Structural and process controls of AIEgens for NIR-II theranostics

Aggregation-induced emission (AIE) is a cutting-edge fluorescence technology, giving highly-efficient solid-state photoluminescence. Particularly, AIE luminogens (AIEgens) with emission in the range of second near-infrared window (NIR-II, 1000–1700 nm) have displayed salient advantages for biomedical imaging and therapy. However, the molecular design strategy and underlying mechanism for regulating the balance between fluorescence (radiative pathway) and photothermal effect (non-radiative pathway) in these narrow bandgap materials remain obscure. In this review, we outline the latest achievements in the molecular guidelines and photophysical process control for developing highly efficient NIR-II emitters or photothermal agents with aggregation-induced emission (AIE) attributes. We provide insights to optimize fluorescence efficiency by regulating multi-hierarchical structures from single molecules (flexibilization) to molecular aggregates (rigidification). We also discuss the crucial role of intramolecular motions in molecular aggregates for balancing the functions of fluorescence imaging and photothermal therapy. The superiority of the NIR-II region is demonstrated by fluorescence/photoacoustic imaging of blood vessels and the brain as well as photothermal ablation of the tumor. Finally, a summary of the challenges and perspectives of NIR-II AIEgens for in vivo theranostics is given.


Introduction
Light is one of the most fundamental branches of science. It is essential to decipher the interaction between light and matter for promoting the advances in technologies associated with the utilization of light, such as ber-optic communication, optical storage, optical devices, etc. Recent years have witnessed the establishment of strong ties between light and life by being able to visualize and detect many previously unknown life processes. Understanding photophysical behaviors is crucial for the design and interpretation of light-based applications. According to the classical photophysical diagram (Fig. 1a), matter absorbs light energy into excited energy states before immediately falling back to lower energy states mainly through radiative (R) and non-radiative (NR) decays. 1 R is mostly in the form of uorescence, which can be utilized for bioimaging and biosensing. Heat generation is the main outcome of NR, which has numerous applications in photothermal therapy (PTT), photoacoustic (PA) imaging, laser resurfacing, desalination of seawater, etc. 2,3 Unfortunately, these two critical processes compete against one another: the dominant R will suppress NR and vice versa. So how can we regulate these two processes to attain desirable properties? Among all factors affecting the process of R and NR, the structure is undoubtedly one of the most signicant aspects (Fig. 1a, right). Generally, matter with rigid structures shows a dominated R, resulting in a strong uorescence (Fig. 1b, le). In contrast, exible matter favors molecular motion for an overwhelming NR, giving an intense heat generation (Fig. 1b,  right). Accordingly, simultaneous introduction of rigidication and exibilization into structures endow matters with both uorescence and heat (Fig. 1b, middle). But how should we regulate the structure? Conventionally, structural regulation at the molecular level can precisely control molecular properties. For example, some molecules with rigid fused-ring structures show strong uorescence as isolated species. However, when they aggregate in life mediums such as water, the resultant higher-order hierarchical structure exhibits uorescence quenching, owing to the excimer formation. This is the socalled aggregation-caused quenching (ACQ) phenomenon, which severely hinders in vivo bioimaging. 4,5 On the other hand, some exible molecules, such as tetraphenylethylene (TPE), are almost non-emissive at a molecular level, owing to the dominated NR by active intramolecular motions (Fig. 1c). [6][7][8] Upon molecular aggregation in water, intermolecular hydrophobic interactions restrict the intramolecular motions, giving a signicantly enhanced uorescence. Therefore, the rigidication of molecular structures at a higher-order hierarchical level gives a much stronger uorescence intensity than that of the molecular level. Altogether, by regulation of hierarchical structures (exibilization at a molecular level and rigidication at a morphological level), the same substance may display predesigned photophysical properties to meet various applications.
Aggregation-induced emission (AIE) is an inventive uorescent technique and has the potential to address this issue. 4,33-51 AIE illustrates a unique photophysical phenomenon where molecules are non-emissive in the discrete state because of active intramolecular motions, but emit energetically when aggregated due to the restriction of intramolecular motions (RIM). To drive emission to the NIR-II window, AIE luminogens (AIEgens) are designed as conjugated donor-acceptor (D-A) structures, where they are being twisted with each other (dihedral angle of q, Fig. 2c). Under a polar environment, the intramolecular rotations of D/A units bring the molecules from a locally excited (LE) state to a twisted intramolecular charge transfer (TICT) state (Fig. 2d). [52][53][54] The elevation of the highest occupied molecular orbital level narrows the energy bandgap, giving a red-shied emission. On the other hand, owing to the susceptibility of the TICT state to various nonradiative quenching process, the emission intensity is signicantly weakened. Thus, the TICT process gives a red-shied emission and an excellent photothermal property. According to the RIM mechanism of AIE, the partial restriction of intramolecular motion by molecular aggregation can enhance the uorescence intensity (on a morphological level) while maintaining the TICT property (molecular level). Moreover, introducing molecular motors to molecules further suppresses the intermolecular interactions to trigger the AIE effect. Therefore, NIR-II AIEgens with long-wavelength uorescence and intense brightness can be seized simultaneously.
Based on the above-mentioned structural design (twisted backbone + molecular motor) and process control (AIE + TICT), many outstanding NIR-II AIEgens have been reported. In this feature review, to understand the structural design, process control, as well as involved mechanisms, we have selected some representative examples to analyze and guide further developments of NIR-II AIEgens. The R and NR channels can be opened or blocked by tuning the structure at multi-hierarchy levels. AIE technology endows molecules with high uorescence quantum yield (QY), while the TICT state gives molecules with both red-shied emission and enhanced photothermal property owing to the predominant NR. By controlling structure and process, systems with predominated uorescence intensity, or predominated photothermal property, or a balance of uorescence intensity and photothermal property can be obtained. We hope this review will offer valuable guidance on how to design desirable NIR-II theranostic agents by regulating structure and process.

Design of NIR-II AIEgens
To date, the majority of the existing NIR-II organic chromophores have low quantum yields between 0.3% and 2%. [55][56][57] It will be favorable to design uorophores with inherently longer emission wavelength, as well as much higher QY (>2%) to achieve effective NIR-II uorescence imaging. Given the diversity of designing D-A conjugated molecules, this may be realized through molecular engineering of electron donors, acceptors, spacers, etc. 58,59 On the other hand, AIE features outstanding solid-state photoluminescence. 44,60 These AIEgens naturally own twisted architectures dressed up with several molecular motors, which are valuable for suppressing the robust intermolecular interactions. In general, visible lightemissive AIEgens are relatively easy to achieve through the incorporation of twisted molecular motors. However, this method is not t for NIR-absorbing uorophores, as they suffer from much stronger intermolecular p-p interactions, which results in uorescence quenching when dispersed in a physiological environment. To address this issue, the strategy of "backbone distortion + twisted motors" is generally accepted for designing highly bright NIR-II AIEgens because of the efficient restriction of intermolecular interactions. 61 Accordingly, NIR-II AIEgens are characterized by a motordonor-acceptor-donor-motor (M-D-A-D-M) structure. Strong molecular acceptors, such as 6,7-diphenyl- [1,2,5]thiadiazolo [3,4-g]quinoxaline (DPTQ) and benzobisthiadiazole (BBTD), are generally utilized to drive the emission wavelength to the NIR-II region. 20 The energy gap can be decreased by anchoring molecular donors such as phenyl or substituted-thiophene. The Fig. 3 Representative chemical structure of NIR-II AIEgens annotated with absorption maxima, molar absorption coefficient (specific solvent), emission maxima and NIR-II QY in water (1000-1500 nm, based on QY IR26 ¼ 0.5%). Note: QY in 1 is calculated based on IR820 as reference (QY ¼ 4.2% in ethanol), absorption in 10 is measured in THF.
critical element in molecular design lies in the distortion of the central D-A-D backbone, which is crucial to restrict the intermolecular interactions. Meanwhile, the peripheral twisted motors such as tetraphenylethylene (TPE) and triphenylamine (TPA) are indispensable. On the one hand, molecular motors are benecial for further restricting of intermolecular interactions. On the other hand, they provide intramolecular mobility to access the TICT state. To date, based on the above molecular design philosophy, Tang, Qian and Ding, 61-68 Zheng and Liu, 69 Lu and Xiao, 70 Hong, 71 Chen and Feng, 72 Liu, 73 Xiao, 74 Wu, 75 Wang and Jiang, 76 and Wu 77 et al. developed several types of highly bright NIR-II AIEgens with emission peak >1000 nm, with representative chemical structures shown in Fig. 3. In this review, we will not go into detail for all the NIR-II AIEgens mentioned but select the most representative ones to illustrate the molecular design philosophy and process control clearly.

NIR-II AIEgen with high QY (backbone distortion + molecular motor)
To deeply understand the molecular design strategy, Tang and Ding systematically studied the effects of backbone distortion and molecular motor on the solid-state uorescence efficiency of the dyes. 61 As shown in Fig. 4a, because of the existence of steric hindrance in ortho-substituted alkylthiophene and BBTD core, AIEgen 9 displayed a signicantly distorted backbone (dihedral angle of 48 ). Another critical feature of AIEgen 9 was the presence of twisted molecular motor TPA.
Photoluminescence (PL) spectra were performed to detect emission uctuation upon molecular aggregation using THF/ water mixtures (Fig. 4b). Upon increasing water volume fractions (f w ) from 0% to 50 vol%, the uorescence intensity of AIEgen 9 decreased progressively, due to the forming of a dark TICT state with an increase in polarity. Further increase of f w to 90 vol%, the PL intensity showed a rapid enhancement (AIE effect), due to the suppression of intramolecular rotations decisive to the TICT state through molecular aggregations. Therefore, AIEgen 9 possessed a TICT + strong AIE effect with a high QY of 8.4% when in a nanoparticle state. The a AIE (PL intensity ratio of f w ¼ 90 vol% to f w ¼ 0) is also a clear indicator of solid-uorescence efficiency, with measurements of up to 6.2, which was higher than the other systems (a AIE z 2). 71 To demonstrate the molecular motor effect, AIEgen 13 with phenyl motor while maintaining the highly twisted backbone was used as a control. A typical TICT + AIE effect was observed, however, a AIE sharply decreased to 0.54, suggesting a signicant impact of the molecular motor on the uorescence efficiency. On the other hand, the backbone twisting effect was studied by shiing the alkyl chains from ortho to meta position. In contrast to molecule 9, the resultant molecule 14 adopted a planar backbone (dihedral angle of 1 ), owing to the steric distance of metapositioned alkyl chains to the BBTD core. The PL intensity of molecule 14 decreased with f w because of the TICT effect upon an increase of polarity. Notably, the emission was almost wholly quenched in the aggregate state (f w ¼ 90 vol%), which resulted from the strong intermolecular interactions in the planar D-A-D core. Moreover, removing the twisted TPA motor in molecule 15 further intensied the ACQ effect. Overall, backbone distortion and motor twisting have an equivalent effect on the solid-state uorescence efficiency of NIR-II AIEgens.
Beneting from high QY of AIEgen 9 nanoparticles (AIE9 NPs) in the NIR-II region, in vitro uorescence property was examined. As shown in Fig. 4c, a linear relationship between uorescence intensity with AIE9 NPs concentration was observed, demonstrating the advantages of AIE: the more it gathers, the brighter it glows. To display the in vivo uorescence imaging quality of AIE9 NPs, blood vessel imaging of mouse hindlimb was carried out via intravenous tail injection of AIE9 NPs. Indocyanine green (ICG), a NIR-I dye with clinical approval, is served as a comparison. The imaging quality from AIE9 NPs outperformed that of ICG (Fig. 4d), indicating that highly bright NIR-II AIEgens can provide a higher degree of clarity in imaging.
Inammation is a sign of many major diseases such as cancer, diabetes, cardiopathy etc. Noninvasive detection of inammation is crucial for the early diagnosis of pathogenic processes. In particular, the diagnosis of brain inammation is viewed as the Holy Grail for bioimaging, as it is well-protected by the blood-brain-barrier (BBB). Tang and Ding ingeniously utilized immune cells, neutrophils (NEs), as "living" carriers to deliver AIE9 NPs to penetrate the BBB to arrive at the brain inammation site. 61 As displayed in Fig. 4e, the NIR-II uorescence signal at the inamed area injected with neutrophilcarrying AIE9 NPs (AIE9@NE) was much stronger than that of ICG@NE. The signal-to-background ratio (SBR) at the inammation site reached up to 30.6 for AIE9@NE treated group, while it was only 5.6 for the mice administrated with ICG@NE. Notably, 30.6 is among the one of the highest SBR reported so far. Despite a relatively high QY, it is worth noting that nonradiative decay is still the dominant relaxation pathway in NIR-II materials, as AIE9 NPs displayed excellent photothermal conversion properties (Fig. 4f). Therefore, NIR-II AIEgens have the potential for uorescence-guided photothermal therapy.

NIR-II AIEgen with long-wavelength emission (AIE + TICT process)
Due to the reduced photon scattering and almost zero auto-uorescence, in vivo uorescence imaging in the NIR-IIb region (1500-1700 nm) can provide higher resolution and deeper tissue penetration in comparison with conventional NIR-II window. [78][79][80] While inorganic NIR-IIb uorophores have been developed, the study on organic counterparts is still in its infant, due to the lack of long-wavelength materials. To address this dilemma, Tang and Qian proposed a molecular design guideline for the manipulation of TICT and AIE at molecular and agminated levels, respectively, to explore purely organic NIR-IIb uorophores. 66 As shown in Fig. 5a, in line with the molecular design strategy of NIR-II AIEgens, 7 and 16 were synthesized using BBTD as a strong acceptor, with orthosubstituted alkylthiophene as the donor, and TPA as the motor. Notably, the keystone of the molecular guidelines is engaged in the selection of secondary carbon-branched alkyl chains, as they supplied a tunable steric barrier not only for restricting intermolecular interactions but also for the activation of intramolecular motions. On a molecular level, the exible intramolecular rotations of D-A units in AIEgen favored the formation of dark TICT state to yield signicantly red-shied emission wavelength. On the other hand, the NPs formation process (nanoprecipitation with so amphiphilic polymer DSPE-PEG) partially restricted the intramolecular motions of AIEgens, giving signicantly enhanced uorescence signal. Thereof, dyes with both long-wavelength emission and adequate efficiency can be obtained concurrently by linking ostensibly contradicting individuals. The optimized AIEgen 7 with 2-ethylhexyl unit presented an emission tail extending to 1600 nm with a high NIR-II QY of 11.5% in the nanoparticle state.
To glean the advantage of NIR-IIb uorescence imaging of AIE7 NPs, long-pass (LP) lters with 1100 and 1500 nm were examined. As shown in Fig. 5c, distinct vascular anatomy with a negligible background is observed in the NIR-IIb window (1500 nm LP), while ambiguous vessels are found in the conventional NIR-II region (1100 nm LP lter). In particular, the vessels can hardly be identied in the conventional NIR-II region for the liver region, owing to the strong auto-uorescence of the liver. Through-skull high-magnication microscopic imaging of cerebral vasculature was also carried out. Small vessels with a width of 10 mm could be identied (Fig. 5d). To further demonstrate the merits of penetration depth in the NIR-IIb window, uorescence imaging of deeplylocated intestinal tract was performed. The ileum structure could be distinctly visualized aer the gavage of AIE7 NPs at 0.5 h. In contrast to the blurry images observed under 1100 nm LP lter, a clear resolution of tissue features was distinguished in the NIR-IIb region (Fig. 5e). Moreover, NIR-II AIEgen NPs displayed desirable photothermal properties, which were enhanced with elongation of alkyl chains due to the activation of intramolecular motions (Fig. 5f). Therefore, based on structural modulations at molecular (TICT) and agminated levels (AIE), NIR-IIb-based NPs with red-shied emission wavelength and appropriate QY were attained simultaneously.
Very recently, based on the mechanism of AIE + TICT, Xiao et al. developed a series of highly bright NIR-II AIEgen 10 (QY ¼ 11.9%) and 11 (QY ¼ 11.7%) with a uorescence tail extending to the NIR-IIb region. The resultant AIE NPs could be utilized as excellent NIR-IIb imaging agents for superior spatial-temporal resolution of whole-body, cerebral, tumor vasculature, lymphatic drainage systems, etc. 74,81 On the other hand, NIR-IIa (1300-1400 nm) window is also appealing owing to the reduced light scattering and decreased water absorption. Wu et al. reported an AIE-active NIR-IIa conjugated polymer with a high QY of 1.7% in aqueous solution, realizing clear through-skull visualization of the cerebral vasculature. 77 NIR-II AIEgen with tunable photothermal therapy (AIE + TICT process) In a D-A-based NIR-II AIEgen, the equilibrium between uorescence (R) and photothermal effect (NR) is hard to control. To address this issue, Lu and Xiao 70 reported a strategy of tuning AIE and TICT properties to balance the uorescence and photothermal effect through the coordination of human serum albumin (HSA) with NIR-II AIEgen 6 (Fig. 6a). In a polar solvent, the active intramolecular rotation drove AIEgen 6 into the dark TICT state, resulting in enhanced NR and photothermal conversion effect (Fig. 6b). On the other hand, when aggregated in poor solvents, restriction of intramolecular motion and suppression of TICT state increased uorescence (AIE effect). Altogether, the TICT state weakened the uorescence intensity but elevated the photothermal property. At the same time, AIE increased uorescence intensity at the cost of its photothermal property, which was balanced by the nature of intramolecular rotations. Enhancement of HSA ratio in the AIE6@HSA NPs activated the intramolecular motions of AIE6 through the coordination with HSA, displaying an enhanced photothermal conversion property at the cost of uorescence (Fig. 6c). Thus, the prepared AIE6@HSA NPs exhibited both high uorescence efficiency and high photothermal conversion property, providing high specic imaging of not only primary orthotopic mouse colon tumors but also metastatic lesions with a diameter of 0.5 mm Â 0.3 mm (Fig. 6d). Directed by NIR-II uorescence signal, AIE6@HSA NPs allowed precise PTT of both primary and metastatic lesions, attaining a complete cure of tumors in mice. Considering a large NIR-I (700-900 nm) absorptivity, Zheng and Qian et al. also utilized NIR-II AIEgens 4 for in vivo photothermal applications, effectively achieving uorescence-guided photothermal therapy or uorescence/photoacoustic dual model imaging. 67,69 Activation of intramolecular motion for enhanced photothermal therapy Increasing the photothermal conversion property of materials is crucial for their potential biological applications, as it reduces the incident light intensity, allowing for an efficient and safe therapy. Considering the dominated nonradiative decay in the TICT state, Tang and co-workers proposed a strategy for manipulating the TICT state to boost photothermal conversion property via activating molecular motion in aggregates. 82 The molecular design guideline for an efficient photothermal agent is displayed in Fig. 7a. The central planar thiophene-BBTDthiophene core (dihedral angle of 1 ) was expected to quench the emission via strong intermolecular interactions when aggregated. The twisted TPA unit acted as molecular motors to assure intramolecular rotations and as a molecular donor to red-shi the absorption. The critical element of molecular  design lies in the adoption of long-branched alkyl chains to provide the necessary room for molecular motions, which are the prerequisites for the formation of the TICT state. 83,84 As expected, AIEgen 17 with 2-decylmyristyl unit displayed quenched uorescence upon molecular aggregation (Fig. 7b). AIEgen 17based NPs showed a high photothermal conversion efficiency (PCE) of 31.2%, better than the hexyl-decorated counterpart (22.6%), demonstrating a positive effect of long-branched alkyl chains for improving photothermal properties. Besides, the photothermal temperature was positively related to the concentration of NPs (Fig. 7c), which provided a personalized temperature selection for specic applications. Overall, activation of the intramolecular motions of D-A conjugated molecule in aggregates by twisted molecular motors and long-branched alkyl chains facilitated the formation of the dark TICT state to boost nonradiative pathway for heat generation.
To further illustrate the concept of intramolecular motioninduced photothermy (IMIPT), a molecule, 18, possessing a TPE molecular motor and long-branched alkyl chains in the pconjugated planar backbone, was designed. 85 Long-branched alkyl chains were aimed to achieve spatial isolation of molecules for enhanced molecular motions. The combination of pexpanded and strong electron-withdrawing acceptors with TPE (donor and twisted motor) contributed to not only a narrow energy gap benecial for light-harvest ability, but also to trigger the TICT state giving a uorescence quenching effect. Molecule 18 displayed excellent photothermal conversion property with temperatures rising to 81 C (Fig. 8b). The PCE could even reach up to 54.9%, which is much higher than previous reports. 86 The application of 18 NPs for in vivo PA imaging was then investigated. Aer intravenous injection of 18 NPs, a strong PA signal could be identied at the tumor site at 4 h post-injection, which is $2.7-fold higher than that of 0 h. As a control, the PA signal of the muscle tissues remained almost unchanged at the tested time points and was much lower than in the tumor.
Recently, based on the understanding of the strategies for activation of intramolecular motions within NPs, many excellent PTT/PA systems have been developed. Tang and co-workers reported an AIEgen full of molecular motors, allowing efficient intramolecular motions for PA imaging-guided phototherapy. 87 Fan and co-workers boosted nonradiative decay channel through manipulating intermolecular charge transfer, attaining an exceptionally high PCE of 61%. 88 Lee and co-workers reported a kind of p-conjugated oligomer F8-IC linked with A-D-A structure, whose intramolecular exible rotations led to an enhanced non-radiative decay. The resulting F8-IC NPs displayed a high PCE of 82% for high-performance PA imagingguided cancer therapy. 89 Liu and co-workers integrated D-A structure and molecular motors into one molecule, TA1, giving a high PCE of 84.8%. The obtained TA1 NPs could signicantly suppress primary breast tumor growth and metastasis. 90 Peng and co-workers introduced a "barrier-free" motor to the mesoposition of the BODIPY scaffold, allowing efficient dissipation of absorbed photons as heat. The obtained NPs showed a high PCE of 88.3% leading to complete removal of tumors in mice. 91 Li and co-workers reported that the activation of intramolecular rotation around C]N double bond enabled a high PCE of $90%. 92 Altogether, the activation of intramolecular motion within NPs provides a platform for developing efficient photothermal agents.
Despite high PCEs of above photothermal agents, they mainly rely on NIR-I laser. Owing to the deeper tissue penetration and higher maximum permissible exposure to the laser, photothermal agents in superior NIR-II region have received increasing attention. [93][94][95] Nevertheless, pure organic materials with strong absorption in the NIR-II window have been far less reported, calling for high-performance materials with both long-wavelength absorption and high PCE.

Conclusions
In this review, through regulation of hierarchical structures at a molecular level (exibilization) and aggregate level (rigidication), respectively, organic NIR-II uorophores with AIE characteristics for efficient uorescence/photoacoustic imaging and PTT have been summarized. The molecular design strategy of AIEgens, "backbone distortion + motor twisting", has been demonstrated to be indispensable for improving the QY of NIR-II dyes, due to the suppression of strong intermolecular interactions at aggregates. The exibilization in molecular structure brings the uorophores to a TICT state, giving a signicantly red-shied emission with strong NR. By intelligent control of AIE and TICT processes, molecular aggregates with high QY (AIE) and long-wavelength emission (TICT) can be achieved, which nds salient advantages for in vivo NIR-IIb imaging. On the other hand, systems can also display high uorescence efficiency (AIE) with a tunable photothermal effect (TICT), which is suitable for uorescence-guided PTT or uorescence/ photoacoustic dual-mode imaging. To further enhance the photothermal properties, a molecular guideline of "backbone planarization + motor/spacer" is proposed. The resultant uorophores display an ACQ process owing to the intense intermolecular interactions from the planar conformation. However, with the presence of molecular motors and long-branched alkyl chain spacers, a signicantly enhanced photothermal effect is detected owing to the activation of intramolecular motions. Overall, from molecule design to aggregate control, AIE research emphasizes the signicance of studying materials properties at a higher hierarchical level.
While NIR-II AIEgens hold substantial promise for biological theranostics, several concerns need to be addressed for their further development. First, the overall brightness of NIR-II AIEgens can be further improved by increasing the molecular absorptivity while maintaining the high QY, as the backbone twisting in AIEgens may destroy the absorption. Second, pure organic uorophores with peak emission exceeding the golden imaging region (>1500 nm) are seldom reported. Third, investigating ways to design NIR-II AIEgens with targeting ability or in response to external stimuli is of crucial importance. For example, Wu and Zeng reported a nitroreductase-responsive NIR-II AIEgen, realizing the detection of breast cancer metastasis. 96 Last but not least, since many organic NIR-II NPs tend to accumulate in mononuclear phagocyte systems such as in livers and spleens, new NIR-II uorophores should possess the overall dimension within the renal ltration limit ($5 nm) for rapid urine excretion.
In summary, the development of single AIE NPs with balanced uorescence and photothermal therapy functions through structure adjustment at different hierarchical levels is of great interest for the advancement of next-generation NIR-II uorophores.

Conflicts of interest
There are no conicts to declare.