As such, visually‐grounded questions have long been of interest to theoretical linguists and cognitive scientists. Moreover, they have inspired the first attempts to computationally model natural language understanding, where pioneering systems were faced with the highly challenging task—still unsolved—of jointly dealing with syntax, semantics and inference whilst understanding a visual context. Boosted by impressive advancements in machine learning, the task of answering visually‐grounded questions has experienced a renewed interest in recent years, to the point of becoming a research sub‐field at the intersection of computational linguistics and computer vision. In this paper, we review current approaches to the problem which encompass the development of datasets, models and frameworks. We conduct our investigation from the perspective of the theoretical linguists; we extract from pioneering computational linguistic work a list of desiderata that we use to review current computational achievements. We acknowledge that impressive progress has been made to reconcile the engineering with the theoretical view. At the same time, we claim that further research is needed to get to a unified approach which jointly encompasses all the underlying linguistic problems. We conclude the paper by sharing our own desiderata for the future.Answering a question that is  From Quine  could identify a zebra on first acquaintance armed with this symbolic representation alone ’ , interpret negation (‘but is not as wide’) and resolve the anaphora (‘that supports it’).The importance of combining these two main challenges—modelling of symbolic grounding and communication exchanges—was acknowledged by one of the first computational systems about natural language understanding, which focused on visually‐grounded dialogues. Winograd  introducdesiderata, that we use to review recent achievements in a specific subtask tackled by Winograd’s system, namely answering visually grounded questions.From Winograd’s sample dialogue, we extract a list of main linguistic phenomena  and skills (reasoning and pragmatic‐based interpretation) that we believe a multimodal system should be able to model. We will refer to this list as our Thanks to efforts within computational linguistics and computer vision, Visual Question Answering (VQA) has become a widely studied task, and important progress has been made on the development of computational multimodal models. VQA has been treated both as a downstream task and as a pre‐training task to effectively encode multimodal input and trasfer it to other multimodal tasks. In this paper, we will discuss both uses of it. By reviewing how current models handle each of our desiderata, we will highlight where further research is needed to turn VQA from an in‐lab exercise to a real‐life application, and point to what we think is feasible to achieve in the short and medium term.2In the last years, there has been a steep increase of interest in the task of answering visually grounded questions. This revival was motivated by the development of models to assist visually impaired people  which encodes the contexts in which it occurs , each handled by a dedicated module. Whilst NMN  of our list of desiderata extracted from Winograd’s dialogue sample. By so doing, we also emphasize what we believe deserves further attention from the language and vision community.3desiderata we extracted from Winograd’s dialogue sample.As we mentioned above, Winograd looked at the challenges for a visually grounded interactive system solely from the perspective of the language modality (the images were assigned pre‐compiled symbolic representations). On the other hand, most of the work carried out recently on VQA has been driven by the computer vision community. We are now in the fortunate position to promote a joint view on how the long‐standing theoretical questions about grounded language understanding are addressed by computational models. Hence, in what follows, we review where the current visually‐grounded research stands with respect to the 3.1Is there anything which is bigger than every pyramid but is not as wide as the thing thatsupportsit?’. As seen in Section Winograd called for a system that is able to infer from the visual scene the answer to a question of the type: ‘visual reasoning; Andreas et al., non‐relational questions, which require reasoning about the attributes of one particular object instance, to relational questions , and to perform a higher‐level reasoning step over this perceptual knowledge. If one views language from a denotational semantic perspective, it becomes clear that the move from non‐relational to relational skills is also crucial to master language phenomena of increasing complexity, for which yet another step is necessary, namely to deal with questions involving relations between sets of objects. For instance, to properly represent quantifiers a model has to identify the sets of relevant objects; similarly, gradable adjectives require a comparison of the set of entities against which they are interpreted; negation of, for example, a noun points to the alternative sets of the negated noun (the set of other candidate objects), etc. Answering to questions involving these expressions is therefore a higher‐level problem as compared to first‐level relations and non‐relational questions described above.As we have mentioned above, several diagnostic datasets have been released with the aim to assess model abilities to reason over a question grounded in a visual context , in an image, the object to which the expression refers  and gradable adjectives . These expressions can be vague, that is, their interpretation can depend on the context in which they are used. For example, the applicability of words like most or big in a certain context depends on the properties of the set of objects that are relevant for their interpretation. Moreover, their interpretation can be borderline and therefore differ across human speakers.‘at least two or more than half. In contrast, quantifiers such as few or most—whose intepretation largely depends on the  context in which they are uttered—are absent. A strand of work has focused on quantifiers combining formal semantics and cognitive science to propose models to perform grounded quantification in a human‐like manner  image  has been undertaken by the QA community early on. It was shown to be an interesting case‐study in between QA, information retrieval and dialogue systems: users are given the chance to refine their query/question based on the linguistic answer they received (Webb & Webber, Multimodal models have been evaluated also on visual dialogue tasks, in which the agent has to answer a FUQ by grounding it on the dialogue history and on the image the question is about. The most popular dataset, VisDial (Das, Kottur, Gupta, et al., When opening the box of interaction, the next challenge that pops up immediately is question generation. Task‐oriented visual games (de Vries et al., Most multimodal conversational models exploit the encoder‐decoder architecture (Sutskever et al., 4.2The VQA task has been further extended to QA about videos. The largest‐scale Video‐QA dataset currently available is TVQA (Lei et al., 5Reviewing the literature on VQA is, by itself, a stimulating activity since progress is tangible and fast. We hope that this paper will contribute to promote further work and collaboration between experts in the language and vision community, which we have shown to be crucial for the development of fully fledged multimodal models. We agree with the call for more research on contextual language learning promoted by Bisk et al. , and for


After irradiation with UV light, both amides underwent trans to cis isomerization, yielding 40% and 22% of the cis isomer of L1 and L2 amides, respectively. Quantum yields of this process were determined as 6.19% and 2.79% for L1 and L2, respectively. The reverse reaction  could be achieved after thermal or visible light activation. The analysis of the theoretically determined equilibrium structure of the transition-state connecting cis and trans isomers on the reaction path indicated that the trans-cis interconversion is pursued via the flipping of the substituent, rather than its rotation around the N=N bond. The kinetics of thermal back-reaction and the effect of the presence of the selected ions on the half-life of the cis form were also investigated and discussed. In the case of L1, the presence of fluoride ions sped the thermal relaxation up, whereas the half-life time of cis-L2 was extended in the presence of tested ions.Azobenzene derivatives are one of the most important molecular switches for biological and material science applications. Although these systems represent a well-known group of compounds, there remains a need to identify the factors influencing their photochemical properties in order to design azobenzene-based technologies in a rational way. In this contribution, we describe the synthesis and characterization of two novel amides (L1 and L2) containing photoresponsive azobenzene units. The photochemical properties of the obtained compounds were investigated in DMSO by UV-Vis spectrophotometry, as well as  The development of so-called smart materials, whose functions can be activated or inhibited on demand via external stimuli, is of growing interest for material science and biomedical applications . The pretrans (E) isomer and the bent metastable cis (Z) form −): 451.0 for compound C26H19N4O4, calculated 451.5.Azobenzene-4,4′-dicarboxylic acid dichloride  was gradually added to a round-bottom flask containing a magnetic stirred solution of 2-amino-4-nitrophenol  in anhydrous dichloromethane (10 mL) and 4-dimethylaminopyridine  used as a HCl scavenger. Subsequently, the reaction mixture was stirred under a reflux condenser for 12 h (temperature ~40 °C). The obtained precipitate was filtered off under reduced pressure and crystalized from a fresh portion of dichloromethane.L2: orange solid, 155 mg (58%), mp. 322–325 °C; TLC: Rf = 0.50 ; 1H NMR ): 6.99 , 7.14 , 8.01–8.25 , 8.77 , 9.92 , 11.66 , and 13.30 ; 13C NMR ): 107.4, 115.9, 119.9, 122.5, 123.3, 123.3, 126.4, 129.6, 131.2, 133.8, 137.0, 139.5, 139.6, 154.1, 154.6, 156.5, 165.2, and 167.1; IR (KBr pellet) cm−1: 3442, 3096, 1736, 1686, 1637, 1602, 1527, 1421, 1341, 1262, 1185, 870, 783, and 698; HRMS ESI(−), ([M − H]−): 541.1 for compound C26H19N6O8 calculated 541.4.The equilibrium structures and energies of the L1 and L2 compounds in DMSO were determined by employing the Density Functional Theory (DFT) with the hybrid exchange–correlation functional CAM-B3LYP (the long-range-corrected version of B3LYP using the Coulomb-attenuating method ) togethe−5 M) and 1H NMR spectroscopy (c ~ 10−3 M). The reverse isomerization was led in darkness at 333.15 K or using a solar light simulator.UV irradiation experiments were carried out in a quartz cuvette (l = 1 cm) in DMSO. The progress of the photoisomerization was monitored by UV-Vis spectrophotometry  to study their interactions with metal cations and tetra-n-butylammonium (TBA) salts , hydrogen sulfate, thiocyanate, perchlorate, p-toluenesulfonate, benzoate BzO−, acetate AcO−, and dihydrogen phosphate) in order to test the interactions with anions. The stock solutions of ligands (~10−4 M) and metal perchlorates or TBA salts (~10−2 M) were prepared by weighing the respective quantities and dissolving them in DMSO in volumetric flasks. Titrations were carried out in a quartz cuvette with path length of 1 cm with a starting volume of the ligand solution equal to 2.3 mL. To suppress the deprotonation process, titrations were also carried out in the presence of acetic acid . On the basis of the experimental data, the stability constant values were determined using BindFit v.05 software [Complexation studies were performed via the UV-Vis titration of the ligands solution in DMSO with the respective metal perchlorates  and tetra-n-butylammonium fluoride (0.004 mmol) in a 15 mL of acetone: dichloromethane mixture . Subsequently, the mixture was stirred until complete dissolution. After solvent evaporation under reduced pressure, the FTIR spectra of the obtained samples were recorded. For the 1H NMR experiments, the samples were dissolved in 0.7 mL of DMSO-6d before analysis.Complexes for FTIR and trans (E) and cis (Z). According to the calculations performed at the CAM-B3LYP/6-311++G theory level, the most stable structures of L1 and L2 in DMSO solvent , see trans L1 conformers (L1E) and four trans L2 conformers (L2E) with relative energies within 6 kJ/mol , to likely be at room temperatures. As far as the cis isomers are concerned, the most stable cis L1 structure was predicted to be 53 kJ/mol higher in energy than the lowest energy trans L1, while the relative energy of the most stable cis L2 structure was calculated to be 52 kJ/mol (with respect to the lowest energy trans L2). In addition, we found other cis L1 (L1Z) and cis L2 (L2Z) conformers, whose formation from their corresponding, most stable cis structures might be considered likely, as their relative energies are larger by only 1–6 kJ/mol  [trans L1 (1.812), cis L1 (1.946–1.948), trans L2 (1.812–1.815), and cis L2 (1.946–1.951) seem to confirm that the N=N bond order approaches 2 in all considered L1 and L2 systems. In particular, the WBI values indicate that the double-bond character is more profound in the cis isomers, which is consistent with the slightly shorter (by 0.005 Å) N=N separations predicted for those species.In all studied es (WBI)  calculattrans to cis isomerization of azo compounds [trans to cis isomerization of L1.The photochemical behavior of amides was studied by spectroscopic and theoretical methods. In the UV-Vis spectra of the compound solutions in DMSO, the changes upon UV light irradiation (λ = 365 nm) were observed . After 2ompounds . The detompounds ,47) is m0→Si excitation energies with their corresponding oscillator strengths, see In order to support the analysis of the UV-Vis spectra of L1 and L2 solutions in DMSO, we calculated . The observed upfield shifts of OH, NH, and aromatic signals for the cis-enriched mixture of L1 and L2 indicated a characteristic effect of the magnetic shielding of the overlapping phenyl rings of the concaved cis form [On the basis of cis form . The moscis-enriched mixture upon UV-irradiation is consistent with the theoretically predicted energy profiles corresponding to the trans→cis isomerization of L1 and L2 depicted in trans and cis minimum energy structures are separated by the kinetic barriers whose heights (in the trans→cis direction) were evaluated to be 155.7 and 150.5 kJ/mol for L1 and L2, respectively. Clearly, surmounting such significant kinetic barriers would not be possible at room temperature without providing sufficient activation energy (as the rate constants of ca. 10−15–10−14 s−1 are predicted for these processes at T = 298.15 K).The obtainment of the cis to trans isomerization of L1) was possible upon the sample heating at 333.15 K in darkness, whereas the analogous back-isomerization of L2 nitro derivative was achieved after the sample irradiation with visible light. The cis to trans thermal transition of L1 followed the first-order kinetic model , which indicates a larger stability of this form in comparison to the unsubstituted azobenzene . In ordo 2 days .cis-enriched mixture of L2, with visible light, a hyperchromic effect in the region of π→π* band was observed, suggesting cis to trans isomerization. The reverse process , as it was mentioned before, required a longer time (6 min) to achieve the photostationary state.After 4 min of the irradiation of the trans-L2. The 1H NMR spectra of the trans-L2, cis-enriched mixture obtained after irradiation with UV light and solution heated at 333.15 K (previously irradiated with UV light) are shown in cis isomer  of L2, was shifted to lower ppm values (Δδ = −0.17 ppm) at higher temperature, whereas the proton signal related to the second ring (○) was slightly shifted downfield (Δδ = 0.03 ppm), in comparison to the spectrum registered at 298.15 K. The observed shielding effect is likely a consequence of intramolecular CO∙∙∙OH hydrogen bonds breaking, induced by thermal energy provided to the system. Such an interpretation is also supported by the analysis of the UV-Vis spectrum of L2 solution, in which the significant hyperchromic effect observed during the sample heating is associated with heteroatoms, such as oxygen and nitrogen (the region of n→π* transition band).Heating the previously UV-irradiated solution of L2 in darkness caused the increase of the band at ca. 450 nm that corresponds to n→π* transition a. Slight−5 s−1 and the half-life time of the cis isomer estimated as 5.4 h at 298.15 K (15 min at 333.15 K)  . The kinivatives ,51.cis→trans isomerization required overcoming a much smaller, yet substantial, kinetic barrier of 103.0 and 98.4 kJ/mol for L1 and L2, respectively. Since the corresponding Gibbs activation energies (ΔG‡) at T = 298.15 K predicted theoretically  agree relatively well with those obtained experimentally (see −5 s−1 for L1 and 2.0 × 10−4 s−1 for L2) calculated for these reverse isomerization processes using the above mentioned ΔG‡ values seem reliable. In addition, we would like to point out that in both L1 and L2, cases the equilibrium structure of the transition state  and the analysis of the imaginary vibration  indicates that the trans↔cis interconversion is pursued by the flipping of the X substituent, rather than its rotation about the N=N bond, see the transition state structures (labeled TS) depicted in Theoretical calculations showed that ally see , the ratDue to the presence of NH and OH groups serving as the moieties enabling the formation of hydrogen bonds, L1 and L2 compounds may interact with ions. In fact, such potential host-guest interactions may have an influence on the thermal relaxation of azo compounds ,52. Amon1H NMR and FTIR spectra (1H NMR of free amide L1 at 9.47 ppm was broadened and shifted downfield (Δδ ~ 0.33 ppm) in the presence of fluoride anions, thus suggesting the involvement of hydroxyl moieties in the fluoride ion binding (via the formation of H-bonds). The signal of NH protons was slightly shifted to higher ppm values (Δδ = 0.04 ppm), which can be connected with host-guest interactions. In the FTIR spectra of L1 recorded in the presence of fluoride ions, the band at ca. 3400 cm−1 ascribed to the stretching vibrations of the OH and NH groups was broadened and shifted to higher wavenumbers (by 68 cm−1) in comparison to the spectrum of free amide. The amide I band (seen in the L1 spectrum at 1629 cm−1) was split in the complex spectrum and shifted to higher wavenumbers. The shape and intensity of the bands corresponding to the C-O stretching vibrations at 1280–1200 cm−1 were changed in the presence of the anion. The observed changes may be a consequence of the heteroatoms involvement in the interactions with fluoride ions.Among the anions tested, the species exhibiting 1:1 stoichiometry are formed by L2 and fluoride, acetate, or copper(II) ions, whereas 1:2 (S:A) stoichiometry is more probable when L2 interacts with benzoate or dihydrogen phosphate anions. Among the last mentioned, the nitro derivative interacts more strongly with the Y-shaped carboxylic anions. As strongly basic anions in DMSO may cause the deprotonation of amide based ligands, the interactions of L1 and L2 compounds with ions were tested in the presence of the excess of acetic acid, which was used to suppress potential deprotonation ,54. Spec spectra . Namely,1HMR spectrum of the UV-irradiated solution of L1, the shifts of signals ascribed to the protons of the cis form in the presence of fluoride anions were seen , suggesting the involvement of the cis form in the interactions with the anion. On the basis of spectrophotometric titration, it was determined that under measurement conditions, the species of 1:2 stoichiometry (S:A) were formed , with respect to the latter. Recalling that the relative energy of cis L1 (in the absence of F−), with respect to trans L1, was predicted to be substantially larger (53 kJ/mol) than the relative energy of cis L1 (in the presence of F−), with respect to trans L1/F− (3 kJ/mol); one may conclude that the attachment of F− renders the cis and trans isomers of L1 nearly isoenergetic.In the ere seen . The chae formed a. This r−∙∙∙HF complex consists of the negatively charged L2 and HF molecules connected via the hydrogen bond). This finding seems to be supported by the analysis of the FTIR spectrum registered for L2 in the presence of fluoride ions . The stoichiometry of the formed species with fluoride and copper(II) ions has changed at increased temperature, which suggests a different mode of amide-ion interactions  ions stabilized the cis isomer obtained after thermal activation.The presence of tested ions influenced the rate of the thermal olutions . After t1H NMR spectra and theoretical calculations, it was concluded that in DMSO, the most stable form of L1 is a symmetric trans isomer, whereas for nitro derivative L2, the most probable structure corresponds to a trans form, with the peripheral rings rotated by 180 degrees, with respect to each other. The UV-irradiation of both azo-bearing compounds induced trans to cis isomerization. The photoisomerization process of L1 resulted in a higher conversion to the corresponding cis isomer than in the case of L2 (40% vs. 22%). The quantum yield of the trans to cis isomerization of L2 was over two times lower than for L1. The comparison of the half-life times of the respective cis-amides showed the higher stability of cis-L1 than cis-L2 and unsubstituted azobenzene. The return to a pure trans form of L1 was possible after heating the sample in darkness, whereas, in the case of L2, this thermal activation also caused the breaking of intramolecular hydrogen bonds. The rate of thermal back-isomerization could be altered by the presence of ions. Fluoride ions interacting with L1 via hydrogen bridges caused faster cis to trans relaxation. In the case of L2, the presence of tested ions decreased the cis to trans isomerization rate, probably due to the increased affinity of the L2 form obtained after thermal activation towards ions. The results of the theoretical calculations performed for the L1 and L2 species revealed not only the equilibrium structures of their most stable isomers, but also those corresponding to the low energy isomeric forms (together with their relative energies). The analysis of the intrinsic reaction path connecting the trans and cis isomers of both the L1 and L2 systems resulted in the recognizing that the trans-cis interconversion process is pursued by the flipping of a substituent, rather than its rotation about the N=N bond.The new amide derivatives of azobenzene-1,4-dicarboxylic acid were obtained in good (L1) and moderate (L2) yields using a facile synthetic protocol. On the basis of the


TOLLIP polymorphism has been implicated in the development and prognosis of idiopathic pulmonary fibrosis (IPF), mainly in whites. However, ethnic differences in the characteristics of other interstitial pneumonia (non-IPF) subtypes are unclear. We evaluated the association between the rs3750920 genotype and the clinical characteristics of Japanese patients with fibrosing interstitial lung diseases (ILD). We genotyped 102 patients with fibrosing ILD (75 IPF and 27 non-IPF patients) and analyzed the interaction between the rs3750920 genotype distribution and their clinical characteristics. The overall frequencies of the C/C, C/T, and T/T genotypes were 69%, 25%, and 6%, respectively. The proportion of minor T allele carriers was larger in IPF patients than in non-IPF patients . In addition, survival at 3 years was significantly better for carriers than for non-carriers of the T allele. There was no significant association between genotype distribution and change in pulmonary function after introduction of antifibrotic agents. The frequency of the minor T allele of rs3750920 was low in Japanese patients with fibrosing ILD, particularly in non-IPF patients. Carriers of the minor T allele had better survival than non-carriers. Presence of the T allele might thus be an indicator of better outcomes for fibrosing ILD. IPF is distinct from non-IPF ILD; however, IPF and non-IPF ILD share some morphological features and pathological mechanisms2. Findings suggesting a common fibrotic pathogenesis in pulmonary fibrosis have led to recent clinical trials of antifibrotic agents that target not only IPF but also other types of fibrosing ILD4, and to the coining of the term progressive-fibrosing interstitial lung diseases (PF-ILD)5.Idiopathic pulmonary fibrosis (IPF) is a lung disorder characterized by fibrosis of the lung interstitium. Although IPF is the most common and most severe subtype of chronic fibrosing interstitial pneumoniaTERT, TERC, PANR, and RTEL1 had a higher risk of pulmonary fibrosis development and progression and worse outcomes for a variety ILD types6. In addition, shortened leukocyte telomere length was related to poor outcomes for IPF and HP9. In addition to these findings regarding telomere length and telomere-maintenance genes, a genome-wide association study (GWAS) found that single nucleotide polymorphisms (SNPs) in MUC5B and TOLLIP might be associated with IPF susceptibility and clinical outcome11. Interestingly, the rs3750290 genotype—a functional SNP within TOLLIP—interacted with IPF outcome in post hoc analysis of the PANTHER trial: a randomized clinical trial of prednisone, azathioprine, and N-acetylcysteine (NAC) for IPF13. In that study, the T/T genotype of rs3750920 was associated with significantly better survival after NAC therapy, as compared with placebo, while patients with the C/C genotype had worse survival after NAC treatment13. Thus, there may be drug–gene interaction, and genotype-stratified use of antifibrotic agents should be considered in IPF treatment. However, the association of rs3750920 polymorphism with other types of fibrosing ILD was not analyzed. In addition, the current guidelines for IPF1 recommend nintedanib and pirfenidone as the main treatment for fibrosis; NAC treatment is not the current standard. Thus, associations with other antifibrotic agents should be analyzed. Moreover, ethnic differences may exist; however, the present genomic data for IPF have mostly been collected from white populations, and it is unclear if these data are applicable to non-white populations, including Asian populations.Among host factors for ILD, genomic factors have been implicated as risk factors for developing pulmonary fibrosis and were reported to be associated with clinical outcomes. Patients with telomere-related rare variants in TOLLIP rs3750290 SNPs with the clinical characteristics of Japanese patients with fibrosing ILD.This study examined associations of 5. Acute exacerbation of fibrosing ILD was diagnosed by using criteria reported by the international working group on acute exacerbation of IPF14. The data were locked at the end of 2020. The institutional review board of Toho University Graduate School of Medicine approved this study (A18043), and the research was performed in accordance with relevant guidelines/regulation. All patients provided written informed consent.We recruited and collected blood samples from 102 patients who were treated for fibrosing ILD at our institution during the period from 2018 through 2020. The following patient characteristics were extracted from medical records: age, sex, smoking history, laboratory data, pulmonary function test results, gender-age-physiology score, development of acute exacerbation, treatment for fibrosing ILD, and outcome. IPF and other fibrosing ILD were diagnosed by using the guidelines of the American Thoracic Society (ATS)/European Respiratory Society (ERS)/Japanese Respiratory Society/Latin American Thoracic Association and the statement of the ATS/ERSPeripheral whole-blood samples were obtained from patients and stored at − 80 °C. Genomic DNA was extracted with a High Pure PCR Template Preparation Kit , in accordance with the manufacturer’s instructions. The genotype of rs3750290 was determined by real-time PCR using a TaqMan SNP Genotyping Assay . Real-time PCR was conducted with an Applied Biosystems QuantStudio 3D Real-time PCR system .2 test and Fisher exact test. We used the Kaplan–Meier method to analyze outcomes. The log-rank test was used to compare two groups. Cox proportional hazards analysis was used to identify independent predictors of survival. All P values are two-sided, and a P value of less than 0.05 was considered to indicate statistical significance. Statistical analysis was done by using SPSS version 27  and PRISM version 8 .Continuous variables were analyzed with the unpaired t-test when comparing two groups and with one-way ANOVA when comparing more than two groups, as appropriate. Categorical variables were compared with the χThe baseline characteristics of the 102 patients with fibrosing ILD are shown in Table The genotype distribution of rs3750920 in relation to fibrosing ILD subtype is shown in Table Table Kaplan–Meier curves were used to analyze outcome in relation to genotype. Figure Next, we directly compared outcomes for fibrosing ILD in carriers and non-carriers of the T allele of rs3750920. As shown in Fig. Finally, we analyzed the pulmonary function of patients who received antifibrotic agents, including inhaled NAC, during the observation period. Change in FVC during the interval from the start of an antifibrotic agent to 1 year later was compared. We were only able to analyze 22 patients treated with nintedanib Fig. A, 14 patTOLLIP and the clinical characteristics of Japanese patients with fibrosing ILD. The frequencies of the C/C, C/T, and T/T genotypes were 69%, 25%, and 6%, respectively, in the total sample and 63%, 30%, and 7% in IPF patients. The present findings and the respective genotype distribution of 22%, 50%, and 28% for patients in the PANTHER trial13 suggest that Japanese are less likely than whites to be carriers of the rs3750920 minor T allele. Similarly, the genotype distribution of SNPs in MUC5B (rs35705950) were different in Japanese IPF and NSIP patients than in a German cohort15. The rs35705950 minor T allele was present in 3.4% of IPF patients and 1.7% of NSIP patients in Japanese cohorts, as compared with 33.1% and 27.4%, respectively, in German cohorts. Thus, the minor T allele might be less frequent in MUC5B (rs35705950) and TOLLIP (rs3750920) SNPs in Japanese. SNP data provided by the National Center for Biotechnology Information showed a rs3750920 T allele frequency of 43% in Europeans and 32% in east Asians16. A Japanese genetic database (TOGO VAR) indicated that 23% of Japanese had the minor T allele17. The lower frequency of the minor T allele in Japanese subjects compared to European subjects might explain why the proportion of T allele carrier is lower in our IPF patients than in IPF patients in the PANTHER cohort. However, the frequency of the T allele was higher in our patients, especially IPF patients (37%), than in subjects of Japanese database. These findings suggest that there may be an association between IPF development and rs3750920 polymorphism, as indicated by previous GWAS11.We observed an association between the rs3750290 genotype in 18. Although we could not assess genetic distribution in relation to ANA seropositivity in our patients, this tendency was not observed in our patients with CTD-ILD, NSIP, and UCIP.The proportion of T allele carriers was significantly higher in IPF patients than in non-IPF patients in our study. To our knowledge, this is the first report to compare genotype distribution between IPF and other fibrosing ILD subtypes. Oldham et al. reported that the TT genotype of rs3750920 was more frequent in antinuclear antibody (ANA)–positive (ANA titer ≥ 1:320) patients with IPF and interstitial pneumonia with autoimmune features than in those who were ANA-negative13. In contrast, a recent study of 62 white patients with IPF reported no association of survival with rs3750920 genotype19. The present patients with the T allele had significantly better 3-year survival than did those without the T allele, regardless of treatment. This difference remained even when we limited the analysis to IPF patients. However, since the prevalence of the minor T allele was low, especially in non-IPF patients (only 3 patients had C/T and 1 patient had T/T), it was difficult to conduct survival analysis of non-IPF patients. Moreover, because we could not validate the association of the minor T allele of rs3750920 with better outcomes for fibrosing ILD in multivariate analysis, we cannot conclude that rs3750920 genotype is associated with the outcome for fibrosing ILD. We also analyzed the association between decline in FVC and treatment with the antifibrotic agents pirfenidone, nintedanib, and NAC, for each genotype. NAC has been used as inhaled form in Japan21. Although we found no significant interaction between rs3750920 genotype and antifibrotic treatment in our small sample, it is intriguing that FVC improved after inhaled NAC therapy in all 3 patients with the T allele  play a pivotal role in the innate immune system and inflammatory response22. Activation of TLRs by exogenous pathogen–associated molecular patterns triggers inflammatory signaling pathways, which results in suppression of infection. In addition, TLRs recognize host-derived endogenous ligands such as pathogen-associated molecular patterns and are involved in regulating non-infectious tissue injury23. TLR2 and TLR4 are localized on the surface of cells such as alveolar macrophages and lung epithelial cells in the lungs and are crucial in regulating inflammatory response and fibrosis24. TOLLIP encodes toll-interacting protein (TOLLIP), which is a negative regulator of TLRs, including TLR2 and TLR425. TOLLIP is an intracellular adaptor molecule that can bind to IL-1R–associated kinases (IRAK-1) and inhibit activation of nuclear factor kappa B (NFκB), which results in resolution of the inflammatory response caused by the TLRs/ligand signal pathway26. rs3750920 is a functional synonymous variant coding SNPs in TOLLIP exon 3; it was marginally associated with IPF susceptibility in 2 GWAS studies11. TOLLIP mRNA levels were significantly higher in persons with the T/T rs3750920 genotype than in those with the C/C and C/T genotypes in human samples25. Thus, increased TOLLIP expression by the minor allele might be associated with pathogenesis, responsiveness to treatment, and/or ILD prognosis. However, the complexity of immune homeostasis complicates our understanding of pathogenesis. TLR2 signaling was reported to promote pulmonary fibrosis in a model of bleomycin-induced pulmonary fibrosis28. Inhibition of TLR4 exacerbated bleomycin-induced pulmonary fibrosis29, while another study reported that TLR4 enhanced fibroblast activity, thereby promoting wound healing and fibrosis30. In addition, TOLLIP inhibited TLR4 downstream and suppressed inflammation under acute inflammatory conditions. In contrast, in low-grade, chronic inflammation, TOLLIP translocated to mitochondria and facilitated chronic inflammation26. The link between SNPs and the pathogenesis of pulmonary fibrosis should be further investigated.The biological mechanisms underlying the interaction of This study has several limitations. First, it was conducted at a single center and the sample size was small. In addition, we noted no significant difference in overall survival, perhaps because the observational period was short (108 ± 104 months). A nationwide Japanese analysis of genomic factors in idiopathic interstitial pneumonia is ongoing (NEJ036A: UMIN000032117) and might validate our findings. Second, we could not directly compare the genetic distribution of rs3750920 in our patients and healthy Japanese subjects. However, international and Japanese SNP databases provided useful information for analysis of the association of genetic distribution and disease.The present findings indicate that few Japanese with fibrosing ILD, especially those without IPF, are carriers of the rs3750920 minor T allele. Survival might be better for carriers than for non-carriers of the T allele. However, genotype was not associated with change in pulmonary function after treatment with any antifibrotic drug. To our knowledge, this is the first study to investigate associations of clinical characteristics, including outcome for fibrosing ILD, with rs3750920 genotype in a non-white population. Despite the limitations of this study, our results should provide insights regarding IPF and fibrosing ILD treatment. A nationwide Japanese genomic study is ongoing and might validate the present findings.


Chemical vapor deposition (CVD) is regarded as the most promising technique for the mass production of graphene. CVD synthesis under vacuum is the most employed process, because the slower kinetics give better control on the graphene quality, but the requirement for high-vacuum equipment heavily affects the overall energy cost. In this work, we explore the possibility of using electroformed Cu substrate as a catalyst for atmospheric-pressure graphene growth. Electrochemical processes can produce high purity, freestanding metallic films, avoiding the surface defects that characterize the rolled foils. It was found that the growth mode of graphene on the electroformed catalyst was related to the surface morphology, which, in turn, was affected by the preliminary treatment of the substrate material. Suitable conditions for growing single layer graphene were identified. Since its first synthesis, graphene has been a hot topic for many research groups , because2/CH4 gas mixture can be diluted with an inert gas such as argon; two different growth modes have been identified in these atmospheres [Alternatively, the Hospheres . The firAlong with these limitations, synthesis performed at atmospheric pressure has the advantage of excluding high vacuum equipment and reducing largely the production costs, which heavily affect CVD graphene mass production and pose a high barrier to its technological breakthrough . The aimThe quality of the prepared materials was assessed by different characterization techniques: X-Ray fluorescence (XRF), X-Ray diffraction (XRD), and atomic force microscopy (AFM) were used to evaluate substrate purity, crystalline orientation and surface roughness. After electroforming, Cu foils underwent annealing  in hydrov/v phosphoric acid, 25% v/v ethanol, 5% v/v 2-propanol, and 0.1 M urea [2 current density for 30 s. Both substrate annealing and graphene chemical vapor deposition were performed in a tubular furnace using a quartz vial; both treatments were carried out at 1000 °C and atmospheric pressure. In the annealing treatment, pure hydrogen was supplied to the vial at the flow rate of 5 NL/h for two hours, while the CVD atmosphere was a H2-CH4 gas mixture, with CH4 content of 3 vol%. Further experimental details are reported elsewhere [4/H2 flow for 2, 4, 6 and 8 min, while the annealed and the annealed-polished substrates were subjected to process times ranging from 2 to 20 min.Electroformed substrates were prepared by using a stainless-steel mandrel as the cathode and a Cu plate as the sacrificial anode. The cell was set up in the standard two electrodes configuration; the electrolyte used was the Cuproplus bath for Cu plating, supplied by Tecnochimica; plating parameters were used according to the product datasheet. Resulting free-standing Cu foils were 2 cm × 2 cm in size, with a thickness of 50 µm and a good surface finishing. The setup for preparing the Cu foils is illustrated in 1 M urea ; the prolsewhere . To inve2 and a vertical resolution of 0.1 nm. XRD patterns were acquired by means of a Philips PW1830 spectrometer, on a surface area of 2 cm2. Scanning electron microscopy (SEM) analysis was performed using a ZEISS EVO 50 EP instrument equipped with a field-emission gun, under the accelerating potential of 5 kV. Raman spectra were acquired by means of a Horiba LabRam HR800 spectrometer equipped with an argon ion laser operating at 514 nm (2.41 eV). The scattered radiation was collected through a 50 × objective keeping the beam power below 1 mW to avoid sample heating and graphene damage. The intensity correction system (ICS) was always kept active, and a diffraction grating with 600 lines/mm was used; acquisition time was always 45 s.Purity of the obtained samples was assessed by a Fischerscope X-ray Xan instrument, while surface morphology of Cu foils was investigated by using a NT-DMT SOLVER PRO atomic force microscope, with a scanning area of 100 µmGraphene catalytic growth on Cu is generally considered as a surface deposition process, with a typical adsorption-aggregation mechanism . From thSurface morphology of the as-plated foil is characterized by several protrusions roughly globular in shape; somewhere, elongated terraces are also observed a.In the annealing treatment, surface and volume diffusion can produce atomic redistribution, causing the observed reduction in surface roughness. Actually, the annealed surface looks highly smooth, with rare protrusions probably deriving from original large globules flattened only partially during the annealing treatment b.The electropolished surface appears more heterogeneous and consists of terraces and depressions with different sizes delimited by shallow steps c. Electr−1; it is the only first-order Raman process in graphene and is associated to the degenerate E2g phonon at the Γ point of the Brillouin zone. At 1350 cm−1 the D band is usually found, where “D” stands for “defect”: in fact, D Raman peak is originated by a second order process that involves a phonon at the Brillouin zone corner (A1g) and a defected state, treated as a 0 frequency quasi-particle to preserve the formalism [−1, the 2D band is observed; the 2D peak is, indeed, a second-order process where two phonons with opposite wave vectors were involved; hence, the momentum conservation is satisfied without the need of a defect state [G/I2D) is accepted to be lower than 1, and 2D Raman line shape has a FWHM of 24 cm−1 and is fitted by a single component Lorentzian function [−1), which is usually fitted by a single Lorentzian function, as in the case of single layer graphene (SLG). This is due to the decoupling of the layers comprising the turbostratic FLG, each of which retains a high percentage of the SLG transport properties [2D/IG intensity ratio.Raman spectrum of graphene layers features several bands, three of which are of uttermost importance when it comes to describing the main properties of the system of interest. The G band is located, in pristine samples, at 1582 cmormalism ,23,24. Fct state . The 2D function . The speoperties . In the operties : positioRaman spectra on untreated Cu foils showed a luminescence band , typical2D/IG = 0.6) and can be fitted with a single Lorentzian curve, its large FWHM linewidth (58 cm−1) suggests that the film consists of FLG, for the reasons explained above.Intensity of the 2D band progressively increased with increasing deposition time, and the D band was always present, probably related to the small dimension of the graphene domains. The graphene sample with 8 min of deposition time displayed a Raman spectrum d with loSEM observations  revealedSome isolated flakes exhibit hexagonal shape, which is favored by the etching action of hydrogen (97 vol% in the CVD atmosphere) at the edge sites . The ave2/CH4 atmosphere, continuous monolayer graphene was found  compared to pristine and annealed foils, since shorter treatments  led to Raman spectra similar to that in −1, slightly shifted towards higher wavenumbers with respect to pristine graphene. In addition, the position of the 2D peak shifted likewise. On the other side, the D-peak position seems to be consistent in all the reported spectra, remaining within the instrument spectral resolution. Full width at half maximum for the 2D peak is typical of multilayer materials as regards the as-plated and annealed-polished catalysts (>50 cm−1), while it was almost half of that value for the SLG grown on the annealed Cu substrate, in good agreement with the literature. Intensity ratios are always lower than 1 in the case of ID/IG, symptomatic of a good degree of order but small grain size, while the I2D/IG ratio varied depending on the number of layers present (see Results).All the Raman spectra presented show the main peaks typical of graphene. As can be seen from 2D/IG ratio. The growth route is characterized by the formation of several islands with incomplete substrate coverage  crystalline orientation and its surface became highly smooth, thus, promoting monolayer graphene growth. In fact, the 2D band FWHM is close to the value measured on Si/SiOgraphite . Moreoveonolayer . The ID/The existence of monolayer graphene on annealed Cu could be related to its strong crystalline orientation and the high growth rate of graphene on Cu (111) planes ,36. Howe2-CH4 gas mixtures [4 partial pressure of 1 Pa, single layer graphene with rare multilayer domains was produced; the number of these domains increased at higher CH4 concentrations. If results coming from low-pressure CVD experiments [4 partial pressures ranging from 1 Pa to 63 Pa. Hence, the high CH4 partial pressure (3 kPa) used in this work was expected to produce massive multilayer graphene [The formation of FLG regions on the annealed substrate b may be mixtures . Under aeriments ,9,14,33 graphene . On the 2-CH4 atmospheres [It can be noted that a consequence of using high partial pressures of methane is the reduction in the process time, which decreases from 30–60 min for pressures lower than 63 Pa ,14,33 doospheres .Electropolishing altered the smoothness of the original annealed surface, creating a number of defects that were regarded as preferential sites for nucleation and growth of multilayer graphene. As a result, the growth mode on the annealed-polished surface was similar to that observed on the as-plated Cu substrate, although the reaction time needed to reach a good surface coverage was longer (20 min compared to 6 min), probably because of the lower concentration of surface defects a,c.2D/IG ratio; FWHM was quite large; in conclusion, data support the hypothesis of having 4 to 5 graphene layers on top of annealed-polished electroformed Cu substrate.On the other hand, longer deposition time produced the most ordered of all three films, with good surface coverage and larger domains. The 2D band was less shifted but also less intense, as can be seen from the IAs reported at the beginning of the Discussion section, all the G-band positions in In this paper, we reported the study of graphene/FLG nucleation and growth on fully self-developed Cu substrates made by electroforming process. A complete morphological characterization of the substrates was presented, along with related graphene film Raman analysis. Moreover, electroformed Cu foils were treated to change surface conditions, and modifications in deposited film quality were highlighted. In particular, a conventional annealing treatment produced a highly smooth surface, where single layer graphene was grown. According to the existing literature, massive multilayer graphene would be expected to form in place of single layer graphene, under the investigated growth conditions. Conversely, Cu substrates with uneven surface generated graphene films consisted of few-layer/multilayer islands. Future developments will focus on testing the proposed AP-CVD process on different electroformed substrates, to further explore the electroplating potentialities in graphene synthesis. In addition, the CVD process itself can be improved to yield a desired number of graphene monolayers, mainly by optimizing the methane content in the growth atmosphere.


The strategy affords easy access to a broad scope of 2-substituted and 1,2-disubstituted pyrenes. The C1-substituent can be easily transformed into carboxylic acid, iodide, alkynyl, aryl or alkyl functionalities. This approach gives access to arylated pyrene ammonium salts, which outperformed their non-arylated parent compound during aqueous Liquid Phase Exfoliation (LPE) of graphite and compare favourably to state-of-the-art sodium pyrene-1-sulfonate PS1. This allowed the production of concentrated and stable suspensions of graphene flakes in water.A new and diverse family of pyrene derivatives was synthesised  via C–H arylation of pyrene-1-carboxylic acid.Novel pyrene surfactants for aqueous graphene exfoliation were prepared  On this regard, LPE offers a straightforward approach to obtain graphene dispersions directly from graphite.2 LPE is based on exposing directly the material to a solvent with a surface tension that favours an increase in the total area of graphite crystallites.2 Typically solvents used are NMP and DMF. However, LPE in water is also possible by using an exfoliating agent.3 This also allows to non-covalently functionalise the graphene flakes in situ, by tuning their surface chemistry depending on the exfoliating agent used.2During the last few years graphene has emerged as a revolutionary material attracting much attention due to its unique properties and multiple applications.3 In this context, pyrene derivatives have arisen as a privileged scaffold due to its strong affinity for graphene.4 For instance, several types of pyrene derivatives  would offer a flexible platform to that end. The palladium-catalysed C–H ortho-arylation of aryl carboxylic acids offers an oxygen and moisture-compatible way to derivatise these common feedstocks while choosing the fate of the acid moiety, which can be preserved or removed during the arylation reaction by judicious choice of reagents and conditions.15–17 Thus, the use of a silver-salt additive can lead to decarboxylated products,16 whilst replacing it with a tetramethylammonium salt, the ortho-arylated carboxylic acids are obtained.17Although reports on pyrene-functionalised graphene flakes are very abundant, examples of cationic pyrene-derived surfactants used to obtain suspensions of positively charged graphene flakes are scarce and require further investigation.ivatives .11 Notab1) and 3,5-iodoxylene (2a). After screening of the reaction parameters,18 both the 2-arylated pyrenecarboxylic acid 3a and the corresponding decarboxylated 2-arylpyrene 5a could be obtained in good yields (3a and 5a). Arylated carboxylic acids 3 were obtained in very good yields regardless of the substituents in iodoarenes 2. Both electron-withdrawing and electron-donating functionalities were broadly tolerated; haloarenes were also obtained in good yields (entries 3g–j) and the use of trifluoromethylated aryl iodides led to excellent yields (entries 3k and 3m). Ester-substituted products, which are amenable for further derivatisation, were also obtained in good yields (entries 3o and 3p). On the other hand, products of decarboxylative C–H arylation 5 were obtained in moderate to good yields in most cases and only in two particular cases the decarboxylative coupling proved inefficient (entries 5k and 5n). Nonetheless, the desired products could still be obtained in good yields by decarboxylation of the corresponding carboxylic acid.19 Importantly, this decarboxylative coupling can be a useful one-step alternative to access 2-arylpyrenes. Finally, using a modified protocol of our recent iododecarboxylation of carboxylic acids,20 arylated pyrene acids 3 could be further derivatised into the corresponding iodoarenes 4,18 which grant access to a whole new family of unexplored pyrene derivatives (vide infra). In this case yields ranged from moderate to good, although as expected, the best yields were obtained among the most electron-rich substrates (4a–e). To our delight, some substrates bearing electron withdrawing groups and halogen substituents also gave very satisfactory results . In general, products 3–5 were obtained in good yields and reactions could be scaled up with ease .The feasibility of the proposed approach was first assessed using pyrene-1-carboxylic acid .18PS1 is one of the most commonly used negatively charged organic surfactants for aqueous graphene exfoliation and has found application in different graphene-based technologies.2Pyrene cations logue 11 .18 CompoPS1, see  under th10a–d and non-arylated pyrene cation 11. As expected, the more concentrated suspensions, prepared with electron-poor compounds 10b and 10d, were darker than the rest.Aqueous suspensions of exfoliated graphene were diluted (×10) for visual inspection. The pictures in 10 is that steric clashes result in the benzylic ammonium group to be oriented perpendicular to the plane of the pyrene ring, locking the molecule in a favourable conformation  for analysis of lateral size. an 30 mV , confirmIn conclusion, we have developed a versatile strategy for the synthesis of 2-arylated pyrenes, including 1,2-disubstituted derivatives with several functionalities easily introduced in position 1, such as carboxylic acid, iodide, alkynyl, aryl and alkyl . This approach also offers a one-step alternative to the synthesis of 2-aryl pyrenes and opens new possibilities for the design and synthesis of unprecedented functional pyrene-based materials, previously unexplored due to its synthetic inaccessibility. Importantly, the cationic arylated pyrene ammonium salts accessible by this method can be used to obtain stable aqueous graphene suspensions by liquid phase exfoliation of graphite, composed mostly of single- and few-layer graphene flakes, and of concentrations comparable to those achieved with state-of-the-art anionic pyrene exfoliators.The authors declare no conflict of interest.SC-011-C9SC05101E-s001