visualized using a Dako Real Envision Kit , following to the manufacturer’s protocol, after staining with primary antibodies: anti-SPAG5 antibodies (Proteintech) at a dilution of 1:100, anti-p-PI3K (Abcam) at 1:250, anti-p-AKT (Abcam) at 1:500, anti-p-mTOR  at 1:50, and anti-Ki67 (Abcam) at 1:30. To evaluate SPAG5, Ki67, p-PI3K, p-AKT, and p-mTOR IHC staining, we used a semiquantitative scoring system that assessed the staining intensity and positive areas. The staining index  was calculated using the positive staining intensity  combined with the immune-positive cell proportion .Cell viability was tested with MTT kit (Sigma) according to the manufacturer’s instruction. For colony formation assay, a certain number of transfected cells were placed in each well of 6-well plates and maintained in proper media containing 10% FBS for 2 weeks, during which the medium was replaced every 4 days. Colonies were then fixed with methanol and stained with 0.1% crystal violet (Sigma) in PBS for 15 min. Colony formation was determined by counting the number of stained colonies. EdU experiments were performed using a EdU Cell Proliferation Assay Kit  according to the manufacturer’s instructions.g for 5 min, and then fixed overnight at − 20 °C in 75% ethanol. Then, the cells were subjected to RNase treatment for 30 min at 37 °C prior to PI staining  in dark conditions at 4 °C for 60 min. Flow cytometry  was utilized to determine the cell cycle distribution, following the manufacturer’s protocol.Flow cytometry incorporating propidium iodide (PI) was utilized for cell cycle analysis. Briefly, at 72 h after transfection, T24 cells were pelleted and washed by centrifugation in ice-cold PBS at 125×5) twice, which were then suspended in binding buffer (100 µL) and 5 μL of FITC-conjugated annexin-V, and incubated in dark conditions at room temperature for 30 min. Thereafter, PI (100 μL) was added and incubation continued for 5 min, before the addition of Binding Buffer (400 µL). Subsequently, a flow cytometer CANTO™ II (BD Biosciences) was used to examine the cells and the data were examined with the aid of the FlowJo software .A PI kit and Annexin-V-Fluorescein isothiocyanate (FITC) kit  were used following the manufacturer’s instructions. Pre-chilled PBS was used to wash the cells  were injected into the right flank of male BALB/c nude mice. The mice were divided randomly into four groups when the tumor volume was 200 to 300 mm3. For the in vivo imaging study, 400 μL of FeSiNTs/FAM-siSPAG5, the equivalent of free FAM-siSPAG5, FeSiNTs only, or PBS were injected intratumorally. Then, at 0.5, 4, 8, and 16 h post-injection, the Xenogen IVIS Lumina system  was utilized for mouse scanning, with an exposure time of 1 s per image. Living Imaging software  was then used to analyze the images.To generate the xenograft tumor model, T24 cells , an equivalent amount of unloaded FeSiNTs, siSPAG5, or PBS solution, respectively. Injections were repeated once per week for 5 weeks. Meanwhile, every 4 d, the short and the long axial lengths of the tumors were measured and the mice were weighed. Kaplan–Meier survival curves were designed to evaluate mouse survival. The tumor xenografts were excised, weighed, and snap-frozen for cryosectioning, or formalin-fixed for paraffin sectioning. Hematoxylin and eosin (H&E) of tumor histological sections was performed. A TdT-mediated dUTP Nick-End Labeling ) assay of primary tumor sections assessed tumor apoptosis following the supplier’s protocol. A Dako Real Envision Kit  was used to perform IHC staining, utilizing primary antibodies to detect protein expression.To study FeSiNTs/siSPAG5’s tumor suppressive effects, a subcutaneous bladder cancer model comprising BALB mice with T24 tumors was constructed as described previously. At a tumor volume reached about 100 mmN-methyl-Nnitrosurea (MNU)  using a 22-gage angiocatheter one every 14 days five times. After catheterization, the rats remained anesthetized for approximately 45 min to avoid spontaneous micturition [Female SD rats were anesthetized using ether inhalation, and their bladders were infused using 0.2 ml turition , 36.After successful tumor induction (approximately 16 weeks), 60 rats were assigned to 4 groups of 15 rats. Then, the rats were anesthetized and 500 μL of FeSiNTs/siSPAG5, siSPAG5 only, an equal dose of free FeSiNTs, or PBS were instilled into the rats’ bladders. The rats stayed sedated for approximately 45 min after instillation to minimize spontaneous micturition. The treatments were repeated once per week for 5 weeks. At 48 h after therapy termination, the rats were killed humanely. Their bladders were excised, and fixed in 4% paraformaldehyde for 24 h, paraffin-embedded, and subjected to histopathological examination. At the midportion of the bladder, transverse sections were cut, followed by H&E staining.6 cells in 100 µL of cold PBS) were injected into BALB/c-nude mice tail veins. At 4 weeks after injection, siRNA therapy was initiated. The mice were divided randomly into four groups of 5 and treated with 0.3 mg/kg of FeSiNTs/siSPAG5, an equivalent dose of siSPAG5 only, an equivalent dose of free FeSiNTs, or PBS solution once a week for 5 weeks through their tail veins. After treatment for 5 weeks, the mice were euthanized with CO2 and lung tissues were removed for histological examination and imaging. We then counted the macroscopic metastatic nodules in each lung.T24 cells  were injected via their tail vein with 25 μg FeSiNTs/siSPAG5, an equivalent dose of siSPAG5 only, an equivalent dose of free FeSiNTs, or PBS solution. Twenty-four hours later, the animals were sacrificed and their heart, liver, spleen, lungs and kidney were removed and formalin (10%)-fixed. Fixed tissues were embedded in paraffin and H&E stained using standard protocols.Twenty-four hours post-tail vein injection, blood samples were obtained from the mice and subjected to centrifugation to obtain the serum, which was stored at − 20 °C for further analysis. Modular analytics  was used to determine the serum AST, ALT, creatinine (CREA), and blood urea nitrogen (BUN). For the immunotoxicity assay, enzyme-linked immunosorbent assay (ELISA) kits (Abcam) were used to measure serum cytokine levels (interleukin (IL)-1β, IL-6, Iinterferon alpha (IFN-α), and TNF-α).P < 0.05. Data are shown as the mean ± the standard error of the standard deviation (SD).GraphPad Prism VI software  was utilized for statistical analysis. Student’s t test evaluated the statistical significance between groups and one-way analysis of variance (ANOVA) was used for multiple comparisons. Statistical significance was accepted at Additional file 1: Figure S1. Large-scale TEM images of FeSiNTs with different hydrothermal environments.Additional file 2: Figure S2. Partial enlarged TEM images of FeSiNTs with different hydrothermal environments.Additional file 3: Figure S3. (A) DSL data, (B) XRD patterns, (C) Nitrogen adsorption–desorption isotherms, and(D)Zeta-potential of FeSiNTs with different hydrothermal environments.Additional file 4: Figure S4. siRNA-binding efficiency (A) and cytotoxicity (B) of FeSiNTs.Additional file 5: Figure S5. Cytotoxicity of FeSiNTs detected by EdU assays. **P < 0.01.Additional file 6: Figure S6. Relaease analysis of FeSiNTs/siRNA after incubation with medium (A) and serum (B) for different times, as determined by gel electrophoresis.Additional file 7: Figure S7. Lysosomal escape of FeSiNTs/siSPAG5 in T24 cells.Additional file 8: Figure S8. Effects of SPAG5 silencing by FeSiNTs/siSPAG5 on T24 proliferation, detected using a EdU assay.Additional file 9: Figure S9. Effects of SPAG5 silencing by FeSiNTs/siSPAG5 on the colony formation ability of T24 cells, tested using a plate colony assay. **P < 0.01.Additional file 10: Figure S10. Annexin V and propidium iodide staining of T24 cells treated with FeSiNTs/siSPAG5 complexes and other formulations, and analysis of apoptosis using flow cytometry. **P < 0.01.Additional file 11: Figure S11. After FeSiNTs-mediated SPAG5 knockdown in T24 cells, FACS was used to analyze the cell cycle. **P < 0.01.Additional file 12: Figure S12. Tumour tissues analyzed using western blotting from the subcutaneous xenograft model.Additional file 13: Figure S13. SPAG5 and Ki67 immunohistochemistry analysis, and TUNEL staining analysis of the tumors treated with PBS, siSPAG5, FeSiNTs, and FeSiNTs/siSPAG5. **P < 0.01.Additional file 14: Table S1. Tumor suppression effect of different treatment on bladder histopathologic changes in SD rat bladders of different groups.Additional file 15: Figure S14. Biosafety evaluation of FeSiNTs/siSPAG5 in vivo.Additional file 16: Table S2. Biochemical parameters of blood from the mice at 24 h after last injection of various drugsAdditional file 17: Figure S15. IFN-α (A), IL-1β (B), IL-6 (C), and TNF-α (D) analysis of mouse blood at 24 h after PBS, siSPAG5, FeSiNTs, and FeSiNTs/siSPAG5 injection.Additional file 18: Figure S16. Immunohistochemistry analysis of members of the PI3K/AKT/mTOR signaling pathway in tumor tissues treated with PBS, siSPAG5, FeSiNTs, and FeSiNTs/siSPAG5. **P < 0.01.


The abnormal activation of signal transducer and activator of transcription (STAT) protein family is recognized as cause or driving force behind multiple diseases progression. Therefore, searching for potential treatment strategy is pursued by multiple scientific groups. We consider that providing comprehensive, integrated and unified dataset for STAT inhibitory compounds may serve as important tool for other researchers. We developed SINBAD (STAT INhbitor Biology And Drug-ability) in response to our experience with inhibitory compound research, knowing that gathering detailed information is crucial for effective experiment design and also for finding potential solutions in case of obtaining inconclusive results. SINBAD is a curated database of STAT inhibitors which have been published and described in scientific articles providing prove of their inhibitory properties. It is a tool allowing easy analysis of experimental conditions and provides detailed information about known STAT inhibitory compounds. Together, these active STAT complexes regulate gene transcription in the nucleus by binding to specific DNA-response elements in the promoter of their target genes. In this way, STATs facilitate action of interferons, cytokines, interleukins and growth factors and are involved in fundamental cellular processes such as cell growth, proliferation and apoptosis, embryonic development, immune responses and inflammation, and response to viral infections2. As a consequence, abnormal activation of STAT proteins is implicated in many human diseases, including viral and bacterial infections, inflammatory diseases, autoimmune diseases, multiple types of cancer which identifies STATs as highly attractive therapeutic targets.Signal transducers and activators of transcription (STATs) facilitate action of cytokines and growth factors, which are the main tool of the organism to battle any kind of immune challenge like inflammation or cancer. The STAT family consists of seven proteins: STAT1, STAT2, STAT3, STAT4, STAT5A, STAT5B and STAT6. Each STAT protein is composed of 5 domains: N-terminal, ‘coiled-coil’ (CC), DNA-binding (DBD), Src Homology 2 (SH2) and C-terminal transactivation domain. Activity of these proteins depends on Janus Kinase (JAK)-mediated tyrosine phosphorylation of a conserved tyrosine residue (pTyr) flanking the highly conserved SH2 domain. pTyr-SH2 interactions are crucial for STAT dimer formation and may further result in formation of multimeric complexes with other protein families. For a number of STATs, phosphorylation of specific serine residues has also been shown to be important3. STAT inhibitory strategies focus on preventing STAT dimerization by using small molecules identified by in silico 3D modelling and virtual screening of compound libraries. As such, searches for STAT-targeting compounds, especially exploring the interaction area of the SH2 domain and the phosphorylated tyrosine residue, yielded many synthetic small molecules (over 100 compounds). Among the most potent are STA-21, STATTIC, STX-0119 and OPB-311217. Other types of inhibitors include natural products  peptides and peptidomimetics , oligodeoxynucleotide decoys and antisense oligonucleotides12.Different STAT inhibitory strategies have been developed, in which STAT activity may be inhibited in a direct or indirect manner. Directly, by influencing processes such as dimerization (targeting SH2 domain) or DNA binding (targeting DNA binding domain), direct inhibition of phosphorylation (targeting transactivation domain). Or indirectly through inhibition of proteins upstream of STATs, such as the Janus kinases (JAK) JAK1, JAK2, JAK3 or Tyrosine Kinase 2 (TYK2) or of multiple interferon or interleukin receptors mediating STAT activation13. Dysregulated host immune responses and robust production of inflammatory cytokines and interferons, known as the “cytokine storm”, correlate with disease severity and poor prognosis during SARS-CoV2 infection14. Since many of these factors are potent activators of STAT signaling pathways, this identifies STATs as potential therapeutic targets in COVID-19 disease as well. Anti-IL6 antibodies, as well as JAK inhibitors (indirect STAT inhibitors)- Baricitinib, Fedratinib, and Ruxolitinib have already been selected as part of a potential treatment strategy against COVID-19 as a combined antiviral and anti-inflammatory approach19. Ruxolitinib entered phase III clinical trial  and Fedratinib phase II, both were used in pneumonia associated COVID-19 cases.Identification of specific and effective STAT inhibitory strategies could provide a tool to increase our understanding of their functional role in different diseases. Moreover, promising results for several STAT inhibitors in recent clinical trials predicts STAT-inhibiting strategies may find their way to the clinic, and could serve as therapeutic strategies in cancer, inflammation, autoimmunity and viral infections. Since December, 2019, a disease outbreak caused by a novel coronavirus (SARS-CoV-2) was declared a global public health emergency by WHO and named coronavirus infected disease-19 (COVID-19)in silico, in vitro, ex vivo, in vivo methods, in different experimental settings and disease models and the inclusion of a number of these inhibitors in clinical trials. With our database we provide a comprehensive tool for detailed characterization of compounds disrupting STAT signaling in various conditions allowing better understanding of their nature and mode of action. In addition, our database can be a source of information for other groups and function as primary selection tools for potential known inhibitors for further investigation in SARS-CoV-2 research.The relevance of STATs as therapeutic targets is emphasized by the numerous studies and publications of STAT inhibitors, involving multiple 20. An initial inhibitor list was created based on a collection of review articles selected in the search described below. This was followed by manual selection of suitable research manuscripts which further were divided into small groups based on the described inhibitors. Using scientific search engines - National Center for Biotechnology Information (NCBI) - PubMed and Google Scholar, firstly we focused on gathering names of known STAT inhibitory compounds. For this purpose, we initially used available Review articles which summarized STAT inhibitory strategies with a description of exemplary inhibitors, many of which focused on cancer research. This was followed by using more advanced search and phrase-base options: “(“stat1 inhibitor”) OR (“stat2 inhibitor”) OR (“stat3 inhibitor”) OR (“stat4 inhibitor”) OR (“stat5 inhibitor”) OR (“stat5a inhibitor”) OR (“stat6 inhibitor”) OR (“stat1 inhibition”) OR (“stat2 inhibition”) OR (“stat3 inhibition”) OR (“stat4 inhibition”) OR (“stat5 inhibition”) OR (“stat5a inhibition”) OR (“stat6 inhibition”). In this way, from PubMed we extracted 1559 potential literature sources for our database. Additionally, we checked separately every compound by its name in both NCBI PubMed and Google Scholar. This approach allowed us to thoroughly screen available literature, followed by initial manual screening of each publication and further selection and proper grouping , EXPERIMENTS (EXP), VENDORS (VEN), REFERENCES (REF), CLINICAL_TRIALS (CLT), DISEASES (DIS). Each compound was given its own unique ID number (CPD_ID) which allowed us to easily form interactions between the tables. Table COMPOUNDS contains 175 inhibitors with their basic features such as ID number, name (CPD_NAME), weight (CPD_WEIGHT), weight unit (CPD_WEIGHT_UNIT), ZINC ID (CPS_ZINC_ID), SMILES code (CPD_SMILES), CAS number (CPD_CAS), molecular formula (CPD_MOLECULAR_FORMULA), mode of inhibition, (CPD_MODE_OF_INHIBITION)) inhibited target (CPD_TARGETED_PROTEIN). More importantly, the core information of the database is gathered in the table EXPERIMENTS which consist of over 20,500 records providing data about the experimental approach used to characterize each inhibitor. Within the table there are variables such as record number (EXP_ID), compound number at the original list (EXP_COMPOUND originating form CPD_ID) reference number given in the table REFERENCES (EXP_REFERENCE), name of the investigated event/process (EXP_INVESTIGATED_EVENT), type of experiment (EXP_EXPERIMENT), concentrations of tested compounds (EXP_CONCENTRATION), unit of concentration (EXP_CONCENTRATIONS), cell line or tissue types used in experiment (EXP_CELL_LINE/TISSUE), organism that tissues or cell lines originated from (EXP_ORGANISM), animal model used in in vivo testing (EXP_ANIMAL_MODEL), investigated STAT protein (EXP_STAT protein). To make this table searchable and functional we decided to use two variables: INVESTIGATED_EVENT and EXPERIMENT (Fig. The datasets generated and analyzed in this study are available at ENT Fig.  instead For the database creation we used Django web framework, Docker for efficient deployment, Nginx as web server, Elasticsearch as a search engine and finally MariaDB as SQL database.The SINBAD database provides multiple options of filtering or searching depending on the individual users’ preference – it can be used as a dataset downloaded on a personal computer and managed with R, Excel or used online. With SINBAD the user can address multiple questions regarding STAT inhibition and conditions in which it was tested (exemplary webpage layout described in The SINBAD database is constantly being updated by the administrator of our group. What is more, it is possible for external users to upload their own published results, for which we provided a simple procedure. The User can request to add their own published data through a special contact form through which the User will receive access to a dedicated uploading panel. However, unpublished data will first have to be verified and approved by the administrator. One limitation of our dataset is that it does not include publications focusing on non-STAT target proteins. We are aware that there are multiple publications covering inhibitory properties of presented inhibitors that target pathways other than JAK-STAT. We are planning to expand SINBAD with additional experimental data gathered form publications focusing on non-STAT targets, including transcription factors such as IRFs, NF-κB and others.Supplementary Data Figure 1Supplementary Data


The origin of these qualia and their relationship with the material world have long been subject to intense debate. Here, I propose that qualia arise from—and can even determine—the operation of competitive coherence. The idea behind competitive coherence is that (1) the behaviour of a system depends on the activity of only a subset of the elements of the system and (2) the selection of elements belonging to this subset depends on both the history and the present state of the system (which includes its present environment). In arguing that competitive coherence is characteristic of living systems at every level, I contend that it is to bacteria that we should turn if we are to understand qualia. This is because bacteria were here first, constitute much of the biomass, are very complex, made the world as we know it, and help control its denizens. Finally, I propose that qualia are important in the functioning of high-level systems such as ecosystems.Subjective experiences, sensations, or feelings, qualia is underappreciated. Here, I make four proposals. Firstly, living systems traverse sequences of active states that determine their behaviour; these states result from competitive coherence, which depends on connectivity-based competition between a Next process and a Now process, whereby elements in the active state at time n+1 are chosen between the elements in the active state at time n and those elements in the developing n+1 state. Secondly, bacteria should help us link the mental to the physical world given that bacteria were here first, are highly complex, influence animal behaviour and dominate the Earth. Thirdly, the operation of competitive coherence to generate active states in bacteria, brains and other living systems is inseparable from qualia. Fourthly, these qualia become particularly important to the generation of active states in the highest levels of living systems, namely, the ecosystem and planetary levels.The relevance of bacteria to subjective experiences or  Indeed,ia exist , with quiousness . These qat, etc. . In his at, etc. , ChalmerUnlimited associative learning is thought to allow an organism that has passed this threshold to be motivated by a novel stimulus or action that can then serve as the basis for learning and that constitutes the basis of qualia [is [In grappling with this hard problem, it is often argued about how far subjective experience extends “down the phylogenetic scale” ,7,8,9,10f qualia . It requf qualia  nor indef qualia  to tensef qualia  and theralia [is ,19. q’.ualia [qualia is the microtubular network found in neurones [qualia. The “threshold of complexity” argument might be used in an attempt to disqualify bacteria on the basis of their size and their early origin in evolution; the counter-arguments are, respectively, that size and certain sorts of complexity are unrelated (or are even inversely related) and that their early origin has given bacterial complexity billions of years to evolve. A few authors have addressed the question of whether cells are conscious and have q’.ualia ,9,20. Onneurones  (howeverneurones ). It mayneurones , and thaneurones . A relatIntegrated Information Theory, which is based on taking the essential, phenomenological properties of qualia and then requiring them from the physical substrate [if a phenomenon is analyzable in fully structural terms, then explaining the origin and nature of the structure of that phenomenon amounts to explaining the origin and nature of the phenomenon itself”, which is based on the idea that “the components of qualia are unconscious associations and the structures of qualia are the structures of networks of these unconscious associations” [The hard problem has been approached using ubstrate . Accordiiations” . active in determining its behaviour; this selection is achieved via competitive coherence, a scale-free concept that is based on the dynamics of patterns of connectivity in networks. We argue that this concept not only helps explain the behaviour of living systems at all levels but also captures the essential characteristics of living systems [qualia [Integrated Information Theory in proposing that it is the patterns of connectivity resulting from competitive coherence that reveal qualia. In the strong version of the competitive coherence hypothesis, qualia are not just associated with the generation of active states but are partly responsible for it; indeed, I further propose that the qualia at the highest levels of living systems are sufficiently strong so as to have a major effect on the functioning of these systems. In what follows, a program is used to simulate the operation of competitive coherence and thereby show not only how it permits the learning of a difficult task but also how it generates different patterns of connections that could characterise different qualia. I do not claim that this simulation actually generates qualia within the computer as, to quote Alfred Korzybski, “A map is not the territory it represents, but, if correct, it has a similar structure to the territory, which accounts for its usefulness”. Some neuroscientists and philosophers may be unfamiliar with just how sophisticated and important bacteria are and why it is interesting to study them in the context of qualia. This is not just because bacteria instantiate an experimentally accessible version of competitive coherence. It is also because bacteria: (1) exhibit complexity on many scales, (2) are closest in evolution to the first cells to appear on Earth, and (3) are inseparable from “higher” organisms with which they form holobionts. Moreover, bacteria and their accompanying viruses or bacteriophages are extremely abundant and make up a substantial proportion of the Earth’s biomass. Hence, there are good reasons to claim that bacteria are the dominant form of life on Earth and, in proposing that qualia have a function at the highest levels of organisation, I therefore propose that it is to bacteria we should turn. The inverse approach to the hard problem, which I undertake here, is to try to understand how living systems work by asking how they solve the most important problems that face them and then by asking how subjective experience might be involved. In the case of cells, it has been argued that one of these problems is how to generate, and how to proceed between, a very limited number of states or phenotypes when almost infinite choices seem to be available to them . We have systems . One of  systems ,30. Cert [qualia  and, herqualia. The hypothesis is made up of two parts. Firstly, competitive coherence characterises many living systems including bacteria. Secondly, competitive coherence generates Active subset selected from all the constituents or elements of the system at a particular level of organisation. At the level of societies, organisations are constrained by the need to reconcile (1) coherence with their present environment and (2) coherence with their past states; for example, to grow and survive, research laboratories (1) must select an Active subset of PhD and post-doctoral students in response to new discoveries and to new funding initiatives but (2) must reconcile this selection with the research history of the laboratory and, in particular, with its historical skills, experience, and interests. There are a myriad of psychological, social and economic factors operating in this context but, from the point of view of a selection for the Active subset based on connectivity, these factors can be considered as links with different strengths. Note that connections between elements is an abstraction that subsumes all the interactions  between the elements of the system that affect whether an element becomes a member of the Active subset. Biological systems on all scales are confronted with the challenge of how to be in a state that is coherent with both (1) present environmental conditions and (2) the system’s previous states . These sActive subset must be related in a meaningful way to the previous subset . At the level of cells, a bacterium such as Escherichia coli has evolved to generate Active subsets in the form of cell states; as for other living systems, (1) a state must contain members that work coherently together and with the environment and (2) the states themselves must occur in a coherent sequence. This is because incoherence within and between cell states is counter-selected: a cell that simultaneously induces the genes for rapid growth in good conditions and for stress resistance in bad conditions is likely to be outcompeted by other cells; a cell that divides before replicating its DNA will die (see below). At the level of brains, only a subset of neurones are active at any one time ; if brain activity is to generate coherent behaviour, this Active subset of a system’s elements that determines the state of that system at a particular time [next week; this is a Next process at work. This is because, for example, it is easier to discuss shared transport arrangements with those already present than with those who are absent as they not playing that week. Hence, those who play this week are mostly likely to play again next week. Another important factor is the coherence of the team. Suppose there are two candidate goalkeepers available next week, the choice of one of them rather than the other may have consequences on the choice of other players since players must be chosen who can play together (and perhaps travel together); this is a Now process at work. In this analogy, the composition of the team each week is determined by a competition between the Next and Now processes so as to satisfy the demands of both the status quo and coherence.The concept of competitive coherence is that competition for two sorts of coherence underlies the generation of the lar time ,33,34. IActive subset via the Next and Now processes of the factors governing the behaviour. Suppose that this animal realises that the alpha male is racing towards it. This realisation corresponds to a Now process forcing the perception of the threat into the Active subset; the subsequent loading of the subset and resultant behaviour depend on a competition between a Next process to continue mating and a developing Now process to flee. Another example of competitive coherence, this time operating at a different level, occurs in the behaviour of individual animals. Consider a subordinate pack animal  that has begun a series of behaviours that should result in “forbidden” mating; each of these behaviours corresponds to the loading into the Active subset ensures that when the Next and Now processes are trying to load two different groups of elements into the Active subset (each resulting in a different behaviour), this subset does not contain some incoherent compromise. The importance of competition can be shown by a different principle—“collaborative” coherence—in which selection for inclusion in the Active subset is based on adding the Next and Now scores together for each element and then ranking them. Learning the simple task described below then becomes impossible (data not shown).It should be emphasized that the “competition” in competitive coherence is between two sorts of coherence, one reflecting the system’s history and the other its present situation. The connectivity-based competition for inclusion into the Active subset of the elements of the system. Here, I propose that attaining each of these states at every level in the real world via competitive coherence corresponds to a quale, that is, a quale cannot be separated from the processes of attaining the state. Attaining these states is the result of a competition between the system’s elements. This competition is based on the connectivities of these elements in two networks, one, the Next connectivity, representing the history of the system, and the other, the Now connectivity, representing the present state of the system in its environment. Insofar as each quale corresponds to the generation of a particular state of a living system by competitive coherence, this means that qualia are intimately associated with particular patterns of connections and vice versa. In the competitive coherence hypothesis, a coherent sequence of coherent states is fundamental to the functioning and indeed survival of living systems. As proposed above, each of these states is an Now and Next processes operate together in competitive coherence . To grasp this idea, it may be helpful to consider electric currents flowing in the same direction through two adjacent, parallel conducting wires; in this situation, the currents generate magnetic fields that tend to force the wires towards one another . In an analogous way, when the two processes, the oherence , they geActive subset according to selection for these different types of connection. One possibility is to have an Active subset for each type of connection and then to have a second level of competitive coherence between the Active subsets themselves. In essence, each Active subset becomes an object in its own right and a second round of competitive coherence is performed on these objects using the connections between them; this corresponds to the transition to a higher level.Each level of a living system, from the molecular to the planetary scale, has a corresponding complexity (that may or may not subsume the complexity of the levels below it) . Competiqualia that accompany the generation of the states via competitive coherence themselves become correspondingly complex and intense at higher levels such that, at the level of the planet, they have significant functions. These functions may result in actions on the material world or on something else. In which case, given the likely difficulty of denizens at one level in understanding those at a higher level, it is possible that the functions of qualia will remain forever obscure to us. In the proposal here, the Active subsets are selected at different times that determine the state of the system in response to environmental inputs . Firstly, a Next process operates to count and rank the occurrences of the addresses in the Next fields of elements 7, 29 and 31; this results in the addresses at the top of the Next ranking being 14, 45 and 72. Since address 14 has the highest score, the corresponding element 14 is selected as the first member of the developing Active state, S+1. As element 14  has a Now field, a Now process can start to act. This Now process counts and ranks the occurrences of the addresses in the Now field of element 14; this results in address 59 having the highest score and therefore being at the top of the Now ranking. Competitive coherence then entails a comparison between the Next score of address 45 (which is now at the top of the Next ranking) and the Now score of address 59 (which is now at the top of the Now ranking) and, since 45 has the higher score, element 45 is selected as the second element for the developing ActiveS+1 state. The Now process now counts and ranks the occurrences of the addresses in the Now fields of both element 14 and element 45 with the result that the score of address of element 59 increases (and remains at the top of the Now ranking). Competitive coherence then compares the Next address score of element 72 with the Now address score of element 59 and, because the address score of element 59 is higher, element 59 is selected as the final member of the Active state, which becomes the current state of the system. To illustrate the hypothesis, a computer program was written based on competitive coherence  see Sup. The essl inputs . Each elActive subset of three of these elements; each element has a Now and a Next field; each field contains the addresses of a maximum of 30 different elements to which the element could be connected. The program was given the task of learning to respond to an input sequence of n with outputs n, respectively. These inputs could be considered as representing different environments that require incompatible responses; for example, input 1 could represent a nutrient-rich environment suitable for growth with 100 as the appropriate response , input 3 could represent a nutrient-poor environment unsuitable for growth with 98 or 99 as the appropriate responses , and input 2 could represent a transitional state with 99 or 100 as the responses  being present in a following state of the Active subset. A maximum of three states after the input is permitted before the program is forced to give an output. Learning depends on the changes in connectivity that result from the changes in the frequency of the addresses in the fields of the elements; for example, the contents of three of the network’s elements before and after learning are shown in Active subset shown in Now and Next fields of elements 5, 15 and 16 contain a random selection of the addresses of the elements in the network whilst, after learning, these fields no longer have a random content: the Now field of element 5, for example, mainly contains the addresses of elements 15 and 16  being present in the 5 and 16 b. Put diNow and Next processes for the selection of elements to be present in the Active subset. This selection depends on the number of times  the address of an element is present in the Now and Next fields of certain elements; the elements considered are those that have already been selected in the developing or in the preceding state of the Active subset, respectively. For example, to select progressively the elements for a new state of the Active subset  are considered and the numbers of occurrences of the addresses in these fields are counted to give Next scores .There is a fundamental difference between the lists used by the Next and Now processes is easier to see in an example with a bigger Active subset, as in 495 to 251) are selected from the Next scores  of their addresses because their scores are higher than the Now scores of the other elements. It is important to note that the Next scores in Active subset whilst the Now scores increase from left to right as there are more and more addresses to be scored (which is because there are more elements in the Active subset and hence more Now fields). This change in the influence of the two processes results in an element from the Now process, 215, being selected in preference to 40, an element from the Next process because the address of element 215 has the higher score (The nature of the competition between the er score a.Active subset follows a clear pattern (Next ranking (which is obtained from the Next fields of the elements in the previous state of the Active subset), the curve of the scores of the elements in this ranking decreases whilst the curve of those from the Now fields increases. In the case of the Next field, this trend is simply due to the element with the highest score being selected first. In the case of the Now field, this trend is due to the pool from which the scores are obtained and increasing with each element selected. The order of the elements contending for selection to the  pattern b. As eleNow and Next fields of all the elements. Initially, the fields of all the 1000 elements were filled with the addresses of these elements chosen at random to give the set of all connections. Hence, a total of 30,000 addresses was chosen for the initial set (1000 elements x 30 addresses in each field). For each element, the number of times its address was present in the set was counted to give its rank . Then, the number of different addresses in the set with the same rank was calculated to give the distributions shown in Now and Next fields, respectively. These initial distributions, which are essentially binomial , changed as a result of learning and acquired a long tail  may become connected to generate new perceptions and behaviours. The equivalent in the program would be for each pattern in the Active subset to be represented by one of a new class of elements that could then be linked to one another and selected via competitive coherence to join a separate, higher level Active subset. In the competitive coherence hypothesis, the significance of these patterns of connectivity to E. coli can make use of a wide variety of different sources of carbon, nitrogen, phosphorus and sulphur to grow at a correspondingly wide range of rates, and growth in each of these different conditions is associated with a different set of cell states. E. coli has to contend with changes in oxygen tension, temperature, and osmotic and pH conditions, and may have to survive exposure to uv, desiccation or the presence of heavy metals, free radicals or antibiotics. To survive such vicissitudes, E. coli has many networks of genes controlled in part by transcriptional regulators of which there are over 300 [E. coli can therefore produce a very large number of different cell states. In addition, even if the environment is constant and allows steady state growth, a vegetatively growing bacterium goes through the cell cycle, which comprises the events of the replication and the segregation of the chromosomes followed by division to give daughter cells. Each of these cell cycle events corresponds to a particular sequence of cell states. How exactly does competitive coherence pan out at the level of a single bacterium? The bacterium over 300 . E. coliBacterial cells have been selected with a capacity to grow and survive . Put differently, bacteria have evolved to respond to both external and internal conditions. Such responses entail both the generation of a cell state, in which the cell’s contents work together efficiently and harmoniously, and the generation of a coherent sequence of cell states. Contradiction and incoherence are punished since, for example, a cell that simultaneously induces the expression of a set of genes encoding heat shock proteins and a set encoding cold shock proteins is likely to be outcompeted by rival cells that induce each set of genes only when needed. A similar fate would befall a cell that proceeded from one cell state to another very different one without good environmental reason; for example, cells that quit the spore state and germinate in hostile conditions are punished by death. There is therefore a strong selective pressure to evolve a mechanism for producing coherent cell states and for passing smoothly from one state to another state.Active subset can be selected include macromolecules  and small molecules and ions . A gene that it is being transcribed, an mRNA that is being translated, and an enzyme that is catalysing a reaction all belong to the Active subset. The bacterial equivalents of the Now and Next fields are the binding sites in the upstream regions of genes for transcriptional activators and repressors, terminators in mRNA, motifs in proteins that determine post-translational modifications , monovalent and divalent ion binding, association with lipids, and association with other enzymes. The Now process corresponds, for example, to RNA and proteins that have affinities for one another assembling synergistically into a functional structure whilst the Next process corresponds to a transcription factor generated in one cell state leading to the transcription in the next cell state of the genes that depend on it. The number of cell states generated by competitive coherence would correspond, in the hypothesis, to the number of qualia, but this simplified picture of a bacterial cell is, however, just too simple . They include: cytoskeletal filaments similar to tubulin [Subjective experience is sometimes considered to have a threshold of complexity below which an organism is “too simple” to have feelings. This might be thought to be the case of bacteria which were once believed to be simple, unstructured bags of enzymes. Now, we know that bacteria are complex and highly structured. Molecules and macromolecules often come together into spatially extended assemblies, unctions  or Self-ructures . Hyperst tubulin , actin [ tubulin , interme tubulin , as well tubulin ,42, CTP  tubulin  and RNas tubulin  (which c tubulin ); conden tubulin  and divi tubulin , microco tubulin , “nucleo tubulin ,50,51, c tubulin , lipid r tubulin ,54 or on tubulin , acidoca tubulin , polypho tubulin , cluster tubulin , the enz tubulin  and of o tubulin , protein tubulin ,63,64,65 tubulin , associa tubulin ,68, and  tubulin  . It Bacterial complexity continues above the level of the single cell. Bacteria possess a wide variety of chemicals that they use to signal both to bacteria of the same and of different species and to their eukaryotic hosts . These iIt has been proposed that “there are two basic factors necessary to be considered a multicellular organism: cell–cell adhesion to form a new evolutionary unit, and intercellular communication leading to coordinated activity” . The mulThere is still more to bacterial complexity than the above. Biofilms, for example, are not just multicellular but often comprise organisms from different species and even different kingdoms. They contain physiologically differentiated distinct subpopulations due to factors that include mutations and a multiplicity of different microenvironments due to the diffusion of intercellular signalling molecules, external stressors, nutrient/oxygen, and waste products; in turn, these microenvironments change due to feedback from the differentiated subpopulations. Hence, the biofilm grows and ages . Biofilmqualia are indeed generated by competitive coherence, this could occur at several levels of bacterial organisation and involve different players, different types of connections and different processes. In the case of genetic organisation at the hyperstructure level, competitive coherence affects a particular hyperstructure as follows: a Next process allows those genes that are already expressed as part of a hyperstructure to help determine which genes are expressed next in that hyperstructure [Now process allows those genes that are starting to be expressed together in a hyperstructure to recruit related genes to the hyperstructure. If tructure . A Now pActive subsets would usually include those for ribosome synthesis, chemotaxis, ATP generation, sugar metabolism and transport, and cell cycle events (see above). At this higher level (as with the generation of a hyperstructure at the lower level), the composition and existence of the cell result from competitive coherence between hyperstructures in which a Next process ensures continuity and a Now process ensures coherence. A new cell state is the result of (1) a Next process, whereby the current set of hyperstructures in the cell determines the next set, and (2) a Now process, whereby the developing set of hyperstructures progressively recruits, maintains or dismisses hyperstructures. Competition between these two processes ensures a sequence of sets of hyperstructures (cell states) that optimise growth and/or survival.In the case of organisation at the level of the cell, the state of a cell at any one time corresponds to the set of hyperstructures present within it. The hyperstructures in these Active subset corresponds to the cells that are growing or moving) and the microbiome within a holobiont  and even ecosystems within the living planet . Competitive coherence also determines the functioning of systems at levels above the cell. In the case of bacteria, these include the biofilm (where the the host ). Competqualia in the functioning of a living system increases with that level of that system such that qualia only play a significant role at the highest levels. As the states generated by competitive coherence become increasingly complex with the increase in the level of the system, the accompanying qualia become correspondingly complex and influential. At some level, these qualia become functionally analogous to the role played by gravity in the organisation of matter, which becomes increasingly important as mass increases . This level may be that of the cell, the collection of cells (like the brain), the ecosystem or the planet . In the case in which the actions of qualia are at a level above that of individual human beings, it is likely that they will remain forever inexplicable, just as the function in perception of the passage of an ion flux in a channel in a neurone is inexplicable to that channel. The fundamental question here is whether the importance of qualia come from? One possibility is that qualia are inseparable from the material world [qualia are only present in living systems; this could be either because living systems are doing something that generates qualia  or because these systems are constructed in a certain way out of certain materials . For eipanspermia [Next process) but some of these new molecules had their own, different preferences for other molecules (which is a Now process). The hypothesis that microbes actually initiated life on Earth via nspermia ,94 is conspermia  and in fnspermia . In the nspermia ,98,99. T30 bacteria [31 bacteriophages, alias bacterial viruses [Apis mellifera, as a superorganism shows that its behaviour resembles that of individual organisms insofar as the swarm obeys psychophysical laws, which include the relationship between the length of time taken to choose a nest site and the number and quality of the sites available [There is a flaw in the neurocentric reasoning that since humans are the dominant species on Earth due to the functioning of their brains they therefore possess consciousness in its highest form. This is because, by many criteria, bacteria were, are, and will be the dominant species on Earth. They were here first, they occupy almost every possible niche from over a kilometre deep in the crust  to the ubacteria  and poss viruses . These b viruses . A supervailable . By one vailable .holobiont [Even if subjective experience were confined to “superior” animals such as humans, we would need to take into account what exactly these animals are. Rather than existing as an independent organism, a multicellular host and its microbiota constitute a olobiont ,112, whiqualia can be discerned. For example, those bacteria in the gut that benefit most from their hosts consuming a particular food would have a selective advantage if they could make such consumption a pleasure for the host [We too are holobionts. In (and on) a human, bacterial cells outnumber human cells even if the total mass of these bacteria is much less than that of the human cells ; moreovethe host ,132; recsmall world networks and self-organised criticality [Active subset of the elements of the system that determines its behaviour at a particular time , competitive coherence reconciles the coherence of behavioural states over time  with the coherence of the present state . The fact that only living systems have been selected—or have learnt—to perform competitive coherence means that the connectivity generated by competitive coherence may have characteristics unique to life. Some of these characteristics can be captured by training a neural net based on competitive coherence to learn a task. From this program, the connectivities of the Next and Now elements can be obtained as they are recruited to the Active subset. These results show that competitive coherence generates states that, in terms of connectivity, have structured, differentiated and unified properties. Such states are required of qualia by Integrative Information Theory [qualia” [Now and Next curves shown in Now and Next fields of each element would vary with the importance of the element in the task that has been learnt whilst the size of the Active subset would also vary with, for example, the growth of the system.The connectivity of many types of systems can be described using concepts such as ticality ,134,135.n Theory . In this[qualia” . Recogniqualia result from competitive coherence to play a role in the functioning of a living system echoes the idea that consciousness plays a role in the functioning of an internal integrating system concerned with representational operations [qualia are considered as a causative factor in my hypothesis too, albeit only at a very high level of living systems (see below). This sounds as if I am saying that qualia are associated with a system when it reaches a certain level of complexity. This idea has been criticised on several grounds [qualia depend on the operation of competitive coherence, which is quantifiable [The idea that erations . Fundameerations ) and qua grounds . It shoutifiable , the protifiable . Moreovetifiable  since a tifiable  (for reftifiable ) and it qualia. This is because bacteria were fundamental to the origins of life and, according to several criteria, are the dominant lifeform on Earth. Where then should one look in the bacterial world for relationships between competitive coherence and qualia?It is particularly interesting to consider bacteria when exploring the possible relationship between competitive coherence, life and quale might accompany or indeed drive such formation. This could be the case, for example, in the assembly of the hyperstructures in which the ribosomes  themselves are made; the richer the growth medium, the larger these hyperstructures. Assuming that pain and pleasure are the most fundamental qualia, the quale corresponding to the assembly of a large ribosomal hyperstructure might be one of contentment or satiety.At the intracellular level, bacteria have a rich, hierarchical and “hypercomplex” organisation that comprises assemblies of molecules and macromolecules termed hyperstructures . Insofarqualia might be sought since they all involve major changes in connectivity. Firstly, chemotaxis in bacteria such as E. coli entails cells moving up a concentration gradient of an attractor or down a gradient of a repellent, swimming continuously whilst the concentration increases or decreases, respectively, and tumbling (to find a new direction) when the concentration ceases to improve [E. coli entails major changes in its hyperstructures to accompany and drive the replication and segregation of its DNA and the subsequent cell division that gives two daughter cells. It has been proposed that initiation of chromosome replication in this bacterium as it grows is related to the changes in hyperstructure dynamics that it would experience if such replication were postponed; in this case, the quale corresponding to attaining the maximum size of the ribosomal hyperstructure might be the feeling of satiety, whilst the quale corresponding to the assembly of the replisome might be one of excitement. Moreover, cell division is a risky process and involves the assembly of the divisome and its action on the cell wall, which puts the cell at risk of lysis, and in this case the quale might be anxiety. These qualia, along with others, might form part of the overall cell cycle experience at the level of the bacterium itself.At a higher level of the individual bacterium itself, there are three areas in particular where  improve ; this be improve . It is cqualia and the activities of these bacterial populations is no more absurd than a similar association with the activities of collections of neurones.At the level of large bacterial populations, it is instructive to compare them with the ensembles of neurones that make up brains. Bacterial cells contain similar macromolecules to those found in neurones and these macromolecules exist in similarly structured intracellular environments. Whilst neuronal connections are limited to synapses on dendrites, bacterial connections are very diverse and include nanotubes  and nanoqualia? At the level of interactions between bacteria, it might be imagined that the pain experienced by a bacterium as it undergoes lysis by bacteriophages is sensed by other bacteria, but I know of no evidence that resistance to bacteriophages can be transmitted other than genetically [qualia and cell division, again, I know of no evidence that division in one cell affects division in others. In the case of chemotaxis and other organised movements, a function for qualia is conceivable given that E. coli cells can move collectively as a swarm [qualia changes to become increasingly complex and intense. This results in high-level qualia having significant functions. In these functions, bacteria are likely to play a major role since they constitute a substantial part of life at the levels of holobionts, ecosystems and planet Earth. The task of understanding such high-level functions may fall into the domain of theologians but risks being unknowable.In the context of competitive coherence, what—if anything—might be the functions of etically . In the  a swarm . At stilqualia. Qualia are generated by—and can help generate (in higher level systems)—the competitive coherence that determines the state of a living system. Indeed, the different patterns of connectivity generated by competitive coherence are the connectivity correlations of different qualia.Of all the ways of generating connections, competitive coherence is special because its operation is characteristic of living systems at all hierarchical levels. The fundamental proposal of this paper is that the particular dynamics and structures of connections involved in competitive coherence are inseparable from


Fel d1 is an important allergen produced by cats that causes IgE reactions in up to 95% of cat-allergic adults. Immunotherapy to reduce human allergy to cats has demonstrated that people have the capacity to produce allergen-specific neutralizing antibodies that block IgE-mediated allergic responses. We wished to determine if “blocking” antibodies could be used to reduce the IgE binding ability of cat allergens prior to their exposure to humans. Here, we describe the characterization of Fel d1-specific antibodies. We demonstrated the efficacy of a rabbit polyclonal and an allergen-specific chicken IgY to bind to Fel d1 in cat saliva and block Fel d1-IgE binding and IgE-mediated basophil degranulation. Fel d1 blocking antibodies offer a new and exciting approach to the neutralization of cat allergens. Human sensitivity to allergens released by cats is common –3. Cats Crosslinking of IgE to receptors on mast cell and basophil surfaces causes rapid cellular degranulation and release of chemical mediators that are responsible for clinical symptoms of allergies. Therapies against IgE-mediated allergy include (1) avoidance of the instigating allergen, (2) symptomatic therapies such as antihistamines, steroids, and bronchodilators, and (3) allergen-specific immunotherapy (SIT). All these options have downsides. It is very difficult to achieve 100% avoidance especially given the ubiquitous nature of allergens such as Fel d1 . Symptom4 “blocking antibodies” that could interact with the allergen, thereby inhibiting its ability to bind to IgE [Given the limitations of current allergy reduction strategies, we wanted to investigate a novel approach to neutralizing cat allergens. It has been reported that patients receiving SIT therapy developed allergen-specific IgGd to IgE –16. To dd to IgE  and thend to IgE .Fel d1 is a tetramer composed of two noncovalently linked heterodimers , 20. EacThe purified cat major allergen protein Fel d1 and polyclonal antibodies against purified native Fel d1 made in rabbit serum were obtained from Indoor Biotech . The monoclonal rabbit anti-Fel d1 antibody FGI was obtained from FabGennix, Inc. . The chicken egg anti-Fel d1 IgY antibody was harvested from egg yolks from hens inoculated with purified Fel d1. Human plasma samples from allergy-free patients or those with known allergies were purchased from Plasma Lab International . Cat saliva was collected from healthy cats at the Nestlé Purina Petcare Center  using a commercially available Salivette® . The cats were allowed to chew on the Salivette for about 10-15 seconds, and then, the Salivettes were centrifuged (1000 g for two minutes) at room temperature to obtain the saliva. Following centrifugation, samples were transferred to microtubes and frozen immediately at −80°C.μl biotinylated anti-Fel d1 monoclonal Ab (clone# 3E4 C4 C10) was added and incubated for 1 hour at room temperature. Wells were washed again three times with PBS-T, and 100 μl diluted streptavidin-peroxidase  was added before incubation for 30 minutes at room temperature. After washing wells, three additional times with PBS-T, 100 μl 1 mM ABTS in 70 mM citrate phosphate buffer (pH 4.2 and 1/1000 dilution of H2O2) was added for color development and read at 405 nm.Fel d1 concentrations in saliva were determined using a commercial sandwiched ELISA kit from Indoor Biotech , using the manufacturer's directions. Briefly, a monoclonal Fel d1 Ab (clone# 6F9 A4 H1) was coated onto 96-well plates and maintained at 4°C overnight; then, the coated wells were blocked with 1% bovine serum albumin (BSA) in phosphate-buffered saline with Tween-20 (PBS-T) for 30 minutes at room temperature. Diluted Fel d1 control and saliva samples were added to individual wells and incubated for 1 hour at room temperature. After washing three times with PBS-T, 100 This ELISA is modified from a model described by others . BrieflyThe humanized rat basophilic leukemia (RBL) cell line used in this study was kindly provided from Dr. Vogel's laboratory . In his lab, RBL-2H3 cells  were transfected with cDNA coding for the human high affinity IgE receptor (FcɛR1) chains. The surface expression of the IgE binding alpha-chain was detected by fluorescence-activated cell sorting, and the functional integration of the humanized receptors into the signal transduction cascade was addressed by intracellular calcium mobilization. Mediator release was measured in response to human IgE and a variety of crosslinking allergen preparations. Several clones were obtained that were able to bind to allergen-specific human IgE, and clone RBL-703/9 was used in this study.β-hexosaminidase release assay included Tyrode's buffer (20x stock) made with 0.1370 M NaCl, 0.0027 M KCl, 0.0004 M NaH2PO4, and 0.0005 M MgCl2 × 6H2O in 1 l double distilled water, used fresh or stored at 4°C. The 1x diluted Tyrode's buffers were made by diluting Tyrode's buffer 20x stock solution 1 : 20 in double-distilled water with or without 50% D2O . Release buffer was made to contain 1.4 mM CaCl2 × 2H2O, 100 mM HEPES, 5.6 mM glucose, and 0.1% BSA in 1x diluted Tyrode's buffer with D2O, and pH was adjusted to 7.45 (stable only 2 days when stored at 4°C). Total release buffer was made with 1% Triton X-100  in phosphate-buffered saline. Release stop solution was made with 0.2 M glycine in double-distilled water, adjusted to pH 10.7, and stored at -20°C. Substrate solution was made by dissolving 1.3 mg/ml p-nitrophenyl-N-acetyl-β-D-glucosamine  in 0.1 M Na2HPO4, then adjusted to pH 4.5 with 0.4 M citric acid, and stored at -20°C.Buffers for the β-hexosaminidase release assay to detect the degree of reduction. The allergen stock solution was prepared from saliva from the same cat preincubated with buffer PBS only.Polyclonal antibodies against purified native Fel d1 were obtained from Indoor Biotech , and the chicken egg anti-Fel d1 IgY antibody (harvested from egg yolks obtained from hens inoculated with purified Fel d1) were diluted in 1x PBS . Cat saliva as a source of Fel d1 was added and mixed well and then incubated at room temperature for at least 1 h or overnight at 4 degrees before being used in chimeric ELISA or For the hexosaminidase release assay, RBL-703/9 cells were cultured in Minimum Essential Medium (MEM) supplemented with 5% fetal calf serum (FCS) and 5% glutamine  for 2 weeks before being used. Purified polyclonal human myeloma IgE was obtained from Biodesign International  and used to sensitize the RBL cells. Goat anti-human Fc-specific IgE Ab  was used as the positive control.6 cells/ml in cell culture media and aliquoted to 50 μl/wells in a sterile cell culture-grade 96-well plate. Purified human IgE solutions were added to all wells except wells designed as “no-stimulate” control, and plates were then incubated overnight in a cell culture incubator . Incubated plates were washed three times with 1x Tyrode's buffer without D2O. Total release buffer (100 μl 1% Triton X-100) was added to control wells designated for total release, and 100 μl Tyrode's buffer without D2O was added to wells designated as spontaneous release wells.The assay was conducted as previously described  with minμl of anti-IgE or allergen dilutions were added, and the plate was incubated for 1 hour in an incubator . After the incubation, culture supernatants were harvested. In a fresh microtiter plate, 30 μl supernatant was added to 50 μl substrate solution and incubated for 60 min at 37°C. Finally, 100 μl stop solution was added and extinction (OD) determined at 405 nm (reference filter at 620 nm) in an ELISA reader.For the test wells, 100 Relative release was calculated according to the following formula:p value of <0.05 was considered significant. Finally, Cohen's D test was used to determine the size of treatment effects. A value of 0.5 on Cohen's D test indicates a medium effect while a value of 0.8 or greater indicates a large effect.The Shapiro-Wilk test was used to evaluate normal distribution of data. Subsequently, a one-way analysis of variance (ANOVA) was applied to all data, followed by Dunnett's test when overall treatment differences were significant. A p < 0.001), but no inhibition was noted with the FGI antibody .We selected two commercially available rabbit anti-Fel d1 antibodies for initial assessment: Indoor Poly  and “FGI” antibodies . Indoor Poly is an antiserum containing rabbit IgG antibodies to multiple Fel d1 epitopes. “FGI” is a monospecific rabbit anti-Fel d1 antibody raised against a peptide sequence covering amino acids 23-40 in chain 1, which is a known IgE binding site . Human pantibody . Cohen'sεR1. The binding of antigen-specific IgE to FcεR1 sensitizes effector cells to release mediators in response to subsequent exposure to that specific antigen. This humanized basophil cell line was previously used to test allergen potency [εR1 crosslinking in the presence of cat saliva containing Fel d1 induced RBL cell degranulation which was measured by quantification of β-hexosaminidase release (mediator release). Cat saliva was preincubated with increasing dilutions of the Indoor Poly antibody, FGI antibody, control rabbit serum, or monoclonal anti-Fel d1 (clone 6F9). Results were expressed as a percentage reduction in maximum release measured in a Triton X-100 control in which all cells are lysed by the detergent. Mediator release was dose-dependently blocked  in saliva samples preincubated with Indoor Poly, but not with the FGI anti-Fel d1 antibody or control rabbit serum  cell line stably transfected with human Fc potency . RBL celit serum . Monocloit serum . The effp < 0.001)  .Chicken egg yolk-derived immunoglobulin Y (IgY) is an alternative high-yield approach to generate antibodies. Multiple studies have proven the efficacy and safety of oral administration of IgY in reducing diarrhea in domesticated animals . We geneHere, we report the characterization of antibodies: the first is a commercially available Fel d1-specific polyclonal antibody (Indoor Poly) and the second is partially purified egg yolk-derived polyclonal IgY antibodies raised against Fel d1, which could effectively reduce cat saliva-derived Fel d1-to-human IgE binding and IgE-mediated basophil degranulation. The results presented provide evidence of a novel strategy to reduce IgE accessible Fel d1 levels in cat saliva. These blocking antibodies offer a new and novel approach to the reduction of cat allergens. Fel d1 is relatively abundant in many indoor environments and can be transported on clothing to environments in which no cats are present ; therefoThe failure of the monospecific polyclonal antibody (FGI) to block IgE binding as compared to the polyclonal Indoor Poly gives insights into the mechanism of action of the effective blocking antibodies. At least three IgE-specific binding sites have been identified on Fel d1, and binding is conformational, requiring correct orientation of chains 1 and 2 for maximal IgE response , 28–31. in vitro nature of these studies is acknowledged. These in vitro studies reported here must be followed with appropriate in vivo studies in cats and studies in cat-allergic people to determine if clinically meaningful benefits can be provided. Polyclonal antibodies, such as egg yolk-derived IgY that is known to be well tolerated and effective, offer the potential of a safe and noninvasive way to reduce the allergenicity of cats.This study provided important findings about blocking antibodies to Fel d1 and demonstrated the potential of a new approach for reducing the allergenicity of Fel d1; nevertheless, the


Common to several allergic diseases is the generation of immunoglobulin E (IgE) by plasma cells, when exposed to an innocuous antigen. Asthma and chronic obstructive pulmonary disease (COPD) are two prevalent chronic airway inflammatory diseases. Asthma is mediated in some patients through eosinophilic inflammatory mechanisms that include allergic sensitization and Th2-mediated immune airway response. COPD, on the other hand is mainly considered a Th1-mediated inflammatory process with neutrophilic predominance or a non-Th2 inflammation, occasionally associated with the presence of airway bacteria or viruses. IgE production appears to play an important role in the development of both COPD and asthma, as it has been associated to respiratory symptoms, lung function, bacterial and viral infections, airway remodeling and bronchial hyperreactivity in both diseases. The aim of this review is to summarize all current data concerning the role of specific and total IgE in COPD and asthma and to highlight similarities and differences in view of possible therapeutic interventions. T lymphocytes are distinguished by the presence of cell membrane molecules known as cluster of differentiation 4 (CD4) and CD8. The CD4 T-lymphocytes are also known as helper T cells and can be further subdivided into Th1 and Th2, producing, respectively, Th1- and Th2-type cytokines  by plasma cells, when exposed to an innocuous antigen . Upon inAsthma and chronic obstructive pulmonary disease (COPD) are two common chronic airway inflammatory diseases. Asthma is often mediated through an eosinophilicinflammatory mechanism that includes allergic sensitization and Th2-mediated immune airway responses . COPD, o1 reversibility, a higher sputum eosinophil count, higher FENO levels and more exacerbations in the past year  from the SPIROMICS cohort and 36% of the COPD patients (N = 696) from the COPDGene cohort presented with atopy, defined as positive sensitization to any of the 10 indoor and outdoor allergens measured in the study  when compared to controls (n = 51)  and grass pollens (pooids and panicoids) presented the highest specific IgE-binding intensities in COPD  and house dust mites   and the cockroach allergen Bl. germanica, but no significant specific IgE-binding to pollens, house dust mites or the cockroach allergen Pr. americana (P < 0.01), poorest lung function  and increased exacerbation rate   in COPD patients when compared to controls and an increased number of exacerbations was detected in the COPD patients that were sensitized against air-fungi   and negatively with lung function , suggesting that indoor air and surfaces represent a potential source of fungal allergen exposure  were increased in COPD ((n = 51) . House d in COPD . A signiOPD n = 2, 55.8% aOPD n = 2, 55.8% aOPD n = 2, 55.8% aOPD n = 2, 55.8% aOPD n = 2, 55.8% aOPD n = 2, 55.8% aOPD n = 2, 55.8% a1/FVC), independent of smoking and asthma status  and this was in turn related to the reduction of asthma and COPD with asthmatic characteristics exacerbations following omalizumab treatment, as omalizumab suppresses this receptor –15. In vD. farinae and D. pteronyssinus), German cockroach and mold mix] they were found to be more frequently positive at least to one specific IgE when compared to controls  (p > 0.05). A higher proportion of current smokers had elevated total IgE levels and at least one positive specific IgE, when compared to former smokers  . However < 0.05) . Moreove smokers . This is smokers .Several observational studies have associated allergic sensitization by 1 year of age with respiratory viral infections in infancy –59, and A viral infection can attract several cell types, such as Th1 cells, CD8 cells, neutrophils and DC, to the site of inflammation, and they can all participate in the allergic response of the patient  Figure . MoreoveThe etiology of IgE production during viral infections is not yet clearly understood, as IgE production is normally a result of Th2-biased responses . In seveSpecifically, the Respiratory Syncytial Virus (RSV) was suggested to enhance Th2 sensitization to aeroallergens . Some stHuman rhinovirus (HRV) is also known to be a major cause of asthma exacerbations in infants and infection with HRV is associated with a higher incidence of asthma onset , 89–91. Children with parainfluenza infection in the first year of life had higher odds of developing asthma in their second year of life . In anotA growing body of evidence implicates viral respiratory tract infections as the predominant risk factor associated with exacerbations of COPD and the development of chronic airway disease , 100. MoStreptococcus pneumoniae has been related to adult asthma exacerbation  triggers histamine release through both IgE- and non-IgE-dependent mechanisms  and of fibronectin (24 h) are stimulated , as well as by stimulating their proliferation  Figure . When ASimulated . All theimulated .in vitro  was sufficient to stimulate ASMC remodeling by upregulating microRNA-21-5p, which in turn downregulated phosphatase and tensin homolog (PTEN) expression and supported mammalian target of rapamycin (mTOR) signaling  in vitro . They reignaling . Interesignaling . Therefoignaling .1 (Airway hyperreactivity is a general term to describe the increased response of the airways to bronchoconstricting agonists, such as methacholine or histamine, a hallmark of asthma . In COPD1 . Moreove1  and also1 . However1  (Table 11 (Remodeling of the airways is a well-recognized feature of COPD, depended on COPD severity based on post-bronchodilator FEV1 . There a1 –144. HowA recent study demonstrated that IgG antibodies play a key role in controlling IgE-mediated inflammatory responses in patients with nasal polyps, by interfering with allergens potentially binding to cell-bound IgEs . DepletiAllergen-specific IgGs are competing with IgEs for allergen binding, thereby decreasing the allergen-induced effector cell activation . This suIn this review we summarized current data concerning the role of IgE in COPD and asthma . It appeMK and DS: conception of the review topic. MK, EP, AG, and DS: literature review, writing of the manuscript, finalization of the manuscript, and approval of the submitted article. All authors contributed to the article and approved the submitted version.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.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.