The design of new hemostatic materials to mitigate uncontrolled bleeding in emergencies is challenging. Chitosan-based hemostatic hydrogels have frequently been used for hemostasis due to their unique biocompatibility, tunable mechanical properties, injectability, and ease of handling. Moreover, chitosan (CS) absorbs red blood cells and activates platelets to promote hemostasis. Benefiting from these desired properties, the hemostatic application of CS hydrogels is attracting ever-increasing research attention. This paper reviews the recent research progress of CS-based hemostatic hydrogels and their advantageous characteristics compared to traditional hemostatic materials. The effects of the hemostatic mechanism, effects of deacetylation degree, relative molecular mass, and chemical modification on the hemostatic performance of CS hydrogels are summarized. Meanwhile, some typical applications of CS hydrogels are introduced to provide references for the preparation of efficient hemostatic hydrogels. Finally, the future perspectives of CS-based hemostatic hydrogels are presented. Uncontrolled bleeding caused by trauma can lead to various complications such as hypothermia, decreased blood pressure, bacterial infection, and even shock ,2. If noTraditional hemostatic materials are usually bandages and gauze dressings, which stop bleeding by direct compression ,6. They Hydrogels are prepared from natural or synthetic polymers and have promising applications in the field of hemostatic materials ,16. DiffChitosan (CS) can be made from chitin by a deacetylation reaction . As showHemostasis, as a complex physiological process, involves the cells, extracellular matrix components, and signaling compounds . It can 2+, which forms a complex with activated VIIIa, PF3, and Ca2+ to further activate X. The activated partial thromboplastin reaction time is mainly used clinically to reflect the degree of the rapidity of the endogenous coagulation pathway [The coagulation factors involved in hemostasis are indicated by Roman numerals. The endogenous coagulation pathway plays a key role in the maintenance and consolidation phase after the initiation of the coagulation process. The endogenous coagulation is initiated by the activation of factor XII to produce XIIa after foreign body exposure, which in turn activates XI to XIa. Then, XIa activates IXa in the presence of Ca pathway . In the  pathway . The hem pathway .CS plays an important role in all processes of hemostasis. During hemostasis, CS can induce the aggregation of platelets and plasma proteins to help the blood clotting and vasoconstriction at the injured site ,65,66,67p = 0.045). This is attributed to the electrostatic interaction between CS and the erythrocytes that form an agglutination network. In addition, CS can also bind to hemoglobin and change its microstructure to form complexes through hydrogen interactions [Attracting and gathering red blood cells is the first step in CS to stop bleeding. The CS-acetic acid salt can cause red blood cells to adhere, aggregate, deform, and finally, form a firm blood clot to block the bleeding point. Many hemostatic agents are ineffective or limited in coagulopathies where coagulation is lost . Howeverractions . 2+ in platelets was significantly increased, which elevated the expression of glycoprotein IIb/IIIa  in the platelet membrane. For example, a sodium tripolyphosphate cross-linked CS-based soft hemostatic hydrogel was synthesized by Patil et al. [3, or CS alone. As a positive control, cells treated with SFLLRN peptide and ADP also showed platelet aggregation. In contrast, this phenomenon did not occur in heparin-treated cells because heparin inhibited the thrombin-mediated platelet aggregation.The second step of CS hemostasis is the induction of platelet aggregation and activation ,73. Aftel et al. . CS-baseMoreover, the physical properties of CS itself endow it with the property of excellent non-specific membrane adhesion. CS can adhere to bleeding wounds and combine with hemoglobin through hydrogen bonds, electrostatic interactions, and hydrophobic interactions to form a CS/hemoglobin complex, resulting in changes in the microstructure of hemoglobin and an increase in viscosity . In addiEscherichia coli and Staphylococcus aureus with minimum inhibitory concentrations of 6 mg/mL and 8 mg/mL, respectively. The biocompatibility and in vitro wound healing activity of N-SuC were investigated using human dermal fibroblasts. It was shown that N-SuC significantly accelerated wound healing by inducing a higher level of angiogenesis and granulation tissue formation [p < 0.01). In order to assess hemostatic performance, six rats were used in each experimental group to measure blood loss with and without hydrogel at the bleeding site. The total blood loss in the untreated liver was approximately 106.7 ± 27 mg in 3 min compared to 28.3 ± 5 mg in the sulfhydryl-modified CS. Sulfhydryl-modified CS had a significant hemostatic effect (* p < 0.01).The hemostatic mechanism of CS derivatives is consistent with CS. For instance, introducing quaternary ammonium groups increases the number of positively charged centers of CS, therefore, the platelet aggregation is higher than in CS. Furthermore, it has been found that platelet aggregation increases with the degree of substitution . Howeverormation . Anotherormation  The phosormation . As showormation  preparedStudies have shown that hemolysis is a major challenge to the preparation of hemocompatible hemostatic materials ,83,84,85p < 0.05). Some studies reported that CS with deacetylation <80% is conducive to the formation of a network space structure [The hemostatic effect of CS is affected by the degree of deacetylation, relative molecular weight, and chemical modifications. With a decrease in deacetylation, the coagulation effect tends to increase . In a retructure ,90. Aftetructure . Meanwhitructure . Other structure . Moreover, the relative molecular weight also affects the coagulation-promoting effect of CS. Hattori et al.  investigCS can react with many functional groups to graft or cross-link. In addition, amino groups can undergo alkylation, and quaternization, and react with aldehydes and ketones. Moreover, after introducing hydrophobic or hydrophilic groups into the polymer backbone, the stability of CS can be improved . N-decanal (Schiff base reaction). The lyophilized hydrophobic alkyl-modified CS was used in a porcine arterial transection model to control bleeding from a fatal arterial injury. The results showed that the total blood loss in the hydrophobic alkyl-modified CS gauze treatment group (4.7 mL/kg) was lower than that in the control group (13.4 mL/kg). Additionally, hydrophobic alkyl-modified CS mixed with sodium citrate rapidly converted the human blood into an elastic gel. Its gelling ability was similar to that of fibrin-based sealants [N-alkyl modification is a typical way to introduce hydrophobic groups to the CS backbone. The hydrophobic alkyl side groups can be non-covalently inserted into the erythrocyte membrane, which significantly increases the degree of aggregation of red blood cells . In addisealants .Carboxyl modification is another typical route to adding hydrophilic groups to the CS backbone. Carboxymethyl CS (CMCS) has antibacterial, hemostasis, and healing effects, and shows an enhanced histocompatibility compared to CS. Controlling the appropriate degree of carboxymethyl substitution can increase the hydrophilicity of CS. In addition, the CMCS hydrogel helps to locally absorb the excess water and enrich the procoagulant substances in the blood, which significantly reduces the coagulation time in vitro and in vivo . For ins*, p < 0.05; **, p < 0.01).Hydroxybutyl CS can be prepared by reacting hydroxybutyl with the C-6 hydroxyl and C-2 amino groups of CS. Hydroxybutyl CS is soluble in water and is temperature sensitive. In a study, Wang and co-workers prepared catechol–hydroxybutyl CS (HBCS-C) hydrogels as potential hemostatic material. As shown in N-hydroxysuccinimide as a cross-linking agent. The presence of silk fibroin improved the original porosity, water absorption, and hemostatic ability of CS dressing. The characterization results showed that the CS/SF hydrogel was characterized by a higher swelling ratio under acidic conditions and an excellent compressive mechanical property (50–130 kPa) because of the three-dimensional network structure. In addition, CS/SF hydrogel showed excellent antibacterial and hemostatic properties and was not toxic to human skin cells, showing great potential as wound dressings. CS and succinic anhydride dissolved in pyridine form a solid CS-succinic acid ester (CSS) derivative. The CSS-based hemostatic agent has obvious hemostatic efficacy and safety in the rat model of liver trauma and has potential clinical application value in the treatment of severe internal bleeding by intravenous administration . Pandit The primary amino groups of CS can be chemically modified by pyrogallol. This functional modification is beneficial in the improvement of wet adhesion, facilitating biological interface adhesion and improving hemostatic function . The speCS hydrogels have excellent biodegradability, self-healing, swelling, and mechanical properties. These properties are partially related to the preparation methods , and canChemically cross-linked hydrogels are primarily formed by the reaction of polymer functional groups with cross-linking agents. Cross-linking agents are compounds with two or more reactive functional groups that can cross-link with other polymers. The most commonly used cross-linking agents for the preparation of CS-based hydrogels are glutaraldehyde, formaldehyde, ethylene glycol diglycidyl acid, and genipin . The Schiff reaction is a chemical reaction involving a dy-namic covalent imine bond formation via the crosslinking of amine groups and aldehyde groups . Taking In a recent study, Song et al.  prepared3-based hydrogel by cross-linking CS with glutaraldehyde and being loaded with nano-silver to accelerate wound healing. PEG and CS solution were used to reduce silver nitrate and silver nanoparticles (AgNPs). The prepolymer was then cross-linked with glutaraldehyde to form the hydrogel. The results showed that the hydrogel has a higher porosity and better swelling than the CS-PEG hydrogel. Moreover, it can slowly and permanently release AgNPs, which have excellent properties as an efficient bactericide and antioxidant. Risbud et al. [Glutaraldehyde can be cross-linked with CS. Thus, this chemical cross-linker has been extensively used to prepare CS hydrogels for biomedical applications. Masood et al.  reportedd et al.  preparedGenipin can be used as a cross-linking agent. Under acidic and neutral conditions, it reacts spontaneously with primary amines from the polymer chain. It can also cross-link with CS and other polysaccharides to make artificial bones, wound dressings, etc. Since the aldehyde group is toxic for cells and causes severe inflammation in the human body, the low toxicity of genipin makes it a preferred cross-linker for CS hydrogel preparation. As shown in 3+ showed excellent mechanical properties. The average strength and elongation at the break of the PAM-AA/CNF/CS hydrogels were 11 MPa and 480%, respectively. Fan et al. [Physically cross-linked hydrogels are formed by physical entanglements through hydrogen bonding, hydrophobic association or electrostatic interactions. Physically cross-linked hydrogels are reversible and can be transformed back to a solution state when external conditions changed. Moreover, due to the presence of reversible physical cross-linking bonds, physically cross-linked hydrogels often have self-healing properties. Physical cross-linking can avoid the toxicity of chemical cross-linking agents. However, the main disadvantages of physically cross-linked hydrogels are their poor mechanical properties and the difficulties in controlling their pore sizes . Hydrophn et al.  preparedCS-based hydrogel has a good biocompatibility and can absorb wound exudate, avoid secondary wound infection, induce rapid wound healing, and prevent scarring. In addition, CS-based hydrogels can be loaded with bioactive substances  and is slowly released at the wound site ; therefoA CS-based hemostatic hydrogel can be injected into the body and fill an irregularly shaped defect area with minimal surgical trauma . Self-heThe combination of self-healing properties and hemostatic properties can be used in scenarios such as trauma hemostasis, tissue adhesion, and bone repair . Tang etCS hemostatic adhesives have gained widespread attention as wound dressing adhesives due to their ease of handling, non-invasive bonding, immediate sealing, and hemostatic effect . TherefoRecently, Jung et al.  designedWith the development of science, a wide variety of hemostatic materials have emerged due to the necessity of efficient and safe repair of wounds caused in battlefields, surgical procedures, and traffic accidents. Depending on the wound type, these hemostatic materials also have different applications. For example, bleeding from external wounds caused by sharp objects, traffic accidents, battlefields, etc., can be stopped using gauze, bandages, and tourniquets. There has been a tremendous level of development and commercialization in various types of hemostatic agents in modern medicine. However, in situations of battlefields and surgical bleeding, these hemostatic materials confront many obstacles which may prevent wounds from healing. Despite sustained advances in the use of hemostatic agents in modern medicine, many obstacles limit their successful use in the treatment of uncontrolled bleeding. Therefore, the development of fast, efficient, easy-to-handle, and multifunctional hemostatic materials is of great significance and highly demanded. Hydrogels can be prepared using natural or synthetic polymers and have promising applications in the field of hemostatic materials. CS-based hydrogels are highly favored because of their low cost, use of renewable resources, good biocompatibility, high efficiency, and rapid hemostasis due to their abundant amino groups. Moreover, they can adsorb red blood cells and plasma proteins through electrostatic interactions to promote platelet adhesion and achieve rapid hemostasis. In addition, CS can stimulate collagen deposition and self-assembly near the wound. CS derivatives obtained by the modification of CS confer additional functions to CS while enhancing the hemostasis. In addition, CS-based hydrogels can be loaded not only with bioactive substances such as growth factors and antibiotics but also with stem cells as wound dressings for the treatment of larger skin defects. However, these hydrogels also have poor mechanical properties and it is difficult to control the gelling time. Therefore, the preparation of mechanically durable CS hemostatic hydrogels with controllable gelation behaviors constitutes a priority of future research. Moreover, the combination of homeostasis and other functionalities may also promote the development of CS-based hemostatic hydrogels ,135. In


Introduction: Successful aging is a multidimensional construct covering behavioral, social, and psychological domains of well-being. We aimed to identify well-being profiles and their association with mobility-limitation-free survival.Methods: A total of 1488 healthy individuals aged 60+ from the Swedish National study on Aging and Care in Kungsholmen (SNAC-K) were followed-up for 15 years. Mobility limitation was defined as a walking speed <0.8m/s and vital status information was obtained from the National Cause of Death Register. Well-being profiles were derived from different behavioral, social and psychological indicators using latent class analysis among men and women. Cox and Laplace regression models were applied to examine the association with the incidence of a composite endpoint of mobility limitation or death.Results: At baseline, three well-being profiles  were identified, which followed a clear gradient in all behavioral, social and psychological indicators. Compared to those in the worst profile, men and women in the intermediate profile had 27%  and 23%  lower hazard of developing mobility limitation/death. An even greater protective effect was seen among individuals in the best versus worst profile . Men in the intermediate and best profiles survived 1 and 3 years longer without mobility limitation, respectively; these figures were 2 and 3 years for women.Conclusions: Better profiles of behavioral, social and psychological well-being may prolong mobility-limitation-free survival by at least one year among older adults. Our findings strengthen the evidence-base to achieve successful aging through multi-domain interventions. Successful aging is a multidimensional construct covering behavioral, social, and psychological domains of well-being, all amenable to individual actions and public health interventions –4. Even Nevertheless, none of these factors alone seems sufficient to explain the great heterogeneity in aging phenotypes, and successful aging may rather result from simultaneously adhering to several of these protective factors , 19. In In this study, we used latent class analysis to detect data-driven subgroups of people with similar well-being profiles according to behavioral , social  and psychological  well-being indicators, as defined by the Centers for Disease Control and Prevention (CDC) . Given tMost studies looking at indicators of well-being in old age have focused on single outcomes such as frailty, disability or death , 32. UnlThe specific aims of this study were: 1) to identify distinct well-being profiles among men and women separately, by using latent class analysis; 2) to determine which of these profiles are associated with the greatest benefit in terms of mobility-limitation-free survival; and 3) to quantify these potential benefits in absolute terms by calculating differences in median age at onset of mobility limitation or death across profiles.https://www.snac-k.se/), an ongoing population-based study. SNAC-K includes a random sample of adults aged 60 years or older living at home or in institutions in the Kungsholmen district of Stockholm, Sweden, between 2001 and 2004. A random sample of individuals from 11 age cohorts  were invited to participate in the study. The baseline SNAC-K population included 3363 individuals (73.3% participation rate) who have been followed up regularly: every six years for the young-old cohorts (<78 years) and every three years for the older cohorts (≥78 years). At each study wave, physicians, nurses, and psychologists conducted extensive clinical examinations, interviews, and assessments following standard procedures. SNAC-K data has been linked to the National Patient Register and the Swedish Cause of Death Register to obtain information on medical history and vital status.Data were gathered from the Swedish National Study on Aging and Care in Kungsholmen  and 55 individuals (1.8% of the remaining sample) with a Mini-Mental State Examination (MMSE) score <24 because of the limited reliability of their self-reported exposures. We further excluded 704 individuals (23.6% of the remaining sample) with mobility limitation at baseline, i.e., walking speed <0.8 meters per second (m/s), according to an established clinical cut-off [This study included data from baseline and four follow-ups of SNAC-K, extending until December 2015 (whereas the death register coverage extended until December 2016). Out of the 3363 participants from SNAC-K baseline, we first excluded 322 individuals (9.6%) with a definite or questionable dementia diagnosis according to the Diagnostic and Statistical Manual of Mental Disorders . It was reported as meters per second (m/s), reflecting the speed for whichever length walked . MobilitA composite endpoint, considered to be an indicator of mobility-limitation-free survival, was operationalized by taking into account the time from study entry until the development of mobility limitation  or death, whichever occurred first.Usual dietary intakes were assessed using a validated self-administered 98-item semi-quantitative food frequency questionnaire (SFFQ). Participants were asked to report their average intake frequency of each food and beverage over the past 12 months on a fixed 9-level scale ranging from never to ≥4 times per day. Zero imputation was applied to replace SFFQ missing items . This isInformation was obtained from baseline data and smokers and former smokers were asked how long they had smoked and the number of cigarettes smoked per day. Former smokers were also asked at what age they had stopped smoking. We further categorized smoking into current, former and never smokers.Participants were asked to specify the type and frequency  of the leisure activities they regularly engaged in. We grouped the reported activities into physical, mental, and social according to the category they predominantly belonged to, following a procedure described elsewhere . The scoPhysical leisure activities included: gardening, hiking in the forest/pick berries or mushrooms, hunting/fishing, doing home repairs, and carrying out light exercise  or moderate to intense exercise . Mental leisure activities included: using the internet/playing computer games, painting/drawing/working with clay/pottery, carrying out car or mechanical repairs, and reading books.Based on the same procedure described above, the following activities were identified: playing a musical instrument, listening to music, going to sports events, participating in study circles/courses, attending church/revival meetings, participating in volunteer work, going to restaurants/pubs/cafés, dancing, watching TV, playing chess/card games, going to the cinema/theatre/concert, knitting/weaving/sewing, participating in associations/clubs, traveling, reading the newspaper, reading a magazine/journal, going to the bingo and museum/art exhibition. Scores were standardized and further categorized into low, moderate and high according to tertiles.Participants were asked about their marital status, cohabitation status, parenthood, friendships, and the frequency of direct or remote contacts with parents, children, relatives, neighbors, and friends , 41. EacParticipants were asked about the satisfaction concerning their contacts with subjects from their social network, perceived material and psychological support, sense of affinity with potential associations, relatives, and residence area, and feeling part of a group of friends , 41. EacLife satisfaction was assessed through the validated self-reported Life Satisfaction Index A (LSI-A) that captures five different components: zest versus apathy, resolution and fortitude, congruence between desired and achieved goals, positive self-concept, and mood tone. The LSI-A consists of 20 items with an “agree,” “disagree,” or “uncertain” response (range 0-100). A high score indicates that the person takes pleasure from the round of activities that constitutes his or her everyday life, regards life as meaningful and resolutely accepts life as it has been, feels he or she has succeeded in achieving his or her major goals, holds a positive self-image, and maintains happy and optimistic attitudes and mood , 41. We Negative affect reflects the extent to which a person feels guilt, anger, or fear, and it considers the following features: distressed, upset, scared, nervous, and afraid. Respondents were asked to report whether and to what extent they had felt in the above-mentioned affective states during the last four weeks. The response options were “not at all,” “a little,” “somewhat,” “quite a bit,” and “very much” (range 5-25) , 41. We Positive affect considers the following affective features: active, inspired, determined, alert, and enthusiastic. Respondents were asked to report whether and to what extent they had felt in the above-mentioned affective states during the last 4 weeks. The response options were “not at all,” “a little,” “somewhat,” “quite a bit,” and “very much” (range 5-25) , 41. We Several covariates were considered as possible confounders and measured at baseline: age (continuous), highest level of formal education , number of chronic diseases (continuous) , and Mingsem lclass to estimate the latent class models using the maximum-likelihood estimation. Analyses were run separately in men and women. We started with 2 class-solutions, incrementally increasing the number of classes. The ideal number of classes was determined using statistical criteria and interpretations based on theoretical knowledge. Goodness-of-fit indices included Akaike Information Criterion (AIC) and Bayesian Information Criterion (BIC). Participants were assigned to one specific class according to their highest posterior probability. Subsequently, the distribution of all behavioral, social, and psychological factors across the derived classes was examined.Latent class analysis (LCA) was used to derive the well-being profiles based on the 10 indicators of behavioral, social and psychological well-being assessed at baseline. LCA is a statistical person-centered approach which groups individuals into unobserved classes based on responses to manifest variables. This results in subgroups of individuals similar to each other and distinct from those in other classes . We usedThe association between the well-being profiles and the incidence rate of the composite endpoint  was analyzed through multivariate Cox proportional hazards regression models, from which hazard ratios (HRs) and 95% confidence intervals (CIs) were obtained. The proportional hazard assumption was assessed by regressing the scaled Schoenfeld residuals against survival time. No deviation from the proportional hazard assumption was detected. The models were first adjusted for age and education, and additionally by baseline number of chronic diseases and MMSE score in a second phase. We used Laplace regressions to quantify the differences (in years) in median age at onset of mobility limitation or death , according to the well-being profiles. In other words, we analyzed the differences in age when 50% of subjects in each well-being profile had developed the composite endpoint. Population-attributable fractions (PAFs) were calculated to estimate the proportion of the composite outcome that would be avoided if all subjects belonged to the best profile, a measure that is valuable from the public health perspective. Because the interpretation of PAFs is more straightforward with binary exposures, all profiles were compared against the one with the greatest mobility-limitation-free survival.2). Second, we further plotted the original data on a reduced two-dimensional grid, while labelling the observations with their corresponding well-being profiles obtained from LCA. Finally, we repeated the Cox and Laplace regression analyses: a) additionally adjusting by personality traits, b) excluding participants who were only interviewed at baseline, and c) excluding participants who dropped-out without developing mobility limitation or death.We carried out the following sensitivity analyses. First, in order to verify the robustness of the classes derived from LCA, we performed a data dimension reduction using multiple correspondence analysis (MCA), a generalization of principal component analysis (PCA) for categorical data. We used the obtained dimensions to plot the coordinates for every category of the 10 well-being indicators and their corresponding sum of squared Cosine  at baseline. The majority were female (59%) and had at least high-school level education (91%) . At baseA three-latent-class solution was identified to be optimal for both men and women, based on the BIC parameter and the interpretation of the identified profiles . The worMen and women in the best profile were younger, with higher levels of education, had less chronic diseases at baseline, and had a lower incidence of the composite endpoint compared to those in the worst profile . The assIn agreement with the Cox regressions, results from Laplace regressions showed that men in the intermediate and best profiles survived 1 and 3 years longer without mobility limitations, respectively, compared to those in the worst profile after adjustment for potential confounders . Women iIn this large community-based cohort study of Swedish older adults aged ≥60 years, we found strong synergistic effects among different behavioral, social, and psychological indicators of well-being, leading to a clear gradient of well-being both in men and women. We also found that individuals in the intermediate and best well-being profiles lived between one to three years longer without mobility limitation compared to those in the worst profile, independent of potential confounders.The few previous studies looking at multiple indicators of well-being simultaneously have suggested that these tend to be interrelated and to cluster within individuals , 24, 45.Our finding that well-being profiles may considerably prolong mobility-limitation-free survival among older adults has been partially corroborated by previous evidence. Studies carried out with populations from the blue zones constitute a relevant example. These populations are characterized by a high adherence to a healthy diet and physical activity, increased satisfaction with social ties, high social support and psychological well-being, and exceptional longevity without severe disability . A recenAlthough previous research suggests that non-pharmacological multi-domain interventions can help preserve mental health and cognitive function among older adults , there iThe strengths of our study include the longitudinal study design, with a relatively large sample size, and with comprehensive information on a number of behavioral, social and psychological well-being indicators. Thanks to the availability of repeated measures of walking speed and vital status throughout the 15-year follow-up, we were able to create a composite endpoint of mobility-limitation-free survival. Moreover, the use of Laplace regressions allowed for the quantification of the years gained in association with belonging to different well-being profiles, enriching the interpretation of our results.However, several limitations should be considered. Our exposures were assessed only at baseline to avoid reverse causality, which could be challenging among subjects who are less likely to maintain similar levels over time. Additionally, a subsample of the eligible population (35%) lacked data for any study variables, which could lead to selection bias as those with missing data were sicker. Nevertheless, this may be less problematic in our study given that the sample was purposefully selected to be functionally healthy at baseline. Information on all well-being indicators was self-reported, which might have led to misclassification of the exposures. Participants in this study were cognitively healthier and able to self-report their exposure levels, which might have led to an underestimation of the associations in the general population. Finally, our findings, and especially those referring to the well-being profiles, may have a limited transferability beyond the exceptionally healthy and socioeconomically affluent SNAC-K population.While theoretical insights into different models of successful aging are on the rise, empirical evidence from population-based longitudinal data on the complex interplay among the distinct well-being domains and their association with person-centered outcomes, such as mobility-limitation-free survival, is currently lacking. This study addresses such an important gap and provides further evidence to better understand and promote functional independence in community-dwelling older adults through primary prevention multi-domain interventions.Supplementary FiguresSupplementary Tables


XPC gene exhibit strong susceptibility to skin cancer due to defective removal via GG-NER of genotoxic, solar UV-induced dipyrimidine photoproducts. However, XPC is increasingly recognized as important for protection against non-dermatologic cancers, not only through its role in GG-NER, but also by participating in other DNA repair pathways, in the DNA damage response and in transcriptional regulation. Additionally, XPC expression levels and polymorphisms likely impact development and may serve as predictive and therapeutic biomarkers in a number of these non-dermatologic cancers. Here we review the existing literature, focusing on the role of XPC in non-dermatologic cancer development, progression, and treatment response, and highlight possible future applications of XPC as a prognostic and therapeutic biomarker.Xeroderma pigmentosum complementation group C (XPC) is a DNA damage recognition protein essential for initiation of global-genomic nucleotide excision repair (GG-NER). Humans carrying germline mutations in the  Genomic instability from altered DNA repair processes is a hallmark of cancer, playing an important role in both tumor development and treatment response . Importaa]P-7,8-dihydrodiol-9,10-epoxide (BPDE) and aflatoxin B1 (AFB1). Much of our understanding of NER comes from studying the repair of UV-induced lesions, such as pyrimidine-pyrimidone (6-4) photoproduct (6–4PPs) and cyclobutane pyrimidine dimers (CPDs), for which NER serves as the primary repair pathway  light, tobacco smoking, and other chemicals, as well as endogenous sources such as oxidative stress caused by normal cellular metabolism . The nucmidone 6- photopro pathway . The NER pathway . DifferiThere is a clear and established association between defective NER and tumor development, as illustrated by the rare autosomal recessive congenital syndrome xeroderma pigmentosum (XP). XP patients are characterized by defective nucleotide excision repair (NER) of sunlight-induced dipyrimidine photoproducts . DependiXPC is increasingly recognized as an important player in solid organ cancer development and response to cancer therapeutics, both through its canonical role in GG-NER and through other repair pathways. Here we review the most recent updates on the role of XPC in non-dermatologic malignancies.in vivo is found in a heterodimeric form with RAD23B (human orthologue HR23B) which further stimulates XPC’s role in NER repair  partially unwinds the DNA duplex at the site of DNA damage, creating an opened bubble , 19. TFIA number of recent studies highlight that post-translational modifications of XPC, including polyubiquitination, SUMOylation and phosphorylation, likely impact XPC efficiency to detect DNA damage and initiate NER –25. Polyin vitro studies have elucidated a role of Rad4, the yeast homolog of XPC, in the recognition and repair of multiple contiguous mismatched base pairs , leading to subsequent unwinding, DNA bending, and flipping out of the mismatched nucleotides and stabilization of this conformation to allow for subsequent DNA repair  and thymine DNA glycosylase (TDG)  decreased tumor development, further supporting a link between XPC, oxidative damage and cancer development (Mounting evidence points to a role of XPC in base excision repair (BER). BER is the primary repair mechanism of small, base modifications that do not distort the DNA helical structure. Fibroblasts obtained from XP-C patients displayed increased oxidative DNA damage after UVB-irradiation compared to fibroblasts without an and APE1 . Likewisactivity . Numerouse (TDG) –37. XPC se (TDG) . This mase (TDG) . This rose (TDG) . In vivoype mice . Howeverspecific . Urethanelopment . Althougin vitro using a host-cell reactivation assay . In humans, deficient MMR, through both sporadic and inherited genetic disease, is linked to aging and cancer by promoting genomic instability , 44. In on assay . Furtheron assay –52. Specon assay , 54. Addncreases . Furtherand MSH6 . Cells dA damage . Finallye repair , 58. An e repair . These XXPC knock-down in HeLa cells was associated with increased sensitivity to the chemotherapeutic drug, etoposide, the cytotoxicity of which is dependent on replication-induced DSB; gamma-irradiation of these cells lead to cell cycle alterations without altered clonogenic survival  repair. Long-term survival . Furtheradiation . More diular DNA . XPC defular DNA . Overallin vitro and in vivo  have a significantly higher frequency of spontaneous mutations in the hprt gene in splenic T lymphocytes as compared to Xpa-/- and Csb-/- mice; this was also enhanced with aging  and has not been observed with an increased frequency in other XP patients  were associated with variable responses to imatinib in BCR-ABL driven chronic myelogenous leukemia (CML)  . In regaXPC, were associated with longer time to progression- in individuals treated with vincristine-adriamycin-dexamethasone followed by high dose melphalan and stem cell transplantation  due to the reliance on alkylating agents in the treatment of this malignancy; DNA damage caused by alkylating agents are typically repaired by NER. Dumontet et al. found that SNPs in multiple genes, including antation . Similarantation . Though XPC polymorphisms and variations in gene expression have been studied in lung cancer  with advanced age (16-17 months), although development of adenocarcinomas were rare without a co-existing loss of another tumor suppressor gene  and NOH-AAF leads to lung adenocarcinoma development  exposed to the carcinogen, urethane, developed an intermediate number of lung tumors when to compared to urethane-treated Xpc deficient and proficient littermate mice, suggesting a gene-dose effect and further supporting a role for intermediate levels of XPC expression, either through polymorphisms or epigenetic regulation, in lung cancer development  was associated with an increased odds of prostate cancer, associated with a 3.83-fold increased risk in a Tunisian population. In contrast, other XPC polymorphisms, including those heterozygous for Lys939Gln (939Lys/Gln) along with the PAT D/D haplotype are considered protective of prostate cancer (XPC PAT polymorphism (PAT +/+ or PAT +/-) along with tobacco smoking in a Chinese population   is the most common malignancy in males , and XPC studies Table 1.e cancer . One morpulation . Other sorphisms , 135 (Tognosis) , 92. HowXPC polymorphisms have been identified as one factor that may increase or decrease the risk of ovarian cancer as summarized in XPC Ala499Val polymorphism was found to correlate to a decreased odds of ovarian cancer (OR 0.35) while the XPC Lys939Gln polymorphism was associated with an increased risk of ovarian cancer (OR 1.72) in a dominant genetic model   . Going fpression . This fuXPC polymorphisms were associated with low penetrance susceptibility to bladder cancer  in pancreatic cancer risk , as part of a 5-polymorphism panel (high risk genotype) that has a 79% sensitivity and 85.4% specificity of predicting 5 years progression free survival  and rs2733532-C>T, with an increased odds of breast cancer  or homozygous (XPC-GG), is associated with increased risk of HCC; these genotype variants correlated with decreased XPC tumor protein expression by IHC as well as a shorter overall survival  carcinoma development. In a case-control study of hepatocellular carcinoma HCC related to aflatoxin B1 exposure, survival .XPC expression, but the mechanism by which this occurs is less clear. The XPC gene, along with several other tumor suppressor genes, is located on chromosome 3p, a frequently site of chromosomal deletion in human tumors  screen of prostate cancer, tization . Howevertization . Since itization . Howevertization . Additiotization . On the XPC mRNA and protein expression has been described in a number of cancers, with gene polymorphisms, deletions, and transcriptional regulation all active areas of research in the regulation of XPC expression. Additionally, research supports a role of XPC in the prognosis and treatment response in several of these cancers. Although XPC’s essential role in the recognition of bulky DNA lesions and subsequent activation of GG-NER, when altered, is a leading mechanism for development of UV-induced dermatologic malignancies and in modifications of cancer response to chemotherapies including cisplatin, recent data support a non-canonical role of XPC in DNA damage response and repair mechanisms, tumor suppressor transcriptional regulation, and in the development of non-dermatologic malignancies. Future studies would benefit from studying XPC as a biomarker of cancer prognosis and response to treatment in non-dermatologic malignancies.XPC is increasingly recognized as playing an important role in the development of non-dermatologic malignancies. Decreased Conceptualization, NN and CRS. Writing—original draft preparation, NN, BMW and CRS. Writing—review and editing, NN and CRS. Funding acquisition, CRS. All authors have read and agreed to the published version of the manuscript.This work was funded by the U.S. Department of Veterans Affairs BLR&D, Merit Review grant I01-BX005353 to CRS and in part by the National Institutes of Health, U.S.A. T32HL091816 to BMW.CRS is employed by the U.S. Department of Veterans Affairs. The contents of this manuscript do not represent the views of the U.S. Department of Veterans Affairs or the United States Government.CRS has served on Scientific and Medical Advisory Boards for Biodesix, Inc., bioAffinity Technologies, and as a scientific medical consultant for Bristol-Myers Squibb Company; these are not relevant to the topic of this manuscript. The remaining 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 attribute importance analysis revealed that the NM’s type, core-size and dosage are the most important attributes influencing the prediction. Our findings corroborate with those of the prior research landscape regarding the importance of P-chem characteristics. This study expands the application of ML in the nano-domain beyond safety-related outcomes by capturing the functional performance. Accordingly, this study has two objectives: (1) to develop a model to forecast the antioxidant efficiency of NMs to complement conventional in vitro assays and (2) to underline the lack of a comprehensive database and the scarcity of relevant data and/or data management practices in the nanotechnology field, especially with regards to functionality assessments.Reactive oxygen species (ROS) are compounds that readily transform into free radicals. Excessive exposure to ROS depletes antioxidant enzymes that protect cells, leading to oxidative stress and cellular damage. Nanomaterials (NMs) exhibit free radical scavenging efficiency representing a potential solution for oxidative stress-induced disorders. This study aims to demonstrate the application of machine learning (ML) algorithms for predicting the antioxidant efficiency of NMs. We manually compiled a comprehensive dataset based on a literature review of 62 in vitro studies. We extracted NMs’ physico-chemical (P-chem) properties, the NMs’ synthesis technique and various experimental conditions as input features to predict the antioxidant efficiency measured by a 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. Following data pre-processing, various regression models were trained and validated. The random forest model showed the highest predictive performance reaching an R ROS are generated during normal cellular functions as a by-product of aerobic metabolism and are essential to regulate physiological functions in biological systems [Reactive Oxygen Species (ROS) are molecules that contain one or more unpaired electrons in their orbitals and can lead to the formation of free radicals, e.g., superoxide (O systems . However systems . Therape systems .The antioxidant efficiency (anti-ox) is defined as the ability of molecules to scavenge the free radicals. Antioxidant compounds  control autoxidation by preventing the formation of free radicals or interrupting their spread by neutralizing them which delays or inhibits the damage to the target cells, decreasing oxidative stress, enhancing immune function and increasing longevity ,6. NumerNanotechnology has gained tremendous attention due to its potential to treat diseases and it is growing rapidly within the medicinal disciplines ,14,15. NThere are numerous organic and inorganic NMs exhibiting strong ROS-scavenging activities ,23. StudIn recent decades, there has been considerable interest in digitising the information in medical and pharmaceutical disciplines and the use of such data to explain biological phenomena . As more2 = 0.89. They employed important attribute analysis using the F-score method and determined the variables that affect the antioxidant capacity of cherries. Another example is from Idowu and Fatokun [ML models have been used to predict anti-ox activities, for example, Karydas  successf Fatokun , which e Fatokun  demonstr2 = 0.83.Given the time-consuming, expensive and work-intensive nature of NMs’ synthesis, having a computational method that can predict the anti-ox properties is beneficial. In addition, due to the large number of NMs’ P-chem possible combinations and the impact of experimental variables, it is not feasible to evaluate all of them using conventional laboratory tests to determine the ones with the highest capacities. In this work, we manually compiled a dataset derived from a literature review of in vitro studies in different mediums that can be used by other researchers to train models, augment the dataset with additional features, fill data gaps through grouping approaches and predict the efficiency of NMs to scavenge free radicals. Despite the limited data available , we explDue to high missing values among the input-related data collected, we selected NMs’ type, core size, shape, dosage, coating as well as the synthesis process, medium used, absorbance and duration as the input features for the model. This data cleansing resulted in a final dataset consisting of 10 columns and 1027 rows. The HDI, EE, PDI, SA, ZP, temperature, pH and thermal stability columns were excluded due to the presence of numerous missing values. Null data in core size, dosage and shape were replaced with the mean (for numeric features) or mode value  . Additio2 = 0.83.Following data homogenization and pre-processing, we trained and compared several regression models. The models are trained with the training dataset comprising 70% of the data (719 rows). Internal 10-fold cross validation is performed to optimize the hyper parameters and to ensure the models’ robustness and goodness-of-fit. Following the model training, we performed model validation using 308 rows out of 1027 (30% of data) as a test set  to evaluate the final tuned predictivity. As shown Nonlinear regression models (RF and ET) outperform linear models (LASSO and EN) because they exhibit more parameters (properties of training data learned during the learning process). RF is excellent at processing categorical, binary and numeric features, making it ideal for working with high-dimensional data with minimal pre-processing requirements.Depicted in The feature importance in the RF model is calculated using the impurity Gini-based feature importance (or Mean Decrease in Impurity). The duration of exposure, the medium used and the absorbance are relatively important attributes that affect the prediction. The synthesis techniques, shape and coating (presence or absence) have relatively less effect on the prediction.The purpose of this research was initially to (i) gather data from the literature related to the anti-ox efficiency of NMs, (ii) to present the landscape of data gaps related to functionality assessments, (iii) to apply a tool to demonstrate the feasibility of ML for this outcome and iv) to identify significant factors influencing the prediction of the outcome.Tremendous efforts are being put together to create databases with complete datasets and also many initiatives to promote the FAIR  principles are being created in the nano-domain ,50. To t2O2, OD and metal chelating. However, we selected the DPPH assay as the outcome due to substantial missing values among the other techniques. According to the literature, ABTS and DPPH assays are recommended with ABTS considered lengthy and expensive. It is challenging to conclude which approach is the most suitable for assessing the anti-ox activity. Bedlovičová [To evaluate the free radical scavenging capacity, we gathered data from six different methods such as DPPH, ABTS, NO, Hlovičová  examinedRegarding P-chem characteristics, only 35%, 22%, 10%, 18% and 1% of the selected studies reported ZP, HDS, EE, PDI and SA of NMs, respectively. Therefore, due to a substantial number of missing values, we were unable to incorporate all these data. Therefore, for this study we employed the type of NM, medium used, core-size, shape, synthesis techniques, coating, dosage, absorbance, duration and DPPH variables. However, the performance of the models was compared by including ZP values computed with an ML missing values imputation methodology, and/or including shape . These phttps://github.com/mahsa-mirzaei/NMAOP_M-M_wo-ZP-/blob/main/AOP_M%26M_wo%20ZP%20.ipynb, accessed on 18 January 2023), has a well-defined approach and is validated and built to an explicit endpoint derived from one assay [2 value, R2 = 0.83. In the nano domain, ML tools have been implemented to predict and assess the toxicity and functionality of NMs [We utilized a model that adheres to OECD (organization for economic co-operation and development) principles . In addition to complementing conventional in vitro testing, this approach is substantially more cost-effective and time-efficient. The capacities of algorithms can be exploited to fulfill new approach methodologies [Significant numbers of NMs have been evaluated to date, resulting in a large pool of information scattered through the literature; it becomes increasingly challenging to analyze all NMs using traditional test methods. Accordingly, the demand for computational methods increases. We investigated inorganic and organic NMs to develop a model with good predictability  information related to NMs’ P-chem properties such as the NM type, core-size, coating, hydrodynamic size (HDS) and its polydispersity index (PDI), shape, surface area (SA), zeta potential (ZP) and encapsulation efficiency (EE); (2) experimental conditions such as the methods of NMs’ synthesis such as chemical (top-down and bottom-up approach) and green , the culture medium for the anti-ox activity evaluation, information related to the application of the NF , pH, temperature, thermal stability, absorbance and duration of the test. In addition, the characterization methodologies used, i.e., scanning electron microscopy (SEM), field emission scanning electron microscopy (FESEM), transmission electron microscopy (TEM), high-resolution transmission electron microscopy (HRTEM), X-ray Diffraction (XRD), energy dispersive X-ray (EDX), Fourier transform infrared spectroscopy (FTIR), dynamic light scattering (DLS) and Ultraviolet–visible spectroscopy (UV–vis spectroscopy) to evaluate the P-chem properties were collected.Output-related collection: various assays can be used to assess the anti-ox efficiency of NPs including 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS), nitric oxide (NO) or total radical-trapping antioxidant potential (TRAP). After collecting the above information, we selected the DPPH assay that measures the anti-ox efficiency of NMs as the output to be predicted, since it is the most popular, rapid, inexpensive and simple test . In addiData pre-processing: PyCaret executes missing value imputation and categorical one-hot encoding. The missing values in the numerical features are replaced with the mean value of the features in the training dataset, whereas the missing values in categorical features are imputed with the most frequent or the mode value . RafsunjModel development: We trained several regression models including Random Forest (RF), Light Gradient Boosting Machine (LIGHTGBM), Extra Trees (ET), Decision Tree (DT), K-Nearest Neighbors (KNN), Least Absolute Shrinkage And Selection Operator (LASSO), Elastic Net (EN) Least Angle Regression (LAR), passive aggressive regressor (PAR), Lasso Least Angle Regression (LLAR), Huber Regressor (HUBER), Adaboost Regressor (ADA), Bayesian Ridge (BR), Ridge Regression (RIDGE), Gradient Boosting Machine (GBM) and Orthogonal Matching Pursuit (OMP). All models were evaluated and compared to find the best model with good predictivity. The following seven models are selected for reporting in this study: RF is a supervised algorithm that employs both decision trees and bagging techniques. These decision trees organize attributes by arranging them by their values. RF executes feature selection, each tree calculates the significance of a feature based on its ability to improve the integrity of the leaves . LIGHTGB2) were employed to assess the performance of the models. In addition, the external data (30% of data) was used to assess the predictability of the models.Model validation: 10-fold cross-validation was performed on the training set (70%) to optimize the hyperparameters and to ensure robustness. The mean absolute error (MAE), root mean square error (RMSE) and coefficient of determination . Furthermore, this paper emphasizes the significance of a consistent and systematic reporting structure and comparable measurement techniques indicating that their absence can result in an obstacle for modelling purposes. This is an important finding for policymakers and research funding bodies alike. The ability to predict functionalities represents a great opportunity in the healthcare field. Additionally, it will assist insurers and other stakeholders in the healthcare ecosystem to determine whether it is safe and beneficial (profitable) to invest in the development of nano-antioxidant therapies. Future research should therefore attempt to address this challenge through data capturing templates from empirical researchers that facilitate the construction of a comprehensive dataset.


We examined the white matter of patients with and without focal to bilateral tonic-clonic seizures (FBTCS), and control participants. A neural network based tract segmentation model (Tractseg) was used to isolate tract-specific, track-weighted tensor-based measurements from the tracts of interest. We compared the group differences in the track-weighted tensor-based measurements derived from whole and hemispheric tracts. We identified several regions that displayed significantly altered white matter in patients with focal epilepsy compared to controls. Furthermore, patients without FBTCS showed significantly increased white matter disruption in the inferior fronto-occipital fascicle and the striato-occipital tract. In contrast, the track-weighted tensor-based measurements from the FBTCS cohort exhibited a stronger resemblance to the healthy controls (compared to the non-FBTCS group). Our findings revealed marked alterations in a range of subcortical tracts considered critical in the genesis of seizures in focal epilepsy. Our novel application of tract-specific, track-weighted tensor-based measurements to a new clinical dataset aided the elucidation of specific tracts that may act as a predictive biomarker to distinguish patients likely to develop FBTCS. Focal to bilateral tonic-clonic seizures (FBTCS) are a feature of focal epilepsy that can lead to an increased risk of cardiac arrhythmias , seizurevia its role in the synchronization of abnormal cortical-subcortical ictal discharge  measurements. Here, tensor-based images are weighted by reconstructed streamlines traversing a given voxel to generate a TW version of the tensor-based image . TW measSince the subcortical structures involved in FBTCS are also involved in focal seizures , we inclThe innovation in this study is the combination of a neural network based tract segmentation model , with sTwenty-seven adults with focal epilepsy were recruited from the Comprehensive Epilepsy Centre at the Royal Prince Alfred Hospital ; and MRI was performed at the Brain and Mind Centre . Inclusion criteria were adults diagnosed with focal epilepsy, aged 18–60, presenting without surgery, and with or without a cortical brain lesion, who were willing and able to comply with the study procedures for the duration of their participation. Exclusion criteria were pregnant women and individuals with intellectual disabilities. The 20 controls were neurologically normal individuals. Written informed consent was obtained from all participants before study participation. Ethical approval was obtained from the RPAH Sydney Local Health District (RPAH-SLHD) ethics committee . All research and methods were performed in accordance with the Declaration of Helsinki and conducted in accordance with the relevant guidelines and regulations prescribed by the RPAH-SLHD.- “FBTCS-Y”: In this subgroup, the 19 patients were defined as having frequent (more than two per year) or infrequent (one per year) large, homolateral and simultaneous FBTCS. The FBTCS may have occurred during observation at the RPAH Epilepsy Centre or reported by the participant as occurring elsewhere.- “FBTCS-N”: In this subgroup, the 8 patients had never experienced FBTCS.“All patients”: The entire cohort of 27 patients; all diagnosed with focal epilepsy. The “All patients” group was further separated into the following two subgroups:“Controls”: The control group consisted of 20 age and sex matched healthy individuals with no history of epilepsy.The participants were placed into the following groups:TM MR750 3T scanner . For each participant, the following sequences were acquired: Pre-contrast 3D high-resolution T1-weighted image (0.7 mm isotropic) using fast spoiled gradient echo (SPGR) with magnetization-prepared inversion recovery pulse ; and axial diffusion-weighted imaging  with a uniform gradient loading  in 64 directions and 2 b0 s. An additional b0 image with reversed phase-encoding was also acquired for distortion correction  , alongsidhollander algorithm , fractional anisotropy , axial diffusivity  and radial diffusivity  . Next, tTractseg , a convoA neuroimaging specialist (CW) and neuroimaging researcher (CM) conducted quality control checks by visually inspecting each tract for consistency and anatomical correctness. The cerebellum areas in each subject were inspected and registration accuracy was verified. The right TPAR tract failed the quality control check in 10 participants; therefore the whole tract was removed from the analysis. Twenty-four tracts were included in the final statistical analysis.The chi-square test was used to investigate the existence of the following relationships in the patient subgroups: lesion presence and seizure onset side; lesion presence and FBTCS. Chi-square also tested for differences in drug resistance between patient subgroups and gender differences between the patient subgroups and controls. Analysis of variance (ANOVA) tests were conducted to examine differences in age between the patient subgroups and controls, and disease duration between the patient subgroups.LTW−TM × Lcount) + (RTW−TM × Rcount))/(Lcount + Rcount)] . Univariate analysis of covariance (ANCOVA) tests, which are robust to multiple comparisons including covariates  and control participants. Again, the weighted average of each tract's left and right sides was computed (as described above) to investigate the whole-tract differences in TW-TM between the three groups. ANCOVAs were then used to investigate group differences in a given hemisphere. Here, the TW-TM for a given tract from a given hemisphere was compared between the three groups.N = 17) and “drug responsive” (N = 8) groups. To test for group differences, a multivariate analysis of covariance (MANCOVA) was conducted per TW-TM. Another separate MANCOVA was conducted to test for differences between the FBTCS-Y and FBTCS-N groups, with drug resistance as a covariate.It has been shown that drug resistant patients may have widespread alterations . To addrp = 0.05 for all ANCOVAs. Bonferroni correction was used to correct for multiple comparisons as it has been shown to be a robust statistical method to control for multiple comparisons in the specific context of diffusion tensor imaging and tensor based metrics derived from an epilepsy cohort  = 1.506, p = 0.234] or gender [p = 0.402] between the FBTCS-Y, FBTCS-N, and control groups. There was no significant difference in drug resistance classification between the FBTCS-Y and FBTCS-N groups, [p = 0.186]. There was no significant difference in disease duration between the FBTCS-Y and FBTCS-N groups, . There was no significant association between lesion presence and seizure onset side or between lesion presence and FBTCS. Twenty-five patients  and 19 controls  were included in this study after passing the quality control (QC) check described in the Methods section 2.3. There were no significant differences in age [Individual ANCOVA tests were conducted to examine the between-group differences in the two grouping conditions .p-values extracted from the pairwise comparisons); all other results are reported in the p-value is reported as p < 0.05. The exact p-values and confidence intervals for the ANCOVAs (except for Section 3.4) are provided in the To focus on the results that may yield clinical value, only the TW-TM that displayed a mean significant difference between the groups are reported here and in the following sections (p < 0.05). The uncinate fasciculus (UF) had a lower average TW-FA in the “All patients” group compared to controls . Nineteen tracts showed a higher average TW-RD in the “All patients” group compared to controls (p < 0.05). The middle and superior cerebellar peduncles (MCP and SCP) had higher mean TW-AD in “All patients” compared to controls . p-values, effect sizes, and confidence intervals from the ANCOVAs between the “All patients” and control groups are reported in Compared to controls, the mean TW-ADC was higher in the “All patients” group for 16 out of 24 whole brain tracts , and in the TW-FA of the corpus callosum (CC) (p = 0.043), thalamo prefrontal and premotor tracts . When compared to controls, the FBTCS-N group also had higher mean TW-RD in 18 out of 24 tracts (p < 0.05) and higher mean TW-AD in the inferior fronto-occipital fascicle , and the occipital striato and thalamo tracts . Interestingly, compared to the FBTCS-Y group, the FBTCS-N group showed significantly higher mean TW-TM in the inferior fronto-occipital fascicle, and occipital striato and thalamic tracts . The FBTCS-Y group also had significantly higher TW-AD than the controls in the middle cerebellar peduncle . p-values, and confidence intervals from all ANCOVAs between the patient subgroups and control group are reported in Group differences were observed in a number of TW-TM in the whole-tract comparison. Here, we report the significant OVAs see . When cop = 0.047 and p = 0.028, respectively). All mean differences, exact p-values and confidence intervals for the ANCOVAs between the FBTCS-Y, FBTCS-N, and Control groups are provided in Compared to the FBTCS-Y group, the significantly higher TW-ADC and TW-AD of the IFO tract in the FBTCS-N group was retained only in the right side . The tests of between subjects effects showed that as a covariate, drug resistance was only significant in the model for the TW-AD and TW-FA of the commissure anterior and did not change any prior results. Notably, significant TW-TM differences were not observed in the commissure anterior in the other grouping conditions.Compared to the drug responsive group, the drug resistant group had higher mean TW-AD and TW-FA in the commissure anterior (CA) and this difference was significant  taken in the context of the subcortical region, may be relevant to the seizure semiology observed in FBTCS. The region-specific alterations are likely a function of seizure pathology rather than whole-brain differences. The ensuing discussion focuses solely on the compelling results that could yield clinical utility. Of particular interest were the whole brain tracts, where significantly different mean TW-TM were observed between the patient subgroups. Readers may refer to the The first remarkable finding was the significant difference between the two patient subgroups in the TW-ADC and TW-AD of the inferior fronto-occipital fascicle (IFO), and striatal and thalamic occipital tracts (STOCC and TOCC), providing support for the second hypothesis. The thalamocortical circuit has been implicated as a critical mechanism for the genesis of FBTCS  by engagvia thalamostriatal projections may mediate generalized seizures  derived from the thalamo-prefrontal and thalamo-premotor tracts might be explained by premorbid connections in the subcortical structures. A recent application of magnetic resonance fingerprinting in normal appearing MRI scans from a focal epilepsy cohort revealed predominantly ipsilateral changes in the tissue properties of the thalamus and basal ganglia , suggestLastly, given the overall significant differences were observed between the FBTCS-N group and controls, the significantly higher TW-AD of the middle cerebellar peduncle in the FBTCS-Y group is noteworthy. Unlike the thalamic tracts, the middle cerebellar peduncle is cross-hemispheric, connecting the cerebellum to the pons, and is composed entirely of centripetal fibres. Previous work has shown the role of the middle cerebellar peduncle in generalized tonic-clonic seizures , potentiAlthough increased AD has been shown to result from white matter maturation, all groups were age-matched, suggesting the possibility that the higher TW-AD in the middle cerebellar peduncle of the FBTCS-Y group may be due to distinct premorbid FBTCS mechanisms. Since increased AD can also imply better organization of fibre structure, the TW-AD differences between the FBTCS-Y and FBTCS-N groups in the cerebellar region could be a feature of the FBTCS brain whereby those with FBTCS-Y have stronger premorbid fibre connections and the ability to sustain the intrahemispheric flow of ictal discharge. The middle cerebellar peduncle is presented as a compelling region of interest in post-surgical imaging, and future works could investigate this further.It is noted that although the inferior cerebellar peduncle makes up the peduncle group and is anatomically adjacent to the superior and middle cerebellar peduncles, it is neither functionally important in information relay nor inhibition, which may explain the absence of significant, between groups difference in this tract. Finally, there was no significant relationship between lesion presence and seizure onset side, or lesion presence and FBTCS, reinforcing the conjecture that the tract-specific TW-TM differences between the patient subgroups were due to premorbid connections rather than the effects of seizure injury. Our findings promote the possibility that propagation of ictal discharge from the epileptogenic zone to the contralateral hemisphere could be inhibited by premorbid damage to critical information relay and anatomically relevant tracts.Imaging studies of patients with epilepsy are traditionally limited by small sample sizes –21, 53, In summary, we used a robust and reproducible model to obtain track-specific TW-TM to show region-specific white matter alterations in a subgroup of patients with FBTCS, compared to those with focal epilepsy alone, and control participants. Our findings provide mechanistic insights into how structural changes may impact the functional role of the thalamic and striatal regions in individuals with FBCTS. We highlight specific tracts  that may be involved in FBTCS. Our results lay the foundation for a better understanding of seizure propagation in FBCTS and offer potential biomarkers that can help explain disease progression and aid treatment.The datasets generated and/or analyzed in the current study are not publicly available as they are from RPAH patients, and only individuals named on the approved ethics are authorized to access the data. However, de-identified data can be made available on reasonable request to the corresponding author, subject to approval from the relevant governing ethics entities.The studies involving human participants were reviewed and approved by the RPAH Sydney Local Health District (RPAH-SLHD) Ethics Committee. Written informed consent for participation was not required for this study in accordance with the national legislation and the institutional requirements.CM: conceptualization, methodology, data curation, imaging analysis, statistical analysis, manuscript writing, and review and editing. AD'S: imaging and diffusion analysis pipeline implementation, data curation and analysis, and manuscript revision. RZ: imaging pipeline—data acquisition and Tractseg implementation. MB and OK: conceptualization and manuscript revision. AN: clinical data acquisition, clinical advisory, conceptualization, methodology, and manuscript revision. CW: conceptualization, methodology, data curation, imaging analysis, statistical analysis, and manuscript revision. All authors contributed to the article and approved the submitted version.This study received funding from UCB. The funder was not involved in the study design, collection, analysis, interpretation of data, the writing of this article or the decision to submit it for publication.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 conversion of adenosine to inosine at the wobble position of select tRNAs is essential for decoding specific codons in bacteria and eukarya. In eukarya, wobble inosine modification is catalyzed by the heterodimeric ADAT complex containing ADAT2 and ADAT3. Human individuals homozygous for loss of function variants in ADAT3 exhibit intellectual disability disorders. We created a flexible computational tool to scan the human, mouse, nematode, fruit fly, and yeast exomes for genes either enriched or depleted in ADAT-dependent codons as compared to background models of codon bias derived from the exomes themselves. We find that many genes are enriched or depleted for ADAT-dependent codons as compared to the genomic background in all five species. Among those genes enriched for ADAT-dependent codons in humans, we find there is significant Gene Ontology (GO) enrichment for genes involved in diverse neurological processes. This pattern persists in the mouse exome but not the fruit fly or nematode exome. In the nematode exome, genes enriched in ADAT-dependent codons are GO enriched for translation associated genes, and in yeast there is GO enrichment for genes involved in metabolic functions. There is also GO-term overlap between yeast and fruit flies. Importantly, in its generalized form, ADATscan can also be used to scan any exome for genes enriched in any subset of codons specified by the user. E. coli or the ADAT2/3 heterodimeric complex in eukaryotes . Wobble inosine modification by ADAT2/3 is indispensable in eukaryotes for decoding certain codons, though the subsets are slightly variable depending on the specific eukaryotic organism . Human individuals homozygous for loss of function mutations in ADAT3 exhibit intellectual disability disorder , and in extreme cases, microcephaly . Previous studies have identified specific molecular defects associated with pathogenic ADAT3 variants that cause intellectual disability . However, the precise causes for human cognitive disorders caused by wobble inosine deficiency remain elusive.Modification of wobble adenosine to inosine in the anticodon of tRNAs is a conserved enzymatic reaction, catalyzed by the TadA homodimeric complex inTo screen for genes that may be the most (or the least) likely to be translationally impacted by decreased ADAT activity, we created a computational tool to scan exome data for enrichment or depletion of ADAT-dependent codons. Our methodology is to first establish a background model for ADAT-dependent codon usage in the exome (see Methods). This null model is then applied to each nonredundant gene entry in an exome FASTA file whereby an expected number of ADAT-dependent codons is calculated and compared to the observed number via simple chi-square test . These chi-square tests are then corrected for multiple comparisons  to yield a conservative list of genes that are enriched or depleted for ADAT-dependent codons. This tool also outputs the frequency of ADAT-dependent codons in each gene , the counts of these codons in each gene, and the p-values for enrichment or depletion of ADAT-dependent codons for each gene. The tool also outputs a file that allows for plotting ADAT-dependent codon frequencies across gene bodies .Using this tool, we identified 5952 human genes that are significantly depleted for ADAT-dependent codons and 6713 human genes that are significantly enriched for ADAT-dependent codons as compared to the human exomic background. Subsequent GO-term enrichment analysis showed that genes enriched in ADAT-dependent codons are biased for genes involved in neurological associated processes . The human genes identified represent candidates that may underlie the neurodevelopmental phenotypes seen in patients homozygous for loss of function variants in ADAT3. Notably, mouse genes enriched for ADAT-dependent codons also exhibit an overrepresentation of genes involved in neurological processes . In contrast, no enrichment for genes involved in neurological processes was found within the fruit fly or nematode exome . The nematode exome showed GO-term enrichment among those genes enriched for ADAT-dependent codons for translation associated genes. For the yeast exome, genes enriched for ADAT-dependent codons are GO-term enriched for genes involved in metabolic functions, and there is some GO-term overlap between fruit fly and yeast . There was no striking trend in terms of ADAT-dependent codon frequency across gene bodies among enriched, depleted, and nonsignificant genes . Altogether, these studies demonstrate that ADATscan can be used to predict which genes are presumably most dependent on ADAT for efficient translation and serve as a starting point for identifying biological processes linked to wobble inosine modification. Importantly, ADATscan is user-friendly and can be applied to any exome for any set of codons based on user input.Definition of ADAT-dependent codonhttp://gtrnadb.ucsc.edu/(Data Release 19). For humans, mice, nematodes, and fruit flies this codon set is: ACC, GCC, CCC, TCC, CTC, ATC, GTC, CGC. For yeast, the codon set is: TCC, CCC, CGC, CGA, ATC, ACC, GTC, GCC. Of note, humans, unlike most other eukaryotes, have three cognate tRNAs to decode ATC. However, these tRNAs are lowly expressed . As such, this codon was retained in the analysis for humans.The definition of an ADAT-dependent codon is a -C or -A ending codon which lacks a cognate tRNA for decoding it, and is thus dependent on wobble inosine modified tRNAs. This information can be found atBackground modelThe background model for ADAT-dependent codon usage is calculated using the relevant set of codons and the relevant exome data. Every codon in the exome is translated. If a codon corresponds to an amino acid that is encoded by an ADAT-dependent codon, then it is counted and recorded. This is done separately for all relevant amino acids. For each of these amino acids, the number of instances of ADAT-dependent codons are also counted. The latter count is divided by the former count for each relevant amino acid, yielding the background frequency of ADAT-dependent codon usage for each amino acid. This model is output to a file by ADATscan.Testing for enrichment and depletionFor each sequence in the exome file, amino acids that are encoded by ADAT-dependent codons are counted separately. These counts are then multiplied by their respective background frequency of ADAT-dependent codon usage in the exome as calculated in the background model. These numbers are then summed to yield a null estimate for the number of ADAT-dependent codons expected to appear in the protein based on amino acid composition and the background frequency of ADAT-dependent codon usage. This expectation is then compared to the observed number of ADAT-dependent codons in a 2x2 chi-square table that is filled as shown below:These tests are then corrected for multiple comparisons  to yield a conservative list of genes that are enriched or depleted for ADAT-dependent codons. In cases where more than 500 genes were enriched, the 500 genes with the smallest p-values were used for GO-term enrichment.https://ftp.ncbi.nlm.nih.gov/pub/CCDS/, current release: 12/11/22). CCDS IDs were converted to gene names for GO-term analysis using g:Profiler . The nematode exome data were retrieved from WormBase . Fruit fly exome data were retrieved from FlyBase . Yeast exome data were retrieved from theSaccharomycesGenome Database . GO-term enrichment was performed using PANTHER  for all species using a Fisher’s exact test and a Bonferroni correction . ADATscan and the outputs obtained in this study are freely available and can be found athttps://github.com/elonganrochester/ADATscan. Five versions are provided. One tailored to ADAT-dependent codon detection in CCDS, one tailored to ADAT-dependent codon detection in nematodes, one tailored to the files from Flybase, one tailored to ADAT-dependent codon detection in yeast, and a generalized user-friendly version that can take any codon set and any exome FASTA file as input. All versions allow for user input regarding the desired multiple-testing correction as either the Bonferroni correction or the Benjamini-Hochberg procedure with a user-specified FDR. An example of how to run ADATscan is also provided.Human and mouse exome data were retrieved from the CCDS database (10.22002/krvth-eb002Description: This file contains the results of ADATscan when the Bonferroni correction is applied. Resource Type: Dataset. DOI:10.22002/fs9vf-svg51Description: This file contains the results of ADATscan when the Benjamini-Hochberg procedure is applied. Resource Type: Dataset. DOI:10.22002/z1437-thb52Description: Link to github. Resource Type: InteractiveResource. DOI:


Myelin oligodendrocyte glycoprotein (MOG) antibody-associated disease (MOGAD) is an acquired inflammatory demyelinating disease with optic neuritis (ON) as the most frequent clinical symptom. The hallmark of the disease is the presence of autoantibodies against MOG (MOG-IgG) in the serum of patients. Whereas the role of MOG in the experimental autoimmune encephalomyelitis animal model is well-established, the pathogenesis of the human disease and the role of human MOG-IgG is still not fully clear.PubMed was searched for the terms “MOGAD,” “optic neuritis,” “MOG antibodies,” and “experimental autoimmune encephalomyelitis” alone or in combination, to find articles of interest for this review. Only articles written in English language were included and reference lists were searched for further relevant papers.B and T cells play a role in the pathogenesis of human MOGAD. The distribution of lesions and their development toward the optic pathway is influenced by the genetic background in animal models. Moreover, MOGAD-associated ON is frequently bilateral and often relapsing with generally favorable visual outcome. Activated T-cell subsets create an inflammatory environment and B cells are necessary to produce autoantibodies directed against the MOG protein. Here, pathologic mechanisms of MOG-IgG are discussed, and histopathologic findings are presented.MOGAD patients often present with ON and harbor antibodies against MOG. Furthermore, pathogenesis is most likely a synergy between encephalitogenic T and antibody producing B cells. However, to which extent MOG-IgG are pathogenic and the exact pathologic mechanism is still not well understood. Myelin oligodendrocyte glycoprotein (MOG), a minor component of myelin in the central nervous system (CNS), is expressed in the outermost layer of myelin . It is a789MOGAD is a rare disease with an incidence of 0.16/100,000 people , but the1415161718192021Despite the increasing knowledge of clinical MOGAD presentations, the pathophysiology and importantly, the pathogenic role of human MOG-IgG, remains to be fully determined. This review aims to summarize present studies on MOG-IgG pathology and pathogenesis of this rare inflammatory demyelinating disease with a focus on optic pathway involvement.The introduction of state-of-the-art cell-based assays for the detection of human MOG-IgG resulted in the characterization of a novel subset of ADS different from MS and NMOSD ,11,23. I1 and MOGβ1, both of which have this hydrophobic domain. However, most of the samples recognized all or most MOG isoforms tested, despite the lack of this domain  genotype and MOGAD development; importantly, no cause for disease pathogenesis has been found. A recent study found a protective effect of the HLA-C*03:04 allele , whereas4748495051Similar to the EAE animal, in human MOGAD, a synergy between encephalitogenic T cells and B cells is observed Fig. . Under n55565859606263646566686970In both cases, T-cells home to the brain after priming where they most likely enter the brain parenchyma through the meninges or the choroid plexus ,65,78,79Refs. 80818283858687As different T-cell subsets have diverse roles in immunopathogenesis, it is important to understand toward which lineages T cells are polarized. Different studies examining the cytokine/chemokine profiles in patients with MOGAD measured increased levels of T-helper (Th)17-related cytokines/chemokines , granulocyte-colony stimulating factor, Th1-related cytokines , and several B-cell associated factors  in cerebrospinal fluid (CSF) and serum 9293.92Tocilizumab (anti-IL-6 receptor antibody) is used off-label for the treatment of AQP4-IgG seropositive NMOSD and because of increased IL-6 levels in the CSF of MOGAD patients ,93, off-Increased neurofilament light chain levels were observed in the serum of MOGAD patients that also correlated with attack severity and could therefore serve as a potential biomarker . In addiIn MOGAD, the detection of MOG-specific T cells is still challenging. One study stimulated patient-derived peripheral blood mononuclear cells of MOG-IgG-positive patients with different MOG peptides, but could not find any specific proliferation. The authors suggest that the use of peptides could be insufficient for T-cell stimulation . AnotherAs MOGAD is associated with the presence of MOG-IgG, the question arises whether these antibodies are directly pathogenic, or the epiphenomena of a secondary immune response against MOG. Understanding this distinction can help to figure out the role of MOG-specific B cells in disease development. B cells can damage CNS tissue through diverse mechanisms including release of toxic exosomes and cytokines, antigen presentation to T cells, and antibody secretion  thickness revealed higher values in acute MOGAD-ON compared with MS because of optic disc edema. PRNFL thickening could serve as an indicator to distinguish MOGAD and MS in acute ON . After tIn EAE animal models immunized with MOG, the lesion distribution was determined by different influences such as gender, the genetic background and the immunization method used 153154. D153155156157Histopathologic examinations of animals revealed infiltration of inflammatory cells, demyelination with axonal loss, and reactive gliosis in retina and optic nerves ,157,160.167168To summarize, the spectrum of MOGAD-associated symptoms is broad, but most patients present with ON, that is usually associated with a good visual recovery. Histopathology revealed perivenous demyelinating lesions and infiltration of leukocytes. Nevertheless, the role of human MOG-IgG is less clear and different pathogenic mechanisms are discussed. Future studies that aim to define the exact pathogenesis, are needed to further identify targets for efficient treatment strategies.STATEMENT OF AUTHORSHIPConception and design: M. Lerch, M. Reindl; Acquisition of data: Not applicable; Analysis and interpretation of data: Not applicable. Drafting the manuscript: M. Lerch, A. Bauer, M. Reindl; Revising the manuscript for intellectual content: M. Lerch, A. Bauer, M. Reindl. Final approval of the completed manuscript: M. Lerch, A. Bauer, M. Reindl.