Generation and maturation of dendritic cells for clinical application under serum-free conditions

Monocyte-derived dendritic cells (MoDCs) in clinical use for cancer immunotherapy are ideally generated in serum-free medium (SFM) with inclusion of a suitable maturation factor toward the end of the incubation period. Three good manfacturing practice (GMP) grade SFMs (AIM-V, X-VIVO 15, and X-VIVO 20) were compared with RPMI-1640, supplemented with 10% fetal bovine serum or 10% human serum. DCs generated for 7 days in SFM were less mature and secreted less interleukin (IL) 12p70 and IL-10 than DCs generated in 10% serum. DC yield was comparable in SFMs, and a greater proportion of cells was viable after maturation. Toll-like receptor (TLR) ligands were compared for their ability to induce cytokine secretion under serum-free conditions in the presence of interferon (IFN) gamma. With the exception of Poly I:C, TLR ligands stimulated high levels of IL-10 secretion. High levels of IL-12p70 were induced by two TLR4-mediated stimuli, lipopolysaccharide and Ribomunyl, a clinical-grade bacterial extract. When T-cell responses were compared in allogeneic mixed leukocyte reaction, DCs stimulated with Ribomunyl induced higher levels of IFN gamma than DCs stimulated with the cytokine cocktail: tumor necrosis factor-alpha, IL-1 beta, IL-6, and prostaglandin E-2. In the presence of IL-10 neutralizing antibodies, DC IL-12p70 production and T-cell IFN gamma were increased in vitro. Similarly, DCs stimulated with Ribomunyl, IFN gamma, and anti-IL-10 induced high levels of tetanus toxoid-specific T-cell proliferation and IFN gamma secretion. Thus, MoDCs generated ill SFM efficiently stimulate T-cell IFN gamma production after maturation in the presence of a clinical-grade TLR4 agonist and IL-10 neutralization.

Serum-free process development:improving the yield and consistency of human mesenchymal stromal cell production

Background aims: The cost-effective production of human mesenchymal stromal cells (hMSCs) for off-the-shelf and patient specific therapies will require an increasing focus on improving product yield and driving manufacturing consistency. Methods: Bone marrow-derived hMSCs (BM-hMSCs) from two donors were expanded for 36 days in monolayer with medium supplemented with either fetal bovine serum (FBS) or PRIME-XV serum-free medium (SFM). Cells were assessed throughout culture for proliferation, mean cell diameter, colony-forming potential, osteogenic potential, gene expression and metabolites. Results: Expansion of BM-hMSCs in PRIME-XV SFM resulted in a significantly higher growth rate (P < 0.001) and increased consistency between donors compared with FBS-based culture. FBS-based culture showed an inter-batch production range of 0.9 and 5 days per dose compared with 0.5 and 0.6 days in SFM for each BM-hMSC donor line. The consistency between donors was also improved by the use of PRIME-XV SFM, with a production range of 0.9 days compared with 19.4 days in FBS-based culture. Mean cell diameter has also been demonstrated as a process metric for BM-hMSC growth rate and senescence through a correlation (R² = 0.8705) across all conditions. PRIME-XV SFM has also shown increased consistency in BM-hMSC characteristics such as per cell metabolite utilization, in vitro colony-forming potential and osteogenic potential despite the higher number of population doublings. Conclusions: We have increased the yield and consistency of BM-hMSC expansion between donors, demonstrating a level of control over the product, which has the potential to increase the cost-effectiveness and reduce the risk in these manufacturing processes.

SeaWiFS (version R2010.0) Monthly Global 9 km Microplankton Fraction, links to NetCDF files

Phytoplankton cell size is important to biogeochemical and food web processes. The goal of this study is to estimate phytoplankton cell size distribution from satellite imagery of spectral remote sensing reflectance (Rrs(lambda)). Previous studies have indicated phytoplankton size classes have distinctive absorption spectra despite the physiological and taxonomic variability within an assemblage. For this study, the chlorophyll specific absorption spectra for phytoplankton size class extremes, pico- and microphytoplankton, are weighted by the percent microplankton (Sfm) and are the basis of phytoplankton size retrieval from SeaWiFS imagery. Satellite retrievals of Sfm are done through implementation of a forward optical model look-up table (LUT) that incorporates the range of absorption and scattering variability due to phytoplankton size, chlorophyll concentration ([Chl]) and dissolved and detrital matter (acdm(443)) in the global ocean from which Rrs(lambda) is calculated by the radiative transfer software, Hydrolight. The Hydrolight modeled Rrs(lambda) options for a given combination of [Chl] and acdm(443) within the LUT vary only due to Sfm. For a given pixel, the LUT search space was limited by satellite imagery of [Chl] and acdm(443). Within the narrowed search space, SeaWiFS Rrs(lambda) was matched with the closest LUT Rrs(lambda) option and the associated Sfm was assigned. Thresholds at which changes in Rrs(lambda) due to Sfm could be discerned were established in terms of [Chl] and acdm(443). In situ high-precision liquid chromatography-derived estimates of cell size are used in conjunction with matched daily satellite estimates of Sfm for validation and agree well. A single month is displayed as an example of the Sfm retrieval.

Photographs of the Mount Meager, southwestern British Columbia

On August 6, 2010, a large (~50 Mm**3) debris avalanche occurred on the flank of Mount Meager in the southern Coast Mountains of British Columbia, Canada. We studied the deposits to infer the morphodynamics of the landslide from initiation to emplacement. Structure from motion (SfM) photogrammetry, based on oblique photos taken with a standard SLR camera during a low helicopter traverse, was used to create high-resolution orthophotos and base maps. Interpretation of the images and maps allowed us to recognize two main rheological phases in the debris avalanche. Just below the source area, in the valley of Capricorn Creek, the landslide separated into two phases, one water-rich and more mobile, and the other water-poor and less mobile. The water-rich phase spread quickly, achieved high superelevation on the valley sides, and left distal scattered deposits. The main water-poor phase moved more slowly, did not superelevate, and formed a thick continuous deposit (up to ~30 m) on the valley floor. The water-poor flow deposit has structural features such as hummocks, brittle-ductile faults, and shear zones. Our study, based on a freshly emplaced deposit, advances understanding of large mass movements by showing that a single landslide can develop multiple rheology phases with different behaviours. Rheological evolution and separation of phases should always be taken into account to provide better risk assessment scenarios.

Recupero digitale di dati storici e tracce per un turismo culturale nell'area del Centro Cadore: fotogrammetria e GIS nella valorizzazione di un territorio.

La tesi si occupa dell'uso di più tecniche fotogrammetriche e di strumenti GIS nel recupero digitale e nell'integrazione di una molteplicità di dati storici, opportunamente georeferenziati, inerenti l'area del Centro Cadore, ai fini anche della valorizzazione turistico-culturale del territorio. Un ampio spazio viene dato alla caratterizzazione del territorio e delle fonti - cartografiche, fotografiche e testuali - che sono state recuperate ed organizzate in archivio. Le applicazioni fotogrammetriche comprendono la generazione di ortofoto digitali a scala territoriale da immagini storiche e modelli 3D close-range ottenuti con tecniche SfM.

The Accuracy and Reliability of Traditional Surface Flow Type Mapping: Is It Time for A New Method of Characterising Physical River Habitat?

Surface flow types (SFT) are advocated as ecologically relevant hydraulic units, often mapped visually from the bankside to characterise rapidly the physical habitat of rivers. SFT mapping is simple, non-invasive and cost-efficient. However, it is also qualitative, subjective and plagued by difficulties in recording accurately the spatial extent of SFT units. Quantitative validation of the underlying physical habitat parameters is often lacking, and does not consistently differentiate between SFTs. Here, we investigate explicitly the accuracy, reliability and statistical separability of traditionally mapped SFTs as indicators of physical habitat, using independent, hydraulic and topographic data collected during three surveys of a c. 50m reach of the River Arrow, Warwickshire, England. We also explore the potential of a novel remote sensing approach, comprising a small unmanned aerial system (sUAS) and Structure-from-Motion photogrammetry (SfM), as an alternative method of physical habitat characterisation. Our key findings indicate that SFT mapping accuracy is highly variable, with overall mapping accuracy not exceeding 74%. Results from analysis of similarity (ANOSIM) tests found that strong differences did not exist between all SFT pairs. This leads us to question the suitability of SFTs for characterising physical habitat for river science and management applications. In contrast, the sUAS-SfM approach provided high resolution, spatially continuous, spatially explicit, quantitative measurements of water depth and point cloud roughness at the microscale (spatial scales ≤1m). Such data are acquired rapidly, inexpensively, and provide new opportunities for examining the heterogeneity of physical habitat over a range of spatial and temporal scales. Whilst continued refinement of the sUAS-SfM approach is required, we propose that this method offers an opportunity to move away from broad, mesoscale classifications of physical habitat (spatial scales 10-100m), and towards continuous, quantitative measurements of the continuum of hydraulic and geomorphic conditions which actually exists at the microscale.

Improving the Accuracy of 3-D Reconstruction in Robotic Vision Applications

Thesis (Ph.D.)--University of Washington, 2016-08

POSE ESTIMATION AND 3D RECONSTRUCTION USING SENSOR FUSION

A camera maps 3-dimensional (3D) world space to a 2-dimensional (2D) image space. In the process it loses the depth information, i.e., the distance from the camera focal point to the imaged objects. It is impossible to recover this information from a single image. However, by using two or more images from different viewing angles this information can be recovered, which in turn can be used to obtain the pose (position and orientation) of the camera. Using this pose, a 3D reconstruction of imaged objects in the world can be computed. Numerous algorithms have been proposed and implemented to solve the above problem; these algorithms are commonly called Structure from Motion (SfM). State-of-the-art SfM techniques have been shown to give promising results. However, unlike a Global Positioning System (GPS) or an Inertial Measurement Unit (IMU) which directly give the position and orientation respectively, the camera system estimates it after implementing SfM as mentioned above. This makes the pose obtained from a camera highly sensitive to the images captured and other effects, such as low lighting conditions, poor focus or improper viewing angles. In some applications, for example, an Unmanned Aerial Vehicle (UAV) inspecting a bridge or a robot mapping an environment using Simultaneous Localization and Mapping (SLAM), it is often difficult to capture images with ideal conditions. This report examines the use of SfM methods in such applications and the role of combining multiple sensors, viz., sensor fusion, to achieve more accurate and usable position and reconstruction information. This project investigates the role of sensor fusion in accurately estimating the pose of a camera for the application of 3D reconstruction of a scene. The first set of experiments is conducted in a motion capture room. These results are assumed as ground truth in order to evaluate the strengths and weaknesses of each sensor and to map their coordinate systems. Then a number of scenarios are targeted where SfM fails. The pose estimates obtained from SfM are replaced by those obtained from other sensors and the 3D reconstruction is completed. Quantitative and qualitative comparisons are made between the 3D reconstruction obtained by using only a camera versus that obtained by using the camera along with a LIDAR and/or an IMU. Additionally, the project also works towards the performance issue faced while handling large data sets of high-resolution images by implementing the system on the Superior high performance computing cluster at Michigan Technological University.

Assessing the relationship between bee/flower diversity and vegetation structural heterogeneity through UAV photogrammetry

The ecosystem services provided by bees are very important. Factors as habitat fragmentation, intensive agriculture and climate change are contributing to the decline of bee populations. The use of remote sensing could be a useful tool for the recognition of sites with a high diversity, before performing a more expensive survey in the field. In this study the ability of Unmanned Aerial Vehicles (UAV) images to estimate biodiversity at local scale has been analysed testing the concept of the Height Variation Hypothesis (HVH). This approach states that, the higher the vegetation height heterogeneity (HH) measured by remote sensing information, the higher the vertical complexity and the higher vegetation species diversity. In this thesis the concept has been brought to a higher level, in order to understand if the vegetation HH can be considered a proxy also of bee species diversity and abundance. We tested this approach collecting field data on bees/flowers and RGB images through an UAV campaign in 30 grasslands in the South of the Netherlands. The Canopy Height Model (CHM) were derived through the photogrammetry technique "Structure from Motion" (SfM) with resolutions of 10cm, 25cm, 50cm. Successively, the HH assessed on the CHM using the Rao's Q heterogeneity index was correlated to the field data (bee abundance, diversity and bee/flower species richness). The correlations were all positive and significant. The highest R2 values were found when the HH was calculated at 10cm and correlated to bee species richness (R2 = 0.41) and Shannon’s H index (R2 = 0.38). Using a lower spatial resolution the goodness of fit slightly decreases. For flower species richness the R2 ranged between 0.36 to 0.39. Our results suggest that methods based on the concept behind the HVH, in this case deriving information of HH from UAV data, can be developed into valuable tools for large-scale, standardized and cost-effective monitoring of flower diversity and of the habitat quality for bees.

Il ruolo delle tecniche geomatiche nella realizzazione di Digital Twin City: una sperimentazione sulla città di Bologna

L’incessante urbanizzazione e il continuo aumento della popolazione urbana stanno generando nuove sfide per le pubbliche amministrazioni, le quali necessitano soluzioni per la gestione sostenibile di risorse primarie (cibo, acqua, suolo, fonti energetiche) e la corretta pianificazione delle città, allo scopo di salvaguardare le condizioni ambientali, la salute dei cittadini e il progresso economico: in questo complesso panorama si afferma il concetto di Digital Twin City (DTC) o gemello digitale della città, la controparte virtuale di oggetti e processi in ambiente urbano in grado di comunicare con essi e di simularne, replicarne e predirne i possibili scenari. In questo contesto, un elemento essenziale è costituito dal modello geometrico 3D ad alta fedeltà dell'ambiente urbano, e la Geomatica fornisce gli strumenti più idonei alla sua realizzazione. La presente elaborazione si sviluppa in tre parti: nella prima è stata condotta un’analisi della letteratura sui DTC, in cui sono state evidenziate le sue caratteristiche principali come l’architettura, le tecnologie abilitanti, alcune possibili modellazioni, ostacoli, scenari futuri ed esempi di applicazioni reali, giungendo alla conclusione che un accurato modello 3D della città deve essere alla base dei DTC; nella seconda parte è stata illustrata nel dettaglio la teoria delle principali tecniche geomatiche per la realizzazione di modelli 3D ad alta fedeltà, tra cui le tecniche fotogrammetriche di aerotriangolazione e l’algoritmo Structure from Motion (SfM); nella terza e ultima parte è stata condotta una sperimentazione su tre zone campione del comune di Bologna di cui, grazie ad un dataset di fotogrammi nadirali e obliqui ottenuto da un volo fotogrammetrico realizzato nel 2022, sono state ottenute Reality Meshes e ortofoto/DSM. I prodotti della terza zona sono stati confrontati con i medesimi ottenuti da un dataset del 2017. Infine, sono state illustrati alcuni strumenti di misura e di ritocco dei prodotti 3D e 2D.