Increased Multiplexity in Optical Tissue Clearing-Based Three-Dimensional Immunofluorescence Microscopy of the Tumor Microenvironment by Light-Emitting Diode Photobleaching.

Optical tissue clearing and three-dimensional (3D) immunofluorescence (IF) microscopy is transforming imaging of the complex tumor microenvironment (TME). However, current 3D IF microscopy has restricted multiplexity; only 3 or 4 cellular and noncellular TME components can be localized in cleared tumor tissue. Here we report a light-emitting diode (LED) photobleaching method and its application for 3D multiplexed optical mapping of the TME. We built a high-power LED light irradiation device and temperature-controlled chamber for completely bleaching fluorescent signals throughout optically cleared tumor tissues without compromise of tissue and protein antigen integrity. With newly developed tissue mounting and selected region-tracking methods, we established a cyclic workflow involving IF staining, tissue clearing, 3D confocal microscopy, and LED photobleaching. By registering microscope channel images generated through 3 work cycles, we produced 8-plex image data from individual 400 µm-thick tumor macrosections that visualize various vascular, immune, and cancer cells in the same TME at tissue-wide and cellular levels in 3D. Our method was also validated for quantitative 3D spatial analysis of cellular remodeling in the TME after immunotherapy. These results demonstrate that our LED photobleaching system and its workflow offer a novel approach to increase the multiplexing power of 3D IF microscopy for studying tumor heterogeneity and response to therapy.

CX3CR1+ Macrophage Facilitates the Resolution of Allergic Lung Inflammation via Interacting CCL26.

Rationale: The resolution of inflammation is an active process coordinated by mediators and immune cells to restore tissue homeostasis. However, the mechanisms for resolving eosinophilic allergic lung inflammation triggered by inhaled allergens have not been fully elucidated. Objectives: Our objectives were to investigate the cellular mechanism of tissue-resident macrophages involved in the resolution process of eosinophilic lung inflammation. Methods: For the study, we used the institutional review board-approved protocol for human subsegmental bronchoprovocation with allergen, mouse models for allergic lung inflammation, and novel transgenic mice, including a conditional CCL26 knockout. The samples were analyzed using mass cytometry, single-cell RNA sequencing, and biophysical and immunological analyses. Measurements and Main Results: We compared alveolar macrophage (AM) subsets in the BAL before and after allergen provocation. In response to provocation with inhaled allergens, the subsets of AMs are dynamically changed in humans and mice. In the steady state, the AM subset expressing CX3CR1 is a relatively small fraction in bronchoalveolar space and lung tissue but drastically increases after allergen challenges. This subset presents unique patterns of gene expression compared with classical AMs, expressing high C1q family genes. CX3CR1+ macrophages are activated by airway epithelial cell-derived CCL26 via a receptor-ligand interaction. The binding of CCL26 to the CX3CR1+ receptor induces CX3CR1+ macrophages to secrete C1q, subsequently facilitating the clearance of eosinophils. Furthermore, the depletion of CX3CR1 macrophages or CCL26 in airway epithelial cells delays the resolution of allergic lung inflammation displaying prolonged tissue eosinophilia. Conclusions: These findings indicate that the CCL26-CX3CR1 pathway is pivotal in resolving eosinophilic allergic lung inflammation.

Commercially Sexually Exploited Adolescent Girls: The Association Between Externalizing Disorders and Parental Incarceration with Suicide Attempts.

Commercial sexual exploitation (CSE) is a significant public health concern disproportionately affecting ethnoracially minoritized girls. Despite strong associations of CSE with suicide attempts, little is known about correlates of suicide among girls with CSE histories. Elevated rates of externalizing disorders (i.e., attention-deficit/hyperactivity disorder [ADHD], disruptive behavior disorders [DBD], conduct disorder [CD]) were observed among CSE youth, particularly in ethnoracially minoritized samples. Youth with CSE histories are frequently affected by parental incarceration, which is correlated with risk for suicide attempts. We tested cross-sectional simultaneous associations of externalizing disorders and parental incarceration with number of suicide attempts among 360 ethnoracially diverse girls affected by CSE (Mean age = 18.86). ADHD, DBD, and maternal incarceration were positively associated with number of suicide attempts. Findings implicate clinical/familial correlates of suicide attempts in this marginalized group, suggesting early suicide prevention efforts may improve traction on this problem by focusing on individual and family level factors.

Multiresolution 3D Optical Mapping of Immune Cell Infiltrates in Mouse Asthmatic Lung.

Asthma is a chronic inflammatory airway disease driven by various infiltrating immune cell types into the lung. Optical microscopy has been used to study immune infiltrates in asthmatic lungs. Confocal laser scanning microscopy (CLSM) identifies the phenotypes and locations of individual immune cells in lung tissue sections by employing high-magnification objectives and multiplex immunofluorescence staining. In contrast, light-sheet fluorescence microscopy (LSFM) can visualize the macroscopic and mesoscopic architecture of whole-mount lung tissues in three dimensions (3D) by adopting an optical tissue-clearing method. Despite each microscopy method producing image data with unique resolution from a tissue sample, CLSM and LSFM have not been applied together because of different tissue-preparation procedures. Here, we introduce a new approach combining LSFM and CLSM into a sequential imaging pipeline. We built a new optical tissue clearing workflow in which the immersion clearing agent can be switched from an organic solvent to an aqueous sugar solution for sequential 3D LSFM and CLSM of mouse lungs. This sequential combination microscopy offered quantitative 3D spatial analyses of the distribution of immune infiltrates in the same mouse asthmatic lung tissue at the organ, tissue, and cell levels. These results show that our method facilitates multiresolution 3D fluorescence microscopy as a new imaging approach providing comprehensive spatial information for a better understanding of inflammatory lung diseases.

Three-dimensional spatial quantitative analysis of cardiac lymphatics in the mouse heart.

OBJECTIVE: Three-dimensional (3D) microscopy and image data analysis are necessary for studying the morphology of cardiac lymphatic vessels (LyVs) and their association with other cell types. We aimed to develop a methodology for 3D multiplexed lightsheet microscopy and highly sensitive and quantitative image analysis to identify pathological remodeling in the 3D morphology of LyVs in young adult mouse hearts with familial hypertrophic cardiomyopathy (HCM). METHODS: We developed a 3D lightsheet microscopy workflow providing a quick turn-around (as few as 5-6 days), multiplex fluorescence detection, and preservation of LyV structure and epitope markers. Hearts from non-transgenic and transgenic (TG) HCM mice were arrested in diastole, retrograde perfused, immunolabeled, optically cleared, and imaged. We built an image-processing pipeline to quantify LyV morphological parameters at the chamber and branch levels. RESULTS: Chamber-specific pathological alterations of LyVs were identified, and significant changes were seen in the right atrium (RA). TG hearts had a higher volume percent of ER-TR7+ fibroblasts and reticular fibers. In the RA, we found associations between ER-TR7+ volume percent and both LyV segment density and median diameter. CONCLUSIONS: This workflow and study enabled multi-scale analysis of pathological changes in cardiac LyVs of young adult mice, inviting ideas for research on LyVs in cardiac disease.

Genome sequence and genetic diversity of European ash trees.

Ash trees (genus Fraxinus, family Oleaceae) are widespread throughout the Northern Hemisphere, but are being devastated in Europe by the fungus Hymenoscyphus fraxineus, causing ash dieback, and in North America by the herbivorous beetle Agrilus planipennis. Here we sequence the genome of a low-heterozygosity Fraxinus excelsior tree from Gloucestershire, UK, annotating 38,852 protein-coding genes of which 25% appear ash specific when compared with the genomes of ten other plant species. Analyses of paralogous genes suggest a whole-genome duplication shared with olive (Olea europaea, Oleaceae). We also re-sequence 37 F. excelsior trees from Europe, finding evidence for apparent long-term decline in effective population size. Using our reference sequence, we re-analyse association transcriptomic data, yielding improved markers for reduced susceptibility to ash dieback. Surveys of these markers in British populations suggest that reduced susceptibility to ash dieback may be more widespread in Great Britain than in Denmark. We also present evidence that susceptibility of trees to H. fraxineus is associated with their iridoid glycoside levels. This rapid, integrated, multidisciplinary research response to an emerging health threat in a non-model organism opens the way for mitigation of the epidemic.

Peer correlates of conduct problems in girls.

Conduct problems are increasingly prevalent in girls and they uniquely predict negative outcomes. Yet, few reliable risk factors for aggression and violence in girls and women have been identified. Although preliminary evidence suggests peer relationships may be central to the development of youth conduct problems, especially in girls, rigorous interactive models of peer risk and protective factors for conduct problems are lacking. Based on 3104 10-13-year-old girls in the Adolescent Brain and Cognitive Development study, we tested the independent associations of separate peer risk factors (i.e., relational aggression victimization, physical aggression victimization, and deviant peer affiliation) with multidimensional conduct problems, including their moderation by peer support. Being the victim of relational aggression, being the victim of physical aggression, and deviant peer affiliation were each positively associated with conduct problems and perpetration of aggression whereas peer support was negatively associated with youth report conduct problems and perpetration of physical aggression. Further, elevated peer support significantly attenuated the association of being the victim of relational aggression with teacher-rated conduct problems. These results highlight the sensitivity of conduct problems to peer risk factors and suggest that peer support designates important configurations of risk that differentially relate to conduct problems in girls.

Referral Patterns Associated With Delayed Care for Traumatic Brachial Plexus Injuries.

PURPOSE: Traumatic brachial plexus injuries (BPI) are devastating, time-sensitive conditions that often require definitive treatment at academic tertiary care centers. Delays to presentation and surgery have been associated with inferior outcomes. In this study, we evaluate referral patterns associated with delayed presentation and late surgery in traumatic BPI patients. METHODS: Patients diagnosed with a traumatic BPI at our institution from 2000 to 2020 were identified. Medical charts were reviewed for demographic characteristics, prereferral work-up, and referring provider characteristics. Delayed presentation was defined as greater than 3 months from date of injury to initial evaluation by our brachial plexus specialists. Late surgery was defined as greater than 6 months from date of injury. Multivariable logistic regression was used to identify factors associated with delayed presentation or surgery. RESULTS: A total of 99 patients were included, 71 of whom underwent surgery. Sixty-two patients presented delayed (62.6%), with 26 receiving late surgery (36.6%). There were similar rates of delayed presentation or late surgery by referring provider specialty. Patients whose initial diagnostic EMG was ordered by the referring provider prior to initial presentation at our institution were more likely to have a delayed presentation (76.2% vs 31.3%) and undergo late surgery (44.9% vs 10.0%). CONCLUSIONS: Delayed presentation and late surgery in traumatic BPI patients were associated with initial diagnostic EMG ordered by the referring provider. CLINICAL RELEVANCE: Delayed presentation and surgery have been associated with inferior outcomes in traumatic BPI patients. We recommend that providers direct patients with clinical concern of traumatic BPI directly to a brachial plexus center without further work-up before referral and encourage referral centers to accept these patients.

The Influence of Psychosocial Factors on Disability and Expected Improvement Before Surgery for Adult Traumatic Brachial Plexus Injury.

PURPOSE: Psychosocial factors influence pain and recovery after extremity trauma and may be targets for early intervention. This may be of particular interest for patients with adult traumatic brachial plexus injury (BPI), given the broad and devastating impact of the injury. We hypothesized that there would be an association between depressive symptoms, anxiety, and pain interference with preoperative disability and expectations for improvement after BPI surgery. METHODS: We enrolled 34 patients into a prospective multicenter cohort study for those undergoing surgery for adult traumatic BPI. Before surgery, participants completed Patient-Reported Outcome Measurement Information System scales for pain interference, anxiety, and depressive symptoms, and a validated BPI-specific measure of disability and expected improvement. We performed Pearson correlation analysis between pain interference, anxiety symptoms, and depressive symptoms with (A) disability and (B) expected improvement. We created separate linear regression models for (A) disability and (B) expected improvement including adjustment for severity of plexus injury, age, sex, and race. RESULTS: Among 34 patients, there was a moderate, statistically significant, correlation between preoperative depressive symptoms and higher disability. This remained significant in a linear regression model adjusted for severity of plexus injury, age, sex, and race. There was no association between severity of plexus injury and disability. Depressive symptoms also were moderately, but significantly, correlated with higher expected improvement. This remained significant in a linear regression model adjusted for severity of plexus injury, age, sex, and race. CONCLUSIONS: Depressive symptoms are associated with greater disability and higher expected improvement before BPI surgery. Screening for depressive symptoms can help BPI teams identify patients who would benefit from early referral to mental health specialists and tailor appropriate expectations counseling for functional recovery. We did not find an association between severity of BPI and patient-reported disability, suggesting either that the scale may lack validity or that the sample is biased. LEVEL OF EVIDENCE: Prognostic II.

Attention Bias and Anxiety: The Moderating Effect of Sociocultural Variables in Rural Latinx Youth.

Attention bias confers risk for anxiety development, however, the influence of sociodemographic variables on the relationship between attention bias and anxiety remains unclear. We examined the association between attention bias and anxiety among rural Latinx youth and investigated potential moderators of this relationship. Clinical symptoms, demographic characteristics, and a performance-based measure of attention bias were collected from 66 rural Latinx youth with clinical levels of anxiety (33.3% female; Mage = 11.74; 92.4% Latinx, 7.6% Mixed Latinx). No moderating effects for age or gender were found. Youth below the poverty line displayed an attention bias away from threat in comparison to youth above the poverty line, who displayed an attention bias towards threat. Among youth below the poverty line, this bias away from threat was associated with increased anxiety. Findings highlight the importance of economic adversity in understanding the relationship between attention bias and anxiety.

Targeting Tumor Microenvironment Akt Signaling Represents a Potential Therapeutic Strategy for Aggressive Thyroid Cancer.

Effects of the tumor microenvironment (TME) stromal cells on progression in thyroid cancer are largely unexplored. Elucidating the effects and underlying mechanisms may facilitate the development of targeting therapy for aggressive cases of this disease. In this study, we investigated the impact of TME stromal cells on cancer stem-like cells (CSCs) in patient-relevant contexts where applying in vitro assays and xenograft models uncovered contributions of TME stromal cells to thyroid cancer progression. We found that TME stromal cells can enhance CSC self-renewal and invasiveness mainly via the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) pathway. The disruption of Akt signaling could diminish the impact of TME stromal cells on CSC aggressiveness in vitro and reduce CSC tumorigenesis and metastasis in xenografts. Notably, disrupting Akt signaling did not cause detectable alterations in tumor histology and gene expression of major stromal components while it produced therapeutic benefits. In addition, using a clinical cohort, we discovered that papillary thyroid carcinomas with lymph node metastasis are more likely to have elevated Akt signaling compared with the ones without metastasis, suggesting the relevance of Akt-targeting. Overall, our results identify PI3K/Akt pathway-engaged contributions of TME stromal cells to thyroid tumor disease progression, illuminating TME Akt signaling as a therapeutic target in aggressive thyroid cancer.

Beating the Stoner criterion using molecular interfaces.

Only three elements are ferromagnetic at room temperature: the transition metals iron, cobalt and nickel. The Stoner criterion explains why iron is ferromagnetic but manganese, for example, is not, even though both elements have an unfilled 3d shell and are adjacent in the periodic table: according to this criterion, the product of the density of states and the exchange integral must be greater than unity for spontaneous spin ordering to emerge. Here we demonstrate that it is possible to alter the electronic states of non-ferromagnetic materials, such as diamagnetic copper and paramagnetic manganese, to overcome the Stoner criterion and make them ferromagnetic at room temperature. This effect is achieved via interfaces between metallic thin films and C60 molecular layers. The emergent ferromagnetic state exists over several layers of the metal before being quenched at large sample thicknesses by the material's bulk properties. Although the induced magnetization is easily measurable by magnetometry, low-energy muon spin spectroscopy provides insight into its distribution by studying the depolarization process of low-energy muons implanted in the sample. This technique indicates localized spin-ordered states at, and close to, the metal-molecule interface. Density functional theory simulations suggest a mechanism based on magnetic hardening of the metal atoms, owing to electron transfer. This mechanism might allow for the exploitation of molecular coupling to design magnetic metamaterials using abundant, non-toxic components such as organic semiconductors. Charge transfer at molecular interfaces may thus be used to control spin polarization or magnetization, with consequences for the design of devices for electronic, power or computing applications (see, for example, refs 6 and 7).

Role of high-resolution peripheral nerve magnetic resonance imaging in diagnosing median nerve tethering in a case of both-bone forearm fracture in a child.

Forearm fractures are common injuries in pediatric patients. We present a case of median nerve tethering as a complication of both-bone forearm fracture in a child, with an emphasis on MRI as an appropriate and important complement to clinical and electrodiagnostic examination. Early intervention is essential because delayed surgical management of median nerve tethering can result in poor clinical outcomes as a result of irreversible muscle denervation. In this case, we highlight the importance of MRI to facilitate management, including early surgical intervention when appropriate, in median neuropathy following forearm fractures.

Long-term radiographic outcomes and functional evaluation of ulnar shortening osteotomy in patients with ulnar impaction syndrome and reverse oblique sigmoid notch: a retrospective case series study.

BACKGROUND: Ulnar shortening osteotomy (USO) is an effective treatment for ulnar impaction syndrome. However, there have been reports of osteoarthritis (OA) at the distal radioulnar joint (DRUJ) when USO was performed on patients with a reverse oblique sigmoid notch. This study aimed to evaluate the radiographic and functional outcomes following USO in patients with a reverse oblique sigmoid notch. METHODS: We retrospectively reviewed patients having a reverse oblique sigmoid notch who underwent USO for ulnar impaction syndrome between 2002 and 2013. We evaluated radiographic changes of the DRUJ and functional outcomes of patients. RESULTS: We enrolled 22 patients (22 wrists) with an average age of 49.6 years and a mean follow-up of 93.2 (range, 36-179; standard deviation [SD], 38.2) months. We found that there were changes in the inclination angle of the sigmoid notch, from an average reverse oblique of 14.9o (range, 11o-23o; SD, 3.4o) preoperatively to a more parallel 5.1o (range, 0o-11o; SD, 3.2o) at the final follow-up. The functional results at the final follow-up were good, with a mean visual analogue scale for pain of 0.2 (range, 0-1; SD, 0.4) at rest and 1.3 (range, 0-3; SD, 0.9) during activity, QuickDASH of 15.1 (range, 2.3-34.1; SD, 8.8), and modified Mayo Wrist Score of 91.6 (range, 70-100; SD, 6.4). Seven wrists (31.8%) had changes compatible with OA, but the wrists did not exhibit a significantly worse function when compared to wrists without OA changes, except for supination motion and grip strength. CONCLUSIONS: For patients with a reverse oblique sigmoid inclination following USO, we observed that the inclination angle had a tendency to become parallel and some patients developed OA at the DRUJ. However, long-term functional outcomes could still be good. The reverse oblique sigmoid inclination does not seem to be an absolute contraindication for USO.

Gene x responsive parenting interactions in social development: Characterizing heterogeneity in autism spectrum disorder.

Emerging research suggests that caregiving environments and genetic variants independently contribute to social functioning in children with typical development or autism spectrum disorder (ASD). However, biologically plausible interactive models and complimentary assessment of mechanisms are needed to: (a) explain considerable social heterogeneity, (b) resolve inconsistencies in the literature, and (c) develop and select optimal treatments based on individual differences. This study examined the role of child genotypes and responsive parenting in the social development of 104 children with ASD (ages 4-7 years). We utilized a longitudinal, multi-informant design and structural equation models to evaluate: (a) the additive and interactive effects of biologically plausible candidate genes (5-HTTLPR, OXTR, DRD4) and responsive parenting in predicting prospective social development in ASD across three time points spanning 1.5 years, and (b) whether child emotion regulation mediated observed gene x environment interactions (GxEs). Responsive parenting positively predicted prospective change in child social skills; these associations were moderated by 5-HTTLPR and DRD4 in teacher-report models, and DRD4 in parent-report models. No GxE effects were found for OXTR. Emotion regulation did not significantly mediate the GxEs involving 5-HTTLPR and DRD4. Acknowledging the complexities of GxE research, implications for future research, and targeted intervention efforts are discussed.

Paternal expressed emotion influences psychobiological indicators of threat and safety learning in daughters: A preliminary study.

This preliminary study examined the association of children's anxiety, paternal expressed emotion (EE), and their interaction with psychophysiological indices of children's threat and safety learning. Participants included 24 father-daughter dyads. Daughters (ages 8-13 years, 100% Latina) self-reported their anxiety levels and completed a differential threat conditioning and extinction paradigm, during which psychophysiological responding was collected. Fathers completed a Five-Minute Speech Sample, from which paternal EE (i.e., criticism, emotional overinvolvement) was assessed. Anxiety-dependent associations emerged between paternal EE and individual differences in daughters' psychophysiological responding to safety signals during threat conditioning. Paternal EE was positively associated with psychophysiological responding to safety in daughters with high and mean, but not low, levels of anxiety. Although previous work suggests that chronic harsh maternal parenting is a potential risk factor for children's general threat and safety learning, these preliminary findings implicate milder forms of negative parenting behavior in fathers, particularly for highly anxious children.

Endonasal resection of orbital cavernous venous malformations with septal preservation.

PURPOSE: Multiple surgical approaches have been described to maximize visualization and accessibility for resection while minimizing morbidity in the patient with orbital intraconal tumors. Transnasal endoscopic approaches have become increasingly standard in select orbital cavernous venous malformations but often require a partial septectomy. The purpose of this manuscript is to communicate a septal preserving modified transseptal approach. METHODS: A 37-year old male was found to have an inferomedial intraconal orbital mass, measuring up to 2.6 cm on magnetic resonance imaging. Binarial transseptal access with septal preservation was obtained with a Killian incision on the right and a small incision in the midseptum on the left. RESULTS: Successful tumor delivery through the nasal cavity resulted in orbital relaxation. Postoperative evaluation of the septum demonstrated an intact septum with nearly no evidence of septal trauma from surgical manipulation. CONCLUSION: This technique is easily performed and affords adequate visualization and freedom of movement as traditional binarial transseptal approaches without the disadvantages of partial septal loss such as increased crusting, olfactory disturbance, and loss of nasoseptal flaps.

Parenting behavior and growth of child conduct problems: Moderation by callous-unemotional traits.

Although positive parenting behavior is central to efficacious interventions for child conduct problems (CP), studies of youth CP have focused mostly on negative parenting behavior. That is, few studies have examined dimensions of positive parenting behavior (e.g., positive reinforcement, involvement) as independent predictors of CP and even fewer have investigated their potential moderation by callous-unemotional (CU) traits. A sample of 184 6-9 year-old children with and without attention-deficit/hyperactivity disorder (ADHD) was followed prospectively for two years. Controlling for baseline ADHD diagnostic status, initial CP, and negative parenting (i.e., corporal punishment), we examined CU traits, positive reinforcement and involvement, and their interactions as predictors of two-year change in CP. Positive reinforcement and CU traits independently predicted increased rule breaking behavior whereas parental involvement inversely predicted aggressive behavior. A significant positive reinforcement x CU traits interaction suggested that positive reinforcement predicted a decrease in aggressive behavior, but only in children with low CU traits; conversely, positively reinforcement marginally predicted increased aggressive behavior among children with high CU traits. No other significant parenting x CU traits interaction was observed. We consider these findings within a developmental psychopathology framework where interactive exchanges underlie the development of CP.

Social skills moderate the time-varying association between aggression and peer rejection among children with and without ADHD.

Although childhood aggression is typically associated with peer rejection, some children concurrently employ coercive and socially skilled behavior and successfully avoid negative peer outcomes. However, research on children's dual use of coercive and social behavior has largely employed cross-sectional designs with nonclinical populations and, as a result, little is known about the covariation of aggression with social skills, particularly among high-risk samples. We directly addressed this limitation by testing childhood aggression and social skills as separate time-varying predictors of prospective change in peer rejection in a sample of children with and without attention-deficit/hyperactivity disorder (ADHD). Two hundred and two 5-10-year-old children (M = 7.9 years, SD = 1.2) with and without ADHD were followed prospectively for 6 years. Key constructs, including children's overt aggression, social skills, and peer rejection, were collected at each of the three waves using multiple methods and informants. Controlling for demographic factors and time-varying ADHD symptoms, longitudinal change in child-, parent-, and teacher-reported aggression positively predicted prospective change in parent- and teacher-reported peer rejection. Importantly, predictions were moderated by parent- and teacher-reported social skills, such that aggression inversely predicted peer rejection for children with high social skills. These results demonstrate that social skills meaningfully alter trajectories of peer rejection predicted from cross-time variation in aggression. We discuss the theoretical and empirical implications of these findings within a developmental psychopathology framework, including recommendations for directions for future research.

Comparative Analysis of 3 Techniques of Scapholunate Reconstruction for Dorsal Intercalated Segment Instability.

PURPOSE: The combination of scaphoid rotatory subluxation, dorsal intercalated segment instability, and dorsal scaphoid translation (DST) constitutes stage 5 scapholunate dissociation in the modified classification system of Garcia-Elias. The purpose of this study was to compare the static radiographic outcomes of reduction and association of the scaphoid and lunate (RASL), three-ligament tenodesis (TLT), and anatomic front and back (ANAFAB) reconstructions for stage 5 scaphoid and lunate malrotation and translation. METHODS: Stage 5 scapholunate dissociation was created in 15 fresh-frozen specimens by cutting the scapholunate interosseous ligament, long radiolunate ligament, dorsal intercarpal ligament, and scaphotrapeziotrapezoid ligament complex. Specimens were randomized to receive 1 of the 3 reconstructive techniques. Radiolunate angle (RLA), scapholunate angle, scapholunate gap, and DST were measured in static fluoroscopic posteroanterior and lateral views in 3 conditions: baseline, injured, and reconstructed. RESULTS: Reduction and association of the scaphoid and lunate showed an improved correction of scapholunate gap compared with TLT and ANAFAB. Anatomic front and back demonstrated an improved correction in RLA and scapholunate angle compared with RASL but not TLT. When the reconstruction was compared with baseline, there was a persistent lunate extension for TLT (RLA and scapholunate angle) and RASL (RLA); however, no significant difference in any parameter was found with ANAFAB. Anatomic front and back significantly improved DST, whereas TLT and RASL did not. CONCLUSIONS: In this cadaveric model of scapholunate dissociation with dorsal intercalated segment instability and DST, each of the 3 repairs had different effects on carpal posture and alignment. While only RASL statistically improved scapholunate gap, only ANAFAB significantly improved DST. Both TLT and RASL improved radiographic parameters; however, each had a persistent increase in the lunate extension following reconstruction. CLINICAL RELEVANCE: It may be prudent to consider different reconstructive techniques for different stages of scapholunate instability. Anatomic front and back, a repair that addresses both dorsal and volar ligament stabilizers, provides improved reduction of dorsal intercalated segment instability and DST in stage 5 scapholunate dissociation.