Injury and degenerative disease of the central nervous system (CNS) are among the major causes for disabilities in humans. They result in permanent damage that is not repaired by regenerative processes. In contrast, anamniotes like fish and amphibia display a striking potential for successful regeneration in the CNS. The zebrafish (Danio rerio) has been established as a model for successful regeneration after spinal cord injury. However, it is yet unknown which factors are involved in regeneration after spinal lesions and other insults to the CNS. Focusing on motor neurons, I asked whether regeneration can also be observed in larval zebrafish. This would allow to take advantage of their accessibility to live imaging, pharmacological and genetic manipulation. It is unknown, whether the loss of a specific cell type in the absence of injury, which is reminiscent of the pathological change observed in neurodegenerative diseases, would be sufficient to induce regeneration. Comparing the regenerative response after spinal lesion to that after selective neuronal cell loss would allow to identify factors that act as a trigger for regeneration, e.g. mechanical injury signals, the extent of cell death or microglia activation. To address these questions, an experimental paradigm in which motor neurons can be selectively ablated without the need to inflict tissue damage would prove useful. Key findings of this work are: · Motor neuron generation ceases during early larval developmental stages. · The Nitroreductase system can be used for successful ablation of motor neurons in the larval spinal cord. · New motor neurons are generated in a regenerative response to both targeted ablation of motor neurons and spinal lesion in larval zebrafish after cessation of developmental generation of MNs. To test whether larval zebrafish can be used to analyse motor neuron regeneration, I carried out a birthdating study to establish a developmental time line for motor neuron generation in the spinal cord. The end of developmental motor neuron generation at an early time point, at around 54 hours post fertilisation, allows for the use of larval zebrafish to assess the regenerative response after insults to the spinal cord. In addition, I could show a time dependent role for Hedgehog signalling during the generation of a motor neuron subpopulation. The influence of Hedgehog is diminished before the end of motor neurogenesis. Utilizing the Gal4/UAS system to combine the Nitroreductase‐mCherry fusion protein expressing Tg(UAS:nfsB‐mCherry) with the motor neuron specific driver Tg(hb9:Gal4), I generated a new transgenic zebrafish line for the genetically targeted ablation of motor neurons. In the resulting transgenic fish, the administration of the prodrug Metronidazole induces apoptotic cell death in ~25% of spinal motor neurons leading to impaired motor performance and increased numbers of microglia in the spinal cord. My work shows that larval animals subjected to motor neuron ablation or spinal lesion display a regenerative response detected by increased numbers of newborn motor neurons. Importantly, this happens after developmental production of motor neurons has ceased, suggesting that progenitor cells are reverting to the generation of motor neurons. The data presented shows that in larval zebrafish, the selective loss of motor neurons is sufficient to induce a regenerative response in the spinal cord. The increased numbers of microglial profiles in the spinal cord after both spinal lesion and targeted cell ablation indicates a role for the immune system in mediating a regenerative response. This new targeted cell ablation paradigm in larval zebrafish will allow to identify and characterize the progenitor cell population forming new motor neurons. One can then further investigate how specific loss of motor neurons is sensed and which factors contribute to the activation of the endogenous stem cell populations. Using larval zebrafish has many benefits, as they are accessible to pharmacological testing with small molecules and live imaging. Moreover, the combination of additional transgenic reporter lines will allow for the investigation of single cell behaviour during regeneration.

Atrial Fibrillation (AF) is a debilitating heart rhythm disorder affecting over 2.7 million people in the US and over 30 million people worldwide annually. It has a high correlation with causing a stroke and several other risk factors, resulting in increased mortality and morbidity rate. Currently, the non-pharmocological therapy followed to control AF is catheter ablation, in which the tissue surrounding the pulmonary veins (PVs) is cauterized (called the PV isolation - PVI procedure) aims to block the ectopic triggers originating from the PVs from entering the atrium. However, the success rate of PVI with or without other anatomy-based lesions is only 50%-60%. A major reason for the suboptimal success rate is the failure to eliminate patientspecific non-PV sources present in the left atrium (LA), namely reentry source (a.k.a. rotor source) and focal source (a.k.a. point source). It has been shown from several animal and human studies that locating and ablating these sources significantly improves the long-term success rate of the ablation procedure. However, current technologies to locate these sources posses limitations with resolution, additional/special hardware requirements, etc. In this dissertation, the goal is to develop an efficient algorithm to locate AF reentry and focal sources using electrograms recorded from a conventionally used high-resolution multi-pole diagnostic catheter. / Includes bibliography. / Dissertation (Ph.D.)--Florida Atlantic University, 2019. / FAU Electronic Theses and Dissertations Collection

No description available.

<p>The purpose of the diploma work is to investigate the possibilities to combine three different areas; reflectometry, microfluidics and laser ablation in a microstructured single-mode fiber, thus obtaining a controlled technique for positioning for ablation and collection of liquids from small inclusions.</p><p>Each of the three areas is thoroughly described in different sections of this report. The first part of the experiments in this diploma work consisted of combining reflectometry and microfluidics, the second part combining reflectometry with laser ablation and the final experiment setup consisted of a combination of all three areas. An artificial system for liquid collection was then designed for that purpose.</p><p>The results obtained from experiments and measurements clearly demonstrate that combining reflectometry, laser ablation and fluid collection in a single optical fiber is promising. Future work will include improvements of the technique towards a medical application for bone marrow transplantation.</p>

No description available.

No description available.

Prolonged periods of non-weightbearing activity result in a significant loss of bone mass which increases the risk of fracture with the initiation of mechanical loading. The loss of bone mass is partially driven by declines in bone formation yet the mechanisms responsible for this decline are unclear. To investigate the limitations of osteoblasts during disuse, marrow ablation was superimposed on hindlimb unloaded mice. Marrow ablation is a useful model to study osteoblast functionality as new cancellous bone is rapidly formed throughout the marrow of a long bone while hindlimb unloading is the most common method used to produce skeletal unloading. The specific hypotheses of this study were aimed at determining if changes in osteoblast functionality, differentiation, and/or proliferation were compromised in non-weightbearing bone in response to a bone formation stimulus. Additionally, the influence of having compromised osteoblast functionality at the time of stimulation was assessed in non-weightbearing bones. Key outcome measures used to address these hypotheses included static and dynamic cancellous bone histomorphometry, bone densitometry, and real-time polymerase chain reaction (PCR) analyses of gene expression. The results document similar ablation-induced increases of cancellous bone in both weightbearing and unloaded animals. Similarly, there was no influence of load on ablation-induced increases in cancellous bone forming surface or mineral apposition rate. Unloading did significantly attenuate the ablation-induced increase in bone formation rate, due to reduced levels of total surface mineralization. When osteoblast functionality was compromised prior to marrow ablation, bone formation rate increases were also attenuated in ablated animals due to reduced mineralization. Additionally, increases in forming surface were attenuated as compared to unloaded animals having normal osteoblast function at the time of ablation. Collectively, these data identify mineralization as the limiting step in new bone formation during periods of disuse. The caveat, however, is that when bone formation is stimulated after a period of unloading sufficient to compromise osteoblast functionality, increases in osteoblast recruitment to the bone surface are compromised.

The purpose of the diploma work is to investigate the possibilities to combine three different areas; reflectometry, microfluidics and laser ablation in a microstructured single-mode fiber, thus obtaining a controlled technique for positioning for ablation and collection of liquids from small inclusions. Each of the three areas is thoroughly described in different sections of this report. The first part of the experiments in this diploma work consisted of combining reflectometry and microfluidics, the second part combining reflectometry with laser ablation and the final experiment setup consisted of a combination of all three areas. An artificial system for liquid collection was then designed for that purpose. The results obtained from experiments and measurements clearly demonstrate that combining reflectometry, laser ablation and fluid collection in a single optical fiber is promising. Future work will include improvements of the technique towards a medical application for bone marrow transplantation.

No description available.

No description available.