Quantitative Analysis Of Distribution Research Articles

Quantitative analysis of spatial distribution of land surface temperature (LST) in relation Ecohydrological, terrain and socio- economic factors based on Landsat data in mountainous area

Land surface temperature (LST) is considered as one of the most significantly effective factors on the regional climate and ecology, playing an important role in connecting surface energy and water exchange. In mountainous regions, LST reveals lots of inconsistencies due to the effect of such factors as topography, vegetation, solar radiation, etc. We sought to investigate the the temporal and spatial variation LST in different years and its relationship with effective factors in 5 dimensions using Multiple statistical methods, the sepidan region in northwest Iran. The multi-factorial land use, topographic (elevation, slope, aspect), biophysical indices (normalized difference vegetation index (NDVI), normalized difference moisture index (NDMI), normalized difference built up index (NDBI), and modified of normalized difference water index (MNDWI)), socio-economic (fossil fuel CO 2 emissions (FFCOE) and road density(RD)), and climate (temperature and solar radiation) was studied in the current research. To this end, Images of July 1998 and 2017 were extracted from Thematic Mapper (TM5) and Operational Land Imager/Thermal infrared sensors (OLI/TIRS8). Moreover, ordinary least squares regression (OLS), Best subset regression, and Hierarchical Partitioning Analysis (HP) were used to investigate the relationship between LST and relevant effective factors. The results indicated that the temperature range varied from 10 to 53 °C in the time period mentioned. The highest amount of LST was observed in barren land use and the lowest one was found in garden lands. An negative correlation was found between LST and elevation. On the other hand, the highest value of the Laps rate of surface temperature was observed in the southern aspects and the lowest one was observed in the western aspects. Furthermore, the highest and lowest values of lase rate were found in slopes less than 10°, and in 50 to 60-degree slopes, respectively. The results of the OLS correlation indicated a negative correlation between LST and NDVI, NDMI, and MNDWI, and a positive correlation of LST with climatic and socio-economic indicators. LST’s highest and lowest correlations were found to be with vegetation (R 2 = 0.95) and road density (R 2 = 0.1). Finally, while in 1998 temperature and vegetation were identified as the most influential factors on LST, it was the elevation that was found to be the most effective factor on LST in 2017 with the effective rate of 82.72%. This study offers a valuable viewpoint on the temporal and spatial variations of LST, their complexity, and the environmental factors that affect them. The viewpoint could, therefore, be used for prospective studies on the analysis of the ecosystem's reaction to climate changes.

Quantitative analysis of size and regional distribution of corpora amylacea in the hippocampal formation of obstructive sleep apnoea patients

Corpora amylacea (CoA) are spherical aggregates of glucose polymers and proteins within the periventricular, perivascular and subpial regions of the cerebral cortex and the hippocampal cornu ammonis (CA) subfields. The present study quantified the distribution of CoA in autopsied hippocampi of patients with obstructive sleep apnoea (OSA) using ethanolamine-induced fluorescence. CoA were observed in 29 of 30 patients (96.7%). They were most abundant in periventricular regions (wall of lateral ventricle, alveus, fimbria and CA4), rarely found in the CA3 and CA1, and undetectable in the CA2 or subiculum. A spatiotemporal sequence of CoA deposition was postulated, beginning in the fimbria and progressively spreading around the subpial layer until they extended medially to the wall of the lateral ventricle and laterally to the collateral sulcus. This ranked CoA sequence was positively correlated with CoA packing density (count and area fraction) and negatively correlated with CoA minimum diameters (p < 0.05). Although this sequence was not correlated with age or body mass index (BMI), age was positively correlated with the mean and maximum diameters of CoA. These findings support the view that the spatiotemporal sequence of CoA deposition is independent of age, and that CoA become larger due to the accretion of new material over time.

Quantitative Analysis of Concentration Distribution in Non-Evaporative Diesel Spray with Laser Induced Fluorescence Method

Quantitative Analysis of Concentration Distribution in Non-Evaporative Diesel Spray with Laser Induced Fluorescence Method

Determining the spatial distribution of laser-induced plasma by laser-induced voltaic measurement

Distribution characteristics of laser-induced plasma (LIP) plays a significant role in the mechanism studies and applications. Owing to various technologies and limitations, the quantitative analysis of plasma channel distribution has not been fully expounded. Here in this work, laser-induced voltage (LIV) measurement was employed to assess the distribution of LIP channels. An exponential relationship was observed between the energy and the voltage, indicating that the laser energy has a controllable influence on the plasma channel. The distribution of plasma channels is captured by moving the electrode position in two directions. LIV is provided as an innovative method to describe the distribution of plasma channels, which is a promising means for characterizing the formation, diffusion and expansion of plasma.

A quantitative analysis of extension and distribution of lung injury in COVID-19: a prospective study based on chest computed tomography

BackgroundTypical features differentiate COVID-19-associated lung injury from acute respiratory distress syndrome. The clinical role of chest computed tomography (CT) in describing the progression of COVID-19-associated lung injury remains to be clarified. We investigated in COVID-19 patients the regional distribution of lung injury and the influence of clinical and laboratory features on its progression.MethodsThis was a prospective study. For each CT, twenty images, evenly spaced along the cranio-caudal axis, were selected. For regional analysis, each CT image was divided into three concentric subpleural regions of interest and four quadrants. Hyper-, normally, hypo- and non-inflated lung compartments were defined. Nonparametric tests were used for hypothesis testing (α = 0.05). Spearman correlation test was used to detect correlations between lung compartments and clinical features.ResultsTwenty-three out of 111 recruited patients were eligible for further analysis. Five hundred-sixty CT images were analyzed. Lung injury, composed by hypo- and non-inflated areas, was significantly more represented in subpleural than in core lung regions. A secondary, centripetal spread of lung injury was associated with exposure to mechanical ventilation (p < 0.04), longer spontaneous breathing (more than 14 days, p < 0.05) and non-protective tidal volume (p < 0.04). Positive fluid balance (p < 0.01), high plasma D-dimers (p < 0.01) and ferritin (p < 0.04) were associated with increased lung injury.ConclusionsIn a cohort of COVID-19 patients with severe respiratory failure, a predominant subpleural distribution of lung injury is observed. Prolonged spontaneous breathing and high tidal volumes, both causes of patient self-induced lung injury, are associated to an extensive involvement of more central regions. Positive fluid balance, inflammation and thrombosis are associated with lung injury.Trial registration Study registered a priori the 20th of March, 2020. Clinical Trials ID NCT04316884.

Study of Organic Matter of Unconventional Reservoirs by IR Spectroscopy and IR Microscopy

The study of organic matter content and composition in source rocks using the methods of organic geochemistry is an important part of unconventional reservoir characterization. The aim of this work was the structural group analysis of organic matter directly in the source rock in combination with a quantitative assessment and surface distribution analysis of the rock sample by FTIR spectroscopy and FTIR microscopy. We have developed new experimental procedures for semi-quantitative assessment of the organic matter content, composition and distribution in the source rocks and applied these procedures for the study of the samples from the Bazhenov shale formation (West Siberia, Russia). The results have been verified using the data from the study of organic matter obtained by Rock-Eval pyrolysis and differential thermal analysis. The obtained results demonstrate the prospects of FTIR spectroscopy and FTIR microscopy application for non-destructive and express analysis of the chemical structure and distribution of organic matter in rocks.

GANDA: A deep generative adversarial network conditionally generates intratumoral nanoparticles distribution pixels-to-pixels

Intratumoral nanoparticles (NPs) distribution is critical for the success of nanomedicine in imaging and treatment, but computational models to describe the NPs distribution remain unavailable due to the complex tumor-nano interactions. Here, we develop a Generative Adversarial Network for Distribution Analysis (GANDA) to describe and conditionally generates the intratumoral quantum dots (QDs) distribution after i.v. injection. This deep generative model is trained automatically by 27,775 patches of tumor vessels and cell nuclei decomposed from whole-slide images of 4 T1 breast cancer sections. The GANDA model can conditionally generate images of intratumoral QDs distribution under the constraint of given tumor vessels and cell nuclei channels with the same spatial resolution (pixels-to-pixels), minimal loss (mean squared error, MSE = 1.871) and excellent reliability (intraclass correlation, ICC = 0.94). Quantitative analysis of QDs extravasation distance (ICC = 0.95) and subarea distribution (ICC = 0.99) is allowed on the generated images without knowing the real QDs distribution. We believe this deep generative model may provide opportunities to investigate how influencing factors affect NPs distribution in individual tumors and guide nanomedicine optimization for molecular imaging and personalized treatment.

The Distribution of Mineral Elements and Protein in Soft Rice was Determined and Analyzed by SEM-EDS Technology

Soft rice is a kind of high-quality rice between glutinous rice and sticky rice. It has low amylose content, crystal clear grains, sweet taste, soft glutinous, and is suitable for cooking and porridge. Chalkiness in soft rice is a white opaque part formed by loose endosperm. It is an important character that affects the appearance quality, processing, and cooking quality of rice, and also an important limiting factor that restricts the standard rate of high-quality rice in China. The combination of scanning electron microscope and energy dispersive spectrometer (SEM-EDS) can be used for in-depth analysis of rice, visualization, and quantitative analysis of element distribution in rice. The results showed that there were many kinds of mineral elements in soft rice seeds, among which C and O were the most abundant, followed by N and P, and Mg, Al, P, S, K, Ca, Mn, and Zn were less. The contents of C, N, P, and S in the non-chalky area were significantly higher than those in the chalky area. Especially N and S were the best indicators of protein, and the contents in the chalky area were higher than those in the non-chalky area. It means that the protein content in the chalky part of soft rice seed is less than that in the non-chalky part, which affects the nutritional quality of soft rice. Therefore, the results of this study laid a solid foundation for the in-depth analysis of the distribution of mineral elements and protein in soft rice and their effects on the quality of soft rice, which also provided important information for the cultivation of new high-quality rice varieties in the future.

Quantitative analysis of in-vivo microbubble distribution in the human brain

Microbubbles (MB) are widely used as contrast agents to perform contrast-enhanced ultrasound (CEUS) imaging and as acoustic amplifiers of mechanical bioeffects incited by therapeutic-level ultrasound. The distribution of MBs in the brain is not yet fully understood, thereby limiting intra-operative CEUS guidance or MB-based FUS treatments. In this paper we describe a robust platform for quantification of MB distribution in the human brain, allowing to quantitatively discriminate between tumoral and normal brain tissues and we provide new information regarding real-time cerebral MBs distribution. Intraoperative CEUS imaging was performed during surgical tumor resection using an ultrasound machine (MyLab Twice, Esaote, Italy) equipped with a multifrequency (3–11 MHz) linear array probe (LA332) and a specific low mechanical index (MI < 0.4) CEUS algorithm (CnTi, Esaote, Italy; section thickness, 0.245 cm) for non-destructive continuous MBs imaging. CEUS acquisition is started by enabling the CnTI PEN-M algorithm automatically setting the MI at 0.4 with a center frequency of 2.94 MHz–10 Hz frame rate at 80 mm—allowing for continuous non-destructive MBs imaging. 19 ultrasound image sets of adequate length were selected and retrospectively analyzed using a custom image processing software for quantitative analysis of echo power. Regions of interest (ROIs) were drawn on key structures (artery–tumor–white matter) by a blinded neurosurgeon, following which peak enhancement and time intensity curves (TICs) were quantified. CEUS images revealed clear qualitative differences in MB distribution: arteries showed the earliest and highest enhancement among all structures, followed by tumor and white matter regions, respectively. The custom software built for quantitative analysis effectively captured these differences. Quantified peak intensities showed regions containing artery, tumor or white matter structures having an average MB intensity of 0.584, 0.436 and 0.175 units, respectively. Moreover, the normalized area under TICs revealed the time of flight for MB to be significantly lower in brain tissue as compared with tumor tissue. Significant heterogeneities in TICs were also observed within different regions of the same brain lesion. In this study, we provide the most comprehensive strategy for accurate quantitative analysis of MBs distribution in the human brain by means of CEUS intraoperative imaging. Furthermore our results demonstrate that CEUS imaging quantitative analysis enables discernment between different types of brain tumors as well as regions and structures within the brain. Similar considerations will be important for the planning and implementation of MB-based imaging or treatments in the future.

Distribution characteristics of elemental grade and mineralization grade of Jiama copper deposit

The change of metallogenic element grade in borehole has strong randomness and nonlinearity. Quantitative analysis of metallogenic element grade distribution is one of the important ways to reveal the degree of mineralization enrichment. Taking Jiama porphyry copper deposit as an example, the distribution structure of Cu grade series in exploration lines 32 and 36 is analyzed by lacunarity method, and this paper discusses the relationship between lacunarity index and mineralization intensity. It is found that both the lacunarity index and parameters of Cu element in different boreholes exist distinction, the higher the mineralization intensity, the smaller the lacunarity index and the smoother the curve, pointing that the high-grade Cu element is more enriched and uniform. The size of lacunarity index and the variation trend of lacunarity curve have a certain indicative effect on mineralization grade.

Characteristics of a gold-doped electrode for application in high-performance lithium-sulfur battery

Bulk sulfur incorporating 3 wt% gold nano-powder is investigated as possible candidate to maximize the fraction of active material in the Li-S battery cathode. The material is prepared via simple mixing of gold with molten sulfur at 120 °C, quenching at room temperature, and grinding. Our comprehensive study reports relevant electrochemical data, advanced X-ray computed tomography (CT) imaging of the positive and negative electrodes, and a thorough structural and morphological characterization of the S:Au 97:3 w/w composite. This cathode exhibits high rate capability within the range from C/10 to 1C, a maximum capacity above 1300 mAh gS−1, and capacity retention between 85% and 91% after 100 cycles at 1C and C/3 rates. The novel formulation enables a sulfur fraction in the composite cathode film as high as 78 wt%, an active material loading of 5.7 mg cm−2, and an electrolyte/sulfur (E/S) ratio of 5 μL mg−1, which lead to a maximum areal capacity of 5.4 mAh cm−2. X-ray CT at the micro- and nanoscale reveals the microstructural features of the positive electrode that favor fast conversion kinetics in the battery. Quantitative analysis of sulfur distribution in the porous cathode displays that electrodeposition during the initial cycle may trigger an activation process in the cell leading to improved performance. Furthermore, the tomography study reveals the characteristics of the lithium anode and the cell separator upon a galvanostatic test prolonged over 300 cycles at a 2C rate.

Quantitative analysis of moisture distribution and transfer in firefighter protective clothing exposed to low-intensity radiation with/without hot steam

Objectives. This study aimed to examine moisture distribution and transfer in firefighter protective clothing when moisture from the atmosphere and skin sweat were considered simultaneously.Methods. A self-developed test apparatus was used to simulate moisture transfer through the protective clothing under exposure to thermal radiation. The weights of each layer of fabric before and after heat exposure were measured to analyze the moisture distribution and transfer. Results. The moisture level in each layer of fabric before the exposure presented an increase over the initial moisture content. After dry heat exposure, the moisture content in each layer of fabric reduced gradually. However, the existence of hot steam increased the moisture content stored in the fabric system and accelerated the moisture transmitting to the skin surface. In addition, the amount of outward water evaporation for dry heat exposure was moderately more than inward water evaporation, while the amount of inward water evaporation was greatly more than outward water evaporation for wet heat exposure. Conclusion. Moisture transfer in the firefighter protective clothing was a two-way transmission during both heat exposures. Understanding moisture transfer helps to provide proper guidance to improve the thermal protection of clothing and reduce steam burns.

Approaches to Biomass Kinetic Modelling: Thermochemical Biomass Conversion Processes

Modeling of biomass pyrolysis kinetics is an essential step towards reactors design for energy production. Determination of the activation energy, frequency factor, and order of the reaction is necessary for the design procedure. Coats and Redfern's work using the TGA data to estimate these parameters was the cornerstone for modeling. There are two significant problems with biomass modeling, the first is the determination of the kinetic triplet (Activation energy, Frequency factor, and the order of reaction), and the second is the quantitative analysis of products distribution. Methods used in modeling are either One-step or Multistep methods. The one-step techniques allow the determination of kinetic triplet but fail to predict the product distribution, whereas multistep processes indicate the product's distribution but challenging to estimate the parameters. Kissinger, Coats, and Redfern, KAS, FWO, Friedman are one-step methods that have been used to estimate the kinetic parameters. In this work, after testing more than 500 data points accessed from different literature sources for coal, oil shale, solid materials, and biomass pyrolysis using one-step global method, it was found that the activation energy generated by KAS or FWO methods are related as in the following equations: 𝐸𝐾𝐴𝑆 = 0.9629 ∗ 𝐸𝐹𝑊𝑂 + 8.85, with R² =0.9945 or 𝐸𝐹𝑊𝑂 = 1.0328 ∗ 𝐸𝐾𝐴𝑆 − 8.0969 with R2= 0.9945. The multistep kinetic models employed the Distributed Activation Energy Model (DAEM) using Gaussian distribution, which suffers from symmetry, other distributions such as Weibull, and logistic has been used. These multistep kinetic models account for parallel/series and complex, primary and secondary biomass reactions by force-fitting the activation energy values. The frequency factor is assumed constant for the whole range of activation energy. Network models have been used to account for heat and mass transfer (diffusional effects), where the one-step and multistep could not account for these limitations. Three network models are available, the Bio-CPD (Chemical Percolation Devolatilization) model, Bio-FLASHCHAIN, and the Bio-FGDVC (Functional Group Depolymerization Vaporization Crosslinking models). These models tried to predict the product distributions of the biomass pyrolysis process

Bone Density Micro-CT Assessment during Embedding of the Innovative Multi-Spiked Connecting Scaffold in Periarticular Bone to Elaborate a Validated Numerical Model for Designing Biomimetic Fixation of Resurfacing Endoprostheses.

Our team has been working for some time on designing a new kind of biomimetic fixation of resurfacing endoprostheses, in which the innovative multi-spiked connecting scaffold (MSC-Scaffold) that mimics the natural interface between articular cartilage and periarticular trabecular bone in human joints is the crucial element. This work aimed to develop a numerical model enabling the design of the considered joint replacement implant that would reflect the mechanics of interacting biomaterials. Thus, quantitative micro-CT analysis of density distribution in bone material during the embedding of MSC-Scaffold in periarticular bone was applied. The performed numerical studies and corresponding mechanical tests revealed, under the embedded MSC-Scaffold, the bone material densification affecting its mechanical properties. On the basis of these findings, the built numerical model was modified by applying a simulated insert of densified bone material. This modification led to a strong correlation between the re-simulation and experimental results (FVU = 0.02). The biomimetism of the MSC-Scaffold prototype that provided physiological load transfer from implant to bone was confirmed based on the Huber–von Mises–Hencky (HMH) stress maps obtained with the validated finite element (FE) model of the problem. The micro-CT bone density assessment performed during the embedding of the MSC-Scaffold prototype in periarticular bone provides insight into the mechanical behaviour of the investigated implant-bone system and validates the numerical model that can be used for the design of material and geometric features of a new kind of resurfacing endoprostheses fixation.

Fluorescence Correlation Spectroscopy Combined with Multiphoton Laser Scanning Microscopy—A Practical Guideline

Multiphoton laser scanning microscopy (MPM) has opened up an optical window into biological tissues; however, imaging is primarily qualitative. Cell morphology and tissue architectures can be clearly visualized but quantitative analysis of actual concentration and fluorophore distribution is indecisive. Fluorescence correlation spectroscopy (FCS) is a highly sensitive photophysical methodology employed to study molecular parameters such as diffusion characteristics on the single molecule level. In combination with laser scanning microscopy, and MPM in particular, FCS has been referred to as a standard and highly useful tool in biomedical research to study diffusion and molecular interaction with subcellular precision. Despite several proof-of-concept reports on the topic, the implementation of MPM-FCS is far from straightforward. This practical guideline aims to clarify the conceptual principles and define experimental operating conditions when implementing MPM-FCS. Validation experiments in Rhodamine solutions were performed on an experimental MPM-FCS platform investigating the effects of objective lens, fluorophore concentration and laser power. An approach based on analysis of time-correlated single photon counting data is presented. It is shown that the requirement of high numerical aperture (NA) objective lenses is a primary limitation that restricts field of view, working distance and concentration range. Within these restrictions the data follows the predicted theory of Poisson distribution. The observed dependence on laser power is understood in the context of perturbation on the effective focal volume. In addition, a novel interpretation of the effect on measured diffusion time is presented. Overall, the challenges and limitations observed in this study reduce the versatility of MPM-FCS targeting biomedical research in complex and deep tissue—being the general strength of MPM in general. However, based on the systematic investigations and fundamental insights this report can serve as a practical guide and inspire future research, potentially overcoming the technical limitations and ultimately allowing MPM-FCS to become a highly useful tool in biomedical research.

Quantitative analysis of inulin distribution in the brain focused on nose-to-brain route via olfactory epithelium by reverse esophageal cannulation

This study aimed to determine the effect of intranasal dosing speed and administrating volume of nose-to-brain delivery on candidates for peptide drugs (molecular weight ca. 1–10 kDa). Using inulin as the model molecule of a peptide drug, intranasal administration by cannulation from the airway side through the esophagus was tested in mice. This was done to determine the quantitative distribution levels of the drug in the brain and cerebral spinal fluid (CSF). Distribution levels were increased with slower and constant speed (5 μL/min), with higher dosing volume equivalent to nasal volume per body weight in mice (25 μL), and were recorded 0.27% injected dose per gram of tissue (ID/g) in the brain, and 0.24% injected dose per milliliter (ID/mL) in the CSF at 60 min. Then, brain distribution resulting from reverse cannulation was two times more than that of the typical intranasal administration method using a micropipette. In addition, the percentage of inulin estimated to reach the brain via direct transport (%DTP) during reverse cannulation was estimated to be 93%, suggesting that ~95% of the total dose was transferred directly to the brain via the olfactory mucosa. These results show that distribution of the peptide drug in the brain was increased through constant administration at a slow and constant speed.

Nucleation site potency distributions in thermoelastic martensitic transformation in Ni43Co7Mn39Sn11 particles

The relationship between the nucleation process and thermal hysteresis width in reversible thermoelastic martensitic transformations remains unclear, particularly as the volume of transforming material decreases. Understanding the number density and nature of defects which serve as nucleation sites in this class of materials requires a quantitative analysis of nucleation site potency distributions in different classes of materials with different intrinsic barriers to nucleation. Here, we investigate the size dependence of hysteresis in microscale ${\mathrm{Ni}}_{43}{\mathrm{Co}}_{7}{\mathrm{Mn}}_{39}{\mathrm{Sn}}_{11}$ alloy particles (radius $4.4\text{--}19.0\phantom{\rule{0.28em}{0ex}}\ensuremath{\mu}\mathrm{m}$) during reversible martensitic transformations by collecting temperature-dependent magnetization of 126 individual alloy particles. Size-dependent hysteresis is quantified by a power law model and attributed to friction-induced energy dissipation. In samples with ideal nucleation-limited transformations, martensitic transformation temperatures on cooling decreased with decreasing particle volume due to the low probability of including relatively sparse high-energy nucleation sites. Nucleation site potency distributions are quantified as a function of thermodynamic driving force and compared against potency distributions for thermoelastic martensitic transformations in other classes of materials and for burst martensitic transformations. Across different classes of materials, as the energy barrier associated with the martensitic transformation increases, number densities of defects with sufficient potency to nucleate the transformation decrease dramatically. This finding suggests that very different kinds of defects may be responsible for nucleation of martensitic phase transformations in different material systems.

Nitrogen selectivity upon catalytic ozonation of ammonia with CTAB-NiO catalyst

In this work, CTAB-NiO catalyst was prepared by solid-state method and employed for oxidizing ammonia (NH4+) in aqueous solution by catalytic ozonation. In order to obtain high N2 selectivity, the effects of operating parameters on catalytic ozonation of ammonia were investigated in detail. Experimental results revealed that initial pH played a crucial role both in single O3 process and CTAB-NiO/O3 process, especially in the decomposition of O3 and the pathway of ammonia being oxidized into final products. Meanwhile, CTAB-NiO catalyst exhibited good performance in treating low concentration NH4+-containing wastewater from rare earth smelting. The results of qualitative and quantitative analysis of the reaction product distribution showed that in the CTAB-NiO/O3 system, about 98 % ammonia mainly converted to N2 at 293 K and ambient atmospheric pressure, with a selectivity of 76 %. The mechanism of catalytic ozonation of ammonia in aqueous solution was investigated by adding tert-butanol to the CTAB-NiO/O3 system. The catalytic ozonation of ammonia in aqueous solution was primarily due to the synergistic effect between the active ingredient from CTAB-NiO catalyst and hydroxyl radicals (·OH) generated by ozonolysis.

Abstract 16862: Culture of Cardiac Mesenchymal Cells (CMCs) at Physiologic Oxygen Tension Greatly Enhances Cell Retention In Vivo After Myocardial Infarction

A major obstacle to using cell therapy for heart failure is the poor survival of transplanted cells. We have found that CMCs (a promising type of reparative cells) grown at 5% O 2 (the physiologic O 2 tension in the heart) are more resistant to severe hypoxia in vitro than CMCs grown at 21% O 2 , but the survival of these cells in vivo is unknown. Thus, female mice subjected to a 60-min coronary occlusion and 30 days of reperfusion received 1x10 6 male CMCs cultured at 21% O 2 or 5% O 2 (21%O 2 CMCs or 5%O 2 CMCs) via echo-guided injection into the LV cavity. Quantitative real-time PCR for single copy of a Y-chromosome-specific Sry gene sequence was performed using genomic DNA isolated from heart (right ventricle [RV], risk region [RR] and non-risk region [NR] of LV), lung, liver, spleen and kidney 24 h or 7 days after cell administration (Figure). Compared with 21% O 2 CMCs, culture at 5% O 2 markedly increased cell retention in RR &gt; NR &gt; RV at 24 h and up to 7 days after CMC administration (Figs. C-D). At 24 h, although 25% of all transplanted cells were present in the liver (Fig. E), the number of Sry + cells per mg of tissue was strikingly greater in RR (2131±248) &gt; NR (1447±250) &gt; RV (1047±156) compared with liver (274±131, P &lt;0.01) and kidney (619±140, P &lt;0.05) (n=6/group). These differences persisted up to 7 days (820±156 in RR, 667±156 in NR, 327±33 in RV vs. 103±45 in liver, P &lt;0.01; 179±50 in kidney, P &lt;0.05) (n=4-5/group). In summary, we provide a systematic, time-dependent, and highly quantitative analysis of cell retention and distribution after infusion of CMCs into the LV cavity of failing hearts. The results demonstrate that, compared with 21% O 2 , physiologic 5% O 2 tension greatly enhances the retention of CMCs in the heart at 24 h and 7 days after cell transplantation, especially in the scarred regions where O 2 is very low (1-2%). Thus, culture at 5% O 2 may improve the reparative properties of CMCs (and possibly other cell types) used for cell therapy, providing an approach that may be applicable in the clinic.

Integration and Iteration: Using Advanced, High-Content Imaging and Single-Cell Gene Expression Analysis to Uncover Unique Aspects of Follicular Lymphoma Biology

BACKGROUND: Follicular lymphoma (FL) is an indolent malignancy of germinal center B-cell origin. FL patients experience remarkable heterogeneity in their disease trajectory, with many patients slowly progressing over several years, and a subset of patients experiencing an aggressive clinical course. Uncovering the cell-intrinsic and -extrinsic factors that govern differential progression and outcome in FL patients is thus essential. Beyond the genetic and epigenetic aberrations that contribute to FL oncogenesis, the tumor microenvironment (TME) plays an integral role in supporting the proliferation and survival of malignant cells. In a process described as "re-education", FL tumor cells may actively subvert the normal functions of non-malignant cells present in the TME, including T cells, follicular dendritic cells (FDCs), macrophages, dendritic cells, and stromal cells to support their survival and growth. METHODS: To address the importance of the TME in FL, we molecularly profiled excisional lymph node biopies from untreated FL patients using multiple platforms: bulk RNA sequencing (RNAseq), single-cell RNA sequencing (scRS), and a unique high content imaging method, Iterative Bleaching Extends MultipleXity (IBEX), which utilizes chemical bleaching to image 40+ proteins in the same tissue section by antibody staining. In combination with advanced computational tools for the quantitative analysis of cell types and distribution in tissues, we have used this approach to evaluate the TME:FL interaction within 8 FL samples, with 4 normal lymph nodes as controls. RESULTS: Both FL and TME components reconstructed from bulk RNA-seq were similar to the cellular composition revealed by scRS and IBEX analyses. Moreover, the bulk RNAseq and scRS identified the expression of genes involved in tumorigenesis and oncogenic signaling, often unique to each case. However, RNAseq-based approaches often miss important cellular and acellular components not readily extracted from dense tissues. For example, IBEX imaging can trace the pattern of blood vessels within a section, which cannot be achieved with non-imaging methods. In one case, our multi-parameter imaging studies revealed the close spatial interaction between clonal FL B cells, expressing a B-cell receptor (BCR) possessing a de novo N-linked glycosylation site introduced by somatic hypermutation, and cells expressing dendritic cell-specific intercellular adhesion molecule-3-grabbing non-integrin (DC-SIGN). This contact may activate pro-survival signaling in the malignant B cells. CONCLUSIONS: Integration of bulk RNAseq, scRS, clonotype analysis, and IBEX reveals both shared and unique aspects of different FL tumors. These data highlight the importance of integrating direct tissue analysis by high-content imaging with methods examining aspects of isolated cells. This approach may provide a more complete understanding of tumor biology, which in turn will identify patients at risk for developing aggressive disease and rationally improve treatment strategies for FL. This research was supported in part by the Intramural Research Program of the NIH, NIAID and NCI Figure Disclosures Bagaev: BostonGene: Current Employment, Current equity holder in private company, Patents &amp; Royalties. Plotnikova:BostonGene: Current Employment, Current equity holder in private company, Patents &amp; Royalties. Galkin:BostonGene: Current Employment, Patents &amp; Royalties. Postovalova:BostonGene: Current Employment, Current equity holder in private company. Svekolkin:BostonGene: Current Employment, Current equity holder in private company, Patents &amp; Royalties. Isaev:BostonGene: Current Employment, Current equity holder in private company, Patents &amp; Royalties. Lozinsky:BostonGene: Current Employment, Current equity holder in private company, Patents &amp; Royalties. Meerson:BostonGene: Current Employment. Varlamova:BostonGene: Current Employment, Current equity holder in private company, Patents &amp; Royalties. Ovcharov:BostonGene: Current Employment, Current equity holder in private company, Patents &amp; Royalties. Polyakova:BostonGene: Current Employment, Current equity holder in private company, Patents &amp; Royalties. Nomie:BostonGene: Current Employment, Current equity holder in private company. Kotlov:BostonGene: Current Employment, Current equity holder in private company, Patents &amp; Royalties. Tsiper:BostonGene: Current Employment, Current equity holder in private company, Patents &amp; Royalties. Frenkel:BostonGene: Current Employment, Current equity holder in private company, Patents &amp; Royalties. Attaulakhanov:BostonGene: Current Employment, Current equity holder in private company, Patents &amp; Royalties. Fowler:BostonGene: Current Employment, Current equity holder in private company, Patents &amp; Royalties.