Caliper Measurements Research Articles

Tensile properties of SS316L produced by LPBF: Influence of specimen dimensions and surface condition

This study is part of the global effort to develop precise and accurate mechanical testing methodologies for metallic parts in as-built condition produced by powder-bed additive manufacturing processes. Miniature 316L stainless steel specimens with a 7 × 1 × 1.5 mm3 reduced section were produced by laser powder bed fusion. Three distinct surface conditions were considered. Standard specimens were also produced for comparison. Special attention was given to the non-destructive measurement of the cross-sectional area of small as-built specimens. The ultimate tensile strength of the as-built miniature specimens was 12% below standard-sized or machined specimens. Using caliper measurements lead to an overestimation of the section by as much as 20% compared to CT-scan values, resulting in an underestimation of UTS. Point micrometer measurements yielded more accurate estimations of cross-sectional area. With the experimental error taken into account, the specimen dimensions and surface condition influenced UTS. A coarser average microstructure for miniature as-built specimens was linked to a reduction of UTS by about 4% compared to standard specimens produced in the same conditions. Despite a larger scatter obtained with miniature specimens, no effect of specimen dimensions on the average value of elongation at break could be observed. Higher surface roughness of the order of Ra = 15 μm resulted in an elongation at break reduced from 52.7 ± 5.2 to 39.2 ± 5.0% for as-built miniature specimens. It was concluded that using large-scale or machined specimens may produce less accurate tensile characterization of thin 316L sections produced by laser powder bed fusion. Mechanical coupons with dimensions comparable with the structural part of interest should be employed. Accurate measurement of the cross-sectional area of small as-built specimens is critical for accurate measurements of mechanical strength. These results rationalize the scatter on tensile properties of LPBF 316L available in the literature.

Ice Accretion: Image Post-Processing Measurement Techniques for 2D Ice Shapes

Ice accretion poses substantial safety hazards for the manned and unmanned aviation industries. Its study is essential for icing events risk assessment and for the development of efficient ice protection systems. The existing ice accretion measurement techniques—casting, molding, and laser-scanning—are time-consuming, sometimes cumbersome to use, and highly expensive, while hand tracing is inexpensive, but has lower accuracy and time-consuming post-processing. This work presents two low-cost, fast, and easy-to-use measurement techniques for 2D ice accretion profiles. Both employ algorithms of automatic ice shape detection, one based on unmediated image-processing, another based on the processing of manual ice tracings. The techniques are applied to ice accretion experiments conducted in an icing wind tunnel at low Reynolds numbers, and their results are validated against ice thickness caliper measurements. A comparison of the results shows that both techniques accurately measure the leading-edge ice thickness and the 2D shape of the ice accretion profiles. One technique is faster, with higher measurement accuracy, but produces interrupted-line 2D ice profiles and requires good lighting conditions, while the other generates continuous-line 2D profiles and has no application restriction, but it is slower, with lower accuracy. A discussion is conducted, aiming to help one determine the best applications for each ice accretion measurement technique presented.

Phantom-based assessment of motion and needle targeting accuracy of robotic devices for magnetic resonance imaging-guided needle biopsy.

The current study proposes simple methods for assessing the performance of robotic devices intended for Magnetic Resonance Imaging (MRI)-guided needle biopsy. In-house made agar-based breast phantoms containing biopsy targets served as the main tool in the evaluation process of an MRI compatible positioning device comprising a needle navigator. The motion accuracy of mechanical stages was assessed by calliper measurements. Laboratory evaluation of needle targeting included a repeatability phantom test and a laser-based method. The accuracy and repeatability of needle targeting was also assessed by MRI. The maximum error of linear motion for steps up to 10mm was 0.1mm. Needle navigation relative to the phantom and alignment with the various biopsy targets were performed successfully in both the laboratory and MRI settings. The proposed biopsy phantoms offered tissue-like signal in MRI and good haptic feedback during needle insertion. The proposed methods could be valuable in the process of validating the accuracy of MRI-guided biopsy robotic devices in both laboratory and real environments.

Introduction and validation of an automatic and low-cost three-dimensional facial imaging system: a comparison to direct anthropometry and Vectra H1.

Three-dimensional (3D) imaging is a powerful tool for the analysis of soft tissue morphology. 3D photogrammetry outperforms conventional photogrammetric methods and gains popularity among plastic surgeons. However, commercial 3D imaging systems bundled with analytical software are costly. This study intends to introduce and validate an automatic, low-cost, and user-friendly 3D facial scanner. An automatic and low-cost 3D facial scanning system was developed. The system consisted of a 3D facial scanner running automatically on a sliding track and a 3D data processing tool. Fifteen human subjects underwent 3D facial imaging by the novel scanner. Eighteen anthropometric parameters were measured on the 3D virtual models and compared with caliper measurements (the gold standard). Further, the novel 3D scanner was compared to the commonly used commercial 3D facial scanner Vectra H1. Heat map analysis was used to evaluate the deviation between the 3D models obtained by the two imaging systems. The 3D photogrammetric results were highly correlated with the direct measurement results (P<0.001). The mean absolute differences (MADs) were less than 2 mm. Bland-Altman analysis indicated that, for 17 of the 18 parameters, the largest differences within the 95% limits of agreement margin were within the 2.0 mm clinical acceptance. Heat map analysis showed the average distance between the 3D virtual models was 0.15 mm, with a root mean square of 0.71 mm. The novel 3D facial scanning system is proven to be highly reliable. It provides a good alternative to commercial 3D facial scanners.

Thermal expansion and boundary excess free volume of electroformed nanocrystalline Ni-Co from dilatometer and digital caliper measurements

The coefficient of thermal expansion (CTE) as measured by dilatometry of electroformed nanocrystalline Ni-32at%Co macro-defect free sheet metal up to 500 °C can be separated into three regions. Below 185 °C, the nanocrystalline material's CTE is similar to its polycrystalline counterpart. Between 185 and 310 °C, the CTE of the nanocrystalline material plateaus, then drops at temperatures above 310 °C due to volume shrinkage induced by different grain growth stages. The grain boundary excess volume (BEV) was determined to be between 0.20–0.24 nm. In addition, it is shown that simple digital caliper measurements are a reliable technique to estimate BEV for specimens with grain boundaries being the main defect.

D113. Comparison of Methodologies for Craniofacial Soft-tissue Cephalometrics: The Value of Virtual Reality

PURPOSE: Myriad options are available for soft-tissue analysis, which is vital to perioperative evaluation and research. Our objective is to compare accuracy, precision, and costs of available cephalometric modalities using facial soft-tissue measurements. METHODS: Twenty soft-tissue facial measurements were performed by 5 measurers of varying experiences on 5 adult subjects, using 6 methods - manual calipers, cone-beam CT, virtual reality (VR), 3D photogrammetry, iPad-based 3D photogrammetry, and 2-dimensional photos. Measurement sessions were timed and performed in triplicate, for a total of 9000 measurements. Intraclass correlation coefficient (ICC) was calculated for accuracy and one-way ANOVA was used for comparison. Coefficient of variation was compared among groups to evaluate precision of different methods, considering caliper measurements as the gold standard. RESULTS: ICC among raters was 0.932, indicating excellent reliability. VR was significantly faster than others (137 s vs 217 s for caliper, p<0.001). Coefficient of variation was highest for 2D photographs and lowest for VR (6.4 vs 11.0, p<0.001). The coefficient of variation of caliper was similar to all methods, except 2D photography which was significantly worse. Measurements with the greatest absolute difference from caliper measurements, across modalities, were those around the eyes (left to right exocanthion), tragion to antitragion, and tragion to exocanthion. CONCLUSION: 2D photography is not an accurate way for cephalometric measurements. Virtual reality is the fastest, has the lowest variation between measurements, and is equivalent to caliper measurements. For studies involving a large amount of cephalometrics, virtual reality measurements may be a good option to improve study throughput.

Relationship of Pelvic Positional Change with Leg Length and Offset Measurement in Experimental Total Hip Arthroplasty

During total hip arthroplasty (THA), both pelvic and femur positions affect leg length (LL) and offset (OS) measurements because LL and OS calipers depend on the fixed reference points on the pelvis and femur, respectively. However, LL and OS measurement errors because of pelvic positional changes have not been described. This study aimed to clarify the effects of pelvic positional changes on LL and OS measurements in relation to the pelvic reference using a THA simulator. We developed an experimental THA simulator using Sawbones models of the hemipelvis and femur that facilitated modification of the obliquity, tilt, and rotation of the pelvis. Using an LL and OS caliper, LL and OS measurement errors due to pelvic positional changes were determined with the femoral position fixed. Measurements were performed from two pelvic reference positions: the iliac tubercle (P1) and the top of the iliac crest intersecting the line of the femoral long axis (P2). Concerning pelvic obliquity, the total error of LL was 25.0 mm in P1 and 26.5 mm in P2, while the total error of OS was 13.0 mm in P1 and 10.9 mm in P2. For pelvic tilt, the total error of LL was 9.0 mm in P1 and 3.8 mm in P2, while the total error of OS was 0.5 mm in P1 and 1.0 mm in P2. Regarding pelvic rotation, the total error of LL was 13.8 mm in P1 and 3.2 mm in P2, while the total error of OS was 3.8 mm in P1 and 4.0 mm in P2. Pelvic positional changes alter LL and OS measurements. The acceptable range (error <2 mm) on LL and OS measurement errors of pelvic obliquity was only 2°, regardless of the pelvic reference position. The pelvic reference position should be at the top of the iliac crest intersecting the line of the long axis of the femur because of a small LL measurement error with pelvic tilt and rotation.

Cranial Shape Measurements Obtained Using a Caliper and Elastic Bands Are Useful for Brachycephaly and Deformational Plagiocephaly Screening.

We assessed a method for screening the cranial shape of 1-month-old infants using a simple measuring instrument instead of a three-dimensional scanner. The Mimos craniometer was used to measure cranial length, cranial width, and two diagonal lengths to calculate the cranial index (CI) and cranial asymmetry (CA). We defined a CI > 90% as brachycephaly and CA > 5 mm as deformational plagiocephaly (DP). Intra- and inter-examiner accuracy analyses were performed on a dummy doll and 1-month-old infants. The measurements of healthy 1-month-old infants were compared with previously reported three-dimensional scanner measurements. Intra- and inter-rater measurements showed good accuracy; diagnostic accuracy comparisons of brachycephaly and DP using a three-dimensional scanner showed kappa values of 1.0 and 0.8, respectively. Comparisons were made among 113 infants matched for day-age at the date of measurement; there were no significant differences in the CI (85.0% vs. 85.2%, p = 0.98) and CA (5.9 mm vs. 6.0 mm, p = 0.48) between the scanner and caliper measurements, nor in the prevalence of brachycephaly (12.4% vs. 17.7%, p = 0.35) or DP (58.4% vs. 56.6%, p = 0.89). This simple measurement method using calipers and bands was useful in screening for brachycephaly and DP in 1-month-old infants.

Tracing timing of growth in cultured molluscs using strontium spiking

IntroductionGrowth experiments present a powerful tool for determining the effect of environmental parameters on growth and carbonate composition in biogenic calcifiers. For successful proxy calibration and biomineralization studies, it is vital to identify volumes of carbonate precipitated by these organisms at precise intervals during the experiment. Here, we investigate the use of strontium labelling in mollusc growth experiments.MethodsThree bivalve species (Cerastoderma edule, Mytilus edulis and Ostrea edulis) were grown under monitored field conditions. The bivalves were regularly exposed to seawater with elevated concentrations of dissolved strontium chloride (SrCl2). In addition, the size of their shells was determined at various stages during the experiment using calliper measurements and digital photography. Trace element profiles were measured in cross sections through the shells of these molluscs using laser ablation ICPMS and XRF techniques.ResultsOur results show that doses of dissolved strontium equivalent to 7-8 times the background marine value (~0.6 mmol/L) are sufficient to cause reproducible peaks in shell-incorporated strontium in C. edule and M. edulis shells. No negative effects were observed on shell calcification rates. Lower doses (3-5 times background values) resulted in less clearly identifiable peaks, especially in M. edulis. Strontium spiking labels in shells of O. edulis are more difficult to detect, likely due to their irregular growth.DiscussionStrontium spiking is a useful technique for creating time marks in cultured shells and a reproducible way to monitor shell size during the growing season while limiting physical disturbance of the animals. However, accurate reconstructions of growth rates at high temporal resolution require frequent spiking with high doses of strontium.

Abstract 5017: Simultaneous targeting of EphA3 on glioblastoma and tumor microenvironment to overcome resistance to CART cell therapy in brain cancer

Abstract Glioblastoma multiforme (GBM) is the most aggressive and common form of brain tumor, and it is characterized by an immunosuppressive tumor microenvironment (TME) that supports tumor growth and pathology. An integral part of the GBM stroma is the mesenchymal stem cells (MSC), which promote GBM growth. Receptor tyrosine kinase EphA3, a member of the Eph family, is overexpressed in GBM tumors and in MSCs within TME, making it an attractive target for Chimeric antigen receptor T-cell (CART) therapy. CART has proven incredible promise in certain hematological malignancies therapy but remained short in solid tumors, particularly GBM. We hypothesize that concurrent targeting of cancers and TME by EphA3-CART can be a potential therapeutic option in GBM. We first showed the expression of the EphA3 receptor by Immunohistochemistry and western blot on GBM patient-derived xenograft (PDX), as well as on the SNB-19 and U87 human GBM cell lines and adipose-derived MSCs using flow cytometry. We then generated EphA3-CAR construct derived from a clinically validated anti-EphA3 scFv cloned in a lentiviral vector containing CD28 and CD3ζ, and generated EphA3 CART cells with over 90% efficiency. EphA3 CART cells demonstrated potent antigen specific effector functions. To demonstrate the antitumor activity in vivo, we subcutaneously grafted 1 × 106 GBM cells per NOD-SCID-γ-/- mice and at ~150-300 mm3 tumor volume, mice were then treated with 5 × 106 EphA3-CART or un-transduced (UTD) T cells control. Tumors were monitored by bioluminescence imaging (BLI) and caliper measurements, and mice were followed for survival. Mice treated with EphA3 CART cells exhibited a significant reduction in tumor volume (p&amp;lt;0.005), and prolonged overall survival (p=0.0015). Finally, we tested the ability of EphA3-CART cells to overcome MSC induced CART cell suppression by co-culturing EphA3-CART cells with MSC and EphA3+ GBM target cells while using the CD19+ mantle cell lymphoma cell line JeKo-1 and CART19 as control. MSC conditioned coculture of GBM and CART showed significant suppression of CAR19 in the CART19 - JeKo-1 system (p=000.3), but no significant suppression of EphA3-CART cells. In summary, our results indicate that EphA3 CART cells exhibited potent and specific antitumor activity in vitro and in vivo and ameliorated MSC induced inhibition of CART cell functions, representing a potentially promising therapeutic option in GBM. Citation Format: Ekene J. Ogbodo, Michael W. Ruff, Reona L. Sakemura, Claudia ManriquezRoman, Truc Huynh, James H. Girsch, Olivia L. Sirpilla, Kun Yun, Carli M. Stewart, Ismail Can, Lionel K. Fonkoua, Mehrdad Hefazi, Elizabeth L. Siegler, Saad S. Kenderian. Simultaneous targeting of EphA3 on glioblastoma and tumor microenvironment to overcome resistance to CART cell therapy in brain cancer. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5017.

Abstract 1789: Pre-clinical studies to advance anti-ROR1 CAR-T cell therapy for metastatic prostate cancer

Abstract Background: Alterations in WNT signaling are frequently associated with tumorigenesis and metastasis in many cancers including prostate cancer (PCa). The Wnt ligand, WNT5A, is required for normal prostate gland development and is increased in bone metastatic, castration resistant PCa (CRPC) patients. WNT5A signaling is mediated in part through ROR1, a non-canonical Wnt receptor and fetal oncoprotein for which the therapeutic inhibitory antibody, Zilovertamab, has been developed. Its safety has been clinically proven in trials for chronic lymphocytic leukemia (CLL) and metastatic breast cancer. We sought to investigate Zilovertamab-based anti-ROR1 therapies for metastatic prostate cancer.Our Hypothesis is that WNT5A may activate a stem-cell-like program via ROR1 which leads to therapy resistance in metastatic prostate cancer. The anti-ROR1 biologic, Zilovertamab, may inhibit this mechanism of resistance. The expression of ROR1 on CRPC and NEPC tumors and its lack of expression on normal adult tissues makes it a promising CART cell target. Methods: We used the patient-derived xenograft (PDX), PCSD13, small cell bone metastatic prostate cancer model and the neuroendocrine PCa cell lines, PC3 and DU145, to test the effect of Zilovertamab and Zilovertamab-CART cells in vitro using the real time cell viability, proliferation, and cell cycle tracking assays in an Incucyte S3. We used in vivo bioluminescence and tumor caliper measurements to monitor effects in vivo. Results: Studies using RNASeq, qRT-PCR, FACS and Westerns showed high expression of ROR1 in PC3, DU145, and in PCSD13. CRISPR-Cas9 Knock out of ROR1 in PC3 and DU145 cells showed increased inhibition of proliferation at lower docetaxel concentrations. Treatment of PCSD13 PDX in vivo with Zilovertamab increased docetaxel-mediated tumor growth inhibition. Mice bearing PC3 xenografts injected intravenously with Zilovertamab anti-ROR1 CAR-T cells showed durable, tumor ablation in 67% of mice compared to 22% of mice injected with activated T cells from the same donor and 0% of untreated mice. Survival of mice at Day 70 injected with CART was 78% compared to 0% of mice injected with control donor T cells and 0% of untreated mice. Zilovertamab antibody could inhibit Zilovertamab CART cell tumor cell killing. Conclusions: ROR1 was expressed at high levels on castration resistant small cell PCa and neuroendocrine PCa cell lines and PDX models. Zilovertamab synergized with docetaxel to inhibit tumor growth in patient derived xenograft in vivo and in organoid cultures. Zilovertamab-based CART cells durably eradicated ROR1+ prostate cancer xenograft tumors. These studies supported the recently launched Phase 1b clinical trial of Zilovertamab plus docetaxel in metastatic CRPC patients. Clinical development of GMP Zilovertamab CART cells for a clinical trial in CLL is in progress and may lead to rapid progression to a clinical trial for metastatic CRPC and NEPC. Citation Format: Christina A.M. Jamieson, Jamillah Murtadha, Christopher S. Oh, Michelle Muldong, Evodie Koutouan, JOngwook Kim, Niloofar Etemadfard, Hae Soo Choo, Navyaa Sinha, Sanghee Lee, Christina Wu, Gabriel Pineda, Kathleen Lennon, Karl Willert, Catriona H. Jamieson, Terry Gaasterland, Rana Mckay, Christopher J. Kane, Thomas J. Kipps, Anna A. Kulidjian, Nicholas A. Cacalano, Charles Prussak. Pre-clinical studies to advance anti-ROR1 CAR-T cell therapy for metastatic prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1789.

Abstract 4140: Oncogenic KRAS inhibition with ADT-007 primes T cell responses in pancreatic ductal adenocarcinoma

Abstract Background: Pancreatic ductal adenocarcinoma (PDAC) has low (&amp;lt;11%) 5-year survival due to the presence of advanced disease at diagnosis. Currently standard of care provides a median survival of 1 year in the majority of patients presenting with advanced disease. Targeting oncogenic KRAS, which is mutated in &amp;gt;90% of human PDAC tumors, provides an opportunity to inhibit a critical pathway and significantly impact standard of care in patients diagnosed with PDAC. Furthermore, evidence indicates that inhibiting-oncogenic KRAS signaling modulates anti-tumor immune responses and demonstrates synergy with immune checkpoint blockade in pre-clinical models. The purpose of this study is to evaluate efficacy and impact on anti-tumor immunity of a novel inhibitor, ADT-007 that demonstrates pan-RAS inhibitory activity. Methods: Female C57BL/6J, C57BL/6J. Ifng/Thy1.1 KI;Cg-Tg-IL2tm1/eGFP/Weav (IFNg-Thy1.1/IL-2-GFP reporter mice), or immune deficient (T/B cells) Rag 1 −/− mice (n=5-10/group) were implanted SQ with 106 mouse derived PDAC cell lines (7160c2 and 2838c3) bearing KRASG12D mutation, and then randomized (day 7), into vehicle (5% DMSO/5% Kolliphor/90% H2O) or ADT-007 treatment, which was administered SQ (peritumoral), BID at 5 mg/kg for 2 - 3 weeks. Tumor growth in vitro (IC50), KRAS-MAPK/AKT signaling, in vivo tumor volumes (tv = L x W2/2), and tumor immune responses were assessed by CellTiter Glo, phosphoflow/western blotting, bi-weekly caliper measurements, and multi-parameter flow cytometry, respectively. Results: ADT-007 inhibited KRAS-MAPK/AKT signaling in human and mouse PDAC cell lines inducing G2-M phase arrest and apoptosis at low nanomolar concentrations. Peritumoral administration of ADT-007 inhibited tumor cell growth in vivo, modulating T cells in the tumor microenvironment (TME), specifically, an increase in tumor infiltrating, CD44+ CD62L− CD4+ and CD8+ T cells and increases in production of TNFα, IFN-γ, IL-2, and granzyme B upon stimulation with PMA/ionomycin. Conclusion: The small molecule inhibitor, ADT-007 blocks oncogenic KRAS signaling and modulates T cell activation and function in the TiME of immune competent, syngeneic mouse models of PDAC. More broadly, these findings indicate that a pan-RAS inhibitor is capable of modulating anti-tumor immune responses extending previous observations that targeted inhibition of oncogenic KRASG12C modulates tumor immunity in animal models of colon and lung cancer. Citation Format: Jeremy B. Foote, Tyler E. Mattox, Adam B. Keeton, Ganji N. Purnachandra, Yulia Maxuitenko, Xi Chen, Jacob Valiyaveettil, Donald J. Buchsbaum, Gary A. Piazza, Bassel F. El-Rayes. Oncogenic KRAS inhibition with ADT-007 primes T cell responses in pancreatic ductal adenocarcinoma. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4140.

Abstract 6033: RA123, a new GLS1 allosteric inhibitor demonstrates in vitro and in vivo activity in multiple myeloma models

Abstract The conditional essential amino acid glutamine is utilized by tumors to sustain bioenergetic requirements in a nutrient-poor microenvironment. Based on this specific tumor need, efforts have been made to target the glutamine metabolism using glutaminase inhibitors. In this study, we present in vitro and in vivo studies demonstrating activities and target engagement of a new specific GLS1 allosteric inhibitor (RA123). In vitro, RA123 was tested in imid-resistant, CD38 low RPMI8226 multiple myeloma cell line using the ATP Cell titer Glo assay over a 72h treatment period. In vivo, RA123 was tested in mice xenografted with RPMI8226 multiple myeloma cell line. Compound was given at three different under a BID (twice a day) regimen. Tumor growth under treatment was monitored by caliper measurement for 20 days. A terminal PK/PD study on the same model was setup measuring impact of the drug on the glutamine and glutamate tumor content post 24 hours after the last administration. Over the same period, PK measurements were planned to document drug exposure in the plasma and in the tumor. Finally, in a stand-alone study, target engagement was documented using imaging modality. [18F]FSPG PET radiotracer was used to visualize and quantify impact of RA123 on tumor (RPMI8226) glutamate pool. RA123 was given at three doses under a BID regimen. Treatment was administered for 3 days, and PET signal was recorded at baseline and at end of day 3. In vitro study demonstrated that RA123 was able to induce cell killing of RPMI8226 after 72 hours of treatment. These data translated in vivo by a dose-dependent effect on cell growth. In a terminal PK/PD study, RA123 confirmed a dose-dependent impact on glutamine and glutamate tumor levels which correlated with both plasma and tumor exposure. Further confirmation of target engagement was observed with reduced PET uptake post- RA123 treatment at the three doses tested. In conclusion, this study demonstrated that RA123, a new specific GLS1 allosteric inhibitor was able to impact multiple myeloma tumor cell growth both in vitro and in vivo. This effect was associated with reduced glutamate pools and accumulation of glutamine levels within the tumor cells. Citation Format: Christophe Henry, Dimitri Gorge-Bernat, Pascal Pannier, Isabelle Meaux, Jane Cheng, Fangxian Sun, Olivier Pasquier, Philippe Lienard, Erwan Jouannot, Thierry Gouyon, Geneviève Estenne-Bouhtou, Bailin Zhang, Bérangère Thiers, Laurent Debussche, David Machnik. RA123, a new GLS1 allosteric inhibitor demonstrates in vitro and in vivo activity in multiple myeloma models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6033.

PD04-10 PRE-CLINICAL STUDIES TO ADVANCE ZILOVERTAMAB-BASED ANTI-ROR1 DUAL SPECIFICITY CAR T-CELL THERAPY FOR METASTATIC PROSTATE CANCER.

PD04-10 PRE-CLINICAL STUDIES TO ADVANCE ZILOVERTAMAB-BASED ANTI-ROR1 DUAL SPECIFICITY CAR T-CELL THERAPY FOR METASTATIC PROSTATE CANCER.

Effect of joint line orientation parameters on initial bone resection in mechanically aligned total knee arthroplasty: a retrospective clinicoradiological correlation study

BackgroundDiscrepancies in bone resection between the medial and lateral compartments are very common in total knee arthroplasty (TKA) when mechanical alignment (MA) is used. The purpose of this study was to explore whether and how joint line orientation affects the initial bone resection in mechanically aligned TKA.MethodsA total of 194 patients (225 knees) diagnosed with osteoarthritis (OA) were included. Virtual bone resection was conducted in the coronal view using full-length weight-bearing radiographs according to the technical requirements of MA, and the reliability of the virtual resection was verified via intraoperative caliper measurements. Correlation and regression analyses were conducted between the initial bone resection within the extension gap (EG) and various parameters, including the hip-knee-ankle (HKA) angle, mechanical lateral distal femoral angle (mLDFA), joint line congruence angle (JLCA), and medial proximal tibial angle (MPTA). Moreover, the correlation between intraoperative bone resection adjustments and joint line orientation parameters was also investigated.ResultsAll knees in the current case series were artificially divided into 4 subgroups: subgroup 1, containing 148 varus knees (65.8%) with valgus femurs; subgroup 2, containing 48 varus knees (21.3%) with varus femurs; subgroup 3, containing 17 valgus knees (7.6%) with varus tibias; and subgroup 4, containing 12 valgus knees (5.3%) with valgus tibias. In subgroup 1, the mLDFA and MPTA were positively correlated with the initial bone resection with regression coefficients of 0.670 and 0.089, respectively. Moreover, in all varus knees, intraoperative bone resection adjustments were negatively correlated with mLDFA and MPTA, with categorical regression coefficients of -0.426 and − 0.230, respectively.ConclusionWhen MA-TKAs are performed in varus knees with valgus femurs, the initial bone resection within the EG is mainly positively correlated with mLDFA, while the intraoperative bone resection adjustment is significantly correlated with mLDFA and MPTA in all varus knees.

Cross-sectional reliability and concurrent validity of a quantitative 2-dimensional ultrasound image analysis of effusion and synovial hypertrophy in knee osteoarthritis

ObjectiveEffusion-synovitis is related to pain and progression in knee osteoarthritis (OA), but current gold standard ultrasound (US) measures are limited to semi-quantitative grading of joint distension or 1-dimensional thickness measures. A novel quantitative 2-dimensional image analysis methodology is applied to US images of effusion-synovitis; reliability and concurrent validity was assessed in patients with knee OA. MethodsCross sectional analysis of US images collected from 51 patients with symptomatic knee OA were processed in ImageJ and segmented in 3DSlicer to produce a binary mask of the supra-patellar synovitis region of interest (ROI). Area measures (mm2) of total synovitis, effusion and hypertrophy components were exported. Intra-rater reliability and test-retest reliability (1–14 days washout) were estimated with intra-class correlation coefficients (ICCs). Concurrent validity was measured by Spearman correlations between quantitative measures and gold standard OMERACT and caliper measurements of synovitis. ResultsIntra-rater reliability for hypertrophy area was estimated at 0.98, 0.99 for effusion area, and 0.99 for total synovitis area. The test-retest reliability for total synovitis area was 0.63 (SEM 87.8 ​mm2), 0.59 for hypertrophy area (SEM 21.0 ​mm2), and 0.64 for effusion area (SEM 73.8 ​mm2). Correlation between total synovitis area and OMERACT grade was 0.84, 0.81 between total synovitis area and effusion-synovitis calipers, and 0.81 between total effusion area and effusion calipers. ConclusionThis new research tool for image analysis demonstrated excellent intra-rater reliability, good concurrent validity, and moderate test-retest reliability. Quantitative 2D US measures of effusion-synovitis and its individual components may enhance the study and management of knee OA.

MACGRAN-IJ: AN EFFICIENT METHOD TO ANALYZE DIMENSIONS AND COLOR OF WHEAT GRAINS

The dimensions, shape, and color of wheat grain are important in plant breeding, although their manual measurement is difficult and time-consuming. While it is true that digital image analysis (DIA) with specific software has made it easier, some are semiautomatic, exclude color, and require the digitization of individual grains. The goal of this study was to compare the precision, accuracy, and efficiency of MacGran-IJ versus SmartGrain, GrainScan, and manual caliper measurement. The morphological characteristics of durum wheat (Triticum durum Desf.) and bread wheat (Triticum aestivum L.) grains were measured using DIA with MacGran-IJ, including color (in RGB and CIELab color spaces), size (mm2), lateral length (LLG, mm), ventral length (VLG, mm), lateral (LWG, mm) and ventral (VWG, mm) width, perimeter (mm), and circularity. Length, LWG, and VWG were measured with a digital caliper. The time (s) taken in each method was recorded. Size, LLG, VLG, and VWG were the same between methods for both species, but the LWG measurement obtained with vernier showed differences between the three programs (p ≤ 0.01), as well as low accuracy and precision. VWG obtained manually had R2 values of 0.92, 0.91, and 0.89 (p ≤ 0.01) for MacGran-IJ, SmartGrain, and GrainScan, respectively. The ventral and lateral color red of wheat grain differed between MacGran-IJ and GrainScan; the lateral part of the bread wheat grain showed differences for red, green, and blue (p ≤ 0.05). DIA in CIELab color required 11.57 s, and in RGB it needed 7.25 s (p ≤ 0.01). Manual measurement of 50 wheat grains required 1528.07 s, whereas SmartGrain took 101.84 s, MacGran-IJ 45.4 s, and GrainScan 54.11 s (H = 36.59, p ≤ 0.01). MacGran-IJ is an automated, simple, and efficient method for accurately measuring the size, length, and width of wheat grains using digitized images

Intralesional Treatment of Basal Cell Carcinoma in a Genetically Inducible Mouse Model

Introduction: Patients with nevoid basal cell carcinoma syndrome (NBCCS), also termed Gorlin syndrome, develop numerous basal cell carcinomas (BCCs). Current treatment options include systemic hedgehog pathway inhibitors, with significant side effects when used long term, and multiple surgeries with the risk of scarring and sometimes even disfigurement over the long term. We have developed an intralesional treatment for BCC based on the intracellular delivery of an antisense oligonucleotide (ASO) targeting Gli1, the main protein active in the hedgehog pathway responsible for BCC. To test the efficacy of our treatment, we developed an inducible NBCCS-like mouse model, genetically modified at the level of the expression of the Ptch1 and p53 genes in the skin.&#x0D; Methods: The inducible mouse model uses Ptch1- and P53-knockdown mice irradiated with UVB three times a week until BCCs appear. These BCCs were treated once or twice a week with injections of an antisense oligonucleotide (ASO) targeting the production of the Gli1 protein, with and without the use of a peptide (the “Feldan shuttle") designed to introduce the ASO into the tumor cells. The evolution of the tumors was monitored several times a week with caliper measurements and photography. Following the treatments, the tumor site tissue was harvested and analyzed by both routine (H&amp;E) and specialized (IHC, IF) histologic methods .&#x0D; Results: Intralesional treatment of BCCs in the inducible mouse model using the ASO and peptide combination (FLD103) resulted in rapid and marked tumor regression, and sometimes complete eradication, over a few weeks. In addition, healthy perilesional skin appeared unaffected by the treatment, demonstrating targeted treatment specificity. The healed skin appeared grossly normal. Histologic examination of the treated sites after recovery showed no residual BCC as well as complete healing of the epidermis, dermis and subcutaneous tissue. We also observed that intralesional treatment with the ASO alone is not associated with tumor regression and in fact permits continued tumor growth.&#x0D; Conclusion: We have demonstrated the utility of a specific intralesional treatment for BCC in a genetically inducible mouse model. This alternative and novel treatment produces rapid results with normal healing of the treated site. This treatment could offer an attractive option for patients with multiple BCCs who wish to avoid systemic treatments or multiple surgeries.

Effects of methylphenidate on femoral bone growth in male rats.

The use of Methylphenidate (MP) can have adverse effects on bone growth and mineralization. This study aimed to investigate the underlying pathophysiology of MP-induced skeletal deficits in growing rats using stereological and immunohistochemical methods. Male rats, aged 4weeks, were orally treated with MP through an 8-h/day water drinking protocol. The rats (n=30) were randomly divided into three groups: MP-High Dose (30/60mg/kg/day MP), MP-Low Dose (4/10mg/kg/day MP), and control (water only). After 13weeks, the femoral bones were assessed using calliper measurements, dual-energy X-ray absorptiometry, and biomechanical evaluation. The total femur volume, cartilage volume, growth zone volume, and volume fractions were determined using the Cavalieri method. Immunohistochemical analyses were conducted using alkaline phosphatase and anti-calpain antibody staining. Rats exposed to MP exhibited significant reductions in weight gain, femoral growth, bone mineralization, and biomechanical integrity compared to the control group. The total femoral volume of MP-treated rats was significantly lower than that of the control group. The MP-High Dose group showed significantly higher ratios of total cartilage volume/total femoral volume and total growth zone volume/total femoral volume than the other groups. Immunohistochemical evaluation of the growth plate revealed reduced osteoblastic activity and decreased intracellular calcium deposition with chronic MP exposure. The possible mechanism of MP-induced skeletal growth retardation may involve the inhibition of intracellular calcium deposition in chondrocytes of the hypertrophic zone in the growth plate. In this way, MP may hinder the differentiation of cartilage tissue from bone tissue, resulting in reduced bone growth and mineralization.

Abstract PD2-03: Targeting TGF-β and over-expressing GM-CSF in the Tumor Microenvironment (TME) with AMUN-003 Inhibits Tumor Growth and Metastases and Augments Immune Checkpoint Inhibitor (ICI) Response in Triple Negative Breast Cancer (TNBC)

Abstract Introduction: Despite several therapeutic advancements, metastatic TNBC remains incurable for nearly all patients and is a frequent cause of cancer-related deaths worldwide. We tested the hypothesis that inhibiting suppressive signals sustained by TGF-β and concurrently stimulating recruitment of inflammatory cells with GM-CSF within the TME would result in improved anti-tumor responses. We report here pre-clinical single agent and ICI combination data for a newly developed oncolytic adenovirus rAd.sT.GM (AMUN-003) that expresses both sTGFβRIIFc (a TGF-β protein decoy), and GM-CSF tested in an immunocompetent mouse model. Methods: In addition to manufacturing Amun-003 which is a replication competent adenovirus encoding both sTGFβRIIFc and GM-CSF transgenes, we generated multiple replication competent control viruses coding for sTGFβRIIFc alone, GM-CSF alone or no transgene. We implanted 4T1 cells in the flanks of BALB/c mice, a syngeneic tumor model that is immune competent. On day 7 and day 10 post tumor cell inoculation, we performed saline or single agent virus injections into the tumor. We monitored the health of the animals, performed periodic tumor caliper measurements and collected sera for analysis of cytokines and markers of tissue injury. Animals were sacrificed on Day 25 and tissues collected. Similarly, with the same animal model and using reduced dose Amun-003, we tested combinations with systemic anti-CTLA-4, anti-PD-1 monoclonal antibodies, or both. Animals were sacrificed on Day 25 with specimens collected as above but also to report the metastatic burden in distant tissues. Results: In single agent experiments, all adenovirus constructs were similarly tolerated by the animals with no notable differences in animal weights. General safety as assessed by serum LDH and IL-6 demonstrated no difference between animal groups treated with the different virus constructs. All adenovirus resulted in delay of cancer growth and on day 25, excised tumors from sacrificed animals weighed less than tumors from animals treated with buffer. However, a single dose of Amun-003 was the most effective at delaying tumor progression than any other tested constructs (Day 21 and Day 25: P &amp;lt; 0.0001 vs the buffer group by two-way ANOVA analysis). Next, we conducted experiments evaluating a single dose of Amun-003 alone or in combination with anti-CTLA4, anti-PD1 or both ICI. Combinations of Amun-003 with ICI delayed progression better than ICI treatments alone or Amun-003 alone. Moreover, the combination with both ICI and Amun-003 was most effective at delay of tumor progression (Day 22 and Day 25: P &amp;lt; 0.0001 vs the buffer group; D25: P=0.0119 vs dual ICI therapy). Lung surface metastatic nodule counts and lung tissue luciferase assay for 4T1-luc2 cells both showed treatment consisted of Amun003 and both ICI was most potent in inhibiting lung metastasis (Lung nodules: P &amp;lt; 0.0001 vs the buffer group; Relative luminescence: P=0.0036 vs the buffer group, both by one-way ANOVA analysis). Finally, the metastatic burden in the lung was least in animals treated with Amun-003 and both ICI (42% metastases free) (P = 0.0373 vs the buffer group by fisher’s exact test). Conclusions: In syngeneic immune competent animals harboring an aggressive TNBC tumor, single agent Amun-003 appeared safe and was more effective at controlling tumor progression following an intratumoral injection when compared to other tested adenovirus constructs. In the same model, combinations of ICI with Amun-003 resulted in delayed tumor growth and control of metastatic spread. These results with Amun-003 support advancement to human clinical testing. Citation Format: Sooncheon Shin, Beniamin Filimon, Yuefeng Yang, Zebin Hu, Prem Seth, Vijayakrishna K. Gadi, Weidong Xu. Targeting TGF-β and over-expressing GM-CSF in the Tumor Microenvironment (TME) with AMUN-003 Inhibits Tumor Growth and Metastases and Augments Immune Checkpoint Inhibitor (ICI) Response in Triple Negative Breast Cancer (TNBC) [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr PD2-03.