Micro-magnetic Resonance Research Articles

Alterations in Adipose Tissue and Adipokines in Heterozygous APE1/Ref-1 Deficient Mice.

The role of apurinic/apyrimidinic endonuclease 1/redox factor-1 (APE1/Ref-1) in adipose tissue remains poorly understood. This study investigates adipose tissue dysfunction in heterozygous APE1/Ref-1 deficiency (APE1/Ref-1+/-) mice, focusing on changes in adipocyte physiology, oxidative stress, adipokine regulation, and adipose tissue distribution. APE1/Ref-1 mRNA and protein levels in white adipose tissue (WAT) were measured in APE1/Ref-1+/- mice, compared to their wild-type (APE1/Ref-1+/+) controls. Oxidative stress was assessed by evaluating reactive oxygen species (ROS) levels. Histological and immunohistochemical analyses were conducted to observe adipocyte size and macrophage infiltration of WAT. Adipokine expression was measured, and micro-magnetic resonance imaging (MRI) was used to quantify abdominal fat volumes. APE1/Ref-1+/- mice exhibited significant reductions in APE1/Ref-1 mRNA and protein levels in WAT and liver tissue. These mice also showed elevated ROS levels, suggesting a regulatory role for APE1/Ref-1 in oxidative stress in WAT and liver. Histological and immunohistochemical analyses revealed hypertrophic adipocytes and macrophage infiltration in WAT, while Oil Red O staining demonstrated enhanced ectopic fat deposition in the liver of APE1/Ref-1+/- mice. These mice also displayed altered adipokine expression, with decreased adiponectin and increased leptin levels in the WAT, along with corresponding alterations in plasma levels. Despite no significant changes in overall body weight, microMRI assessments demonstrated a significant increase in visceral and subcutaneous abdominal fat volumes in APE1/Ref-1+/- mice. APE1/Ref-1 is crucial in adipokine regulation and mitigating oxidative stress. These findings suggest its involvement in adipose tissue dysfunction, highlighting its potential impact on abdominal fat distribution and its implications for obesity and oxidative stress-related conditions.

Analysis of wood anatomy via high-resolution micro-MRI and micro-CT: first approaches and results from Pozzo di Bellafonte (Montaione, Florence, Italy)

Several artefacts were found during the excavation performed at “Pozzo di Bellafonte”, located in an archaeological area in the province of Florence (Italy). The material found in the archaeological site included a large amount of wood samples; therefore, a method for understanding the positioning, the shape and the orientation of the wood sample vessels was necessary for identification of botanical taxa. In recent years, the study of ancient remains has benefited greatly from the use of imaging techniques commonly used for medical diagnosis and biomedical research, such as the micro-Computed Tomography (micro-CT) and high-field micro-Magnetic Resonance Imaging (micro-MRI). In this paper such imaging methodologies were employed for assessing the internal structure of representative wood samples extracted from the above mentioned. The complementarity of micro-CT and micro-MRI for the investigation of wood specimen anatomy is also discussed, by underlining what micro-MRI provides as compared with micro-CT and by proposing a tool constituted by the combination of micro-CT/micro-MRI for the study of such wood samples in a multimodal approach.

Metabolic modulation to improve MSC expansion and therapeutic potential for articular cartilage repair

BackgroundArticular cartilage degeneration can result from injury, age, or arthritis, causing significant joint pain and disability without surgical intervention. Currently, the only FDA cell-based therapy for articular cartilage injury is Autologous Chondrocyte Implantation (ACI); however, this procedure is costly, time-intensive, and requires multiple treatments. Mesenchymal stromal cells (MSCs) are an attractive alternative autologous therapy due to their availability and ability to robustly differentiate into chondrocytes for transplantation with good safety profiles. However, treatment outcomes are variable due to donor-to-donor variability as well as intrapopulation heterogeneity and unstandardized MSC manufacturing protocols. Process improvements that reduce cell heterogeneity while increasing donor cell numbers with improved chondrogenic potential during expansion culture are needed to realize the full potential of MSC therapy.MethodsIn this study, we investigated the potential of MSC metabolic modulation during expansion to enhance their chondrogenic commitment by varying the nutrient composition, including glucose, pyruvate, glutamine, and ascorbic acid in culture media. We tested the effect of metabolic modulation in short-term (one passage) and long-term (up to seven passages). We measured metabolic state, cell size, population doubling time, and senescence and employed novel tools including micro-magnetic resonance relaxometry (µMRR) relaxation time (T2) to characterize the effects of AA on improved MSC expansion and chondrogenic potential.ResultsOur data show that the addition of 1 mM L-ascorbic acid-2-phosphate (AA) to cultures for one passage during MSC expansion prior to initiation of differentiation improves chondrogenic differentiation. We further demonstrate that AA treatment reduced the proportion of senescent cells and cell heterogeneity also allowing for long-term expansion that led to a > 300-fold increase in yield of MSCs with enhanced chondrogenic potential compared to untreated cells. AA-treated MSCs with improved chondrogenic potential showed a robust shift in metabolic profile to OXPHOS and higher µMRR T2 values, identifying critical quality attributes that could be implemented in MSC manufacturing for articular cartilage repair.ConclusionsOur results suggest an improved MSC manufacturing process that can enhance chondrogenic potential by targeting MSC metabolism and integrating process analytic tools during expansion.

P-112 Magnetic resonance spectroscopy at the micro scale (Micro-MRS) for non-invasively assessing individual mammalian embryos’ metabolism

Abstract Study question Can Micro-Magnetic Resonance Spectroscopy (micro-MRS) reveal the metabolic fingerprint of mammalian pre-implantation embryo? Summary answer Our research demonstrates that rapid, non-invasive analysis of embryos using Micro-Magnetic Resonance Spectroscopy (micro-MRS) can effectively uncover the significance of embryonic metabolites during development. What is known already Non-invasive selection of optimal embryos for transfer is a crucial challenge in assisted reproduction for both humans and animals. Magnetic Resonance Spectroscopy (MRS) emerges as a promising solution due to its chemical sensitivity and non-invasive nature. Despite its effectiveness in analysing large organisms, MRS faces obstacles in small sample handling and sensitivity, particularly with embryos, limiting its clinical and research applications. Our study overcomes these challenges using innovative microchip-based MRS sensors. This approach enabled successful MRS at the embryo scale, revealing metabolic biomarker linked to developmental potential in pre-implantation bovine embryos and opening new possibilities in assisted reproduction technology. Study design, size, duration Multiple MRS measuring sessions were conducted on two distinct embryo cohorts. The first cohort involved naturally arrested IVP embryos at early (8-cell) and late (morula/blastocyst) stages, establishing metabolic biomarkers in mammalian pre-implantation embryos. The second cohort involved 8-cell embryos which were further cultured over 10 days post-MRS with imaging at specific intervals (3 hours post-thawing, days 5, 8, and 10). This step was crucial to back-correlate biomarkers with in-vitro development based on bright field microscopy. Participants/materials, setting, methods Both studies involved vitrified samples that were subjected to a 50-minute MRS measurement following a 3-hour post-warming culture period. Single bovine embryo analysis was performed using our 1nL micro-MRS probe. Pre-implantation bovine embryos were obtained through IVF (n = 32 samples for cohort one and, n = 61 for cohort two). Post-MRS, the embryos were cultured in equilibrated IVC medium in a humidified incubator with 5% CO2 and 6% O2 at 38 °C. Statistical significance was set at p < 0.05. Main results and the role of chance Micro-Magnetic Resonance Spectroscopy (Micro-MRS) was utilized to identify metabolic markers in single bovine embryos, providing insights into their pre-implantation developmental stages. The spectroscopy data revealed distinct peak regions in the spectra of both cohorts, indicative of essential life-supporting fatty acids. Notably, up to six lipid markers were identified in the first cohort, five of which displayed statistically significant differences (p < 0.05). However, the majority of these biomarkers in the first cohort did not show significant expression differences in the groups of embryos assigned to the second cohort. An exception was noted in the case of the Principal Lipid Component (PLC) marker, which was significantly less prevalent in the arrested embryos than in those progressing to the blastocyst stage (p = 0.0016). Further data analysis based on AI-classification algorithms revealed up to 80% precision in predicting the development to the blastocyst stage solely based on their metabolic profiles at the 8-cell stage. Limitations, reasons for caution Magnetic Resonance Spectroscopy (MRS) shows promise for clinical and research use, requiring reliable signal detection from cellular components. While suitable for research, clinical deployment demands further safety assessments and protocol refinement. Additionally, larger sample sizes and data on post-MRS implantation and pregnancy rates are needed to validate AI-classification algorithms. Wider implications of the findings Micro-Magnetic Resonance Spectroscopy (Micro-MRS) holds potential as a reliable and non-invasive method for embryo screening prior to transfer. This innovative tool can shed light on the role of embryonic metabolites during development making Micro-MRS a key instrument in advancing both embryology research and human assisted reproduction techniques. Trial registration number not applicable

Moxibustion Promoted Axonal Regeneration and Improved Learning and Memory of Post-stroke Cognitive Impairment by Regulating PI3K/AKt and TACC3

BackgroundThis study aimed to investigate whether moxibustion could affect PI3K/Akt pathway to regulate Transforming acidic coiled-coil containing protein 3 (TACC3) and promote axonal regeneration to improve learning and memory function in middle cerebral artery occlusion (MCAO) rats. MethodsSixty SD rats were randomly divided into 4 groups: sham-operated control group (SC), model control group (MC), model + moxibustion group (MM), and model + inhibitor + moxibustion group (MIM). The rats in MC, MM, and MIM groups were made into MCAO models, and PI3K inhibitor LY294002 was injected into the rats in MIM group before modeling; while the rats in SC group were only treated with artery separation without monofilament inserting. After that, the rats in MM and MIM groups were intervented with moxibustion. We used the Zea-Longa scale, micro-Magnetic Resonance Imaging (micro-MRI), Morris water maze (MWM), TUNEL, western blot (WB), immunofluorescence and immunohistochemistry to evaluate the neurological deficits, cerebral infarct volume, learning and memory, apoptotic cell percentage in the hippocampal, the expression level of axonal regeneration and PI3K/AKt related proteins, the expression level of TACC3. The detection of 2 h after surgery showed the result before moxibustion and 7 days after the intervention showed the results after moxibustion. ResultsAfter 7 d of intervention, the scores of Zea-Longa and the cerebral infarct volume, the escape latency, the percentage of apoptosis cells of MM group were lower than that of MC and MIM groups; the frequency of rats crossed the previous platform location, PI3K, p-Akt/t-Akt and TACC3, the level of GAP-43 in MM group was more than MC and MIM groups (P < 0.05). While no statistical difference existed between MIM group and MC group (P > 0.05). ConclusionMoxibustion can promote axonal regeneration and improve learning and memory of Post-stroke cognitive impairment via activating the PI3K/AKT signaling pathway and TACC3.

Abstract 5285: DKK1 blockade inhibits progression of bone metastases via multiple mechanisms and synergize with zoledronate treatment

Abstract Background: Despite the important breakthroughs of immune checkpoint blockade (ICB) therapy in multiple primary cancers, responses of ICB therapy in bone metastases remain extremely poor, largely due to the highly immunosuppressive bone metastasis microenvironment. However, the current understanding of bone metastasis microenvironment is still incomplete, and specific treatment targets for patients with bone metastasis are still lacking. Methods: Transcriptome sequencing data of gastric and breast cancer bone metastasis samples from The TCGA and GEO databases were obtained. The correlation between DKK1 expression and prognosis, intertumoral immune infiltration of bone-metastatic patients was analyzed. Mouse bone metastasis models from gastric, breast and lung cancer were established, and in vivo treatment efficacy of DKK1 and/or zoledronic acid was evaluated by micro-computed tomography (CT), micro-magnetic resonance imaging (MRI) and immunohistochemistry (IHC) staining. The changes of immune components wthin bone-tumor microenvironment were explored via flow cytometry, CyTOF and scRNA-seq analysis. The impact of DKK1 on different immune cell types was analyzed via in vitro co-culture models. Results: Serum detection of gastric cancer patients from our center showed that compared with non-metastatic patients, DKK1 concentration in patients with bone metastasis was the significantly increased (p&amp;lt;0.0001); and compared with other metastatic sites of breast cancer patients, bone metastases had the highest level of DKK1 mRNA expression (p&amp;lt;0.05). Using multiple bone metastasis mouse models, we found that DKK1 blockade significantly controlled the tumor burden of bone metastases. Results of micro-CT and TRAP staining showed that the bone destruction and osteoclast activity within bone lesions were significantly inhibited after DKK1 blockade. In addition, results of flow cytometry and immunofluorescence staining on bone metastasis samples showed that both innate and adaptive anti-immune responses were obviously improved after DKK1 blockade, including increased infiltration of CD8+ T cells, M1 macrophages and decreased infiltration of M2 macrophages. Mechanistically, DKK1 directly induced inhibition of CD8+ T cell activation, M2 polarization of macrophages and maturation of osteoclasts, which all contributes to the immune suppression in the bone metastasis microenvironment. Finally, combined therapy of DKK1 blockade and zoledronic acid had synergistic treatment effects in bone metastases. Conclusion: Our study provides novel insights into the multiple role of DKK1 in the bone metastasis immune microenvironment. DKK1 is a potential immunotherapeutic target for bone metastases, and dual blockade of DKK1 and zoledronic acid is a promising combination immunotherapy strategy for patients with bone metastasis. Citation Format: Tao Shi, Kaijie Liang, Yipeng Zhang, Xiaoyu Zhou, Baorui Liu, Jia Wei. DKK1 blockade inhibits progression of bone metastases via multiple mechanisms and synergize with zoledronate treatment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5285.

Unraveling Spatially Dependent Hydrophilicity and Reactivity of Confined Carbocation Intermediates during Methanol Conversion over ZSM-5 Zeolite.

Carbocations play a pivotal role as reactive intermediates in zeolite-catalyzed methanol-to-hydrocarbon (MTH) transformations. However, the interaction between carbocations and water vapor and its subsequent effects on catalytic performance remain poorly understood. Using micro-magnetic resonance imaging (μMRI) and solid-state NMR techniques, this work investigates the hydrophilic behavior of cyclopentenyl cations within ZSM-5 pores under vapor conditions. We show that the polar cationic center of cyclopentenyl cations readily initiates water nucleus formation through water molecule capture. This leads to an inhomogeneous water adsorption gradient along the axial positions of zeolite, correlating with the spatial distribution of carbocation concentrations. The adsorbed water promotes deprotonation and aromatization of cyclopentenyl cations, significantly enhancing the aromatic product selectivity in MTH catalysis. These results reveal the important influence of adsorbed water in modulating the carbocation reactivity within confined zeolite pores.

Yanggan Jiangmei Formula alleviates hepatic inflammation and lipid accumulation in non-alcoholic steatohepatitis by inhibiting the NF-κB/NLRP3 signaling pathway

The role of NF-κB and the NLRP3 inflammasome in the chronic inflammatory microenvironment of non-alcoholic steatohepatitis (NASH) has been posited as crucial. The Yanggan Jiangmei Formula (YGJMF) has shown promise in ameliorating hepatic steatosis in NASH patients, yet its pharmacological mechanisms remain largely unexplored. This study was conducted to investigate the efficacy of YGJMF in NASH and to elucidate its pharmacological underpinnings. To simulate NASH both in vivo and in vitro, high-fat-diet (HFD) rats and HepG2 cells stimulated with free fatty acids (FFAs) were utilized. The severity of liver injury and lipid deposition was assessed using serum indicators, histopathological staining, micro-magnetic resonance imaging (MRI), and the liver-to-muscle signal intensity ratio (SIRL/M). Furthermore, a combination of enzyme-linked immunosorbent assay (ELISA), immunohistochemistry (IHC), immunofluorescence, real-time quantitative polymerase chain reaction (RT-qPCR), and Western blotting analyses was employed to investigate the NF-κB/NLRP3 signaling pathway and associated cytokine levels. The results from liver pathology, MRI assessments, and biochemical tests in rat models demonstrated YGJMF's significant effectiveness in reducing liver damage and lipid accumulation. Additionally, YGJMF markedly reduced hepatocyte inflammation by downregulating inflammatory cytokines in both liver tissue and serum. Furthermore, YGJMF was found to disrupt NF-κB activation, consequently inhibiting the assembly of the NLRP3 inflammasome in both the in vitro and in vivo models. The preliminary findings of this study suggest that YGJMF may alleviate hepatic steatosis and inhibit the NF-κB/NLRP3 signaling pathway, thereby exerting anti-inflammatory effects in NASH.

The High Potential of Micro-Magnetic Resonance Imaging for the Identification of Archaeological Reeds: The Case Study of Tutankhamun

This study explores the potential of micro-magnetic resonance imaging (μ-MRI) for identifying archaeological reeds found in the tomb of Tutankhamun. Reed plants had various historical uses in the past, with ancient Egyptians extensively employing them for crafting a wide range of items. The distinct cross-sectional characteristics of Arundo donax (giant reed) and Phragmites australis (common reed) are observed and described via optical microscopy and μ-MRI in this study. While optical microscopy offers higher resolution, μ-MRI provides advantages for studying archaeobotanical specimens, as it eliminates the need for mechanical sectioning and potentially damaging fragile samples. The application of μ-MRI on a selected archaeological reed allowed us to identify it as Phragmites australis, showing that μ-MRI can yield clear images, maintaining the integrity of the sample. In contrast, diagnostic features appeared greatly deformed on the thin section observed via optical microscopy. Despite the limitations related to the sample size and the need for sample soaking, μ-MRI presents a valuable tool for analyzing archaeological remains in the field of cultural heritage, with the potential for broader applications. Overall, this study contributes to expanding the toolkit available to researchers studying plant remains, providing insights into reed identification and preservation in archaeological contexts.

U-Net based vessel segmentation for murine brains with small micro-magnetic resonance imaging reference datasets.

Identification and quantitative segmentation of individual blood vessels in mice visualized with preclinical imaging techniques is a tedious, manual or semiautomated task that can require weeks of reviewing hundreds of levels of individual data sets. Preclinical imaging, such as micro-magnetic resonance imaging (μMRI) can produce tomographic datasets of murine vasculature across length scales and organs, which is of outmost importance to study tumor progression, angiogenesis, or vascular risk factors for diseases such as Alzheimer's. Training a neural network capable of accurate segmentation results requires a sufficiently large amount of labelled data, which takes a long time to compile. Recently, several reasonably automated approaches have emerged in the preclinical context but still require significant manual input and are less accurate than the deep learning approach presented in this paper-quantified by the Dice score. In this work, the implementation of a shallow, three-dimensional U-Net architecture for the segmentation of vessels in murine brains is presented, which is (1) open-source, (2) can be achieved with a small dataset (in this work only 8 μMRI imaging stacks of mouse brains were available), and (3) requires only a small subset of labelled training data. The presented model is evaluated together with two post-processing methodologies using a cross-validation, which results in an average Dice score of 61.34% in its best setup. The results show, that the methodology is able to detect blood vessels faster and more reliably compared to state-of-the-art vesselness filters with an average Dice score of 43.88% for the used dataset.

P-127 Micro-Magnetic Resonance Spectroscopy of individual mammalian embryos: a safe and non-invasive diagnostic tool for embryo screening in assisted reproduction

Abstract Study question Is micro-magnetic resonance (micro-MRS) a safe tool to non-invasively unravel the metabolic fingerprint of single mammalian embryos? Summary answer We successfully tested the safety of micro-MRS in a mouse model. No long or short-term adverse effect was found in vitro and in vivo. What is known already Non-invasive selection of the best embryo to transfer is one of the most significant challenges for ART professionals. The chemical sensitivity, resolving power, and, more importantly, the non-invasive nature of MRS makes it an excellent candidate to investigate the building blocks of complex organisms. Although MRS is a well-established technique for the biochemical profiling of large organisms, handling small samples like embryos and 3D cell cultures and spheroids alongside sensitivity issues has prevented its adoption for clinical and research applications. Our group has overcome these limitations with microchip-based sensors to leverage non-invasive MRS technology down to the embryo scale. Study design, size, duration This safety study was divided into two main phases in order to test all the components needed to operate the micro-MRS analysis. In phase 1 we used &amp;gt; 800 2-cell embryos to test a) radiofrequency exposure, b) device activation, and c) static magnetic field exposure. In phase 2 we confirmed in-vivo that MF exposure was not affecting live animals (n267) over 3 generations of mice by assessing different IVF outcomes, natural mating, live parameters and histopathology. Participants/materials, setting, methods Cryopreserved 2-cell B6D2F1 mouse embryos were purchased cryopreserved from JanvierLabs. In phase1 embryos were thawed and used for testing micro-MRS main components' safety. Mouse embryo assay (MEA) was used to first assess micro-MRS adverse effects in-vitro. In phase 2 embryos were exposed to magnetic field in a 9.4T Magnet for 1h at 37 °C, then surgically transferred to surrogate mothers. Both mothers and progeny were tracked up to F3. Statistical significance was set at p &amp;lt; 0.05. Main results and the role of chance This is the first study ever conducted to assess the safety and efficacy of micro-MRS on pre-implantation mammalian embryos. Our results show, with statistical significance, that micro-MRS is safe for use in mammalians. In particular, we have de-risked the three main aspects/components that could affect the embryos: 1) Materials to which the embryos are exposed when loaded into the device, 2) Radiofrequency energy required to activate the device and perform a measurement, and 3) Magnetic field required to analyze the embryos biochemistry in a non-invasive way. Overall no adverse effect was observed when the primary IVF outcomes were assessed, such as implantation rate and live birth rate. In addition, no visible alteration of the overall appearance of ED14.5 foetuses, F1, F2 and F3 mice was observed when comparing the mice obtained from treated or control embryos. The surrogate mothers did not display any significant alteration or visible health issues after embryo transfer. F1 and F2 mice were able to naturally mate and conceive new progeny suggesting that the reproductive potential of the animals is not affected by the magnetic field exposure. Furthermore, histopathological analysis of 5 representative organs did not reveal adverse effects on both surrogate mothers or F1 pups. Limitations, reasons for caution A vital step towards establishing MRS as a clinical and research tool is the reliable detection of a wide range of signals from cellular components. Our method is ready for R&amp;D studies, while its clinical application requires further safety studies and protocol optimization. Wider implications of the findings A non-invasive quick assay would provide the means to reveal the role of embryonic lipids throughout development. Micro-MRS can further develop into a safe embryo assay for selection before embryo transfer. This would apply to both human and animal ART, whose success rate is relatively low. Trial registration number NA

Evaluation of the efficacy of micro-Magnetic Resonance Imaging compared with light microscopy to investigate the anatomy of modern and ancient waterlogged wood

Light microscopy is the conventional method used to investigate wood anatomy, identify the wood taxon, and assess its conservation state. It generally requires the mechanical cut of thin sections from a sample to obtain informative images. When dealing with wooden artworks or ancient remains (e.g., archaeological waterlogged wood), it is important to avoid sample destruction. In this work the efficacy of micro-magnetic resonance imaging (μ-MRI) to investigate the anatomy of waterlogged wood is assessed in comparison with light microscopy. Images along the three anatomical directions (transverse, tangential and radial) of six modern wood species and one archaeological specimen of waterlogged wood (from the Neolithic site “La Marmotta”) were obtained both by μ-MRI and light microscopy. μ-MRI images were acquired virtually selecting 2D slices along the three wood anatomical directions. A 3D reconstruction was derived from 2D μ-MRI images. Conventional light microscopy histology was obtained by manually cutting thin sections. To the best of our knowledge, this is the first study in which high-resolution MR images and light microscopy images of the three anatomical directions of seven wood species are compared. The non-destructive μ-MRI approach allows to investigate the 2D and 3D topological organization of the whole waterlogged wood sample up to a resolution of 8 μm. Although the optical microscope attains higher image resolutions and remains superior in the observation of wood diagnostic characters, multi-parametric μ-MRI provides physiological investigation complementary to light microscopy, giving information concerning both a single section and the whole volume of the sample. The presented study may represent a starting point for further improvements of μ-MRI techniques applied to the non-destructive investigation of waterlogged wood samples, especially those of interest for cultural heritage.

Experimental assessment of cardiovascular physiology in the chick embryo.

High resolution assessment of cardiac functional parameters is crucial in translational animal research. The chick embryo is a historically well-used in vivo model for cardiovascular research due to its many practical advantages, and the conserved form and function of the chick and human cardiogenesis programs. This review aims to provide an overview of several different technical approaches for chick embryo cardiac assessment. Doppler echocardiography, optical coherence tomography, micromagnetic resonance imaging, microparticle image velocimetry, real-time pressure monitoring, and associated issues with the techniques will be discussed. Alongside this discussion, we also highlight recent advances in cardiac function measurements in chick embryos.

Label-free atherosclerosis diagnosis through a blood drop of apolipoprotein E knockout mouse model using surface-enhanced Raman spectroscopy validated by machine learning algorithm.

The direct preventative detection of flow-induced atherosclerosis remains a significant challenge, impeding the development of early treatments and prevention measures. This study proposes a method for diagnosing atherosclerosis in the carotid artery using nanometer biomarker measurements through surface-enhanced Raman spectroscopy (SERS) from single-drop blood samples. Atherosclerotic acceleration is induced in apolipoprotein E knockout mice which underwent a partial carotid ligation and were fed a high-fat diet to rapidly induce disturbed flow-induced atherosclerosis in the left common carotid artery while using the unligated, contralateral right carotid artery as control. The progressive atherosclerosis development of the left carotid artery was verified by micro-magnetic resonance imaging (micro-MRI) and histology in comparison to the right carotid artery. Single-drop blood samples are deposited on chips of gold-coated ZnO nanorods grown on silicon wafers that filter the nanometer markers and provide strong SERS signals. A diagnostic classifier was established based on principal component analysis (PCA), which separates the resultant spectra into the atherosclerotic and control groups. Scoring based on the principal components enabled the classification of samples into control, mild, and severe atherosclerotic disease. The PCA-based analysis was validated against an independent test sample and compared against the PCA-PLS-DA machine learning algorithm which is known for applicability to Raman diagnosis. The accuracy of the PCA modification-based diagnostic criteria was 94.5%, and that of the machine learning algorithm 97.5%. Using a mouse model, this study demonstrates that diagnosing and classifying the severity of atherosclerosis is possible using a single blood drop, SERS technology, and machine learning algorithm, indicating the detectability of biomarkers and vascular factors in the blood which correlate with the early stages of atherosclerosis development.

066 A novel single embryo micro-magnetic resonance spectroscopy device to improve cattle ART outcomes

066 A novel single embryo micro-magnetic resonance spectroscopy device to improve cattle ART outcomes

Lossy Image Compression in a Preclinical Multimodal Imaging Study.

The growing use of multimodal high-resolution volumetric data in pre-clinical studies leads to challenges related to the management and handling of the large amount of thesedatasets. Contrarily to the clinical context, currently there are no standard guidelines to regulate the use of image compression in pre-clinical contexts as a potential alleviationof this problem. In this work, the authors study the application of lossy image coding to compress high-resolution volumetric biomedical data. The impact of compression on the metricsand interpretation of volumetric data was quantified for a correlated multimodal imaging study to characterize murine tumor vasculature, using volumetric high-resolution episcopic microscopy (HREM), micro-computed tomography (µCT), and micro-magnetic resonance imaging (µMRI). The effects of compression were assessed by measuring task-specific performances of several biomedical experts who interpreted and labeled multiple data volumes compressed at different degrees. Wedefined trade-offs between data volume reduction and preservation of visual information, which ensured the preservation of relevant vasculature morphology at maximum compression efficiency across scales. Using the Jaccard Index (JI) and the average Hausdorff Distance (HD) after vasculature segmentation, we coulddemonstrate that, in this study, compression that yields toa 256-fold reduction of the data size allowed to keep the error induced by compression below the inter-observer variability, with minimal impact on the assessment of the tumor vasculature across scales.

Novel micro-MRI approach for subchondral trabecular bone analysis in patients with hip osteoarthritis is comparable to micro-CT approach

To test the agreement between a newly developed micro-magnetic resonance imaging (MRI) analysis of the subchondral bone and the micro-computed tomography (CT) approach. Samples obtained from 10 patients with osteoarthritis undergoing total hip arthroplasty were scanned with a 7.0 T micro-MRI. Proton density-weighted images and proton density-weighted images with fat suppression were obtained. The results were validated with a micro-CT device. Micro-MRI and micro-CT scans of the same sample were aligned, and regions of interest were delineated on equal areas of the sample. Bone volume fraction was calculated by using in-house plugins. The agreement between the methods was tested with Bland-Altman analysis. The agreement between the methods was good, with average difference of 2.167%. The differences between the methods were not significant (P=0.272, t test). The novel micro-MRI approach could be used for subchondral bone analysis. With further optimization for clinical MRI machines, the approach can be also used in the diagnostics of hip osteoarthritis.

Establishment of a visualized mouse orthotopic model of nasopharyngeal carcinoma.

e18048 Background: Nasopharyngeal carcinoma, one of the most common head and neck tumors, is particularly prevalent in Southeast Asia and is characterized by high rates of metastasis and recurrence. Although mouse orthotopic tumor models are commonly employed in studies investigating the mechanisms underlying tumor development and progression, as well as preclinical treatment, currently no such model exists for nasopharyngeal carcinoma. The aim of the current study is to, therefore, establish an orthotopic murine model for nasopharyngeal carcinoma. Methods: To this end, human nasopharyngeal carcinoma C666-1-luc cells, stably expressing the firefly luciferase gene, were injected subcutaneously into the right axilla of BALB/C nude mice. Four weeks later, the resulting subcutaneous tumors were cut into small blocks and grafted into the nasopharynx of immunodeficient BALB/C nude mice to induce tumor formation. Tumor growth was monitored by bioluminescence imaging and small animal magnetic resonance imaging. The histological and immunological antigen expression associated with orthotopic nasopharyngeal carcinoma were analyzed by tissue section analysis and immunohistochemistry (IHC). Results: The tumor formation rate was over 90%. Fluorescence signal detection, micro-magnetic resonance imaging and hematoxylin and eosin staining revealed the successful growth of tumors in the nasopharynx of nude mice. Moreover, IHC analysis detected positive CK, CK5/6, P40 and P63 expression in the mouse orthotopic tumors, which is consistent with the reported antigen expression in the nasopharyngeal tumors of patients. Conclusions: Hence, we successfully established a mouse orthotopic model of nasopharyngeal carcinoma that is highly reproducible, and simple in operation, with low mortality and high tumor formation rates. Moreover, this model facilitates real-time monitoring of tumor growth via in vivo imaging technology, thus, providing a platform for researching nasopharyngeal carcinoma that is more conducive to preclinical research.

Human Obesity Attenuates Cardioprotection Conferred by Adipose Tissue-Derived Mesenchymal Stem/Stromal Cells.

To explore the impact of obesity on reparative potency of adipose tissue-derived mesenchymal stromal/stem cells (A-MSC) in hypertensive cardiomyopathy, A-MSC were harvested from subcutaneous fat of obese and age-matched non-obese human subjects during bariatric or kidney donation surgeries, and then injected into mice 2weeks after inducing renovascular hypertension (RVH) or sham surgery. Two weeks later, left ventricular (LV) function and deformation were estimated in vivo by micro-magnetic resonance imaging and myocardial damage ex vivo. Blood pressure and myocardial wall thickening were elevated in RVH + Vehicle and normalized only by lean-A-MSC. Both A-MSC types reduced LV mass and normalized the reduced LV peak strain radial in RVH, yet obese-A-MSC also impaired LV systolic function. A-MSC alleviated myocardial tissue damage in RVH, but lean-A-MSC decreased oxidative stress more effectively. Obese-A-MSC also showed increased cellular inflammation in vitro. Therefore, obese-A-MSC are less effective than lean-A-MSC in blunting hypertensive cardiomyopathy in mice with RVH.

Intervertebral Disc Degeneration Induced by Needle Puncture and Ovariectomy: A Rat Coccygeal Model.

Objectives To establish a novel animal model of intervertebral disc degeneration (IVDD) in rats and to investigate the effect of 17β-estradiol (E2) intervention in this model. Methods This study was divided into two parts: animal model (four groups: Sham, ovariectomy (OVX), Puncture, and OVX+Puncture; three-time points: 4, 8, and 12 weeks; three female rats/group/time point) and drug intervention (Sham, OVX+Puncture+corn oil, and OVX+Puncture+E2; three female rats/group). The rats were analyzed by micromagnetic resonance imaging (MRI), hematoxylin and eosin (HE) staining, and safranin-O staining. Results MRI and histological scores significantly differed among the four groups at the three-time points (all P < 0.05). IVDD progressed with time in the OVX, Puncture, and OVX+Puncture groups (all P < 0.05). The changes were the most obvious in the OVX+Puncture group. In the E2 intervention part, the Veh group had the worst MRI signals and histological scores (P < 0.05). The MRI scores in the E2 group were less obvious compared to the Sham group (P > 0.05). Also, the histological scores were significantly different between the Sham and E2 groups (P < 0.05). Conclusions The combination of ovariectomy and needle puncture can synergically induce IVDD in rat coccygeal discs. Estrogen treatment can effectively ameliorate IVDD progression in the newly established IVDD models.