Quantitative lipoprotein analytics using nuclear magnetic resonance (NMR) spectroscopy is currently commonplace in large-scale studies. One methodology has become widespread and is currently being utilized also in large biobanks. It allows the comprehensive characterization of 14 lipoprotein subclasses, clinical lipids, apolipoprotein A-I and B. The details of these data are conceptualized here in relation to lipoprotein metabolism with particular attention on the fundamental characteristics of subclass particle numbers, lipid concentrations and compositional measures.The NMR methodology was applied to fasting serum samples from Northern Finland Birth Cohorts 1966 and 1986 with 5651 and 5605 participants, respectively. All results were highly consistent between the cohorts. Circulating lipid concentrations in a particular lipoprotein subclass arise predominantly as the result of the circulating number of those subclass particles. The spherical lipoprotein particle shape, with a radially oriented surface monolayer, imposes size-dependent biophysical constraints for the lipid composition of individual subclass particles and inherently restricts the accommodation of metabolic changes via compositional modifications. The new finding that the relationship between lipoprotein subclass particle concentrations and the particle size is log-linear reveals that circulating lipoprotein particles are also under rather strict metabolic constraints for both their absolute and relative concentrations.The fundamental structural and metabolic relationships between lipoprotein subclasses elucidated in this study empower detailed interpretation of lipoprotein metabolism. Understanding the intricate details of these extensive data is important for the precise interpretation of novel therapeutic opportunities and for fully utilizing the potential of forthcoming analyses of genetic and metabolic data in large biobanks. Key MessagesThe circulating particle number of a lipoprotein subclass is the defining measure for its lipid concentrations; the particle lipid composition plays only a minor role. The relationship between circulating lipoprotein subclass particle concentrations and the particle size is log-linear.biomolecular constraints for their lipid composition.The overall structure of lipoprotein subclass particles with a spherical shape and an oriented surface monolayer poses strong size-dependent metabolically constrained for both elemental absolute and relative concentration ranges.The circulating lipoprotein subclass particle concentrations in humans are The smallest high-density lipoprotein (HDL) particle concentrations are negatively associated with those of large HDL particles and generally the associations of the smallest HDL particles are similar to those of apolipoprotein B-containing particles.The apolipoprotein B-containing particles constitute <10% of all lipoprotein particles but carry around two-thirds of the circulating lipoprotein lipids. Low-density lipoprotein and intermediate-density lipoprotein particles amount to almost 90% of all apolipoprotein B-containing particles.The supplemental role of lipoprotein subclass data in cardiometabolic risk assessment is slight.In the current era of biobanks and big data, the combination of lipoprotein subclass data with drug-target Mendelian randomization analyses provides great scientific synergy, intricate details and potential cost savings in drug development.The role of cholesterol in the development of human atherosclerosis has been known for >100 years. After the original findings in relation to cholesterol, it took almost 50 years to link cholesterol to circulating low-density lipoprotein (LDL) particles and reveal the association of LDL cholesterol with coronary heart disease (CHD) in the 1950s. LDL cholesterol and a few other lipoprotein measures, particularly triglycerides and high-density lipoprotein (HDL) cholesterol, have been in the spotlight of CHD and related pharmacological research since then.,Broader research on the molecular characteristics of circulating lipoprotein particles began after the invention of using sequential preparative ultracentrifugation to physically isolate native lipoprotein particles according to their densities and then further characterizing them chemically.,Although ultracentrifugation became the workhorse in lipoprotein analytics, the tediousness of the process, particularly with a tight density scale, prevented large studies with a high resolution of lipoprotein fractions.,Almost all of the epidemiological and genetic applications of the lipoprotein data have focused on concentration measures only, even though the compositional data are automatically incorporated into the analytical results for each serum sample.,,,,The separation of lipoprotein subclasses by proton NMR spectroscopy is based on particle size. In more detail, the anisotropy of the magnetic susceptibility, due to the radially oriented surface monolayer of the spherical particles, inherently leads to a size-dependent frequency shift for the NMR resonances of lipoprotein lipids.IJE online) for clinical characteristics. The selection or the traits of the cohorts are not important in this study focusing on overall lipoprotein subclass particle characteristics and associations as the existing literature already affirms them to be highly consistent in and between various large-scale studies.,,,,,The Northern Finland Birth Cohort 1966 (NFBC66) with data for 5651 participants (at 46 years of age) were studied as the primary cohort and the NFBC86 with data for 5605 participants (at 16 years of age) as a replication cohort. More details on the NFBCs can be found in the circulating particle concentrations (the particle numbers are within the range of 0.1 nmol/L and 10 µmol/L), (ii) circulating lipid concentrations (within the range of 0.1 and 10 mmol/L) and (iii) particle lipid compositions as the percentage of a certain lipid class of total lipids (mol%) per subclass particle. Apolipoprotein B is the key protein component in VLDL, IDL and LDL particles, and they are often called apolipoprotein B-containing particles. HDL particles do not contain apolipoprotein B but are abundant in apolipoprotein A-I. All lipoprotein particles also contain other apolipoprotein molecules, but they cannot be quantified by NMR.P-value threshold of 0.002 (i.e. 0.05/24) was thus used to denote evidence in favour of an association.The lipoprotein subclass information can be partitioned into three key categories of variables: (i) IJE online, for replication in the NFBC86). The compositional data are also visualized in the upper part of IJE online). The VLDL subclasses are the most triglyceride-enriched with variation from 69% to 17% of total lipids from XXL-VLDL to XS-VLDL. The IDL and LDL particles are all rich in cholesteryl esters (∼50%) as well as free cholesterol (∼20%). All HDL particles are rich in phospholipids (∼50%) and very poor in triglycerides.IJE online). Roughly 50% of circulating cholesterol (both free and esterified) travels in IDL and LDL particles and some 20–30% in HDL particles. Around 50% of lipoprotein phospholipids in the circulation are in HDL particles. The size-dependent opposite compositional behaviour of triglycerides and cholesteryl esters in the apolipoprotein B-containing subclass particles is pronounced.Over 50% of circulating triglycerides are transported in VLDL particles, some 20% in IDL and LDL, and <20% in HDL particles . The HDL particles are by far the most abundant with concentrations in a micromolar range, the IDL and LDL particles being at the range of 0.1–0.3 micromolar and the largest VLDL particles at the range of 0.1–10 nanomolar. Thus, on average, there are some 60 000 S-HDL and 1500 IDL particles per one XXL-VLDL particle in the circulation; 92% of all lipoprotein particles are HDL particles (The circulating subclass lipid concentrations depend on both the lipid composition of a subclass particle and the concentration (i.e. the number) of those particles in the bloodstream. The distributions for subclass particle concentrations are shown in articles . TherefoIJE online, for the NFBC86) together with a summary comparison between apolipoprotein B-containing particles and HDL particles. In absolute concentration terms, apolipoprotein B-containing particles transport roughly double the amount of cholesterol than HDL particles. The circulating pool of phospholipids is very similar in HDL and apolipoprotein B-containing particles, and very few triglycerides are transported in HDL.An important distinction must be made between the concentration, i.e. number, of circulating particles (‘the particle view’) and the number of lipids that are transported within these particles (‘the cargo view’). This is due to large differences in the subclass particle transport volumes. As illustrated in IJE online, for the NFBC86 data).IJE online). On the other hand, associations between apolipoprotein A-I and B concentrations (g/L) as well as between HDL particle and apolipoprotein B-containing lipoprotein particle concentrations (mol/L) are very weak. This is explained by negative associations between apolipoprotein B and large and medium HDL particles and a counterbalancing effect due to a positive association between apolipoprotein B and the smallest HDL particles (IJE online).The standard lipid measures  behave as expected as illustrated in IJE online). The associations of apolipoprotein B with all VLDL, IDL and LDL subclass concentration measures are positive and strong. VLDL subclass concentration measures associate positively with serum triglycerides and negatively with HDL cholesterol. Associations between LDL cholesterol and IDL and LDL subclass concentration measures are positive and strong. The associations with HDL subclass measures are heterogeneous, although HDL cholesterol and apolipoprotein A-I correlate positively with all HDL subclass particle concentrations.The associations between the 14 lipoprotein subclass measures and the standard lipid measures are all logical (IJE online). The cholesteryl ester composition of all LDL subclass particles is strongly positively associated with circulating total and LDL cholesterol as well as apolipoprotein B concentrations. Circulating HDL cholesterol and apolipoprotein A-I concentrations are negatively associated with the triglyceride composition of all VLDL and IDL particles, positively associated with the cholesteryl ester composition of S-, M- and L-HDL particles, but the association is negative for XL-HDL particles.The triglyceride composition of all lipoprotein subclasses is positively associated with the circulating triglyceride concentration, except in the case of XXL-VLDL ((IJE online). For example, high circulating particle and triglyceride concentrations for apolipoprotein B-containing lipoprotein particles associate with triglyceride enrichment and diminished composition of cholesteryl esters in M- and L-HDL particles. Similar strong relationships exist between the concentrations as well as triglyceride and cholesteryl ester compositions of XS-, S-, M-, L- and XL-VLDL particles. The triglyceride composition of all HDL subclass particles is consistently positively associated with all apolipoprotein B-containing lipoprotein particle and triglyceride concentrations. The circulating concentrations of M-, L- and XL-HDL particles are negatively associated with their triglyceride compositions.An opposite compositional behaviour is typical between triglycerides and cholesteryl esters IJE online). Positive associations between adjacent subclasses are high and there is a general trend of weakening positive associations along the line of the metabolic direction from large VLDL to LDL particles. The association pattern appears different for XL- and L-HDL in comparison to M- and S-HDL subclasses. The subclass particle concentrations of large and medium HDL subclasses associate negatively with all VLDL subclasses but the associations for the smallest HDL subclass are in the opposite direction. Similarly, S-HDL particle concentrations are negatively associated with those of XL- and L-HDL. The circulating triglyceride concentration in every apolipoprotein B-containing subclass associates consistently positively with the corresponding measure in both M- and S-HDL.The associations between all the apolipoprotein B-containing lipoprotein subclass concentration measures form a coherent pattern and reflect the continuous metabolism from VLDL to LDL particles in the bloodstream ((IJE online) are generally a lot weaker and more heterogeneous than the corresponding associations between the concentration measures (IJE online). A global feature is that the triglyceride compositions of almost all lipoprotein subclass particles are strongly positively associated. These compositional triglyceride associations are strong in each major fraction of particles, i.e. within VLDL, IDL and LDL as well as HDL, but also mostly between these fractions with only a few triglyceride compositions in the largest VLDL subclasses showing either very weak or negative associations. Overall, triglyceride and cholesteryl ester compositions tend to associate negatively, particularly within the major fractions of particles.The associations between lipoprotein subclass lipid compositions IJE online) for the NFBC66 and in IJE online) for the NFBC86. The numerical data for these correlations are available in IJE online), respectively. IJE online) illustrate the comprehensive lipoprotein subclass data and the intertwined correlation structure in a metabolic network model.IJE online).A correlation heat map between all the lipoprotein subclass concentration, composition and standard lipid measures as well as apolipoprotein B and A-I are shown in Key results and the fundamental characteristics of lipoprotein subclass particles and metabolism are highlighted in a summary in p-value] in the NFBC66 for all the 168 lipoprotein subclass measures are given in IJE online) (composition measures) and IJE online) (concentration measures). Corresponding sex-specific data for a few clinically relevant lipoprotein measures are shown in IJE online). The current data replicate the well-known sex differences for clinical lipoprotein measures; females tend to have higher circulating HDL cholesterol and lower concentrations of apolipoprotein B-related measures.Sex-specific data [mean , ,,,It has been shown earlier that the circulating lipid concentrations as well as the relative lipid compositions for the lipoprotein subclasses with this methodology are very well concordant with those from density-based ultracentrifugation analyses,As far as we are aware, the log-linear relationship between circulating lipoprotein subclass particle concentrations and the particle size has not been indicated before. The scale of difference in the subclass particle numbers is huge, e.g. 60 000 S-HDL particles per one circulating XXL-VLDL particle . On aver,,The key metabolic role of lipoprotein subclass particle concentrations is not unexpected. Within the particles, the size and structural molecular constraints fundamentally define how many non-polar and polar lipid molecules can be accommodated in both the core and the surface, respectively. Although the core and surface regions are not rigid, the spherical shape with an oriented monolayer poses strong biophysical constraints within the small number of key lipid classes—triglycerides, phospholipids, free and esterified cholesterol—to be integrated into the particles.The log-linear relationship between lipoprotein subclass particle concentrations and the particle size also suggests the existence of fundamental metabolic constraints. This means not only that typical particle and lipid concentration ranges are elemental for humans, but also that only certain relative lipoprotein subclass particle concentrations are possible in human lipoprotein metabolism. We are not aware of previous discussion directly in relation to this, but we find it implicit in the lipoprotein literature and e.g. via the traditional clinical definitions of abnormalities in lipoprotein metabolism and their drug treatment.Altogether, it is likely that these metabolic constraints are just simple reflections of normal metabolic pathways on how the human body handles triglycerides, cholesteryl esters and other dietary lipids. A key corollary, however, is that when evaluating a risk related to a lipoprotein subclass, it would only be relevant to estimate it within the physiologically realistic concentration area.The associations between the standard lipid measures and the subclass concentration measures are intuitive based on how the subclass particle categories are included in the standard lipid measures. For example, HDL cholesterol is the cholesterol carried in all the HDL subclass particles in the circulation and the triglyceride concentration refers to the sum of triglycerides in all the lipoprotein subclass particles. In the case of LDL cholesterol, the situation is clear within the NMR analysis, i.e. it represents the sum of the cholesterol in all LDL subclass particles. Thus, the LDL cholesterol based on the NMR platform is size-specific and does not include cholesterol in any other particles. Generally, however, ‘LDL’ as a concept is far from clear since multiple ‘LDL’ cholesterol assays exist based on remarkably varying methodologies, leading to between-assay heterogeneity in the values of ‘LDL’ cholesterol. The assay-dependent ambiguity can lead to either under- or overestimation of the true (size-specific) LDL cholesterol. We have recently discussed these issues and their potential implications in detail.The overall association pattern for the lipoprotein subclass concentration measures is structured and characterized by strong correlations between adjacent apolipoprotein B-containing lipoprotein subclasses with clearly diminishing associations along the metabolic continuum from VLDL to LDL particles. On the other hand, the association pattern for lipoprotein subclass composition measures is varied and lacks pronounced patterns. Overall, triglyceride and cholesteryl ester compositions tend to associate negatively, particularly within the major lipoprotein fractions VLDL, IDL and LDL, and HDL. A consolidated interpretation of all the associations between the concentration and composition measures is the consistent behaviour of triglycerides. High circulating triglyceride concentrations are reflected by high concentrations of triglyceride-enriched VLDL particles that appear to cause a metabolic spillover effect and result consistently in triglyceride-enriched particles in all lipoprotein subclasses. The smallest HDL subclass behaves similarly to apolipoprotein B-containing subclasses regarding various associations, in contradiction to the other HDL subclasses.,et al. in 2017,This particular NMR platform has been applied in a few large-scale prospective cardiovascular outcome studies with the focus on lipoprotein subclass concentrations.et al.Only a few large-scale epidemiological studies have exploited the compositional lipoprotein subclass data from this NMR platform or in general. Laakso and co-workers studied lipoprotein subclass profiles in individuals with varying degrees of glucose tolerance in a population-based study of >9000 Finnish men.,,,,,,,,The lipoprotein subclass data have recently been actively utilized in drug-target Mendelian randomization analyses.,,,,,,,,The high size resolution offered by NMR-based lipoprotein profiling has resulted in a detailed understanding of how the various stages and remnant particles in the apolipoprotein B-containing lipoprotein cascade relate to cardiometabolic issues and outcomes.The separation of lipoprotein subclasses in NMR is solely based on particle size. Fundamentally, the resolution via NMR chemical shifts is best for the smallest particles and weakest for the largest particles .16 In po,,,,,The highly correlated nature of lipoprotein subclass concentration measures, particularly for adjacent subclasses, is sometimes identified as a limitation or a problem. However, this is a direct representation of human lipoprotein metabolism; it is a continuum, particularly for the apolipoprotein B-containing lipoprotein subclasses. Thus, the adjacent subclass particles, in both concentrations and composition, are inevitably very similar with highly correlated attributes. Particularly at high size resolution of the particles, any other correlation behaviour between measures for adjacent or closely related lipoprotein subclasses would most likely be an indication of methodological hurdles. Nevertheless, the high correlations make part of the information redundant from a strictly statistical perspective but, on the other hand, in the case of continuous metabolic processes, high resolution allows intricate interpretations and findings as already demonstrated in various publications.The various analyses presented here reveal fundamental characteristics and differences in lipoprotein subclass concentration and composition data. We would urge clear nomenclature in relation to these measures and noting consistently whether circulating particle (mol/L) or lipid concentrations (mmol/L) are meant or whether referring to a relative lipid composition in a particular particle subclass . For example, a triglyceride measure could refer to a circulating concentration in the bloodstream of a particular subclass of particles (mmol/L) or to a relative composition of triglycerides in a particular particle subclass . These measures have a very different biological meaning and interpretation. The overall spherical structure of these particles, with an oriented surface monolayer, results in strong size-dependent biomolecular constraints for their lipid compositions. In addition, the human lipoprotein metabolism appears to be fundamentally controlled in such a way that only certain lipoprotein subclass particle and lipid concentrations are possible in the circulation. Lipoprotein subclass-related risk analyses are thus relevant only within physiologically feasible conditions. Comprehensive data on lipoprotein subclass particles in large-scale studies provide opportunities for detailed understanding of lipoprotein metabolism in health and disease. Combining these data with modern genetics in exceedingly extensive studies as in the UK Biobank will facilitate grasping what features might represent causal, and thus potentially treatable and druggable, metabolic risk factors.IJE online.The studies comply with the Declaration of Helsinki, were approved by the local ethics committee  and written informed consents were obtained from each participant.M.A.K. is supported by a research grant from the Sigrid Juselius Foundation, Finland. P.O. is supported by the Emil Aaltonen Foundation. The Northern Finland Birth Cohorts have received funding from the Academy of Finland, Novo Nordisk Foundation and EU.NFBCprojectcenter@oulu.fi) and visit the website (www.oulu.fi/nfbc) for more information.The NFBC data sets are available through an application process for researchers who meet the criteria for access to confidential data. Please contact the project centre (None declared.dyab156_Supplementary_DataClick here for additional data file.


Personalized Therapies in ARDS” we encouraged the submission of articles discussing ARDS pathophysiology and classification, pharmacological and supportive therapies, chest imaging, as well as identification of biological sub-phenotypes as a target for potential novel treatment strategies.In the last decade, research on acute respiratory distress syndrome (ARDS) has made progress in better understanding the variety of presentations, the importance of targeting specific pathophysiological processes, and how these can influence the individual response to treatments within the context of personalized medicine . ParticuWildi et al. validated an ARDS ovine model in 23 sheep on mechanical ventilation (MV), comparing the mRNA expression of biomarkers in leukocytes, which displayed different sub-phenotypes (hyperinflammatory and hypoinflammatory). The authors found that similar activated pathways might be involved -κB pathway) resulting in specific sub-phenotypes, suggesting the importance of omic approaches in ARDS . Interleukin (IL)-10 showed the best performance when differentiating secondary fungal/bacterial infections from C-ARDS and/or ECMO associated inflammation.Superinfections are common in COVID-19 ARDS (C-ARDS) . In a prHagens et al. classified 401 ARDS patients according to the Berlin definition based on case records using both a dichotomous assessment and an 8-grade confidence scale. Classification was based on confidence grades of three experts through the interclass correlation (ICC). The best ICC was found using an 8-grade confidence scale for lung ultrasound (LUS)  and computed tomography (CT) scan . Using the 8-score system instead of dichotomous assessment, the ICC of chest X-ray and CT scan increased by 0.022 and 0.065. Adding information from LUS or chest CT increased ICC by 0.25 when using the 8-grade confidence assessment. This study suggests that ARDS diagnosis requires more than one operator and 8-grade confidence scale.Another challenge is to classify and identify ARDS severity . Hagens Worku et al. evaluated the relative changes in RALE score upon venous-venous ECMO initiation in severe ARDS patients. The RALE score demonstrated excellent inter-rater agreement. RALE increased from baseline during the first day of ECMO and this increase was associated with higher baseline APACHE-III scores and greater reductions in tidal volume. This study suggested that an appropriate ventilator setting for patients with ECMO is yet to be defined, and that the RALE score can be considered for the assessment of the optimal MV target to mitigate the risk of atelectrauma.Even though radiographic assessment of lung edema (RALE) score can be used to evaluate both the extent of edema and ARDS severity , its proSoydan et al. performed a cross-over trial in 30 critically ill children comparing automated ventilation with closed–loop control of the fraction of inspired oxygen (FiO2) to automated ventilation using manual titrations of the FiO2 focusing on time spent in predefined pulse oximetry (SpO2) zones. They found that adaptive support ventilation (ASV) using a close-loop control of FiO2 titration increased the percentage of time spent within optimal SpO2 zones and increased the total number of FiO2 changes per patient, suggesting potential advantages with this strategy as compared to manual titration of FiO2.In contrast with pharmacological therapies, which failed to provide clear beneficial effects in ARDS, some supportive therapies present better results. Nowadays research is focusing on novel supportive strategies, especially MV, to improve patient outcome . Soydan Sandal et al. in a single-blinded randomized cross-over trial, confirmed a better oxygen control using a close-loop oxygen strategy in 23 pediatric patients even when patients were treated with high flow nasal oxygen (HFNO). According to these findings, we can assume that closed-loop oxygen control improves oxygenation in pediatric patients receiving HFNO and ASV for ARDS, thus potentially leading to a more efficient oxygen use, and reduction of the number of manual adjustments.Humayun et al.. They confirmed that acute brain injury is a possible consequence of ARDS, thus requiring specific lung protective MV strategies to limit secondary damage to brain. The authors suggest the use of permissive hypercapnia with personalized Carbon dioxide (CO2) goals in those without high intracranial pressure. Optimal oxygen targets should be higher than those recommended by guidelines (55–80 mmHg) in case of ARDS and concomitant acute brain injury, while the use of positive end-expiratory pressure around 12 cmH2O seems to be safe for the brain. Given the paucity of data in this population, more studies are required to better evaluate MV strategies and blood-gas threshold so as to reduce the risk of brain damage.The need for personalized MV was even more enlightened by a narrative review from Tobar et al. in a randomized double-blind trial, found that the use of a multiparameter electroencephalogram (EEG) protocol to guide sedation in C-ARDS as compared to no-EEG monitoring did not increase 30-day ventilator-free days, but reduced propofol administration and deeper sedation 5 days after randomization. These data suggest the importance of investing in future trials testing the impact of multimodal neuromonitoring strategies during sedation and their implications in ARDS patients.Sedatives and anesthetics may impact patients' spontaneous breathing and MV duration. During the pandemic, metanalyses suggested that neuromonitoring patients with C-ARDS can be useful to guide sedation and detect those at risk of neurological complications , 8. TobaPersonalized Therapy in ARDS” highlighted the relevance, current advances, and rationale for personalized approaches in ARDS, including the role of lung imaging, sub-phenotype identification, and need for physiologically based MV individualized approaches in different sub-populations of ARDS patients, including those with acute brain injury.Our Research Topic on “DB wrote the manuscript. PR edited the manuscript. EB and AT intellectually contributed and revised the manuscript. All authors approved the submitted version.


Merocyanine dyes are of great interest amongst researchers due to their nonlinear optical (NLO) properties and solvatochromism. Molecular structure of these dyes constitutes conjugated pathway between the donor and acceptor substituents, with lowest energy transition of  Merocyanine dyes with donor--2-((E)-2--ylidene)ethylidene)-3-oxo-2,3-dihydro-1H-inden-1-ylidene) malononitrile, which has been employed for solar cell application26. A simple layer stack small molecule based bulk heterojuction solar cell based on the HB194 delivered a promising photon conversion efficiency of 6.1%. This efficiency seems very low as compared to the current state-of-the-art solar cells containing dyes such as dye-sensitized solar cells, but in case of stacked small molecule based bulk heterojunction cells 6.1% efficiency is noteworthy. The measurement of solar cell power conversion efficiencies does not capture the molecular basis for success or failure of the dye. This further motivates us to understand the underlying photophysics of HB194 molecular system, which can direct rational modification of the dye to obtain higher efficiency in bulk heterojuction solar cells. Here, we have employed steady state absorption and emission measurements along with the computational calculations to highlight the solvatochromic behaviour in HB194. To unravel the interplay of different excited states and their lifetimes of HB194 dye in various solvents, we have employed ultrafast transient absorption measurements in visible region of the spectrum (450–750 nm). Detailed analysis of the time-resolved data reveals the role of the solvents in determining the lifetime of the underlying excited states. Also, HB194 shows uniquely red-shifted emission in ethylene glycol, which is assigned to the presence of molecular dimers in the solution using computational calculations. Excited state dynamics of the molecular dimer in ethylene glycol has also been described.In recent years, merocyanine dyes have also been shown as promising substitutes for ruthenium complex based dyes in Graetzel-type dye-sensitized solar cells. This has further gained attention towards understanding the photophysics of these merocyanine dyesThe solution of HB194 with different solvents are prepared in a cuvette cell with optical pathlength of 1 mm. The steady-state absorption and fluorescence spectra are measured on a commercial optical setup. For the transient absorption measurements, the cuvette is mounted in a XYZ translation stage to adjust the position on the focusing point between pump and probe beams. Detailed description of HB194 solution is presented in the “The steady-state absorption spectra are measured in a commercial spectrometer . The measured absorption data of HB194 in different solvents are presented in Fig. The electronic transition (deactivation) from the partial charge transfer (CT) state to the electronically ground state can be directly resolved by fluorescence spectra. The fluorescence spectra in different solvents are presented in Fig. On the basis of the steady-state absorption and emission spectra, we can clearly point out that the absorption peaks are very similar while the emission peaks are significantly solvent-dependent. Using these measurements, we can categorize these steady-state optical data into four different cases on the basis of underlying reorganization energies after CT excitation. The reorganization energy usually corresponds to half the energy difference between absorption and emission peaks. The first case, HB194 (CYH) shows absorption spectrum with the most narrow bands, rendering the clear observation of vibrational progression. Additionally, the fluorescence spectrum (dashed red line) also show two resolvable peaks at 580 and 630 nm, respectively. The HB194 (DCM) and (THF) are sorted into the second case. The observed bandwidth of the peaks in this category is relatively larger compare to first case of HB194 (CYH), which is reasonable considering the stronger interaction between HB194 and solvent molecules in second case . In addition, the main peak of fluorescence spectra are red-shifted to 630 nm and the side peaks locate at 680 nm. The HB194 (DMK) and (MeCN) are sorted into the third case, in which, the main transitions of fluorescence spectra are shifted to 650 nm with a side band at 700 nm . The last case is the absorption and fluorescence spectra of HB194 (EG), where the main peak of fluorescence spectrum is dramatically shifted to 750 nm due to a strong interaction to the polar environment . To minimize the effort with time-resolvent measurement and data analysis of redundant solvent systems, we have selected the representative solvents to investigate the transient dynamics of electronically excited states and understand the role of CT states.The excited-state dynamics of HB194 is studied for 4 representative solvent molecules. In the first case, we measure the TA spectrum of HB194 (CYH), which is shown here in Fig. We then plot the measured TA spectrum of HB194 (DCM) in Fig. In addition to the single point kinetic-trace analysis, we employ global fitting approach to study the population dynamics along the complete detection window. For this, we firstly use the global fitting toolbox , which is developed by home-built Matlab code to treat the TA and two-dimensional photon echo spectra. The code analyses the TA data and yields two associated parts: time-decay traces and the decay-associated spectra (DAS). By this, we perform the data analysis of TA data and present the main results in Fig. The DAS of HB194 (DCM) are plotted in Fig. ab-initio calculations of HB194 in different solvents using linear response model. Firstly, we construct molecular structure and perform the optimization by DFT calculations (B3LYP/6-31G(d)) with the GD3BJ1 dispersion correction. The corresponding excited-state properties were studied using the time-dependent DFT (TDDFT)-PCM2-LC-BLYP*3/6-31G(d) method. Their corresponding rang-separation parameters , which induces a weak interaction of CT state with solvent molecules. It is well known that this weak interaction do not dramatically shift energy landscape of CT state and yields a small magnitude of reorganization energy. Furthermore, on the basis of TA measurement, we observed a short lifetime of CT state in the case of HB194 (CYH) (the longest lifetime component 66 ps). However, in the case of HB194 (DCM) and (DMK), the stronger dipoles of DMK and DCM molecules induce a stronger interaction of CT state to polar environment . The relatively stronger dipole-dipole interactions result in an evident red-shift of emission peak, which yields a larger value of reorganization energy. In the schematic diagram of Fig. However, the detailed mechanism of deactivation of CT states is not straightforward. It is known that the strong dipole interaction between the functional CT state and polar environment induces the structural changes of molecule, which play another important role of determining of lifetime of CT states. For this, our theoretical calculations have demonstrated that the polar environmental molecules result in small-angle distortion between donor and acceptor sides inside HB194 molecule, which slightly change the site energy of CT state. More important, this small structural changes reduce the overlap of transition dipole between CT and ground states, which is the key factor to determine the deactivation rate of CT states.25. Formation of dimers or higher structures of merocyanine dyes is one of the problems associated with these dyes to be used for dye-sensitized solar cells. Based on these studies, we hypothesized formation of dimers in ethylene glycol. The initial structure of two HB194 was constructed in the form of J-aggregation. The ground state of HB194 dimers are optimized and then the excited states are obtained on this basis. The optimized dimer structure constitute a inter-planar angle of 25°, as shown in Fig. The observed anomalously red-shifted emission spectrum of HB194 in ethylene glycol poses a challenge to our proposed excited state picture of the molecule. This anomaly can be explained based on known aggregation tendency of the merocyanine dyes. Numerous literature reports point to the aggregation behaviour of the merocyanine dyesIn this paper, we studied the transient population dynamics of HB194 and its deactivation of CT states to ground state. The steady state measurements along with the quantum chemistry calculations clearly enunciate the solvatochromic behaviour of the molecule. The strong electronic interaction between CT states and polar environment induces the systematic shift of emission spectra with the solvent dielectric. To study the transient dynamics of CT states, we performed the transient absorption measurements of HB194 with different solvents. We observed that the lifetime of CT states strongly relate to the dipole-dipole interaction between system and environment. In the case of HB194 (CYH), the weak interaction to non-polar solvent results in a fast deactivation of population from CT state to the ground state. Lifetimes of the CT states are significantly prolonged to several hundreds of picosecond in the case of HB194 (DCM) and (DMK), in which the stronger interaction to the environment stabilize the energy landscape of CT states. In HB194 (EG), the possible formation of molecular dimers induces the experimentally observed anomalously large red-shifted emission peak. Our study provides an understanding towards the impact of solvent bath interactions in the photophysics of donor-bridge-acceptor based merocyanine system, specifically to the underlying CT states. We envision that the novel merocyanine dyes designed with the consideration of the role of bath interactions with CT states will pave the towards enhanced solvatochromic properties. Additionally, the bath interactions with the CT state can be used in conjunction with molecular aggregation property to design long lived red emitting dyes.The merocyanine dye, HB194 was directly procured from Shanghai Daeyeon Chemicals Co., Ltd (homepage: http://www.shdaeyeonchem.com/) and used without further treatment. The solvents: Cyclohexane (CYH), Dichloromethane (DCM), Dimethyl Ketone (DMK), Ethylene Glycol (EG), Acetonitrile (MeCN), Tetrahydrofuran (THF) were purchased from J &K Scientific Ltd.  To prepare the samples for optical measurements, the HB194 was directly dissolved in solvents and the solutions are diluted to optical density of 0.3 at the maximum absorption peak in steady-state absorption spectra. The UV/VIS absorption spectra were measured by UV-Vis spectrophotometer . The fluorescence spectra were detected by FluoroMax-4 spectrofluorometer  after subjecting the samples to excitation at 570 nm. All spectra were background corrected.The transient absorption measurements on HB194 solutions were performed in a commercial spectrometer , which has been pumped by femtosecond laser . All experiments were carried out at room temperature and a fused silica cuvette of 1 mm path-length was used. In the spectrometer, the white light supercontinuum was generated by focusing on sapphire crystal (3 mm), which was used as a probe pulse. The pump pulse was directly generated by a commercial OPA . The pump and probe pulses are then spatially overlapped on the sample with pump focus of 200 μm at the sample. The energy of the pump pulse for transient absorption of the sample was kept at 22.9 μJ/cm27 dispersion correction. The corresponding excited-state properties were studied using the time-dependent DFT (TDDFT)-PCM-LC-BLYP*/6-31G(d) method. Their corresponding rang-separation parameters  of molecules were collected by Multiwfn program32.Density functional theory (DFT) calculations were performed to study the electronic structures and photo-physical properties of molecular dyes. Ground state of the molecule in different solvents were optimized at the level of B3LYP/6-31G(d) with the GD3BJSupplementary Information.


The diagnosis of persistent postural-perceptual dizziness (PPPD) is primarily based on medical history taking. Research on the value of clinical balance and visual dependence tests in identifying PPPD is scarce.(1) to contrast clinical balance and visual dependence tests between PPPD patients, dizzy non-PPPD patients, and healthy persons; and (2) to evaluate whether these clinical tests can help to identify PPPD in patients with chronic dizziness.post-hoc Tukey, or independent samples t-tests. The value of the clinical tests for PPPD identification was evaluated through logistic regression and Partial Least Squares Discriminant (PLS-DA) analyses.Consecutive patients with chronic dizziness (38 PPPD and 21 non-PPPD) and 69 healthy persons underwent Static Balance tests, the Timed Up and Go test, the Tandem Gait test, and the Functional Gait Assessment (FGA). Visual dependence tests included the Visual Vertigo Analog Scale (VVAS), the Rod-and-Disc test (RDT), and postural sway while facing rotating dots. Groups were compared using ANOVA with p = 0.006). Facing rotating dots, PPPD and dizzy non-PPPD patients had increased postural sway compared to healthy persons  velocity p < 0.001, and COP area p < 0.001; but non-PPPD vs. healthy: COP velocity p = 0.116 and COP area p = 0.207). PPPD patients had no significantly increased postural sway compared to dizzy non-PPPD patients. PPPD and dizzy non-PPPD patients also scored significantly worse on balance tests compared to healthy persons . Differences were insignificant in balance scores between PPPD and dizzy non-PPPD patients, or in RDT scores between the three study groups. In patients with chronic dizziness, a higher VVAS score was most associated with PPPD . The cross-validated (CV) PLS-DA model with all clinical tests included, had fair discriminative ability (CVerror = 47%).PPPD patients had significantly higher VVAS scores than dizzy non-PPPD patients (PPPD patients were more visually dependent, but did not have worse postural balance compared to dizzy non-PPPD patients. Elevated VVAS scores characterized PPPD most in patients with chronic dizziness. With a prevalence of up to 20%, persistent postural-perceptual dizziness (PPPD) is among the top five most common causes of vestibular complaints reported in tertiary care hospitals –3. PPPD Patients with PPPD clinically present with non-rotatory vertigo and postural imbalance, which are present almost on a daily basis. The symptoms are worsened by upright posture, active or passive movements, and visual stimuli . Visual The pathophysiological mechanism of PPPD and its four subtypes is still uncertain . It is tThe diagnosis of PPPD is currently primarily based on medical history taking . The oftSeveral clinical tests exist in the literature that allow for evaluation of postural balance and visual dependence in patients with a vestibular disorder , 15. HowThe aim of this study was 2 fold: (1) to contrast clinical balance and visual dependence tests between PPPD patients, dizzy non-PPPD patients, and healthy persons; and (2) to evaluate whether these clinical tests can help to identify PPPD in patients with chronic dizziness.This study is a cross-sectional study consisting of consecutive patients enrolled between March 2019 and July 2020, either at the Department of Otolaryngology of the Antwerp University Hospital or in one of the two participating general hospitals . The control group consisted of healthy persons from the direct (employees) or indirect environment (family and friends) of the MOVANT research team. The study was performed in the M2OCEAN laboratory (Multidisciplinary Motor Centre Antwerp) of the Antwerp University Hospital.The study report is drafted conform the “Strengthening the Reporting of Observational studies in Epidemiology (STROBE)” guidelines for cross-sectional studies .Ethical approval was obtained from the Medical Ethics Committees of the Antwerp University Hospital (reference number 18/586).Patients' eligibility was assessed by an Ear-Nose-Throat (ENT) specialist through medical history taking [using the SO STONED method ], and thEligibility of healthy persons was verified by the researcher. Their inclusion criteria were (1) speaking the Dutch language; and (2) being at least 18 years old. The exclusion criteria were: (1) history of or currently suffering from rotatory vertigo; (2) frequent episodes of non-rotatory vertigo (more than one episode in 3 months); (3) balance problems; and (4) severe visual impairment, not correctable by e.g., wearing glasses.A patient was diagnosed with PPPD if he or she met all five diagnostic criteria for PPPD as established by the Committee for Classification of Vestibular Disorders of the Bárány Society . These aAge (years), gender, dizziness duration (years), and ENT diagnosis were collected from the electronic patient record.Dizziness Handicap Inventory (DHI) evaluates the physical, functional and emotional handicap experienced by patients as a result of their vestibular symptoms. For 25 statements, patients were asked to indicate the extent to which they applied to them . The total DHI score was recorded, ranging from 0 (no impairment) to 100  (The airment) , 19.Hospital Anxiety and Depression Scale (HADS) evaluates patients' emotional state by means of 7 anxiety-related (HADS-A) and 7 depression-related (HADS-D) questions answered on a 4-level ordinal scale. The total HADS-A and HADS-D scores, both ranging from 0 (no anxiety/depression) to 21 , were retained  test measures patients' perception of verticality in the absence of visual reference points. The experimental setup is shown in The Static Balance tests measures the patients' balance while they were standing still and upright in four different foot positions with eyes closed: (1) feet together, combined with Jendrassik maneuver ; (2) feet 5 cm apart standing on a foam plate , combined with Jendrassik maneuver ; (3) heel-to-toe tandem stance; and (4) standing on one leg. For each condition, the patient had three attempts to maintain the respective condition 30 s. Only the best score (in seconds) out of these three attempts was retained. The sum of these best scores for each of the four conditions constitutes the total static balance score, which ranges from 0  to 120   .Timed Up and Go test evaluates how quickly a person can get up from a chair, walk 3 m, turn around, walk back and sit back down on the chair. The fastest performance time (in seconds) of three attempts was retained  evaluates balance control during ten different walking tasks . The total FGA score ranges between 0  and 30   .Visual Vertigo Analog Scale (VVAS) is a questionnaire consisting of nine statements describing different situations in daily life where disturbing visual stimuli are present . For each statement, patients were asked to mark the extent to which they experienced dizziness on a 10 cm visual analog scale. If a situation was not applicable for the patient, it was not marked and not included in the final score. The total VVAS score is the sum of the marks for each relevant situation, rescaled to a percentage where 100% indicates maximal visually induced dizziness (The izziness .Rod-and-Disc test (RDT) measures the influence of visual disturbance on the perception of verticality. The experimental setup and the four test conditions were the same as for the SVV test, but for the RDT each test had to be performed twice with dots rotating clockwise (CW) and twice with dots rotating counter-clockwise (CCW) on the black screen. More information on the test conditions can be found as The Preliminary analyses showed that the Rod-and-Frame test-which was equivalent to the RDT, except that the dots were replaced by a frame tilted left or right-yielded less pronounced results than the RDT, and is therefore not retained for discussion. The test conditions and data are available as Postural sway while facing rotating dots was evaluated by measuring the displacement of patients' center of pressure (COP) first while looking at a black screen, and then during exposure to dots rotating CW and dots rotating CCW. The experimental setup differed from the SVV and RDT that no line was shown in any of the three tests, and that the patient stood on a force plate . More information on the test conditions can be found as Preliminary analyses showed that the postural sway while facing a tilted frame–which was equivalent to the postural sway while facing rotating dots, except that the dots were replaced by a frame tilted left or right-yielded less pronounced postural sway results compared to the rotating dots conditions, and are therefore not retained for discussion. The test conditions and data are available as The researcher is an accredited physiotherapist (Master's degree). The study took 2.5 h per participant, using the following strict assessment order: first the questionnaires, then the clinical balance tests, and finally the visual dependence tests. The visual dependence tests (except the VVAS) were performed in a different randomized order for every participant to avoid a habituation effect on the results. Breaks were allowed between and during the different tests. The next test was only started when the dizziness symptoms had returned to participant's baseline level. The data were collected pseudonymised in an IBM SPSS Statistics data file stored on a secure server of the University of Antwerp.During data collection, the researcher was blinded to the ENT diagnosis of each patient. Blinding to whether a participant was a patient or healthy person was not possible, as the healthy persons were recruited from the environment of the research team.Descriptive statistics included means and standard deviations for quantitative variables, and frequencies and percentages for categorical variables. Shapiro-Wilk tests indicated whether quantitative variables were normally distributed .p < 0.006 accounting for 9 comparisons, followed by post-hoc analysis with Tukey correction for multiple testing. For inter-group comparison of the categorical variables, chi-square (χ2) tests were used. Two-group comparison of dizziness-related variables was carried out through independent samples t-tests.Inter-group comparison of quantitative variables was carried out through one-way analysis of variance (ANOVA), using a Bonferroni-corrected significance level of Univariable logistic regression and Partial Least Squares Discriminant Analysis (PLS-DA) models were fitted to determine the predictors of PPPD in patients with chronic dizziness. The relation between the clinical tests and PPPD was expressed as odds ratios [95% confidence interval (CI)], area under the receiver operating characteristic curve (AUC), and optimal cut-off with corresponding sensitivity and specificity (according to maximization of the Youden index). For the PLS-DA model, the potential of each of the clinical tests to discriminate between PPPD and dizzy non-PPPD patients was evaluated through their loading values. The overall discriminative value of the PLS-DA model was 5-fold cross-validated (CV) . In briep < 0.05, unless otherwise stated. Statistical analyses were performed using the SPSS software version 27.0 , personal reasons (3 patients), or COVID outbreak (4 patients). The results of two other patients had to be excluded after the study because of unreliable test results due to disturbing background noise near the laboratory during the testing.Of the 59 patients (mean age 57.34 ± 12.96) that successfully participated, 38 were diagnosed with PPPD. The 21 patients without PPPD were primarily diagnosed with vestibular hypofunction (4 patients), bilateral vestibulopathy (1 patient), proprioceptive cervicogenic dizziness (8 patients), multiple sensory deficits (3 patients), and mal de débarquement syndrome (1 patient). In four patients no cause could be found.The control group consisted of 69 healthy non-dizzy persons (mean age 51.71 ± 17.24).An overview of the recruited samples and the study aims are shown in p = 0.453) were not significantly different between the three study groups.Gender numbers  and HADS-D scores  compared to healthy persons. PPPD patients had significantly higher SVV scores (p = 0.016) and dizzy non-PPPD patients were significantly older (p = 0.037), both compared to healthy persons.All patients with chronic dizziness had significantly higher HADS-A , but not significantly higher DHI (p = 0.065), HADS  and SVV scores (p = 0.406), compared to dizzy non-PPPD patients.PPPD patients had significantly higher duration of dizziness complaints  and dynamic balance scores  compared to healthy persons.All patients with chronic dizziness had significantly worse static  or dynamic balance scores .Between PPPD and dizzy non-PPPD patients, no significant differences were found in either static (Data are provided in p = 0.431) between the three study groups. The RDT values were positive in all three groups, indicating that in all cases the perceived vertical had an larger offset in the same direction of the rotation of the dots compared to the SVV tests. The COP lean results (p = 0.800) were not significantly different between the three study groups either. The COP lean values were small ( ≤ 1 mm), indicating that the visual disturbance did not cause the participant to tilt more to one side.There was no significant difference in RDT results  compared to healthy persons. The COP velocity values were positive in all three study groups, indicating larger mediolateral sway in visually disturbing conditions compared to the SVV tests. Next to this, PPPD patients also showed a significantly higher COP area (p < 0.001) compared to healthy persons. The COP area values were positive in all three study groups, indicating that the COP displacement was larger in visually disturbing conditions compared to the SVV condition.Patients suffering from chronic dizziness had larger COP velocity values , but not significantly larger COP velocity values (p = 0.475) and COP area values (p = 0.116), compared to dizzy non-PPPD patients.PPPD patients had significantly higher VVAS scores ]. The cut-off value for the VVAS (through maximization of the Youden index) was 21.01 which resulted in a sensitivity of 0.74 and a specificity of 0.67 for identification of PPPD.In addition, a PLS-DA model was built to classify patients with chronic dizziness into PPPD and non-PPPD based upon all clinical tests listed in The first two components of the PLS-DA accounted for 51% of the variance in the data . However, since the same observations were used to build the model and to test its predictive power, the prediction error is probably underestimated. To obtain an unbiased estimate of the prediction error of the PLS-DA model, and to find the optimal number of components to be included, 5-fold cross validation (CV) was carried out. The CVerror was 47%, as presented in Our results show significantly reduced static and dynamic balance scores in patients with chronic dizziness compared to healthy subjects. However, no significant difference in any balance test results could be demonstrated between patients with and without PPPD. The literature indeed confirms reduced balance in all forms of chronic dizziness , 27–29. Furthermore, our results show increased postural sway while facing rotating dots compared to the baseline condition (black screen) in all three study groups, as indicated by the positive values for COP velocity and COP area. The largest increase in postural sway was found in PPPD patients. These results are in line with the literature which states that disturbing visual stimuli can reduce balance control, especially in persons who are less able to rely on their vestibular and proprioceptive systems in visually challenging conditions  . In PPPDThe COP lean deviations were very limited ( ≤ 1 mm), but in line with the low values reported in the literature , and notAnother finding was the larger SVV deviations in all patients with chronic dizziness (both PPPD and non-PPPD patients) compared to healthy persons, although in none of the three study groups the SVV values were pathological . TheIn our results, the SVV misalignment of RDT was not significantly different in PPPD patients compared to non-PPPD patients. This result agrees with earlier findings indicating that SVV misalignment of RDT is independent of the subtype of chronic dizziness, e.g., no significant differences in scores were found between visual vertigo and labyrinth-defective subjects , or betwPreliminary results showed that participants had less postural sway disturbances in presence of a tilted frame compared to the black screen condition. This adds to the literature that any reference frame, even if it deviates from the earth gravitational as in this study, provides the participant with a visual aid to maintain a more stable upright position. The perception of verticality was more disturbed in the RFT than in the SVV test, but less so than in the RDT. The literature confirms that rotating dots are a stronger visually disturbing stimulus than a tilted frame .Lastly, this study aimed to identify PPPD in chronic dizziness patients by means of commonly used clinical tests. Balance and visual dependence tests were chosen, since poor balance and increased visual dependence have been reported in previous studies as indicators of PPPD , 6. The Strengths of this study include (1) the meticulous elimination of visual fixation points during the visual dependence tests , (2) preliminary analysis of the RFT, RDT and postural sway while facing a tilted frame vs. rotating dots, and (3) the randomization of the visual dependence tests to eliminate a habituation effect on the results. Limitations of the study are (1) the limited sample size, (2) not having performed vestibular tests to confirm the absence of vestibular deficits in healthy persons, (3) the slightly younger age of the healthy persons, (4) the use of “best of three attempts” for the Timed Up and Go and Tandem Gait tests which may have induced a ceiling effect, and (4) the administration time of the test protocol (2.5 h) which may have caused fatigue in some patients.The VVAS was the most useful clinical test for the detection of PPPD of the balance and visual dependence tests examined in this study. Both PPPD and dizzy non-PPPD patients had significantly impaired balance and increased visual dependence compared to healthy persons. This shows the importance of evaluating balance and visual dependence in patients with chronic dizziness. The clinical tests can be useful to chart the patient's individual complaint profile. This profile can then be used to accentuate certain therapy elements, for example the degree of visual desensitization training.Further research can investigate the discriminative value of other clinical tests not discussed in this article, preferably in combination with the VVAS results.PPPD patients were more visually dependent, but did not have worse postural balance compared to dizzy non-PPPD patients. The VVAS had the most, but limited, discriminative value for identifying PPPD in chronic dizziness. In clinical practice, evaluation of balance control and visual dependence is indicated in patients with chronic dizziness, and corresponding exercises should be integrated into their exercise program in a patient-tailored way.https://zenodo.org/, doi: 10.5281/zenodo.6077074.The datasets presented in this study can be found in online repositories. The name of the repository and accession number can be found at: Zenodo, The studies involving human participants were reviewed and approved by Medical Ethics Committees of the Antwerp University Hospital (reference number 18/586). The patients/participants provided their written informed consent to participate in this study.All authors contributed to the outline of the research goals, the analysis/interpretation of data or editing of the manuscript, and the final reviewing and approval of the manuscript. All authors contributed to the article and approved the submitted version.This work was supported by the University of Antwerp , Antwerp, Belgium.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.


The proteasome has key roles in neuronal proteostasis, including removal of misfolded and oxidized proteins, presynaptic protein turnover, as well as synaptic efficacy and plasticity. Proteasome dysfunction is a prominent feature of Alzheimer’s disease (AD) (1–3). Artificial impairment of proteasome function can mimic many neurodegenerative phenotypes . We report impaired proteasome function to represent an early-stage marker of AD preceding many other markers of the disease. Significantly, we show that prevention of proteasome dysfunction by genetic manipulation in fly and cell culture models of AD delays mortality, cell death, and cognitive deficits. We developed a transgenic mouse with neuronal-specific proteasome overexpression which, when crossed with a mouse model of AD showed reduced mortality and cognitive deficits. To establish translational relevance, we developed a set of novel TAT-based proteasome-activating peptidomimetics. These agonists stably penetrate the blood-brain-barrier and enhance 20S as well as 26S proteasome activity. We show that treatment with these agonists protects against cell death in a cell culture model of AD as well as both cognitive decline and mortality in fly and mouse models of AD. The protective effects observed from proteasome overexpression in our models appear to be driven at least in part by increased turnover of the amyloid precursor protein (APP) by the proteasome. We conclude that the proteasome plays an important role in AD progression. Furthermore, augmentation of proteasome function is protective against AD-like pathogenesis in diverse models of the disease, representing a new therapeutic target for treatment of AD.


Oral health is essential to general well-being across the lifespan. Recent study on number of natural teeth, denture use and mortality in non-Singaporean elderly indicated the importance of oral determinants. To the best of our knowledge, no study has envisioned the association between biological age and oral health among community-dwelling older Singaporeans. By leveraging the acquired knowledge from Singapore Longitudinal Ageing Study (SLAS), our aim is to examine the association between biological age, oral health, and mortality. Our hypothesis is that older adults with or without dental problems will have a significant difference in mortality and other health outcomes. Our methodology is to combine a more realistic AI approach and a hypothesis-driven multidisciplinary approach to examine the association between oral health and general health outcomes . Cross-sectional data analyses were performed on 2844 community-living Chinese older adults of 65–80 years old in the Singapore Longitudinal Aging Study II (SLASII) cohort. The Keplan-Meier survival analysis showed statistically significant differences in survival time between the group with dental problems and the group without dental problems (p < 0.001) in an 8-year longitudinal study. Results also showed a significant difference between biologically younger, biologically older, and the intermediate group groups (p = 0.0012). Those groups were classified by the computed biological age. Our preliminary results will produce a plethora of new links between oral health, biological age, and mortality. The research outcome may serve as a baseline for policymakers to evaluate and promote subsidized oral health programs.


Empirical evidence for collaborative health promotion practices is still limited. The Cross-Sectoral Cooperation Self-Assessment Tool (CroCo SA) presented here provides a tool for cities to assess their current commitment to promote urban health and well-being across sectors. The SA tool was validated by co-operating with 9 pilot cities and seven countries in the framework of the Healthy Boost project funded by Interreg Baltic Sea Region (2019-2022). Addressing the current complexity, interdependence and emerging challenges of health promotion requires cross-sectoral cooperation and tools to assess cross-sectoral processes and make their impact visible. A digitized CroCo SA tool was developed and validated. An SA evaluation matrix was created, including 162 cells of options for reflective and guiding responses according to 6 scale x 27 items used in the SA tool. After submission of the electronic form, respondents can view anonymized results in a digital form on the Healthy Boost platform. The feedback report summarizes the results for respondents and provides, both numerical and descriptive, verbal feedback on the state of the assessed city's capacity for cross-sectoral cooperation, based on each assessment. The steps the respondents could take in their respective cities to improve cross-sectoral cooperation are outlined. Recommendations are based on the evidence found to be the prerequisites for effective cross-sectoral work for health promotion. SA can be conducted in many languages offered by the Google translation programme. Guiding feedback of CroCo SA defines areas for improvement in strategies and actions, allowing respondents also to do benchmarking and learn from other cities and countries. Self-assessment itself is a learning process.• The CroCo SA tool offers a method for gathering information about the state of cross-sectoral cooperation for urban health promotion.• By identifying strengths and weaknesses in cooperation, the city can build capacity for cross-sectoral cooperation.