On the Origin of Natural Antibody

Natural IgM (nIgM) is constitutively present in the serum, where it aids in the early control of viral and bacterial expansions. nIgM also plays a significant role in the prevention of autoimmune disease by promoting the clearance of cellular debris. However, the cells that maintain high titers of nIgM in the circulation had not yet been identified. Several studies have linked serum nIgM with the presence of fetal-lineage B cells, and others have detected IgM secretion directly by B1a cells in various tissues. Nevertheless, a substantial contribution of undifferentiated B1 cells to nIgM titers is doubtful, as the ability to produce large quantities of antibody (Ab) is a function of the phenotype and morphology of differentiated plasma cells (PCs). No direct evidence exists to support the claim that a B1-cell population directly produces the bulk of circulating nIgM. The source of nIgM thus remained uncertain and unstudied.

In the first part of this study, I identified the primary source of nIgM. Using enzyme-linked immunosorbent spot (ELISPOT) assay, I determined that the majority of IgM Ab-secreting cells (ASCs) in naïve mice reside in the bone marrow (BM). Flow cytometric analysis of BM cells stained for intracellular IgM revealed that nIgM ASCs express IgM and the PC marker CD138 on their surface, but not the B1a cell marker CD5. By spinning these cells onto slides and staining them, following isolation by fluorescence-activated cell sorting (FACS), I found that they exhibit the typical morphological characteristics of terminally differentiated PCs. Transfer experiments demonstrated that BM nIgM PCs arise from a progenitor in the peritoneal cavity (PerC), but not isolated PerC B1a, B1b, or B2 cells. Immunoglobulin (Ig) gene sequence analysis and examination of B1-8i mice, which carry an Ig knockin that prohibits fetal B-cell development, indicated that nIgM PCs differentiate from fetal-lineage B cells. BrdU uptake experiments showed that the nIgM ASC compartment contains a substantial fraction of long-lived plasma cells (LLPCs). Finally, I demonstrated that nIgM PCs occupy a survival niche distinct from that used by IgG PCs.

In the second part of this dissertation, I characterized the unique survival niche of nIgM LLPCs, which maintain constitutive high titers of nIgM in the serum. By using genetically deficient or Ab-depleted mice, I found that neither T cells, type 2 innate lymphoid cells, nor mast cells, the three major hematopoietic producers of IL-5, were required for nIgM PC survival in the BM. However, IgM PCs associate strongly with IL-5-expressing BM stromal cells, which support their survival in vitro when stimulated. In vivo neutralization of IL-5 revealed that, like individual survival factors for IgG PCs, IL-5 is not the sole supporter of IgM PCs, but is likely one of several redundant molecules that together ensure uninterrupted signaling. Thus, the long-lived nIgM PC niche is not composed of hematopoietic sources of IL-5, but a stromal cell microenvironment that provides multiple redundant survival signals.

In the final part of my study, I identified and characterized the precursor of nIgM PCs, which I found in the first project to be resident in the PerC, but not a B1a, B1b, or B2 cell. By transferring PerC cells sorted based on expression of CD19, CD5, and CD11b, I found that only the CD19+CD5+CD11b- population contained cells capable of differentiating into nIgM PCs. Transfer of decreasing numbers of unfractionated PerC cells into Rag1 knockouts revealed an order-of-magnitude drop in the rate of serum IgM reconstitution between stochastically sampled pools of 106 and 3x105 PerC cells, suggesting that the CD19+CD5+CD11b- compartment comprises two cell types, and that interaction between the two necessary for nIgM-PC differentiation. By transferring neonatal liver, I determined that the early hematopoietic environment is required for nIgM PC precursors to develop. Using mice carrying a mutation that disturbs cKit expression, I also found that cKit appears to be required at a critical point near birth for the proper development of nIgM PC precursors.

The collective results of these studies demonstrate that nIgM is the product of BM-resident PCs, which differentiate from a PerC B cell precursor distinct from B1a cells, and survive long-term in a unique survival niche created by stromal cells. My work creates a new paradigm by which to understand nIgM, B1 cell, and PC biology.

Design of Latent Thermal Energy Storage Systems

This dissertation documents the results of a theoretical and numerical study of time dependent storage of energy by melting a phase change material. The heating is provided along invading lines, which change from single-line invasion to tree-shaped invasion. Chapter 2 identifies the special design feature of distributing energy storage in time-dependent fashion on a territory, when the energy flows by fluid flow from a concentrated source to points (users) distributed equidistantly on the area. The challenge in this chapter is to determine the architecture of distributed energy storage. The chief conclusion is that the finite amount of storage material should be distributed proportionally with the distribution of the flow rate of heating agent arriving on the area. The total time needed by the source stream to ‘invade’ the area is cumulative (the sum of the storage times required at each storage site), and depends on the energy distribution paths and the sequence in which the users are served by the source stream. Chapter 3 shows theoretically that the melting process consists of two phases: “invasion” thermal diffusion along the invading line, which is followed by “consolidation” as heat diffuses perpendicularly to the invading line. This chapter also reports the duration of both phases and the evolution of the melt layer around the invading line during the two-dimensional and three-dimensional invasion. It also shows that the amount of melted material increases in time according to a curve shaped as an S. These theoretical predictions are validated by means of numerical simulations in chapter 4. This chapter also shows that the heat transfer rate density increases (i.e., the S curve becomes steeper) as the complexity and number of degrees of freedom of the structure are increased, in accord with the constructal law. The optimal geometric features of the tree structure are detailed in this chapter. Chapter 5 documents a numerical study of time-dependent melting where the heat transfer is convection dominated, unlike in chapter 3 and 4 where the melting is ruled by pure conduction. In accord with constructal design, the search is for effective heat-flow architectures. The volume-constrained improvement of the designs for heat flow begins with assuming the simplest structure, where a single line serves as heat source. Next, the heat source is endowed with freedom to change its shape as it grows. The objective of the numerical simulations is to discover the geometric features that lead to the fastest melting process. The results show that the heat transfer rate density increases as the complexity and number of degrees of freedom of the structure are increased. Furthermore, the angles between heat invasion lines have a minor effect on the global performance compared to other degrees of freedom: number of branching levels, stem length, and branch lengths. The effect of natural convection in the melt zone is documented.

Optimization of Cooling Protocols for Hearts Destined for Transplantation

Design and analysis of conceptually different cooling systems for the human heart preservation are numerically investigated. A heart cooling container with required connections was designed for a normal size human heart. A three-dimensional, high resolution human heart geometric model obtained from CT-angio data was used for simulations. Nine different cooling designs are introduced in this research. The first cooling design (Case 1) used a cooling gelatin only outside of the heart. In the second cooling design (Case 2), the internal parts of the heart were cooled via pumping a cooling liquid inside both the heart’s pulmonary and systemic circulation systems. An unsteady conjugate heat transfer analysis is performed to simulate the temperature field variations within the heart during the cooling process. Case 3 simulated the currently used cooling method in which the coolant is stagnant. Case 4 was a combination of Case 1 and Case 2. A linear thermoelasticity analysis was performed to assess the stresses applied on the heart during the cooling process. In Cases 5 through 9, the coolant solution was used for both internal and external cooling. For external circulation in Case 5 and Case 6, two inlets and two outlets were designed on the walls of the cooling container. Case 5 used laminar flows for coolant circulations inside and outside of the heart. Effects of turbulent flow on cooling of the heart were studied in Case 6. In Case 7, an additional inlet was designed on the cooling container wall to create a jet impinging the hot region of the heart’s wall. Unsteady periodic inlet velocities were applied in Case 8 and Case 9. The average temperature of the heart in Case 5 was +5.0oC after 1500 s of cooling. Multi-objective constrained optimization was performed for Case 5. Inlet velocities for two internal and one external coolant circulations were the three design variables for optimization. Minimizing the average temperature of the heart, wall shear stress and total volumetric flow rates were the three objectives. The only constraint was to keep von Mises stress below the ultimate tensile stress of the heart’s tissue.

Chemical analysis of water samples of station M1_1207, Red Sea (Table 1-3)

Two water samples and two sediment samples taken in 1965 by the R. V. "Meteor" in the area of the hot salt brine of the Atlantis II-Deep were chemically investigated, and in addition the sediment samples were subjected to X-ray and optical analysis. The investigation of the sulfur-isotope-ratios showed the same values for all water samples. This information combined with the Ca-sulfate solubility data leads us to conclude that, for the most part, the sulfate content of the salt brine resulted from mixing along the boundary with the normal seawater. In this boundary area gypsum or anhydrite is formed which sinks down to the deeper layers of the salt brine where it is redisolved when the water becomes undersaturated. In the laboratory, formation of CaS04 precipitate resulted from both the reheating of the water sample from the uppermost zone of the salt brine to the in-situ-temperature as well as by the mixing of the water sample with normal Red Sea water. The iron and manganese delivered by the hot spring is separated within the area of the salt brine by their different redox-potentials. Iron is sedimented to a high amount within the salt brine, while, as evidenced by its small amounts in all sediment samples, the more easily reducible manganese is apparently carried out of the area before sedimentation can take place. The very good layering of the salt brine may be the result of the rough bottom topography with its several progressively higher levels allowing step-like enlargements of the surface areas of each successive layer. Each enlargement results in larger boundary areas along which more effective heat transfer and mixing with the next layer is possible. In the sediment samples up to 37.18% Fe is found, mostly bound as very poorly crystallized iron hydroxide. Pyrite is present in only very small amounts. We assume that the copper is bound mostly as sulfide, while the zinc is most likely present in an other form. The sulfur-isotope-investigations indicate that the sulfur in the sediment, bound as pyrite and sulfides, is not a result of bacterical sulfate-reduction in the iron-rich mud of the Atlantis II-Deep, but must have been brought up with the hot brine.

(Table 1) Age control and temporal resolution of paleoceanographic records of the North Atlantic

Short-term changes in sea surface conditions controlling the thermohaline circulation in the northern North Atlantic are expected to be especially efficient in perturbing global climate stability. Here we assess past variability of sea surface temperature (SST) in the northeast Atlantic and Norwegian Sea during Marine Isotope Stage (MIS) 2 and, in particular, during the Last Glacial Maximum (LGM). Five high-resolution SST records were established on a meridional transect (53°N-72°N) to trace centennial-scale oscillations in SST and sea-ice cover. We used three independent computational techniques (SIMMAX modern analogue technique, Artificial Neural Networks (ANN), and Revised Analog Method (RAM)) to reconstruct SST from planktonic foraminifer census counts. SIMMAX and ANN reproduced short-term SST oscillations of similar magnitude and absolute levels, while RAM, owing to a restrictive analog selection, appears less suitable for reconstructing "cold end" SST. The SIMMAX and ANN SST reconstructions support the existence of a weak paleo-Norwegian Current during Dansgaard-Oeschger (DO) interstadials number 4, 3, 2, and 1. During the LGM, two warm incursions of 7°C water to occurred in the northern North Atlantic but ended north of the Iceland Faroe Ridge. A rough numerical estimate shows that the near-surface poleward heat transfer from 53° across the Iceland-Faroe Ridge up to to 72° N dropped to less than 60% of the modern value during DO interstadials and to almost zero during DO stadials. Summer sea ice was generally confined to the area north of 70°N and only rarely expanded southward along the margins of continental ice sheets. Internal LGM variability of North Atlantic (>40°N) SST in the GLAMAP 2000 compilation (Sarnthein et al., 2003, doi:10.1029/2002PA000771; Pflaumann et al., 2003, doi:10.1029/2002PA000774) indicates maximum instability in the glacial subpolar gyre and at the Iberian Margin, while in the Nordic Seas, SST was continuously low.

Planktic foraminiferal distribution and stable isotope ratios of sediment core MSM05/5_712-1 from the Arctic Ocean

The Arctic is responding more rapidly to global warming than most other areas on our planet. Northward flowing Atlantic Water is the major means of heat advection towards the Arctic and strongly affects the sea ice distribution. Records of its natural variability are critical for the understanding of feedback mechanisms and the future of the Arctic climate system, but continuous historical records reach back only ~150 years. Here, we present a multidecadal scale record of ocean temperature variations during the last 2000 years, derived from marine sediments off Western Svalbard (79°N). We find that early-21st-century temperatures of Atlantic Water entering the Arctic Ocean are unprecedented over the past 2000 years and are presumably linked to the Arctic Amplification of global warming.

Preparation of Polymer Nanoparticles by Ring Opening Metathesis Polymerization in Aqueous Dispersed Systems with Water-Soluble Ruthenium-based Catalyst

Ring opening metathesis polymerization (ROMP) is a variant of olefin metathesis used to polymerize strained cyclic olefins. Ruthenium-based Grubbs’ catalysts are widely used in ROMP to produce industrially important products. While highly efficient in organic solvents such as dichloromethane and toluene, these hydrophobic catalysts are not typically applied in aqueous systems. With the advancements in emulsion and miniemulsion polymerization, it is promising to conduct ROMP in an aqueous dispersed phase to generate well-defined latex nanoparticles while improving heat transfer and reducing the use of volatile organic solvents (VOCs). Herein I report the efforts made using a PEGylated ruthenium alkylidene as the catalyst to initiate ROMP in an oil-in-water miniemulsion. 1H NMR revealed that the synthesized PEGylated catalyst was stable and reactive in water. Using 1,5-cyclooctadiene (COD) as monomer, we showed the highly efficient catalyst yielded colloidally stable polymer latexes with ~ 100% conversion at room temperature. Kinetic studies demonstrated first-order kinetics with good livingness as confirmed by the shift of gel permeation chromatography (GPC) traces. Depending on the surfactants used, the particle sizes ranged from 100 to 300 nm with monomodal distributions. The more strained cyclic olefin norbornene (NB) could also be efficiently polymerized with a PEGylated ruthenium alkylidene in miniemulsion to full conversion and with minimal coagulum formation.

Parametric Analysis of Concrete Solar Collectors

Concrete solar collectors offer a type of solar collector with structural, aesthetic and economic advantages over current populartechnologies. This study examines the influential parameters of concrete solar collectors. In addition to the external conditions,the performance of a concrete solar collector is influenced by the thermal properties of the concrete matrix, piping network andfluid. Geometric and fluid flow parameters also influence the performance of the concrete solar collector. A literature review ofconcrete solar collectors is conducted in order to define the benchmark parameters from which individual parameters are thencompared. The numerical model consists of a 1D pipe flow network coupled with the heat transfer in a 3D concrete domain. Thispaper is concerned with the physical parameters that define the concrete solar collector, thus a constant surface temperature isused as the exposed surface boundary condition with all other surfaces being insulated. Results show that, of the parametersinvestigated, the pipe spacing, ps, concrete conductivity, kc, and the pipe embedment depth, demb, are among those parameterswhich have greatest effect on the collector’s performance. The optimum balance between these parameters is presented withrespect to the thermal performance and discussed with reference to practical development issues.

Temperature fields in a channel partially filled with a porous material under local thermal non-equilibrium condition-an exact solution

This work examines analytically the forced convection in a channel partially filled with a porous material and subjected to constant wall heat flux. The Darcy–Brinkman–Forchheimer model is used to represent the fluid transport through the porous material. The local thermal non-equilibrium, two-equation model is further employed as the solid and fluid heat transport equations. Two fundamental models (models A and B) represent the thermal boundary conditions at the interface between the porous medium and the clear region. The governing equations of the problem are manipulated, and for each interface model, exact solutions, for the solid and fluid temperature fields, are developed. These solutions incorporate the porous material thickness, Biot number, fluid to solid thermal conductivity ratio and Darcy number as parameters. The results can be readily used to validate numerical simulations. They are, further, applicable to the analysis of enhanced heat transfer, using porous materials, in heat exchangers.

The development of dynamic CFD model of the magnetic semi-levitation of liquid metals

In the casting of reactive metals, such as titanium alloys, contamination can be prevented if there is no contact between the hot liquid metal and solid crucible. This can be achieved by containing the liquid metal by means of high frequency AC magnetic field. A water cooled current-carrying coil, surrounding the metal can then provide the required Lorentz forces, and at the same time the current induced in the metal can provide the heating required to melt it. This ‘attractive’ processing solution has however many problems, the most serious being that of the control and containment of the liquid metal envelope, which requires a balance of the gravity and induced inertia forces on the one side, and the containing Lorentz and surface tension forces on the other. To model this process requires a fully coupled dyna ic solution of the flow fields, magnetic field and heat transfer/melding process to account for. A simplified solution has been published previously providing quasi-static solutions only, by taking the irrotational ‘magnetic pressure’ term of the Lorentz force into account. The authors remedy this deficiency by modelling the full problem using CFD techniques. The salient features of these techniques are included in this paper, as space allows.

Three-dimensional free surface modelling in an unstructured mesh environment for metal processing applications

In the casting of metals, tundish flow, welding, converters, and other metal processing applications, the behaviour of the fluid surface is important. In aluminium alloys, for example, oxides formed on the surface may be drawn into the body of the melt where they act as faults in the solidified product affecting cast quality. For this reason, accurate description of wave behaviour, air entrapment, and other effects need to be modelled, in the presence of heat transfer and possibly phase change. The authors have developed a single-phase algorithm for modelling this problem. The Scalar Equation Algorithm (SEA) (see Refs. 1 and 2), enables the transport of the property discontinuity representing the free surface through a fixed grid. An extension of this method to unstructured mesh codes is presented here, together with validation. The new method employs a TVD flux limiter in conjunction with a ray-tracing algorithm, to ensure a sharp bound interface. Applications of the method are in the filling and emptying of mould cavities, with heat transfer and phase change.

PHYSICA: A multiphysics computational framework and its application to casting simulations

Metal casting is a process governed by the interaction of a range of physical phenomena. Most computational models of this process address only what are conventionally regarded as the primary phenomena – heat conduction and solidification. However, to predict other phenomena, such as porosity formation, requires modelling the interaction of the fluid flow, heat transfer, solidification and the development of stressdeformation in the solidified part of the casting. This paper will describe a modelling framework called PHYSICA[1] which has the capability to stimulate such multiphysical phenomena.

Dynamics of an axisymmetric electromagnetic 'crucible' melting

Different industrial induction melting processes involve free surface and melt-solid interface of the liquid metal subject to dynamic change during the technological operation. Simulation of the liquid metal dynamics requires to solve the non-linear, coupled hydrodynamic-electromagnetic-heat transfer problem accounting for the time development of the liquid metal free boundary with a suitable turbulent viscosity model. The present paper describes a numerical solution method applicable for various axisymmetric induction melting processes, such as, crucible with free top surface, levitation, semi-levitation, cold crucible and similar melting techniques. The presented results in the cases of semi-levitation and crucible with free top surface meltings demonstrate oscillating transient behaviour of the free metal surface indicating the presence of gravity-inertial-electromagnetic waves which are coupled to the internal fluid flow generated by both the rotational and potential parts of the electromagnetic force.