Refine
Year of publication
- 2005 (57) (remove)
Document Type
- Doctoral Thesis (57) (remove)
Language
- English (57) (remove)
Has Fulltext
- yes (57)
Is part of the Bibliography
- no (57)
Keywords
- Doxorubicin (2)
- 15d-PGJ2 (1)
- 3D-Modellierung (1)
- 5-lipoxygenase (1)
- ABC-Transporter (1)
- ABC-transporter (1)
- ALICE (1)
- ALTRO (1)
- ATPase (1)
- Absorptionsspektroskopie (1)
Institute
- Biochemie und Chemie (19)
- Physik (15)
- Pharmazie (11)
- Biowissenschaften (7)
- Geowissenschaften (2)
- Biochemie, Chemie und Pharmazie (1)
- Informatik (1)
- Psychologie (1)
Chemokines play a key role in the cellular infiltration of inflamed tissue. They are released by a wide variety of cell types during the initial phase of host response to injury, allergens, antigens, or invading microorganisms, and selectively attract leukocytes to inflammatory foci, inducing both migration and activation. Monocyte chemoattractant protein-1 (MCP-1), a member of the CC chemokine superfamily, functions in attracting monocytes, T lymphocytes, and basophils to sites of inflammation. MCP-1 is produced by monocytes, fibroblasts, vascular endothelial cells and smooth muscle cells in response to various stimuli such as tumour necrosis factor-a (TNF-a), interferon-g (IFN-g), and interleukin-1b (IL-1b). It also plays an important role in the pathogenesis of chronic inflammation, and overexpression of MCP-1 has been implicated in diseases including glomerulonephritis and rheumatoid arthritis. Oligonucleotide-directed triple helix formation offers a means to target specific sequences in DNA and interfere with gene expression at the transcriptional level. Triple helix-forming oligonucleotides (TFOs) bind to homopurine/homopyrimidine sequences, forming a stable, sequence-specific complex with the duplex DNA. Purine-rich sequences are frequent in gene regulatory regions and TFOs directed to promoter sequences have been shown to prevent binding of transcription factors and inhibit transcription initiation and elongation. Exogenous TFOs that bind homopurine/ homopyrimidine DNA sequences and form triple-helices can be rationally designed, while the intracellular delivery of single-stranded RNA TFOs has not been studied in detail before. In this study, expression vectors were constructed which directed transcription of either a 19 nt triplex-forming pyrimidine CU-TFO sequence targeting the human MCP-1 or two different 19 nt GU- or CA-control sequences, respectively, together with the vector encoded hygromycin resistance mRNA as one fusion transcript. HEK 293 cells were stable transfected with these vectors and several TFO and control cell lines were generated. Functional relevant triplex formation of a TFO with a corresponding 19 bp GC-rich AP-1/SP-1 site of the human MCP-1 promoter was shown. Binding of synthetic 19 nt CUTFO to the MCP-1 promoter duplex was verified by triplex blotting at pH 6.7. Underlining binding specificity, control sequences, including the GU- and CA-sequence, a TFO containing one single mismatch and a MCP-1 promoter duplex containing two mismatches, did not participate in triplex formation. Establishing a magnetic capture technique with streptavidin microbeads it was verified that at pH 7.0 the 19 nt TFO embedded in a 1.1 kb fusion transcript binds to a plasmid encoded MCP-1 promoter target duplex three times stronger than the controls. Finally, cell culture experiments revealed 76 ± 10.2% inhibition of MCP-1 protein secretion in TNF-a stimulated CU-TFO harboring cell lines and up to 88% after TNF-a and IFN-g costimulation in comparison to controls. Expression of interleukin-8 (IL-8) as one TNF-a inducible control gene was not affected by CU-TFO, demonstrating both highly specific and effective chemokine gene repression. Furthermore, another chemokine target, regulated upon activation normal T cell expressed and secreted (RANTES), which plays an essential role in inflammation by recruiting T lymphocytes, macrophages and eosinophils to inflammatory sites, was analysed using the triplex approach. A 28 nt TFO was designed targeting the murine RANTES gene promoter, and gel mobility shift assays demonstrated that the phosphodiester TFO formed a sequencespecific triplex with the double-stranded target DNA with a Kd of 2.5 x 10-7 M. It was analysed whether RANTES expression could be inhibited at the transcriptional level testing the TFO in two different cell lines, T helper-1 lymphocytes and brain microvascular endothelial cells (bend3 cells). Although there was a sequence-specific binding of the TFO detectable in the gel shift assays, there was no inhibitory effect of the exogenously added and phosphorothioate stabilised TFO on endogenous RANTES gene expression visible. Additionally, the small interfering RNA (siRNA) approach was tested as another strategy to inhibit expression of the pro-inflammatory chemokines MCP-1 and RANTES. Two different methods were pursuit, describing transient transfection with vector derived and synthetic siRNA. The vector pSUPER containing the siRNA coding sequence was used to suppress endogenous MCP-1 in HEK 293 cells. An empty vector without RNA sequence served as a control. Inhibition due to the siRNA was measured in stimulated and unstimulated cells. In TNF-a stimulated cells MCP-1 protein synthesis was decreased by 35 ± 11% after siRNA transfection. Using a synthetic double-stranded siRNA, the TNF-a induced MCP-1 protein secretion could be successfully inhibited about 62.3 ± 10.3% in HEK 293 cells, indicating that the siRNA is functional in these cells to suppress chemokine expression. The siRNA approach targeting murine RANTES in Th1 cells and b-end3 cells revealed no inhibition of endogenous gene expression. Gene therapy approaches rely on efficient transfer of genes to the desired target cells. A wide variety of viral and nonviral vectors have been developed and evaluated for their efficiency of transduction, sustained expression of the transgene, and safety. Among them, lentiviruses have been widely used for gene therapy applications. In order to improve the delivery of TFOs or siRNAs into the target cells, cloning of the lentiviral transfer vector SEW, the production of lentiviral particles by transient transfection were performed with the aim to generate lentiviral vector-derived TFOs in further experiments. Here, Th1 cells were transduced with infectious lentiviral particles and transduction efficacy was measured. Transduction efficacy higher than 82% could be achieved using the lentiviral vector SEW, opening optimal possibilities for the TFO or siRNA approach.
I derive a general effective theory for hot and/or dense quark matter. After introducing general projection operators for hard and soft quark and gluon degrees of freedom, I explicitly compute the functional integral for the hard quark and gluon modes in the QCD partition function. Upon appropriate choices for the projection operators one recovers various well-known effective theories such as the Hard Thermal Loop/ Hard Dense Loop Effective Theories as well as the High Density Effective Theory by Hong and Schaefer. I then apply the effective theory to cold and dense quark matter and show how it can be utilized to simplify the weak-coupling solution of the color-superconducting gap equation. In general, one considers as relevant quark degrees of freedom those within a thin layer of width 2 Lambda_q around the Fermi surface and as relevant gluon degrees of freedom those with 3-momenta less than Lambda_gl. It turns out that it is necessary to choose Lambda_q << Lambda_gl, i.e., scattering of quarks along the Fermi surface is the dominant process. Moreover, this special choice of the two cutoff parameters Lambda_q and Lambda_gl facilitates the power-counting of the numerous contributions in the gap-equation. In addition, it is demonstrated that both the energy and the momentum dependence of the gap function has to be treated self-consistently in order to determine the imaginary part of the gap function. For quarks close to the Fermi surface the imaginary part is calculated explicitly and shown to be of sub-subleading order in the gap equation.
In this dissertation a non-deterministic lambda-calculus with call-by-need evaluation is treated. Call-by-need means that subexpressions are evaluated at most once and only if their value must be known to compute the overall result. Also called "sharing", this technique is inevitable for an efficient implementation. In the lambda-ND calculus of chapter 3 sharing is represented explicitely by a let-construct. Above, the calculus has function application, lambda abstractions, sequential evaluation and pick for non-deterministic choice. Non-deterministic lambda calculi play a major role as a theoretical foundation for concurrent processes or side-effected input/output. In this work, non-determinism additionally makes visible when sharing is broken. Based on the bisimulation method this work develops a notion of equality which respects sharing. Using bisimulation to establish contextual equivalence requires substitutivity within contexts, i.e., the ability to "replace equals by equals" within every program or term. This property is called congruence or precongruence if it applies to a preorder. The open similarity of chapter 4 represents a new concept, insofar that the usual definition of a bisimulation is impossible in the lambda-ND calculus. So in section 3.2 a further calculus lambda-Approx has to be defined. Section 3.3 contains the proof of the so-called Approximation Theorem which states that the evaluation in lambda-ND and lambda-Approx agrees. The foundation for the non-trivial precongruence proof is set out in chapter 2 where the trailblazing method of Howe is extended to be capable with sharing. By the use of this (extended) method, the Precongruence Theorem proves open similarity to be a precongruence, involving the so-called precongruence candidate relation. Joining with the Approximation Theorem we obtain the Main Theorem which says that open similarity of the lambda-Approx calculus is contained within the contextual preorder of the lambda-ND calculus. However, this inclusion is strict, a property whose non-trivial proof involves the notion of syntactic continuity. Finally, chapter 6 discusses possible extensions of the base calculus such as recursive bindings or case and constructors. As a fundamental study the calculus lambda-ND provides neither of these concepts, since it was intentionally designed to keep the proofs as simple as possible. Section 6.1 illustrates that the addition case and constructors could be accomplished without big hurdles. However, recursive bindings cannot be represented simply by a fixed point combinator like Y, thus further investigations are necessary.
Mitochondial NADH:ubiquinone oxidoreductase (complex I) the largest multiprotein enzyme of the respiratory chain, catalyses the transfer of two electrons from NADH to ubiquinone, coupled to the translocation of four protons across the membrane. In addition to the 14 strictly conserved central subunits it contains a variable number of accessory subunits. At present, the best characterized enzyme is complex I from bovine heart with a molecular mass of about 980 kDa and 32 accessory proteins. In this study, the subunit composition of mitochondrial complex I from the aerobic yeast Y. lipolytica has been analysed by a combination of proteomic and genomic approaches. The sequences of 37 complex I subunits were identified. The sum of their individual molecular masses (about 930 kDa) was consistent with the native molecular weight of approximately 900 kDa for Y. lipolytica complex I obtained by BN-PAGE. A genomic analysis with Y. lipolytica and other eukaryotic databases to search for homologues of complex I subunits revealed 31 conserved proteins among the examined species. A novel protein named “X” was found in purified Y. lipolytica complex I by MALDI-MS. This protein exhibits homology to the thiosulfate sulfurtransferase enzyme referred to as rhodanese. The finding of a rhodanese-like protein in isolated complex I of Y. lipolytica allows to assume a special regulatory mechanism of complex I activity through control of the status of its iron-sulfur clusters. The second part of this study was aimed at investigating the possible role of one of these extra subunits, 39 kDa (NUEM) subunit which is related to the SDRs-enzyme family. The members of this family function in different redox and isomerization reactions and contain a conserved NAD(P)H-binding site. It was proposed that the 39 kDa subunit may be involved in a biosynthetic pathway, but the role of this subunit in complex I is unknown. In contrast to the situation in N. crassa, deletion of the 39 kDa encoding gene in Y. lipolytica led to the absence of fully assembled complex I. This result might indicate a different pathway of complex I assembly in both organisms. Several site-directed mutations were generated in the nucleotide binding motif. These had either no effect on enzyme activity and NADPH binding, or prevented complex I assembly. Mutations of arginine-65 that is located at the end of the second b-strand and responsible for selective interaction with the 2’-phosphate group of NADPH retained complex I activity in mitochondrial membranes but the affinity for the cofactor was markedly decreased. Purification of complex I from mutants resulted in decrease or loss of ubiquinone reductase activity. It is very likely that replacement of R65 not only led to a decrease in affinity for NADPH but also caused instability of the enzyme due to steric changes in the 39 kDa subunit. These data indicate that NADPH bound to the 39 kDa subunit (NUEM) is not essential for complex I activity, but probably involved in complex I assembly in Y. lipolytica.
The challenging intricacies of strongly correlated electronic systems necessitate the use of a variety of complementary theoretical approaches. In this thesis, we analyze two distinct aspects of strong correlations and develop further or adapt suitable techniques. First, we discuss magnetization transport in insulating one-dimensional spin rings described by a Heisenberg model in an inhomogeneous magnetic field. Due to quantum mechanical interference of magnon wave functions, persistent magnetization currents are shown to exist in such a geometry in analogy to persistent charge currents in mesoscopic normal metal rings. The second, longer part is dedicated to a new aspect of the functional renormalization group technique for fermions. By decoupling the interaction via a Hubbard-Stratonovich transformation, we introduce collective bosonic variables from the beginning and analyze the hierarchy of flow equations for the coupled field theory. The possibility of a cutoff in the momentum transfer of the interaction leads to a new flow scheme, which we will refer to as the interaction cutoff scheme. Within this approach, Ward identities for forward scattering problems are conserved at every instant of the flow leading to an exact solution of a whole hierarchy of flow equations. This way the known exact result for the single-particle Green's function of the Tomonaga-Luttinger model is recovered.
The Kaon-Spectrometer (KaoS) at the heavy-ion synchrotron (SIS) at the Gesellschaft für Schwerionenforschung (GSI) in Darmstadt has been used to study production and propagation of K+ and K- mesons from Au+Au collisions at a kinetic beam energy of 1.5 AGeV. This energy for K+ mesons is close to the corresponding production threshold in binary nucleon-nucleon collisions and far below for K- mesons. The azimuthal angular distributions of particles as a function of the collision centrality and particle transverse momenta have been measured. The properties of strange mesons are expected to be modified by the in-medium meson-baryon potential. Theoretical calculations show that the superposition of the scalar and vector potentials leads to a small repulsive K+N and a strong attractive K-N potential. Additionally, the interaction of kaons and antikaons with nuclear matter is different. The strangeness conservation law inhibits the absorption probability of K+ mesons as they contain an s-quark. K- mesons, however, interact with nucleons via strangenessexchange (K- + N ->Y + pion, where Y = lambda, sigma). Moreover, the reverse process (pion + Y -> K- + N) is the dominant production mechanism of K- mesons at SIS energies. The azimuthal angular emission patterns of kaons are expected to be sensitive to the in-medium potentials. An enhanced out-of-plane emission of K+ mesons was observed in Au+Au reactions at 1.0 AGeV and 1.5 AGeV, and also in Ni+Ni at 1.93 AGeV. The out-of-plane emission of K+ mesons in Au+Au reactions at 1.0 AGeV was interpreted as a consequence of a repulsive K+N potential in the nuclear medium, however, recent transport calculations show that the emission patterns obtained in Au+Au at 1.5 AGeV and Ni+Ni at 1.93 AGeV are additionally influenced by the re-scattering of kaons. For K- mesons the calculations predict an almost isotropic emission pattern due to the attractive K-N potential which counteracts the absorption of K- mesons in the spectator fragments. In Ni+Ni collisions at 1.93 AGeV the azimuthal distribution of K- mesons has been found to be isotropic. In this case, however, the spectators are rather small and have large relative velocities. In addition, the delay of antikaon emission due to strangenessexchange reaction minimizes the interaction with the spectators. As a consequence the sensitivity of the K- meson emission pattern to the K-N in-medium potential is reduced. In Au+Au collisions we found a dependence of the K- meson azimuthal emission pattern on the transverse momentum. The antikaons registered with pt < 0.5 GeV/c are preferentially emitted in the reaction plane and the particles with pt > 0.5 GeV/c show strong out-of-plane enhancement. The emission patterns of K- can be explained in terms of two competing phenomena: one of them is indeed the influence of the attractive K-N potential, however, the second one originates from the strangeness-exchange process.
The quinol:fumarate reductase (QFR) is the terminal reductase of anaerobic fumarate respiration, the most commonly occurring type of anaerobic respiration. This membrane protein complex couples the oxidation of menaquinol to menaquinone to the reduction of fumarate to succinate. The three-dimensional crystal structure of the QFR from Wolinella succinogenes has previoulsy been solved at 2.2 Å resolution. Although the diheme-containing QFR from W. succinogenes is known to catalyze an electroneutral process, structural and functional characterization of parental and variant enzymes has revealed active site locations which indicate electrogenic catalysis across the membrane. A solution to this apparent controversy was proposed with the so-called “Epathway hypothesis”. According to this, transmembrane electron transfer via the heme groups is strictly coupled to a parallel, compensatory transfer of protons via a transiently established pathway, which is inactive in the oxidized state of the enzyme. Proposed constituents of the E-pathway are the side chain of Glu C180, and the ring C propionate of the distal heme. Previous experimental evidence strongly supports such a role for the former constituent. One aim of this thesis is to investigate by a combination of specific 13C-heme propionate labeling and FTIR difference spectroscopy whether the ring C propionate of the distal heme is involved in redox-coupled proton transfer in the QFR from W. succinogenes. In addition to W. succinogenes, the primary structures of the QFR enzymes of two other e- proteobacteria are known. These are Campylobacter jejuni and Helicobacter pylori, which unlike W. succinogenes are human pathogens. The QFR from H. pylori has previously been established to be a potential drug target, and the same is likely for the QFR from C. jejuni. The two pathogenic species colonize mucosal surfaces causing several diseases. The possibility of studying these QFRs from these bacteria and creating more efficient drugs specifically active for this enzyme depends substantially on the availability of large amounts of high-quality protein. Further, biochemical and structural studies on QFR enzymes from e- proteobacteria species other than W. succinogenes can be valuable to enlighten new aspects or corroborate the current understanding of this class of membrane proteins.
Biophysical investigation of the ligand-induced assembling of the human type I interferon receptor
(2005)
Type I interferons (IFNs) elicit antiviral, antiproliferative and immunmodulatory responses through binding to a shared receptor consisting of the transmembrane proteins ifnar1 and ifnar2. Differential signaling by different interferons – in particular IFNalpha´s and IFNbeta – suggest different modes of receptor engagement. In this work either single ligand-receptor interactions or the formation of the extracellular part of a signaling complex were investigated referring to thermodynamics, kinetics, stoichiometry and structural organization. Initially an expression and purification strategy for the extracellular domain of ifnar1 (ifnar1-EC) using Sf9 insect cells yielding in mg amounts of glycosylated protein was established. Using reflectometric interference spectroscopy (RIfS) the interactions between IFNalpha2/beta and ifnar1-EC and ifnar2-EC was studied in order to understand the individual energetic contributions within the ternary complex. For IFNalpha2 a Kd of 5 µM for the interaction with ifnar1-EC was determined. Substantially tighter binding of IFNbeta with both ifnar2-EC and ifnar1-EC compared to IFNalpha2 was observed. For neither IFNalpha2 nor IFNbeta stabilization of the complex with ifnar1-EC in presence of soluble ifnar2-EC was detectable. In addition, no direct interaction between ifnar2 and ifnar1 was could be shown. Thus, stem-stem interactions between the extracellular domains of ifnar1 and ifnar2 do not seem to play a role for ternary complex formation. Furthermore, ligand-induced cross-talk between ifnar1-EC and ifnar2-EC being tethered onto solid-supported, fluid lipid bilayers was investigated by RIfS and total internal reflection fluorescence spectroscopy. A very stable binding of IFNalpha2 at high receptor surface concentrations was observed with an apparent kd approximately 200-times lower than for ifnar2-EC alone. This apparent kd was strongly dependent on the surface concentration of the receptor components, suggesting kinetic rather than static stabilization, which was corroborated by competition experiments. These results indicate that signaling is activated by transient cross-talk between ifnar1 and ifnar2, which is by several orders of magnitude more efficiently engaged by IFNbeta than by IFNalpha2. With respect to differential recognition of different IFNs ifnar1-EC was dissected into sub-fragments containing different of the four Ig-like domains. The appropriate folding and glycosylation of these proteins, also purified in mg amounts were confirmed by SDS-PAGE, size exclusion chromatography and CD-spectroscopy. Surprisingly, only one construct containing all three N-terminal Ig-like domains was active in terms of ligand binding, indicating that these domains were required. Competitive binding of IFNalpha2 and IFNbeta to both this fragment and ifnar1-EC was demonstrated. Cellular binding assays with different fragments, however, highlight the key role of the membrane-proximal Ig-like domain for the formation of an in situ IFN-receptor complex and the ensuing signal activation. Even substitution with Ig-like domains from homologous cytokine receptors did not restore high-affinity ligand binding. Receptor assembling analysis on supported lipid bilayer revealed that appropriate orientation of the receptor is required, which is controlled by the membrane-proximal Ig-domain. All results indicate that differential signalling is encoded by the efficiency of signalling complex formation, which is controlled by the binding affinity of IFNs to the extracellular domains of ifnar1 and 2.
Die vorliegende Arbeit beschäftigt sich mit der Charakterisierung des ALTRO Chips (ALICE TPC Readout), der ein integraler und wichtiger Bestandteil der Auslesekette des TPC (Time Projection Chamber) Detektors von ALICE (A Large Ion Collider Experiment) ist. ALICE ist ein Experiment am noch im Bau befindlichen LHC (Large Hadron Collider) am CERN mit der zentralen Ausrichtung, Schwerionenkollisionen zu untersuchen. Diese sind von besonderem Interesse, da durch sie ein experimenteller Zugriff zu dem QGP (Quark Gluon Plasma) existiert, dem einzigen vom Standardmodell vorhergesagten Phasenübergang, der unter Laborbedingungen erreichbar ist. Im Jahr 2004 wurden Messungen an einem Teststrahl am CERN PS (Proton Synchrotron) durchgeführt. Der Prototyp wurde voll mit FECs bestückt, was 5400 Kanälen entspricht und einer anderen Gasmixtur (Ne/N2/CO2 90%/5%/5%) befüllt. Für das optimale Leistungsverhalten der ALICE TPC muß der Digitalprozessor im ALTRO, bestehend aus vier Berechnungseinheiten, mit den passenden Werten konfiguriert werden. Der Datenfluss beginnt mit dem BCS1 (Baseline Correction and Subtraction 1) Modul, das systematische Störungen und die Grundlinie entfernt. Da der ALTRO kontinuierlich das anliegende Signal abtastet, entfernt es automatisch langsame Grundlinienveränderungen, die Beispielsweise durch Temperaturänderungen auftreten können. Gefolgt von dem TCF (Tail Cancellation Filter), der den Schweif des langsam fallenden, vom PASA generierten Signals entfernt. Um die nichtsystematischen Störungen der Grundlinie zu entfernen, folgt die BCS2 (Baseline Correction and Subtraction 2), die auf einer gleitenden Mittelwertsberechnung mit Ausschluß von Detektorsignalen über einen doppelten Schwellenwert basiert. Die finale Einheit für die Signalverarbeitung ist die ZSU (Zero Suppression Unit), die Meßpunkte unterhalb eines definierten Schwellwertes entfernt. Hier wird der weg beschrieben die TCF und BCS1 Parameter aus vorhandenen Detektordaten zu extrahieren. Während der Analyse der Daten von kosmischen Teilchen fiel bei Signalen mit hoher Amplitude (>700 ADC) eine zusätzliche Struktur in dem Schweif auf. Der Monitor wurde deswegen mit einem gleitenden Mittelwertfilter erweitert, worauf sich diese Struktur auch in kleineren Signalen (> 200 ADC) zeigte. Dieses Signal wird von Ionen erzeugt, die zur Kathode oder zu den Pads driften, bisher ist jedoch weder die Streuung der Elektronenlawine an der Anode, noch die Variationsbreite in den erzeugten Elektronlawinen verstanden oder gemessen worden. Eine erfolgreiche Messung, sowie Charakterisierung wird in dieser Arbeit beschrieben. Im Jahr 2005 im Sommer beginnt der Einbau der Gaskammern der TPC in ALICE, die Elektronik folgt am Ende dieses Jahres. Parallel hierzu wurde der Prototyp der TPC wieder in Betrieb genommen und im Frühling wird ein kompletter Sektor mit der Detektorelektronik ausgestattet. An diesen zwei Aufbauten wird die ALTRO Charakterisierung fortgeführt, verfeinert und komplettiert.
Cancer has become one of the most fatal diseases. The Heidelberg Heavy Ion Cancer Therapy (HICAT) has the potential to become an important and efficient treatment method because of its excellent “Bragg peak” characteristics and on-line irradiation control by the PET diagnostics. The dedicated Heidelberg Heavy Ion Cancer Therapy Project includes two ECR ion sources, a RF linear injector, a synchrotron and three treatment rooms. It will deliver 4*10 high 10 protons, or 1*10 high 10 He, or 1*10 high 9 Carbons, or 5*10 high 8 Oxygens per synchrotron cycle with the beam energy 50-430AMeV for the treatments. The RF linear injector consists of a 400AkeV RFQ and of a very compact 7AMeV IH-DTL accelerator operated at 216.816MHz. The development of the IH-DTL within the HICAT project is a great challenge with respect to the present state of the DTL art because of the following reasons: • The highest operating frequency (216.816MHz) of all IH-DTL cavities; • Extremely large cavity length to diameter ratio of about 11; • IH-DTL with three internal triplets; • The highest effective voltage gain per meter (5.5MV/m); • Very short MEBT design for the beam matching. The following achievements have been reached during the development of the IH-DTL injector for HICAT : The KONUS beam dynamics design with LORASR code fulfills the beam requirement of the HICAT synchrotron at the injection point. The simulations for the IH-DTL injector have been performed not only with a homogeneous input beam, but also with the actual particle distribution from the exit of the HICAT RFQ accelerator as delivered by the PARMTEQ code. The output longitudinal normalized emittance for 95% of all particles is 2.00AkeVns, the emittance growth is less than 24%, while the X-X’ and Y-Y’ normalized emittance are 0.77mmmrad and 0.62mmmrad, respectively. The emittance growth in X-X’ is less than 18%, and the emittance growth in Y-Y’ is less than 5%. Based on the transverse envelopes of the transported particles, the redesign of the buncher drift tubes at the RFQ high energy end has been made to get a higher transit time factor for this novel RFQ internal buncher. An optimized effective buncher gap voltage of 45.4KV has been calculated to deliver a minimized longitudinal beam emittance, while the influence of the effective buncher voltage on the transverse emittance can be neglected. Six different tuning concepts were investigated in detail while tuning the 1:2 scaled HICAT IH model cavity. ‘Volume Tuning’ by a variation of the cavity cross sectional area can compensate the unbalanced capacitance distribution in case of an extreme beta-lambda-variation along an IH cavity. ‘Additional Capacitance Plates’ or copper sheets clamped on drift tube stems are a fast way for checking the tuning sensitivity, but they will be replaced by massive copper blocks mounted on the drift tube girders finally. ‘Lens Coupling’ is an important tuning to stabilize the operation mode and to increase or decrease the coupling between neighboring sections. ‘Tube Tuning’ is the fine tuning concept and also the standard tuning method to reach the needed field distributions as well as the gap voltage distributions. ‘Undercut Tuning’ is a very sensitive tuning for the end sections and with respect to the voltage distribution balance along the structure. The different types of ‘plungers’ in the 3rd and 4th sections have different effects on the resonance frequency and on the field distribution. The different triplet stems and the geometry of the cavity end have been also investigated to reach the design field and voltage distributions. Finally, the needed uniform field distribution along the IH-DTL cavity and the corresponding effective voltage distribution were realized, the remaining maximum gap voltage difference was less than 5% for the model cavity. The several important higher order modes were also measured. The RF tuning of the IH-DTL model cavity delivers the final geometry parameters of the IH-DTL power cavity. A rectangular cavity cross section was adopted for the first time for this IH-DTL cavity. This eases the realization of the volume tuning concept in the 1st and 2nd sections. Lens coupling determines the final distance between the triplet and the girder. The triplets are mounted on the lower cavity half shell. The Microwave Studio simulations have been carried out not only for the HICAT model cavity, but also for the final geometry of the IH-DTL power cavity. The field distribution for the operation mode H110 fits to the model cavity measurement as well as the Higher Order Modes. The simulations prove the IH-DTL geometrical design. On the other hand, the precision of one simulation with 2.3 million mesh points for full cross section area and the CPU time more than 15hours on a DELL PC with Intel Pentium 4 of 2.4GHz and 2.096GRAM were exploited to their limit when calculating the real parameters for the two final machining iterations during production. The shunt impedance of the IH-DTL power cavity is estimated by comparison with the existing tanks to about 195.8MOmega/m, which fits to the simulation result of 200.3MOmega/m with reducing the conductivity to the 5.0*10 high 7 Omega-1m-1. The effective shunt impedance is 153 MOmega/m. The needed RF power is 755kW. The expected quality factor of the IH-DTL cavity is about 15600. The IH-DTL power cavity tuning measurements before cavity copper plating have been performed. The results are within the specifications. There is no doubt that the needed accuracy of the voltage distribution will be reached with the foreseen fine tuning concepts in the last steps.