Growth Pattern Research Articles

Cephalometric Evaluation of Children with Short Stature of Genetic Etiology: A Review.

Introduction: A plethora of biological molecules regulate chondrogenesis in the epiphyseal growth plate. Disruptions of the quantity and function of these molecules can manifest clinically as stature abnormalities of various etiologies. Traditionally, the growth hormone/insulin-like growth factor 1 (IGF1) axis represents the etiological centre of final stature attainment. Of note, little is known about the molecular events that dominate the growth of the craniofacial complex and its correlation with somatic stature. Aim: Given the paucity of relevant data, this review discusses available information regarding potential applications of lateral cephalometric radiography as a potential clinical indicator of genetic short stature in children. Materials and Methods: A literature search was conducted in the PubMed electronic database using the keywords: cephalometric analysis and short stature; cephalometric analysis and achondroplasia; cephalometric analysis and hypochondroplasia; cephalometric analysis and skeletal abnormalities; cephalometr* and SHOX; cephalometr* and CNP; cephalometr* and ACAN; cephalometr* and CNVs; cephalometr* and IHH; cephalometr* and FGFR3; cephalometr* and Noonan syndrome; cephalometr* and "Turner syndrome"; cephalometr* and achondroplasia. Results: In individuals with genetic syndromes causing short stature, linear growth of the craniofacial complex is confined, following the pattern of somatic short stature regardless of its aetiology. The angular and linear cephalometric measurements differ from the measurements of the average normal individuals and are suggestive of a posterior placement of the jaws and a vertical growth pattern of the face. Conclusions: The greater part of the existing literature regarding cephalometric measurements in short-statured children with genetic syndromes provides qualitative data. Furthermore, cephalometric data for individuals affected with specific rare genetic conditions causing short stature should be the focus of future studies. These quantitative data are required to potentially establish cut-off values for reference for genetic testing based on craniofacial phenotypes.

Core regions in immunoglobulin heavy chain enhancers essential for survival of non-Hodgkin lymphoma cells are identified by a CRISPR interference screen.

Chromosomal translocations in non-Hodgkin lymphoma (NHL) result in activation of oncogenes by placing them under the regulation of immunoglobulin heavy chain (IGH) super-enhancers. Aberrant expression of translocated oncogenes induced by enhancer activity can contribute to lymphomagenesis. The role of the IGH enhancers in normal B-cell development is well established, but knowledge regarding the precise mechanisms of their involvement in control of the translocated oncogenes is limited. The goal of this project was to define the critical regions in the IGH regulatory elements and identify enhancer RNAs (eRNA). We designed a sgRNA library densely covering the IGH enhancers and performed tiling CRISPR interference screens in three NHL cell lines. This revealed three regions crucial for NHL cell growth. With chromatin-enriched RNA-Seq we showed transcription from the core enhancer regions and subsequently validated expression of the eRNAs in a panel of NHL cell lines and tissue samples. Inhibition of the essential IGH enhancer regions decreased expression of eRNAs and translocated oncogenes in several NHL cell lines. The observed expression and growth patterns were consistent with the breakpoints in the IGH locus. Moreover, targeting the Eμ enhancer resulted in loss of B-cell receptor expression. In a Burkitt lymphoma cell line, MYC overexpression partially rescued the phenotype induced by IGH enhancer inhibition. Our results indicated the most critical regions in the IGH enhancers and provided new insights into the current understanding of the role of IGH enhancers in B-cell NHL. As such, this study forms a basis for development of potential therapeutic approaches.

Modelación de crecimiento de frutos de tangor murcott e incidencia de factores medioambientales sobre los parámetros estimados

Fruit production forecasts are a tool to plan the harvest and improve market strategies. To carry it out, it is essential to have information about the behavior of fruit development over time. The objective of this work was to find the mathematical-statistical model that best describes the growth pattern of tangor murcott fruit (Citrus reticulata x C. sinensis 'Murcott') and analyze how it is affected by environmental conditions. For this, in nine orchards, located in four locations in the province of Corrientes, Argentina, the equatorial diameter of 2,053 fruit from 82 days after full flowering to harvest were periodically registered during five seasons. The nonlinear models were compared: Logistic, Gompertz, Brody, Von Bertalanffy, Weibull, Morgan Mercer Flodin (MMF), Richards, and their respective re-parameterizations. The magnitudes of nonlinearity measures, coefficient of determination and estimates of residual deviation were considered as the main goodness-of-fit criteria. The selected model-parameterization combination was the fifth parameterization of the Logistic model with random effects on its three parameters. An Analysis of Variance model on the estimates of these parameters for each fruit showed that orchard and season factors were an important source of variability, mainly in those related to the initial size of the fruit and their growth rate. These results will allow the construction of growth tables, which in addition to making yield predictions, can be used to estimate fruit size distribution at harvest and improve the cultural practice of manual fruit thinning.

Investigating the Effectiveness of Ceragenins against Acinetobacter baumannii to Develop New Antimicrobial and Anti-Adhesive Strategies.

A growing body of experimental data indicates that ceragenins (CSAs), which mimic the physicochemical properties of the host's cationic antimicrobial peptide, hold promise for the development of a new group of broad-spectrum antimicrobials. Here, using a set of in vivo experiments, we assessed the potential of ceragenins in the eradication of an important etiological agent of nosocomial infections, Acinetobacter baumannii. Assessment of the bactericidal effect of ceragenins CSA-13, CSA-44, and CSA-131 on clinical isolates of A. baumannii (n = 65) and their effectiveness against bacterial cells embedded in the biofilm matrix after biofilm growth on abiotic surfaces showed a strong bactericidal effect of the tested molecules regardless of bacterial growth pattern. AFM assessment of bacterial cell topography, bacterial cell stiffness, and adhesion showed significant membrane breakdown and rheological changes, indicating the ability of ceragenins to target surface structures of A. baumannii cells. In the cell culture of A549 lung epithelial cells, ceragenin CSA-13 had the ability to inhibit bacterial adhesion to host cells, suggesting that it interferes with the mechanism of bacterial cell invasion. These findings highlight the potential of ceragenins as therapeutic agents in the development of antimicrobial strategies against bacterial infections caused by A. baumannii.

Heavy Metal Levels and Length-Weight Dynamics of Anodontia philippiana (Reeve, 1850) from Barobo and Hinatuan, Surigao del Sur, Philippines

Anodontia philippiana is among the commercially valuable edible clams found in Brgy. Wakat, Municipality of Barobo, and Brgy.Loyola, Municipality of Hinatuan, Surigao del Sur, Philippines. However, anthropogenic activities may pose a threat to the abundance and quality of this species. This study aimed to determine the levels of lead (Pb), nickel (Ni), chromium (Cr), and mercury (tHg) in both the sediments and flesh of A. philippiana using Atomic Absorption Spectrophotometry (AAS). Additionally, the length-weight relationship (LWR) of A. philippiana was assessed to determine the size structure of the species’ population. Among the heavy metals tested, only the concentration of Cr (59±0.38 ppm) in Hinatuan exceeded the prescribed tolerable limit of ≥50 ppm (FAO, 1993). Conversely, the heavy metal levels in the flesh of A. philippiana consistently remained below tolerable limits. Furthermore, A. philippiana from Barobo exhibited a positive allometry (B>3), while those from Hinatuan showed a negative allometry (B<3), suggesting the influence of environmental stressors on growth patterns. The LWR in A. philippiana exhibited significant differences (P<0.001) between the two stations. Despite heavy metal concentrations in A. philippiana flesh and sediments were mostly below standard limits, the observed negative allometric growth pattern in Hinatuan indicates the presence of ecological stressors in the sediments. This study underscores the potential impact of heavy metals on the growth patterns of A. philippiana. It is recommended that regular monitoring of heavy metal concentrations be conducted in these areas. Further studies can provide better insights into morphometric patterns in A. philippiana and other commercially significant bivalves.

Growth Characteristics of Ramet System in Phyllostachys praecox Forest under Mulch Management.

The ramet system is a typical structural type in the life history of clonal plants. This massive structure is formed by many similar ramets connected by underground rhizomes, which are independent and mutually influential. Therefore, the ramet system is unique to bamboo forests, and its role in the construction, maintenance, and productivity of bamboo populations is irreplaceable. Mulch management is a high-level cultivation model for bamboo forests that is used to cultivate bamboo shoots. However, the basic conditions of bamboo ramet systems in this managed model are poorly understood. This study analyzed the underground rhizome morphology, bud bank, and branching of bamboo ramets in a Phyllostachys praecox C.D. Chu et C.S. Chao 'Prevernalis' forest to explore the growth patterns of bamboo ramets in high-level management fields. In mulched bamboo forests, the bamboo rhizomes, distributed in intermediate positions of the bamboo ramet system, were long with many lateral buds and branches, and those at the initial and distal ends were short with few lateral buds and branches. The initial end of the ramet system reduced the ramet system, the intermediate part expanded the ramet system, and the distal end promoted ramet system regeneration. Owing to the continuous reduction, expansion, and renewal of ramet systems, the bamboo rhizome system demonstrates mobility and adaptability. This study found that a higher level of bamboo forest management increased the possibility of artificial fragmentation of the ramet system and that improving the efficiency of the ramet system was beneficial for maintaining its high vitality. Thus, this study provides a crucial reference for guiding the precise regulation of bamboo ramet systems in artificial bamboo forests.

An adaptive phase field approach to 3D internal crack growth in rocks

Modeling the 3D internal crack under compression entails complex fracture mechanics (mode I-II-III fracture), resulting in substantial computational costs and challenges in characterizing fracture morphology characterization for Phase Field Method (PFM) simulations. In the present paper, we developed a 3D adaptive crack propagation model integrated into the PFM to study the internal cracks in rocks. The proposed locally refined solution was implemented within the isogeometric analysis (IGA) framework, utilizing PHT-splines and employing the phase field variable as an error indicator to accurately capture non-smooth crack surfaces. The simulation begins with a coarser mesh, and the spatial discretization is adaptively refined in real-time as the computation progresses and the phase variable threshold is reached. By comparing our results with previous numerical tests, we validated the effectiveness of our model in simulating crack growth. Notably, a smaller length scale parameter led to a higher peak force in notched square plates under tension, and our adaptive PFM successfully reduced errors caused by length scale parameters. We further validated our model by comparing it with uniaxial compression experiments on 3D-printed resin samples with internal penny-shaped cracks, achieving good agreement with the experimental data. Using the 3D adaptive PFM, we predicted changes in crack morphology and fracturing mechanisms by varying the inclination angles of internal penny-shaped cracks. Our numerical results showed that as the inclination angle increased, wing cracks initiated closer to the end tips of the original crack, and their length decreased due to the wings wrapping effect. The corresponding fracturing mechanisms transition from mode I-III fracture (0°) to mode I-II-III fracture (30°, 45°, 60°), and eventually to mode I fracture (90°) as inclination angles increase. Comparing 2D and 3D models, we found that the growth of wing cracks in 3D was significantly influenced by the intermediate principal stress. We conclude that the extended 3D adaptive PFM-based model accurately predicted complex mechanical behavior and crack growth patterns in rock.

The effect of birth type and gender on growth performance, live weight gain, growth pattern modeling, and survival rate in Anatolian Merino lambs

In this study, the aim was to determine the effect of birth type and gender on growth performance, live weight gain, growth modeling, and survival rate in Anatolian Merino lambs. Growth performance was determined in 28 singletons (14 females/14 males), 80 twins (38 females/42 males), and 54 triplets (22 females/32 males) lambs in the study. The body weight of the lambs was measured on the day of birth (day 0), and at 10, 20, 30, 45, and 60 days of age. Birth type was effective on growth performance in Anatolian Merino lambs (P

Development of a 45K pepper GBTS liquid-phase gene chip and its application in genome-wide association studies.

Pepper (Capsicum spp.) is a vegetable that is cultivated globally and has undergone extensive domestication, leading to a significant diversification in its agronomic traits. With the advancement of genomics in pepper and the reduction in sequencing costs, the high-throughput detection of single nucleotide polymorphisms (SNPs) and small insertions-deletions (indels) has become increasingly critical for analyzing pepper germplasms and improving breeding programs. As a result, there is a pressing need for a cost-effective, high-throughput, and versatile technique suitable for both foreground and background selection in pepper breeding. In the present study, Python-based web scraping scripts were utilized to systematically extract data from published literatures and relevant sequence databases focusing on pepper genomes. Subsequent to data extraction, SNPs and indels were meticulously identified and filtered. This process culminated in the delineation of core polymorphic sites, which were instrumental in the development of specific probes. Following this, comprehensive phenotypic and genotypic analyses were conducted on a diverse collection of 420 pepper germplasms. Concurrently, a genome-wide association study (GWAS) was conducted to elucidate the genetic determinants of helical fruit shape in peppers. In this study, a 45K pepper Genotyping-By-Target-Sequencing (GBTS) liquid-phase gene chip was developed on the GenoBaits platform. This chip is composed of 45,389 probes, of which 42,535 are derived from core polymorphic sites (CPS) in the background genetic landscape, while 2,854 are associated with foreground agronomic traits, spanning across 43 traits. The CPS probes are spaced at an average interval of 68 Kb. We have assessed the performance of this chip on 420 pepper germplasms, with successful capture of target DNA fragments by 45,387 probes. Furthermore, the probe capture ratio surpassed 70% in 410 of the 420 germplasms tested. Using this chip, we have efficiently genotyped 273 germplasms for spiciness levels and elucidated the genetic relationships among 410 pepper germplasms. Our results allowed for precise clustering of sister lines and C. chinense germplasms. In addition, through a GWAS for helical fruit shape, we identified three quantitative trait loci (QTLs): heli2.1, heli11.1, and heli11.2. Within the heli11.1 QTL, a gene encoding the tubulin alpha chain was identified, suggesting its potential role in the helical growth pattern of pepper fruits. In summary, the 45K pepper GBTS liquid-phase gene chip offers robust detection of polymorphic sites and is a promising tool for advancing research into pepper germplasm and the breeding of new pepper varieties.

Similar climate–growth relationships but divergent drought resilience strategies in coexisting Mediterranean shrubs

Abstract Anticipating future impacts of climate warming and aridification on drylands requires understanding how coexisting woody plant species respond to climate variability. However, we lack knowledge of the growth resilience capacity of Mediterranean shrubs. Do coexisting trees and shrubs differ in their response to climate? Do coexisting shrub species have comparable post‐drought growth resilience? This study was conducted in two Mediterranean shrublands with sparse trees in semi‐arid north‐eastern Spain. We selected sites situated in formerly agricultural or grassland areas in two regions subjected to semi‐arid Mediterranean climate conditions. We sampled six shrubs' species (Juniperus phoenicea, Juniperus oxycedrus, Pistacia lentiscus, Pistacia terebinthus, Rhamnus lycioides and Rhamnus alaternus) and one tree species (Pinus halepensis) to measure their radial growth and to reconstruct their past growth patterns using dendrochronology. We quantified climate–growth relationships of trees and shrubs, as well as the growth resilience capacity after drought events of the six shrubs. Growth patterns differed between species but a prevalent trade‐off between growth and longevity was found. Growth responses to climate were comparable between species but differed between sites. Most species responded positively to precipitation and negatively to temperature. The most negative correlations of growth with drought severity were found at the driest Valcuerna site. Shrubs differed in their resilience capacity. Across sites, species were more resistant and resilient in Alcubierre, the wettest site. The shrubs P. terebinthus and R. lycioides were more resistant and resilient than the rest of species, which required longer growth recovery times and accumulated stronger growth reductions, particularly J. phoenicea. Synthesis. These results highlight the importance of drought as a driver of growth in Mediterranean scrublands. Growth resilience strategies differed between species with junipers being less resilient to drought than other coexisting shrubs. Further research should investigate how this difference in post‐drought resilience is related to functional traits, particularly those related with plant water‐use strategies.

Tracing the Evolution of E-Government: A Visual Bibliometric Analysis from 2000 to 2023

In the continuously evolving field of E-government, understanding the breadth and depth of academic research is crucial for advancing governmental digital transformation and policy development. This study employs visual bibliometric analysis, using the Web of Science database to map the evolution trajectory of E-government research from 2000 to 2023. By utilizing CiteSpace for a comprehensive examination of 4536 academic articles, this paper outlines the growth patterns and thematic evolution within the field. The development of the E-government field is delineated into four distinct phases: the budding period (2000–2003), the bottleneck period (2004–2014), the development period (2015–2018), and the growth period (2019–2023), which are each marked by unique thematic shifts and technological advancements. The research results reveal the transformation of research focus in different periods, from the initial focus on the technological means and the electronic transformation of government services, moving on to more complex issues such as E-government acceptance and government transparency and corruption, and ultimately to the current focus on innovation and smart cities. In addition, the paper also clarifies that the research boom that began in 2019 is driven by technological innovation opportunities, the improvement in infrastructure, and multidisciplinary research. By depicting these developmental stages and emerging trends, this study not only unveils past academic efforts but also forecasts future research directions, thereby providing valuable insights for researchers and policymakers aiming to understand and implement effective E-government strategies.

Energy inputs imprint seasonality and fractal structure on river metabolic regimes

AbstractThe temporal structures of gross primary production (GPP) and ecosystem respiration (ER) vary across time scales in response to complex interactions among dynamic drivers (e.g., flow, light, temperature, organic matter supply). To explore emergent patterns of river metabolic variation, we applied frequency‐domain analysis to multiyear records of metabolism across 87 US rivers. We observed a dominant annual periodicity in metabolic variation and universal fractal scaling (i.e., power spectral density inversely correlated with frequency) at subannual frequencies, suggesting these are foundational temporal structures of river metabolic regimes. Frequency‐domain patterns of river metabolism aligned best with drivers related to energy inputs: benthic light for GPP and GPP for ER. Simple river metabolism models captured frequency‐domain patterns when parameterized with appropriate energy inputs but neglecting temperature controls. These results imply that temporal variation of energy supply imprints directly on metabolic signals and that frequency‐domain patterns provide benchmark properties to predict river metabolic regimes.

DIPG-08. PROBING THE BLOOD-BRAIN-TUMOR-BARRIER WITH A MICROVASCULAR MODEL OF DIFFUSE MIDLINE GLIOMA

Abstract BACKGROUND Pediatric diffuse midline glioma (DMG) is a uniformly fatal brain tumor in children, with most patients surviving less than 12 months. Current treatment for DMG is limited, and outcomes for this tumor have not improved. The blood-brain-barrier (BBB) remains a key challenge for DMG therapeutics, preventing most drugs from entering brain tissue. Functional characterization of the blood-brain-tumor-barrier (BBTB) – the interaction between tumors and BBB vasculature – remains largely unexplored for DMG. There is a need for a preclinical model of DMG that can probe the functional, physiological, and biological properties of the BBTB in vitro as a platform to evaluate therapeutic efficacy that is representative of human physiology. METHODS To generate an in vitro model of DMG with human microvasculature (DMG-BBTB model), we co-cultured DMG neurospheres, human brain microvascular endothelial cells, human brain vascular pericytes, and human astrocytes. Cells are simultaneously seeded into microfluidic devices at varied concentrations within a fibrin matrix. Self-assembled microvascular networks are formed within 7-8 days and used for downstream functional and phenotypic assays. RESULTS Vascular morphology and solute permeability in microvascular networks are influenced by cell identity, growth pattern, and relative concentration. We show that DMG neurospheres exhibit different patterns of growth when co-cultured as uniform spheroids versus single cells in suspension. Devices representing each growth pattern are further characterized for paracellular diffusion and permeability compared to devices without DMG cells through perfusion of high molecular weight dextran. Permeability values are correlated to biologic features such as the abundance and localization of junctional proteins and transporters. CONCLUSIONS We developed a platform to evaluate the microvasculature of the DMG BBTB as a tool to develop new therapies informed by vascular biology. We probed the functional, physiological, and biological properties of the human DMG BBTB in vitro, and in ongoing work are assessing approved and experimental therapeutics.

MODL-06. USING HYDROGEL MODELS TO RECAPITULATE THE TUMOUR-IMMUNE MICROENVIRONMENT OF AGGRESSIVE GROUP 4 MEDULLOBLASTOMA TUMOURS

Abstract BACKGROUND Medulloblastoma is the most common malignant childhood brain-tumour, predominantly affecting children between the ages of one and nine. It is divided into four molecular subgroups amongst which, Group 3 (G3) and Group 4 (G4) subgroups have the worst prognosis. Subgroups possess distinct extracellular-matrix (ECM) and tumour immune microenvironments (TIME) directly influencing therapy-response and metastasis. Current models of medulloblastoma fail to precisely recreate the complex, subgroup-specific TIME and do not mimic intricate immune-tumour heterotypic signalling facilitating immune-evasion. Here we use a HyStem-hydrogel which forms a hyaluronan-rich environment mimicking the natural brain architecture. This overcomes several limitations of current MB models, allowing customisation of subgroup matching of ECM components, adjustable ECM-stiffness and co-culturing of immune cells. METHODS HyStem-hydrogels were co-assembled with ECM components (with/without laminin) and G4 medulloblastoma cell line CHLA-01-MED. Hydrogels were imaged every three-days using an EVOS M7000 system and assessed for cell-viability with PrestoBlue every seven days. Co-culture laminin-supplemented HyStem hydrogels were established with M1-like or M2-like resting/activated macrophages and their growth tracked in the same manner. On day twenty-eight, hydrogels were fixed and stained for markers of cellular proliferation (Ki-67), ECM (laminin, vitronectin) and macrophage markers. Media from hydrogel co-cultures was collected and ELISAs permofmed to assess cytokine production. RESULTS Hydrogels with/without laminin both remained viable and stable for 28 days. Laminin-supplemented hydrogels displayed a mixed laminar-nodular phenotype which mirrored the heterogenous nature of G4 patient-tumours. Co-culture of CHLA-01-MED cells with activated-macrophages led to cytokine production and altered growth patterns. CONCLUSION We show that HyStem hydrogels recapitulate biologically relevant behaviour of aggressive G4-MB tumours in vitro and represent a promising model for G4-tumours. Future research will involve inclusion of additional brain relevant cells/ECM components and comparisons to a fully customisable peptide-amphiphile hydrogel.

New Interpretation of Human–Land Relation: Differentiated Impacts of Global Demographic Transition on Carbon Emissions

Demographic transition and environmental governance are the most prominent focal points of global concern in the 21st century. We quantitatively evaluate the distinct carbon emission impacts of the global demographic transition by regression models based on C–D production function. Our study highlights that while demographic transition boosts per capita economic growth, it adversely affects overall economic output and aggregate economic growth, highlighting significant reductions in carbon emissions as a result of these demographic changes. However, it has a negative contribution to overall economic output and economic growth. Meanwhile, demographic transition eventually reduces carbon emissions to some extent. The relationship between population urbanization and carbon emissions mainly conforms to an inverted U-shaped curve, while some of it does to a linear growth pattern. However, the relationship between population aging and carbon emissions mainly conforms to an inverted U-shaped pattern. The impacts of demographic transition on carbon emissions confirm the universality of the EKC law in the particular production function.

LGG-12. INTEGRATED CLINICAL AND MOLECULAR CHARACTERIZATION OF 200 DISSEMINATED PEDIATRIC LOW-GRADE GLIOMAS

Abstract Pediatric low-grade gliomas (PLGG) have excellent outcomes overall but are a major clinical challenge when disseminated. Diffuse leptomeningeal glioneuronal tumor (DLGNT) is a recognised entity both clinically and pathologically, however many disseminated LGG (DLGG) fall outside this diagnosis. To better understand the clinical and molecular features of DLGG as well as risk factors for dissemination, we assembled an international consortium of 30 sites, contributing over 200 patients with clinical annotation, along with genomic and methylation profiling. DLGG have worse progression-free (PFS) and overall survival (OS) than PLGG overall (p<0.0001). Seventy (35%) presented with a localized mass and secondary dissemination, including some with initial gross-total resection. The most common growth pattern observed (n=77, 40%) is a dominant suprasellar/optic pathway tumor with leptomeningeal drop-metastases involving the brainstem and spinal cord. Only 27 (14%) patients had diffuse tumors (without an identifiable dominant mass), and these had significantly worse OS (p=0.03). The most common pathologic diagnosis was pilocytic/pilomyxoid astrocytoma (n=92; 53%). DLGNT comprised a minority of cases (n=23, 14%), with a trend (p=0.056) towards worse survival compared to other diagnoses. The most frequent molecular alteration was BRAF fusion (78/151 with molecular testing; 52%), followed by FGFR mutations or fusions (18/151; 12%). BRAF V600E was underrepresented (14/151; 9%). 1p deletion was rare (17 patients) but highly associated with DLGNT. Methylation classification (n=70) was highly concordant with histologic diagnoses rather than clinical behavior. Importantly, patients who received upfront targeted therapies (TT; BRAF and/or MEK inhibition, n=9) had better PFS compared to those receiving chemotherapy (n=146, p=0.05). Furthermore, patients who were treated sequentially with chemo then TT had longer PFS on TT (p=0.011). This study presents the largest cohort of DLGG to date, expanding our understanding of the clinical, pathologic, and molecular features of this disease and supports the use of upfront targeted therapy.

DIPG-34. IDENTIFICATION OF CAR-T CELL ENHANCING COMPOUNDS USING HIGH THROUGHPUT SCREENING

Abstract BACKGROUND The infiltrative growth patterns and restrictiveness of the blood brain barrier greatly limit therapeutic options for children diagnosed with diffuse intrinsic pontine gliomas (DIPGs). However, clinical trials using chimeric antigen receptor (CAR)-T cells have shown promise in DIPG patients. Despite some success, the biological limitations plaguing CAR-T cells hinder their overall efficacy. We propose that leveraging chemical compounds in tandem with CAR-T cells may provide an increased therapeutic benefit against this lethal brain tumor. Our study is aimed at using high throughput screening of >2,000 FDA approved and investigational compounds to identify candidates that enhance CAR-T cell mediated cytotoxicity of DIPGs. METHODS In an initial limited screen (32 compounds) used for optimization, compounds were serially diluted using the Echo 650 acoustic liquid handler. Subsequent co-culture conditions of renilla luciferase transduced DIPG007 tumor cells and clinically relevant firefly luciferase transduced CAR-T cells were plated at a 2:1 effector: target (E: T) ratio. In comparison to DMSO control, drug induced changes on cell viability were measured using cell-specific luciferase activity at 24 and 72 hours. RESULTS The limited screen revealed the BET bromodomain inhibitor, GSK-046, as an initial target based on its ability to increase CAR-T cell viability after 72 hours. Preliminary validation of these findings supports this CAR-T enhancing role of GSK-046 via MTS assays. We also identified nonlethal concentrations of GSK-046 for DIPG007 cells that could be further tested in combination with CAR-T cells to determine therapeutic synergy. CONCLUSIONS Our initial findings warrant further investigation into the combinatorial benefit of GSK-046 and CAR-T cells against DIPGs. Currently, we are optimizing and expanding our drug screen to identify additional compounds that may complement CAR-T cell therapy. Furthermore, we intend to validate the cytotoxic benefit of our prioritized combinations in additional in vitro and in vivo settings.

ESMAC Best Paper Award 2023: Increased knee flexion in participants with cerebral palsy results in altered stresses at the distal femoral growth plate compared to a typically developing cohort

IntroductionFemoral deformities are highly prevalent in children with cerebral palsy (CP) and can have a severe impact on patients’ gait abilities. While the mechanical stress regime within the distal femoral growth plate remains underexplored, understanding it is crucial given bone's adaptive response to mechanical stimuli. We quantified stresses at the distal femoral growth plate to deepen our understanding of the relationship between healthy and pathological gait patterns, internal loading, and femoral growth patterns. MethodsThis study included three-dimensional motion capture data and magnetic resonance images of 13 typically developing children and twelve participants with cerebral palsy. Employing a multi-scale mechanobiological approach, integrating musculoskeletal simulations and subject-specific finite element analysis, we investigated the orientation of the distal femoral growth plate and the stresses within it. Limbs of participants with CP were grouped depending on their knee flexion kinematics during stance phase as this potentially changes the stresses induced by knee and patellofemoral joint contact forces. ResultsDespite similar growth plate orientation across groups, significant differences were observed in the shape and distribution of growth values. Higher growth rates were noted in the anterior compartment in CP limbs with high knee flexion while CP limbs with normal knee flexion showed high similarity to the group of healthy participants. DiscussionResults indicate that the knee flexion angle during the stance phase is of high relevance for typical bone growth at the distal femur. The evaluated growth rates reveal plausible results, as long-term promoted growth in the anterior compartment leads to anterior bending of the femur which was confirmed for the group with high knee flexion through analyses of the femoral geometry. The framework for these multi-scale simulations has been made accessible on GitHub, empowering peers to conduct similar mechanobiological studies. Advancing our understanding of femoral bone development could ultimately support clinical decision-making.

Histopathologic Features and Differential Diagnosis in Challenging Cases of Nodular Lymphocyte Predominant B-cell Lymphoma/Nodular Lymphocyte Predominant Hodgkin Lymphoma.

Nodular lymphocyte predominant Hodgkin lymphoma was termed "nodular lymphocyte predominant B-cell lymphoma" in the International Consensus Classification (ICC), to emphasize clinical and biological differences from classic Hodgkin lymphoma (CHL). The abbreviation "NLP" represents both terms in the ICC and World Health Organization classifications. Variations in the growth pattern, originally reported as Fan patterns A-F, are designated as either grade 1 or grade 2 in the ICC. NLP is uncommon, and in some cases an accurate diagnosis is challenging. The objectives of this article were to review the histopathologic features of NLP and the differential diagnosis from other key entities including de novo T-cell/histiocyte-rich large B-cell lymphoma (THRLBL) and lymphocyte-rich classic Hodgkin lymphoma (LRCHL). Histologically, NLP Fan pattern E (THRLBL-like) can be indistinguishable from de novo THRLBL. However, focal nodular areas, clustering of tumor cells, presence of few admixed small B-cells or FDC meshworks, and T-cell rosettes favor NLP Fan pattern E and argue against de novo THRLBL. NLP may also be confused with LRCHL. Patients with NLP are younger than those with LRCHL, and LRCHL may show mediastinal involvement. In LRCHL, the nodular pattern often contains eccentrically located small regressed germinal centers and intact small dense FDC meshworks, in contrast to the expanded, and fragmented FDC meshworks in NLP. Neoplastic cells that are positive for CD30 and CD15 but negative for CD20 and CD79a are characteristic of LRCHL. Additionally, Fascin and Gata3 are commonly positive in LRCHL but usually negative in NLP.

Long-term imaging of three-dimensional hyphal development using the ePetri dish.

Imaging three-dimensional microbial development and behavior over extended periods is crucial for advancing microbiological studies. Here, we introduce an upgraded ePetri dish system specifically designed for extended microbial culturing and 3D imaging, addressing the limitations of existing methods. Our approach includes a sealed growth chamber to enable long-term culturing, and a multi-step reconstruction algorithm that integrates 3D deconvolution, image filtering, ridge, and skeleton detection for detailed visualization of the hyphal network. The system effectively monitored the development of Aspergillus brasiliensis hyphae over a seven-day period, demonstrating the growth medium's stability within the chamber. The system's 3D imaging capability was validated in a volume of 5.5 mm × 4 mm × 0.5 mm, revealing a radial growth pattern of fungal hyphae. Additionally, we show that the system can identify potential filter failures that are undetectable with 2D imaging. With these capabilities, the upgraded ePetri dish represents a significant advancement in long-term 3D microbial imaging, promising new insights into microbial development and behavior across various microbiological research areas.