Tumor lysis syndrome is an oncologic emergency that usually occurs after chemotherapy in patients with hematologic malignancies. Tumor lysis syndrome is rare in cases of solid tumors, especially when it occurs spontaneously. Herein, we present a case of spontaneous tumor lysis syndrome in a 55-year-old woman who presented with dyspnea and was found to have extensive metastatic small cell lung cancer. She developed acute oliguric renal failure and multiple electrolyte abnormalities requiring hemodialysis. The findings of this case suggest that clinicians should maintain a high index of suspicion for patients with malignancies who demonstrate the classic symptom of laboratory abnormalities even in the absence of chemotherapy.

Master of Science / Department of Chemistry / Christopher T. Culbertson / A microfluidic device integrated with valves and a peristaltic pump was fabricated using multilayer soft lithography to analyze single cells. Fluid flow was generated and mammalian cells were transported through the channel manifold using the peristaltic pump. A laser beam was focused at the cross-section of the channels so fluorescence of individual labeled intact cells could be detected. Triggered by the fluorescence signals of intact cells, valves could be actuated so fluid flow was stopped and a single cell was trapped at the intersection. The cell was then rapidly lysed through the application of large electric fields and injected into a separation channel. Various conditions such as channel geometry, pumping frequency, control channel size, and pump location were optimized for cell transport. A Labview program was developed to control the actuation of the trapping valves and a control device was fabricated for operation of the peristaltic pump. Cells were labeled with a cytosolic dye, Calcein AM or Oregon Green, and cell transport and lysis were visualized using epi-fluorescent microscope. The cells were transported at rates of [simular to] 1mm/s. This rate was optimized to obtain both high throughput and single cell trapping. An electric field of 850-900 V/cm was applied so cells could be efficiently lysed and cell lysate could be electrophoretically separated. Calcein AM and Oregon Green released from single cells were separated and detected by laser-induced fluorescence. The fluorescence signals were collected by PMT and sampled with a multi-function I/O card. This analyzing method using microchip may be applied to explore other cellular contents from single cells in the future.

O objeto principal deste projeto é a tradução integral do diálogo Lísis de Platão. A tradução foi feita a partir do Oxford Classical Text, Platonis Opera, III (Oxford, 1903), editado por John Burnet. Além da tradução, também faz parte deste trabalho um texto introdutório ao diálogo. Apesar de ser enquadrado como um dos diálogos aporéticos de definição por muitos estudiosos, o Lísis apresenta uma complexidade filosófica muito grande e uma estrutura que o distancia dos outros diálogos desse grupo (Cármides, Laques, Eutífron, Hípias Maior). A pergunta não aparece no texto, ainda que esteja subjacente ao diálogo; a postura de Sócrates frente aos seus interlocutores é bastante diversa, assim como os próprios interlocutores não apresentam um pretenso saber. Propomos, a partir dessa perspectiva, realizar uma interpretação positiva do diálogo que contemple dois níveis possíveis de leitura: uma no âmbito dramático e/ou dialógico, e a outra no âmbito lógico-argumentativo ainda que, em diversos momentos, essas duas leituras estejam entrelaçadas. A obra oferece uma valiosa lição sobre o fazer discursivo filosófico que podemos apreender nos dois níveis de leitura. Essa lição também está intimamente ligada às aporias que permeiam todo o diálogo. Dessa forma, apresentamos essas duas camadas de leitura relacionadas com a função positiva da aporia no desenvolvimento da atividade filosófica. / The main purpose of this dissertation is the integral translation of Plato\'s Lysis. The translation is based on Oxford Classical Text, Platonis Opera, III (Oxford, 1903), edited by John Burnet. Besides the translation, an introductory study on dialogue\'s issues is also part of this research. Despite being framed as one of the definitional aporetic dialogues by many scholars, the Lysis presents a highly philosophical complexity and structure that distinguish it from other dialogues of that group (Charmides, Laches, Euthyphro, Hippias Major). The question \' \' does not appear throughout the text, although it underlies the dialogue; Socrates\' behavior towards his interlocutors is quite diverse, as well as the speakers themselves do not present an alleged knowledge on the issue of discussion. From this standpoint, we attempt to presnt a constructive interpretation of the dialogue that comprises a twofold reading: one concerning the dramatic or dialogic framework, and the other the logical-argumentative context even though these two levels are eventually intertwined. The work offers a valuable lesson about the philosophical discourse that we can apprehend within the intersection of these two levels of interpretation. Insofar as this lesson is also closely linked to the aporias that permeate the entire dialogue, we contend that this twofold reading is intrinsically related to the positive role of aporia in the development of philosophical activity.

O objeto principal deste projeto é a tradução integral do diálogo Lísis de Platão. A tradução foi feita a partir do Oxford Classical Text, Platonis Opera, III (Oxford, 1903), editado por John Burnet. Além da tradução, também faz parte deste trabalho um texto introdutório ao diálogo. Apesar de ser enquadrado como um dos diálogos aporéticos de definição por muitos estudiosos, o Lísis apresenta uma complexidade filosófica muito grande e uma estrutura que o distancia dos outros diálogos desse grupo (Cármides, Laques, Eutífron, Hípias Maior). A pergunta não aparece no texto, ainda que esteja subjacente ao diálogo; a postura de Sócrates frente aos seus interlocutores é bastante diversa, assim como os próprios interlocutores não apresentam um pretenso saber. Propomos, a partir dessa perspectiva, realizar uma interpretação positiva do diálogo que contemple dois níveis possíveis de leitura: uma no âmbito dramático e/ou dialógico, e a outra no âmbito lógico-argumentativo ainda que, em diversos momentos, essas duas leituras estejam entrelaçadas. A obra oferece uma valiosa lição sobre o fazer discursivo filosófico que podemos apreender nos dois níveis de leitura. Essa lição também está intimamente ligada às aporias que permeiam todo o diálogo. Dessa forma, apresentamos essas duas camadas de leitura relacionadas com a função positiva da aporia no desenvolvimento da atividade filosófica. / The main purpose of this dissertation is the integral translation of Plato\'s Lysis. The translation is based on Oxford Classical Text, Platonis Opera, III (Oxford, 1903), edited by John Burnet. Besides the translation, an introductory study on dialogue\'s issues is also part of this research. Despite being framed as one of the definitional aporetic dialogues by many scholars, the Lysis presents a highly philosophical complexity and structure that distinguish it from other dialogues of that group (Charmides, Laches, Euthyphro, Hippias Major). The question \' \' does not appear throughout the text, although it underlies the dialogue; Socrates\' behavior towards his interlocutors is quite diverse, as well as the speakers themselves do not present an alleged knowledge on the issue of discussion. From this standpoint, we attempt to presnt a constructive interpretation of the dialogue that comprises a twofold reading: one concerning the dramatic or dialogic framework, and the other the logical-argumentative context even though these two levels are eventually intertwined. The work offers a valuable lesson about the philosophical discourse that we can apprehend within the intersection of these two levels of interpretation. Insofar as this lesson is also closely linked to the aporias that permeate the entire dialogue, we contend that this twofold reading is intrinsically related to the positive role of aporia in the development of philosophical activity.

The purpose of the work described in this dissertation is to better understand the role of Rz and Rz1 function with respect to phage lysis. We determined using both a genetic and biochemical approach that the Rz protein is an inner membrane protein containing a single N-terminal transmembrane domain (TMD) with an Nin/Cout topology. Consistent with previous work on Rz1, the Rz1 lipoprotein was found to be localized to the outer membrane (OM). Following localization, both Rz and Rz1 form homodimers in vivo due to intermolecular disulfide formation. Despite being localized to apposing membranes, the two proteins form a complex. A small number of phages encode a potential single protein equivalent of Rz-Rz1. This protein, termed a spanin, is predicted to tether the inner and outer membranes by a single polypeptide chain. Based on complementation, it was concluded that gp11 from the phage T1 is a functional equivalent of Rz-Rz1. Gp11, and by analogy the Rz-Rz1 two-component spanin complex, threads the meshwork of the PG layer. The presence of an Rz-Rz1 complex, which forms in the presence of peptidoglycan (PG), is supported by in vivo results. The soluble periplasmic domains of Rz and Rz1, which are dimeric and monomeric respectively, were purified. Circular dichroism analysis indicates that Rz is structured, with significant α-helical content, whereas Rz1, in which 10 out 39 residues are proline, is unstructured. Mixing the proteins results in the formation of a complex with significant new α-helical content. Negative-stain images reveal ~ 25 nm x ~ 4 nm rod-shaped structures. Holin independent activity of Rz and Rz1 is found to disrupt whole cells. Furthermore, time lapse microscopy of λ and λRzam lysis allows us to conclude that Rz and Rz1 are essential for lysis. These results suggest a model for Rz-Rz1 function which begins with Rz and Rz1 forming a complex through direct interaction prior to holin and endolysin function. Holin-mediated hole formation allows the endolysin to degrade PG which sterically hinders Rz-Rz1 activity. Removal of PG by endolysin degradation thus triggers Rz-Rz1 OM disruption via fusion of the inner and outer membranes.

The N-terminal hydrophobic domain of the phage P1 endolysin Lyz was found to facilitate the export of Lyz in a sec-dependent fashion, explaining the ability of Lyz to cause lysis of E.coli in the absence of the P1 holin. The N-terminal domain of Lyz is demonstrated to be both necessary and sufficient not only for export to the membrane but also for release into the periplasm of this endolysin. We propose that this unusual N-terminal domain functions as a "signal arrest- release" (SAR) sequence, which first directs the endolysin to the periplasm in membrane-tethered form and then allows it to be released as a soluble active enzyme in the periplasm. To understand why release from the membrane is required for the physiological expression of the lytic activity of Lyz, we examined the role of its seven cysteine residues in the biogenesis of the active endolysin. The inactive, membrane-tethered and the active, soluble forms of Lyz differ in their pattern of intramolecular disulfide bonding. We conclude that the release of Lyz from the membrane leads to an intramolecular thiol-disulfide bond isomerization causing a dramatic conformational change in the Lyz protein. As a result, an active site cleft that is missing in nascent Lyz is generated in the mature form of the endolysin. Examination of the protein sequences of related bacteriophage endolysins suggests that the presence of an SAR sequence is not unique to Lyz. Studies on holin and antiholin indicated that P1 encodes two holins, LydA and LydC. The antiholin LydB inhibits LydA by binding to it directly on the membrane. All above results demonstrate a new paradigm for control of phage lysis, which is, upon depolarization of the membrane by holin function at a programmed time, endolysin is released from the bilayer leading to the immediate lysis of the host.

Proefschrift--Utricht. / "Stellingen": p. [i-iii] Bibliography: p. ix-xii.

Gender of animal offspring is of great interest to farmers where gender selection is achieved via the separation of male-bearing from female-bearing sperms prior to performing artificial insemination. A start-up company (Ab Biotech Inc.) has developed a technique for gender selection based on the production of an intracellular single-domain antibody (sdAb) using the bacterium Escherichia coli capable of sexing bovine sperm. The purpose of this research was to provide a recommendation to Ab Biotech Inc. for the lysis of E. coli. An efficient lysis technique was required in order to release the intracellular sdAb. In the dairy industry, sexing for female calves is preferred since male calves are not useful for the purpose of milk production. Multiple lysis techniques were tested in order to provide a feasible recommendation for Ab Biotech Inc. These techniques included high pressure homogenization, sonication, bead milling and enzymatic/chemical lysis using lysozymes and Triton X-100. Required lysis time, extent of lysis and potential operating costs were contributing factors for determining an optimal technique. The extent of lysis was determined by quantifying the total amount of released protein using SDS-PAGE densitometry. It was recommended to choose bead milling for potential process upscaling since a large amount of fractional lysis (0.70) was obtained over a short amount of lysis time (3 min) with an inexpensive ($9.50/kg) 0.3 mm mixture of glass beads.

京都大学 / 新制・課程博士 / 博士(工学) / 甲第23155号 / 工博第4799号 / 新制||工||1750(附属図書館) / 京都大学大学院工学研究科マイクロエンジニアリング専攻 / (主査)教授 横川 隆司, 教授 井上 康博, 教授 中部 主敬 / 学位規則第4条第1項該当 / Doctor of Philosophy (Engineering) / Kyoto University / DFAM

No description available.