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Background: In direct-to-implant breast reconstruction, accurate preoperative breast volume estimation is crucial for surgeons to select the appropriate implant volume, considering the cosmetic outcomes during surgery. We proposed the prediction model for intraoperative implant volume based on the preoperative estimated volume of the contralateral breast obtained through a three-dimensional surface imaging system (3DSI) as surgeons usually choose the implant volume on the breast which should be reconstructed considering symmetricity with the contralateral breast. Methods: We enrolled 97 patients from our single institution who underwent unilateral mastectomy with immediate breast reconstruction using smooth silicone implants between October 2021 and January 2023. Preoperatively, plastic surgeons measured the volume of the contralateral breast using the VECTRA XT 3D imaging system. Data on implant volume and the types of acellular dermal matrix used during surgery, determined by a single surgeon to ensure symmetry, were also collected. Linear regression analysis was utilized to construct the predictive model. Results: In the multiple linear regression analysis with preoperative contralateral breast volume, age, and body mass index as variables, the coefficient of determination of the model expressed as R squared (R2) was 0.554, and except for age, the other variables were statistically significant. When replaced by mastectomy volume instead of age, R2 increased to 0.723 and all variables were significant. Conclusions: 3DSI can be helpful for preoperative surgical planning and postoperative outcome simulation. With our multiple linear regression model, we can predict the intraoperative implant volume using preoperative contralateral breast volume measured by the 3D scans.

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BACKGROUND: Quantitative biomarkers of facial skin aging were investigated in 109 healthy Asian female volunteers, aged 20 to 70 years. MATERIALS AND METHODS: In vivo 3D Line-field Confocal Optical Coherence Tomography (LC-OCT) imaging, enhanced by Artificial Intelligence (AI)-based quantification algorithms, was utilized to compute various metrics, including stratum corneum thickness (SC), viable epidermal (VE) thickness, and Dermal-Epidermal Junction (DEJ) undulation along with cellular metrics for the temple, cheekbone, and mandible. RESULTS: Comparison with data from a cohort of healthy Caucasian volunteers revealed similarities in the variations of stratum corneum and viable epidermis layers, as well as cellular shape and size with age in both ethnic groups. However, specific findings emerged, such as larger, more heterogeneous nuclei in both layers, demonstrated by an increase in nuclei volume and their standard deviation, and increased network atypia, all showing significant age-related variations. Caucasian females exhibited a flatter and more homogeneous epidermis, evidenced by a decreased standard deviation of the number of layers, and a less dense cellular network with fewer cells per layer, indicated by a decrease in cell surface density. CONCLUSION: Ethnicity-wise comparisons highlighted distinct biological features specific to each population. Asian individuals showed significantly higher DEJ undulation, higher compactness, and lower cell network atypia compared to their Caucasian counterparts across age groups. Differences in stratum corneum and viable epidermal thickness on the cheekbone were also significant. LC-OCT 3D imaging provides valuable insights into the aging process in different populations and underscores inherent biological differences between Caucasian and Asian female volunteers.

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A comprehensive understanding of physio-pathological processes necessitates non-invasive intravital three-dimensional (3D) imaging over varying spatial and temporal scales. However, huge data throughput, optical heterogeneity, surface irregularity, and phototoxicity pose great challenges, leading to an inevitable trade-off between volume size, resolution, speed, sample health, and system complexity. Here, we introduce a compact real-time, ultra-large-scale, high-resolution 3D mesoscope (RUSH3D), achieving uniform resolutions of 2.6 × 2.6 × 6 µm3 across a volume of 8,000 × 6,000 × 400 µm3 at 20 Hz with low phototoxicity. Through the integration of multiple computational imaging techniques, RUSH3D facilitates a 13-fold improvement in data throughput and an orders-of-magnitude reduction in system size and cost. With these advantages, we observed premovement neural activity and cross-day visual representational drift across the mouse cortex, the formation and progression of multiple germinal centers in mouse inguinal lymph nodes, and heterogeneous immune responses following traumatic brain injury-all at single-cell resolution, opening up a horizon for intravital mesoscale study of large-scale intercellular interactions at the organ level.

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INTRODUCTION/OBJECTIVES: To compare conventional and three-dimensional (3D) echocardiographic indices of right ventricular (RV) systolic function in dogs with various stages of myxomatous mitral valve disease (MMVD), classified according to the 2009 guidelines of the American College of Veterinary Internal Medicine (ACVIM), with those from normal dogs. ANIMALS: Seventy-eight unsedated dogs (22 healthy controls, 23 ACVIM stage B1 MMVD, 20 ACVIM stage B2 MMVD, and 13 ACVIM stage C MMVD) were included in the study. MATERIALS AND METHODS: All dogs underwent conventional and 3D echocardiography. Three-dimensional RV end-diastolic volume (EDV), end-systolic volume (ESV), stroke volume (SV), and ejection fraction (EF) were recorded. Right ventricular EDV, ESV, and SV were indexed to bodyweight. Echocardiographic variables were compared across groups using a Kruskal-Wallis test with subsequent post hoc analysis using Dunn's method for multiple comparisons between groups. A P-value of <0.05 was considered significant. RESULTS: Right ventricular EDV was smaller in stage B1 (P=0.012), stage B2 (P=0.035), and stage C (P=0.004) dogs than in controls. Stage B2 (P=0.003) and stage C (P<0.001) dogs had smaller RV ESV than controls. Stage B1 dogs had smaller RV SV than controls (P=0.012). Right ventricular EF was greater in stage C dogs than in controls (P=0.003) and in stage B1 (P=0.017) dogs. CONCLUSIONS: Several 3D echocardiographic indices of RV systolic function differ between dogs with advanced MMVD when compared with normal dogs. Further investigation is required to determine if these differences have clinical implications.

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BACKGROUND: Osteonecrosis of the femoral head (ONFH) affects at least 20,000 patients annually in the United States; however, the pathophysiology of disease progression is poorly understood. The purpose of this study was to determine the relative importance of three distinct elements and their relationship to the collapse of the femoral head: (1) identifiable risk factors; (2) femoral head anatomy; and (3) the extent of the necrotic lesion. METHODS: A single-center retrospective cohort study was performed on patients ≥ 18 years old who presented with ONFH. Ficat classification and femoral head anatomic parameters were measured on radiographs. Osteonecrotic lesion size was measured on magnetic resonance imaging using four validated methods. Multivariable regression analyses were performed to identify predictors of femoral head collapse. RESULTS: There were 105 patients and 137 hips included in the final cohort, of which 50 (36.5%) had collapse of the femoral head. Multivariable analyses demonstrated that medical risk factors (adjusted odds ratio (aOR): 1.15), alcohol exposure (aOR: 1.23), and increased alpha angle (aOR: 4.51) were predictive of femoral head collapse. Increased femoral head offset (aOR: 0.54) was protective against collapse. An increased size of the osteonecrotic lesion was significantly predictive of collapse with all four measure methods evaluated: three-dimensional (3D) volumetric (aOR: 3.73), modified Kerboul (aOR: 2.92), index of necrotic extent (aOR: 1.91), and modified index of necrotic extent (aOR: 2.05). CONCLUSIONS: In an analysis of patients who had ONFH, we identified risk factors such as alcohol exposure, high alpha angle, increased lesion size, and decreased femoral offset as increasing the risk of femoral head collapse. Given the challenges of studying this patient population, large prospective studies of patients who have ONFH should seek to identify whether these factors are reliable indicators of femoral head collapse.

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Background: The blood-brain barrier (BBB) is part of the neurovascular unit (NVU) which plays a key role in maintaining homeostasis. However, its 3D structure is hardly known. The present study is aimed at imaging the BBB using tissue clearing and 3D imaging techniques in both human brain tissue and rat brain tissue. Methods: Both human and rat brain tissue were cleared using the CUBIC technique and imaged with either a confocal or two-photon microscope. Image stacks were reconstructed using Imaris. Results: Double staining with various antibodies targeting endothelial cells, basal membrane, pericytes of blood vessels, microglial cells, and the spatial relationship between astrocytes and blood vessels showed that endothelial cells do not evenly express CD31 and Glut1 transporter in the human brain. Astrocytes covered only a small portion of the vessels as shown by the overlap between GFAP-positive astrocytes and Collagen IV/CD31-positive endothelial cells as well as between GFAP-positive astrocytes and CD146-positive pericytes, leaving a big gap between their end feet. A similar structure was observed in the rat brain. Conclusions: The present study demonstrated the 3D structure of both the human and rat BBB, which is discrepant from the 2D one. Tissue clearing and 3D imaging are promising techniques to answer more questions about the real structure of biological specimens.

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OBJECTIVE: Graft failure is a common complication of cranioplasty.Revision cranioplasty is required to overcome this complication. However, no previous studies have reported outcomes in revision cranioplasty with three-dimensional (3D) custom-made titanium implants. We described our experience with 3D titanium implants in patients with revision cranioplasty. METHODS: We evaluated 43 consecutive patients who underwent revision cranioplasty using 3D custom-made titanium implants between January 2011 and December 2019.The 3D image of the patient's cranium and the plan to close the cranium defect were created in a virtual environment using software programs. Demographic and radiological features were compared based on the materials used in the initial cranioplasty. RESULTS: Previous material was autologous graft (AG) in 27 patients and polymethyl methacrylate (PMMA) in 16 patients. The mean time without revision cranioplasty is longer in patients with PMMA implants than in patients with AG. There was no statistically significant difference in the length of hospital stay between patients with PMMA implants and patients with AG. There were no postoperative adverse events such as infection, wound dehiscence, convulsions, and epidural hematoma in 38 patients during hospitalization. Wound dehiscence developedin 5 patients and surgical repair was required in one. CONCLUSION: Initial cranioplasty with PMMA provides a longer time period than the AG before the revision. However, both of them have similar outcomes based on length of hospital stay and cranial defect area. Custom-made 3D titanium implant is a good option for revision cranioplasty to prevent implant failure and reduce patients' cosmetic concerns.

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This prospective study aimed to compare ultra-high molecular weight polyethylene (UHMWPE) with polyetheretherketone (PEEK) in computer-designed patient-specific implants (PSIs) for cranial defect reconstruction, in terms of complications and aesthetic outcomes. Primary or secondary cranioplasty-eligible patients were included, while patients with active infection or hydrocephalus, or unfit for general anesthesia, were excluded from the study. All the implants were designed and fabricated by the same maxillofacial surgeon using CAD/CAM technology. UHMWPE PSIs were used in group 1 and PEEK PSIs in group 2. Technically, UHMWPE could be milled to a thinner margin thickness than PEEK, which resulted in better handling properties and a smoother end finish. All patients were evaluated over a period of 6 months in terms of overall complications or implant failure as the primary outcome, according to Clavien-Dindo (CVD) grading, and cosmetic satisfaction with the aesthetic results, using a Likert scale, as the secondary outcome. In total, 22 cranioplasty patients were included, with a mean age of 30.8 years (SD = 16.3). Across both groups, 17 patients (77.3%) did not develop postoperative complications. These occurred in three patients in group 1 (CVD grade I, II, and IIIb) (27.3%) and in two patients in group 2 (CVD grade II, IIIa, and IIIb) (18.2%), with no statistical difference (p = 0.6). None of the cases in both groups developed any clinical or radiographic signs of infection, or suffered implant failure. The mean satisfaction score was 4.8 in group 1 and 4.5 in group 2 (SD = 0.6). The difference in satisfaction scores between the two was not statistically significant (p = 0.23). Although UHMWPEE was comparable to PEEK in terms of overall complication rates and cosmesis after craniectomy, UHMWPEE as a material exhibited greater resiliency in technically challenging cases with large, complex/midline-crossing designs, previously fitted meshes, or single-stage resection-reconstruction, allowing better marginal adaptation.

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OBJECTIVE: proximal enamel caries lesions (PEC) are believed to initiate and progress to cavitation below the proximal contact area (PCA), but no evidence exists on the location of initial carious cavitation on the proximal surface with functional PCA. This study aimed to test the association of anatomical areas of the proximal surface with the severity of PEC and the frequency of cavitation in PEC in primary molars DESIGN: laboratory, observational, transversal study. Exfoliated primary molars (n = 33) with functional PCA (biofilm-free PCA surrounded by biofilm) had their proximal surfaces (one/tooth) divided anatomically into up to nine areas: 3 areas based on the occlusal/cervical PCA boundaries (areas I, II, and III; occluso-cervically) and 3 areas based on the bucco/lingual PCA boundaries (A, B, and C), with area IIB representing the PCA and area IIIB as the sub-PCA (below the PCA). PEC (ICDAS scores 1 and 2-3) and cavitation in PEC were quantified in all areas using stereomicroscopy and microCT. PEC volume was quantified in areas IIB and IIIB under microCT RESULTS: PEC severity increased occluso-cervically. PCA and sub-PCA presented different PEC severities (higher in sub-PCA) and similar PCE volumes, but the odds of carious cavitation were much higher (Odds ratio = 197.4; 95 % CI: 8.7/4480.7) in the PCA than in the sub-PCA (no cavitation). CONCLUSION: PCA presented lower PEC severity and similar PEC volume compared to sub-PCA, but PCA concentrated all cavitations in PEC, supporting a new model for the pathogenesis of PEC.

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BACKGROUND: Cutaneous neurofibromas (cNFs) are a major cause of disfigurement in patients with Neurofibromatosis Type 1 (NF1). However, clinical trials investigating cNF treatments lack standardised outcome measures to objectively evaluate changes in cNF size and appearance. 3D imaging has been proposed as an objective standardised outcome measure however various systems exist with different features that affect useability in clinical settings. The aim of this study was to compare the accuracy, precision, feasibility, reliability and accessibility of three imaging systems. MATERIALS AND METHODS: We compared the Vectra-H1, LifeViz-Micro and Cherry-Imaging systems. A total of 58 cNFs from 13 participants with NF1 were selected for imaging and analysis. The primary endpoint was accuracy as measured by comparison of measurements between imaging systems. Secondary endpoints included reliability between two operators, precision as measured with the average coefficient of variation, feasibility as determined by time to capture and analyse an image and accessibility as determined by cost. RESULTS: There was no significant difference in accuracy between the three devices for length or surface area measurements (p > 0.05), and reliability and precision were similar. Volume measurements demonstrated the most variability compared to other measurements; LifeViz-Micro demonstrated the least measurement variability for surface area and image capture and analysis were fastest with LifeViz-Micro. LifeViz-Micro was better for imaging smaller number of cNFs (1-3), Vectra-H1 better for larger areas and Cherry for uneven surfaces. CONCLUSIONS: All systems demonstrated excellent reliability but possess distinct advantages and limitations. Surface area is the most consistent and reliable parameter for measuring cNF size in clinical trials.

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OBJECTIVES: To assess skeletal and dental effects and evaluate possible side effects of maxillary expansion with two different appliances, directly after expansion and 1 year postexpansion. MATERIALS AND METHODS: Forty-two patients with unilateral posterior crossbite (mean 9.5 ± 0.9 years) were randomized to either rapid maxillary expansion (RME) banded on the deciduous second molars and bonded to the primary canines or slow expansion with quad helix (QH) on the permanent first molars. Cone-beam computed tomography records were taken at baseline, directly after correction of the posterior crossbite and at follow-up 1 year after expansion. RESULTS: All patients were analyzed. RME opened the midpalatal suture more anteriorly and inferiorly (mean 4.1 mm) and less posteriorly and superiorly (mean 1.0 mm). No effect on midpalatal suture could be shown in the QH group after expansion, P < .001. Buccal bone width had significantly decreased (P < .001) in the QH group compared with the RME group. Buccal fenestrations and root resorption on the left first molar had a higher prevalence directly after expansion finished in the QH group (P = .0086, P = .013) but were not significant at 1-year follow-up (P = .11, P = .22). CONCLUSIONS: Opening of the suture with RME was more anterior and inferior, and the QH did not open the midpalatal suture at all. More buccal bone loss and fenestrations were seen on the permanent first molar in patients treated with conventional QH than RME anchored to deciduous teeth.

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OBJECTIVES: To assess changes in the results of septal myectomy (SM) following introduction of three-dimensional (3D) imaging and 3D printing in surgical interventions planning and performing in the single-centre settings. METHODS: Between January 2007 and March 2022, 268 consecutive symptomatic patients with hypertrophic obstructive cardiomyopathy and peak pressure gradient at obstruction area ≥50 mmHg underwent conventional SM (n = 112) or SM with heart 3D modelling (n = 156). RESULTS: For comparative analysis, we used propensity score matching (PSM) by 14 variables and there were formed group 1PSM (conventional SM, n = 77) and group 2PSM (3D-modelled SM, n = 77). It was noted for group 2PSM: larger mean resected myocardium mass [10.0 (standard deviation 4.3) vs 5.2 (standard deviation 2.7) g], P < 0.001, no mitral valve replacement cases [0 vs 28 (36.4%), P < 0.001], no iatrogenic ventricular septal defects cases [0 vs 6 (7.8%), P = 0.028], lower rate of major complications [6 (7.8%) vs 17 (22.1%), P = 0.011], smaller residual peak systolic gradient at the obstruction level [7.0 (5.0-9.0) vs 11.0 (7.0-16.0) mmHg, P < 0.001]. During the long-term follow-up, it was noted for group 2PSM as compared to group 1PSM: lower 5-year cumulative incidence of major adverse cardiovascular events [3.8% (95% confidence interval 0.7-11.7%) vs 16.9% (9.5-26.1%), P = 0.007] and cardiac-related death [3.8% (95% confidence interval 0.7-11.7%) vs 13% (95% confidence interval 6.6-21.6%), P = 0.05]. CONCLUSIONS: SM based on 3D virtual and printed heart models is more effective than conventional SM.

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AIM: Evaluate constitutional differences in chin morphology and volume, and mandibular tooth size, between different facial divergence patterns. MATERIALS AND METHODS: The sample consisted of 284 pretreatment cone beam computed tomography (CBCT) images of growing and non-growing patients who were stratified into 4 groups based on mandibular plane inclination to cranial base (SN) angle. Linear and angular measurements were made on the lateral CBCT images: mandibular lateral incisor crown (I-C) total (I-A) lengths, the distances between point D (centre of symphysis) and both incisor apex (D-A) and menton (D-Me) and between cemento-enamel junction and menton (CEJ-Me); chin width at the level of the central incisor apex (CWA) and point D (CWD); and the angles of the anterior and posterior symphyseal slopes. The volume of the symphysis was measured using a specialized 3D imaging software. Group differences and associations between parameters were gauged through the three-way ANOVA analysis. RESULTS: I-C, I-A, D-A, D-Me and CEJ-Me were greater in the hyperdivergent group (p < .001). CWA and CWD were wider in the hypodivergent group (p = .003). Volume of the chin and inter-slope angles were similar in all groups (p > .05). The anterior slope angle decreased with hyperdivegence (p < .005) in opposite pattern to the posterior slope angle (p < .005). CONCLUSIONS: Shape differences in mandibular symphysis were observed between opposite divergence patterns. Mandibular incisors were longer with hyperdivergence and shorter with hypodivergence. However, chin volumes were similar across divergence groups. These findings underline the role of genetic and environmental factors that impact facial growth.

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SUMMARY: In this study, we aim to share the volumes of the carpal bone and the ratio of these volumes to the total volume of wrist bones from Computed Tomography (CT) images obtained from individuals of different ages and sex using 3D Slicer. Right wrist CT images of 0.625 mm slice thickness of 42 healthy individuals (21 female, 21 male) of both sexes were included in our study. Volume calculations were made by loading the images into 3D Slicer, an open-source software package. In this study, mean capitate volume was the largest in both sexes (male 3479.9±679.2; female 2207.1±272.1 mm3), while pisiform had the smallest mean volume (male 810.0±141.2; female 566.6±97.7 mm3). This order was ordered from largest to smallest as hamate, scaphoid, trapezium, lunate, triquetrum, trapezoid, and pisiform. According to this study, carpal bone volumes were larger in males than in females (p0.05). In this study, it was seen that carpal bone volume differed according to sex. However, it was observed that the bone volumes of both sexes took up the same amount in the total bone volume. This information will be very useful in sex determination, 3D anatomical material creation, implant applications and reconstructive surgery.

Nuestro objetivo fue determinar los volúmenes de los huesos del carpo y la relación entre estos volúmenes y el volumen total de estos huesos a partir de imágenes de TC obtenidas de individuos de diferentes edades y sexos utilizando 3D Slicer. En el estudio se incluyeron imágenes de TC del carpo de la mano derecha de los participantes en el estudio, con un grosor de corte de 0,625 mm, en 42 individuos sanos (21 mujeres, 21 hombres) de ambos sexos. Los cálculos de volumen se realizaron cargando las imágenes en 3D Slicer, un paquete de software de código abierto. El volumen medio del hueso capitado fue el mayor en ambos sexos (hombre 3479,9 ± 679,2; mujer 2207,1 ± 272,1 mm3), mientras que el hueso pisiforme tuvo el volumen medio más pequeño (hombre 810,0 ± 141,2; mujer 566,6 ± 97,7 mm3). De mayor a menor el volumen de los otros huesos fue: hamato, escafoides, trapecio, lunato, piramidal, trapezoide y pisiforme. Según este estudio, los volúmenes óseos del carpo eran mayores en hombres que en mujeres (p0,05). En este estudio se observó que el volumen de los huesos del carpo difería según el sexo. Sin embargo, se observó que los volúmenes óseos de ambos sexos ocuparon la misma cantidad del volumen óseo total. Esta información será de gran utilidad en la determinación del sexo, creación de material anatómico 3D, aplicaciones de implantes y cirugía reconstructiva.

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OBJECTIVES: Temporomandibular disorders (TMD) are the most common nonodontogenic cause of orofacial pain, leading to morbidity and impairment. TMD presents a diagnostic challenge due to many aetiologies that exhibit comparable symptoms and refer pain to the temporomandibular joint (TMJ) region. Patients may be referred to dental specialists without accounting for all pain sources. This study aims to identify radiographic confounders (RCs) that can be mistaken for TMD in patients undergoing TMJ assessment using cone-beam computed tomography (CBCT). MATERIALS AND METHODS: A review of 369 CBCT oral maxillofacial radiology reports of the TMJ acquired between July 2020 and June 2023 was completed. Pertinent RCs were classified as endodontic lesions, impacted dentition, sinus pathologies, root fractures, soft tissue calcifications, and others. The chi-squared test assessed the significance of the relationship between RCs and patient variables. RESULTS: A total of 283 RCs were identified in 202 of the 369 cases (54.7%). The most frequent findings included sinus abnormalities (32.5%), endodontic lesions (15.2%), impacted dentition (12.7%), and elongated/calcified stylohyoid process (9.2%). Significant associations were found between sinus pathologies with TMD signs (P = .009) and gender (P = .001). CONCLUSION: Our results indicate that RCs that mimic TMD-related symptoms are prevalent in patients referred for TMJ CBCT imaging. CLINICAL RELEVANCE: Clinicians should be aware of these RCs when diagnosing complaints related to the TMJ. We recommend clinicians first obtain dental clearance and investigate all other potential sources of a patient's complaint before initiating referrals to avoid unnecessary costs and delays in patient care.

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Bone-resorbing osteoclasts (OCLs) are formed by differentiation and fusion of monocyte precursor cells, generating large multi-nucleated cells. Tightly-regulated cell fusion during osteoclastogenesis leads to formation of resorption-competent OCLs, whose sizes fall within a predictable physiological range. The molecular mechanisms that regulate the onset of OCL fusion and its subsequent arrest are, however, largely unknown. We have previously shown that OCLs cultured from mice homozygous for the R51Q mutation in the vesicle trafficking-associated protein sorting nexin 10, a mutation that induces autosomal recessive osteopetrosis in humans and in mice, display deregulated and continuous fusion that generates gigantic, inactive OCLs. Fusion of mature OCLs is therefore arrested by an active, genetically-encoded, cell-autonomous, and SNX10-dependent mechanism. In order to directly examine whether SNX10 performs a similar role in vivo, we generated SNX10-deficient (SKO) mice and demonstrated that they display massive osteopetrosis and that their OCLs fuse uncontrollably in culture, as do homozygous R51Q SNX10 (RQ/RQ) mice. OCLs that lack SNX10 exhibit persistent presence of DC-STAMP protein at their periphery, which may contribute to their uncontrolled fusion. In order to visualize endogenous SNX10-mutant OCLs in their native bone environment we genetically labelled the OCLs of wild-type, SKO and RQ/RQ mice with EGFP, and then visualized the three-dimensional organization of resident OCLs and the pericellular bone matrix by two-photon, confocal, and second harmonics generation microscopy. We show that the volumes, surface areas and, in particular, the numbers of nuclei in the OCLs of both mutant strains were on average 2-6 fold larger than those of OCLs from wild-type mice, indicating that deregulated, excessive fusion occurs in the mutant mice. We conclude that the fusion of OCLs, and consequently their size, are regulated in vivo by SNX10-dependent arrest of fusion of mature OCLs.

Osteoclasts (OCLs) are cells that degrade bone. These cells are generated by fusion of monocyte precursor cells, but the mechanisms that regulate this process and eventually arrest it are unknown. We had previously shown that OCLs cultured from mice carrying the R51Q mutation in the protein sorting nexin 10 (SNX10) lose their resorptive capacity and become gigantic due to uncontrolled fusion. To examine whether SNX10 is required for OCL fusion arrest also in vivo, we inactivated the Snx10 gene in mice and fluorescently labelled their OCLs and OCLs of R51Q SNX10 mice, isolated their femurs, and used advanced 3D microscopy methods to visualize OCLs within the bone matrix. As expected, mice lacking SNX10 exhibited excessive bone mass, indicating that their OCLs are inactive. OCLs within bones of both mutant mouse strains were on average 2-6-fold larger than in control mice, and contained proportionally more nuclei. We conclude that OCL fusion is arrested in control, but not SNX10 mutant, mice, indicating that the sizes of mature OCLs are limited in vivo by an active, SNX10-dependent mechanism that suppresses cell fusion.

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The brain atlas is essential for exploring the anatomical structure and function of the brain. Non-human primates, such as cynomolgus macaque, have received increasing attention due to their genetic similarity to humans. However, current macaque brain atlases only offer coarse sections with intervals along the coronal direction, failing to meet the needs of single-cell resolution studies in functional and multi-omics research of the macaque brain. To address this issue, we utilized fluorescence micro-optical sectioning tomography to obtain sub-micron resolution cytoarchitectonic images of the macaque brain at the sagittal plane. Based on the obtained 8000 image sequences, a reference brain atlas comprising 45 sagittal sections was created, delineating 270 brain regions other than the cortex. Additionally, a website was established to share the reference atlas corresponding image data. This study is expected to provide an essential dataset and tool for scientists studying the macaque brain.

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BACKGROUND: The use of 3D imaging techniques, such as X-ray CT, in root phenotyping has become more widespread in recent years. However, due to the complexity of the root structure, analyzing the resulting 3D volumes to obtain detailed architectural root traits remains a challenging computational problem. When it comes to image-based phenotyping of excavated maize root crowns, two types of root features that are notably missing from existing methods are the whorls and soil line. Whorls refer to the distinct areas located at the base of each stem node from which roots sprout in a circular pattern (Liu S, Barrow CS, Hanlon M, Lynch JP, Bucksch A. Dirt/3D: 3D root phenotyping for field-grown maize (zea mays). Plant Physiol. 2021;187(2):739-57. https://doi.org/10.1093/plphys/kiab311 .). The soil line is where the root stem meets the ground. Knowledge of these features would give biologists deeper insights into the root system architecture (RSA) and the below- and above-ground root properties. RESULTS: We developed TopoRoot+, a computational pipeline that produces architectural traits from 3D X-ray CT volumes of excavated maize root crowns. Building upon the TopoRoot software (Zeng D, Li M, Jiang N, Ju Y, Schreiber H, Chambers E, et al. Toporoot: A method for computing hierarchy and fine-grained traits of maize roots from 3D imaging. Plant Methods. 2021;17(1). https://doi.org/10.1186/s13007-021-00829-z .) for computing fine-grained root traits, TopoRoot + adds the capability to detect whorls, identify nodal roots at each whorl, and compute the soil line location. The new algorithms in TopoRoot + offer an additional set of fine-grained traits beyond those provided by TopoRoot. The addition includes internode distances, root traits at every hierarchy level associated with a whorl, and root traits specific to above or below the ground. TopoRoot + is validated on a diverse collection of field-grown maize root crowns consisting of nine genotypes and spanning across three years. TopoRoot + runs in minutes for a typical volume size of [Formula: see text] on a desktop workstation. Our software and test dataset are freely distributed on Github. CONCLUSIONS: TopoRoot + advances the state-of-the-art in image-based phenotyping of excavated maize root crowns by offering more detailed architectural traits related to whorls and soil lines. The efficiency of TopoRoot + makes it well-suited for high-throughput image-based root phenotyping.

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Fluorescence lifetime imaging microscopy (FLIM) is a powerful imaging tool offering molecular specific insights into samples through the measurement of fluorescence decay time, with promising applications in diverse research fields. However, to acquire two-dimensional lifetime images, conventional FLIM relies on extensive scanning in both the spatial and temporal domain, resulting in much slower acquisition rates compared to intensity-based approaches. This problem is further magnified in three-dimensional imaging, as it necessitates additional scanning along the depth axis. Recent advancements have aimed to enhance the speed and three-dimensional imaging capabilities of FLIM. This review explores the progress made in addressing these challenges and discusses potential directions for future developments in FLIM instrumentation.