>Magister Scientiae - MSc / Background: Malnutrition is a risk factor for the development of pulmonary tuberculosis (TB) and may be responsible for the premature deaths of patients with active disease. An adequate nutritional status may therefore be protective in delaying the onset from latent infection to active disease. In South Africa, very little data is available on the nutritional status of adults who present with tuberculosis. This study therefore aims to compare the nutritional status of newly diagnosed pulmonary tuberculosis patients with TB-free controls.Study population & Design: This is a community based case-control study. Forty-three newly diagnosed pulmonary tuberculosis patients were recruited as cases and matched according to age,gender and race to 43 TB-free close contacts. HIV positive subjects were excluded from the study.Methods: Each participant was interviewed and completed a structured questionnaire to obtain demographic information. Weight was measured to the nearest 0.1kg and height to the nearest 1mm. A 24-hr dietary recall method was used to obtain dietary information. Biochemical analysis was carried out to measure concentrations of transferrin, albumin, CRP, ferritin, zinc,copper, vitamin A and E.Results: The median Body Mass Index (BMI) for cases was 18.80kg/m² (IQR 14.35, 32.11) and TB-free contacts 21.17 kg/m² (IQR 16.75, 34.98) with a significant difference between the groups of p=0.001. There was significant difference in weight (p=0.002) and MUAC (p=0.000)between groups. No significant difference in dietary intake of energy (KJ) (p=0.695), protein(p=0.804), CHO (p=0.801) and fat ( p=0.796) was found between groups. There was a statistically significant increase in ferritin (p=0.000) and C-reactive protein (CRP) (p=0.000) in TB patients, while albumin (p=0.000), serum zinc (p=0.000) and serum vitamin A (p=0.000) were statistically significantly lower among cases. Conclusion: There was no significant difference in the macronutrient intake of TB cases and TB-free contacts, although a significant difference was seen in BMI, MUAC and weight between groups, with all these parameters being lower in TB patients. Ferritin and CRP levels were markedly increased in TB cases while serum zinc, vitamin A and albumin are all significantly lower in TB patients than TB free contacts.

The hepatic acute phase response (APR), stimulated by injury or inflammation, is characterized by significant changes in circulating acute phase protein (APP) concentrations. While individual functions of liver-derived APPs are known, the net consequence of APP changes is unclear. Pneumonia and sepsis elicit systemic inflammation and induce a robust APR. Although APR activation is regarded as a hallmark of infection, direct contributions of liver activation to pulmonary defense during pneumonia and sepsis-induced pneumonia remain unclear. Pneumonia causes a pulmonary inflammatory response coordinated largely by alveolar macrophages, and is typified by cytokine production, leukocyte recruitment and plasma extravasation, the latter of can enable delivery of hepatocyte-derived APPs to the infection site. To determine the functional significance of the hepatic APR during pneumonia, we challenged APR-null mice lacking hepatocyte signal transducer and activator of transcription-3 (STAT3) and RelA with 106 colony-forming units (CFU) Escherichia coli intratracheally. HepSTAT3/RelA-/- mice displayed ablated APP induction, significantly increased mortality, tumor necrosis factor-dependent hepatotoxicity, and pulmonary bacterial burdens. Following a lower (4x105 CFU) E. coli inoculum, hepSTAT3/RelA-/- mice had decreased APP concentrations with reduced pulmonary inflammation and diminished airspace macrophage activation. Similar results were obtained in the context of endotoxemia and pneumonia. We employed an endotoxemia/pneumonia model, whereby 18 hours of intraperitoneal E. coli lipopolysaccharide (5 mg/kg) was followed by intratracheal E. coli (106 CFU) in mice lacking hepatocyte STAT3 (hepSTAT3-/-) or control hepSTAT3+/+ mice. Following endotoxemia and pneumonia, hepSTAT3-/- mice, with significantly reduced levels of circulating and airspace APPs, exhibited significantly elevated lung and blood bacterial burdens and mortality. While neither recruited airspace neutrophils nor lung injury were altered in endotoxemic hepSTAT3-/- mice, in vivo production of reactive oxygen species in alveolar macrophage was significantly decreased. Additionally, bronchoalveolar lavage fluid from this group of hepSTAT3-/- mice allowed greater bacterial growth ex vivo. These results identify a lung-liver axis, whereby the liver response enhances macrophage activation and pulmonary host defense during pneumonia and sepsis-induced pneumonia. Taken together, induction of liver acute phase gene expression programs contributes to countering the deleterious consequences of pneumonia, whether it is alone or in the context of sepsis-induced infection.

C-reactive protein (CRP), a component of the innate immune system, is an antipneumococcal plasma protein. Human CRP has been shown to protect mice against infection with lethal doses of Streptococcus pneumoniae by decreasing bacteremia. in vitro, CRP binds to phosphocholine-containing substances, such as pneumococcal C-polysaccharide, in a Ca2+-dependent manner. Phosphocholine-complexed human CRP activates the complement system in both human and murine sera. The mechanism of antipneumococcal action of CRP in vivo, however, has not been defined yet. In this study, we tested a decades-old hypothesis that the complement-activating property of phosphocholine-complexed CRP contributes to protection of mice against pneumococcal infection. Our approach was to investigate a CRP mutant, incapable of activating murine complement, in mouse protection experiments. We employed site-directed mutagenesis of CRP, guided by its three-dimensional structure, and identified a mutant H38R which, unlike wild-type CRP, did not activate complement in murine serum. Substitution of His38 with Arg in CRP did not affect the pentameric structure of CRP, did not affect the binding of CRP to pneumococci, and did not decrease the stability of CRP in mouse circulation. Employing a murine model of pneumococcal infection, we found that passively administered H38R CRP failed to protect mice against infection. Infected mice injected with H38R CRP showed no reduction in bacteremia and did not survive longer, as opposed to infected mice treated with wild-type CRP. Thus, the hypothesis that complement activation by phosphocholine-complexed CRP is an antipneumococcal effector function was supported. We can conclude now that complement activation by phosphocholine-complexed CRP is indeed essential for CRP-mediated protection of mice against pneumococcal infection.

The miniaturization of optical systems has been a longstanding interest for physicists. By facilitating the design of smaller optical systems, we can improve their versatility and cost-effectiveness. This aim applies to macroscopic imaging systems, technologies that implicitly image, and micrometer-scale optics. Parallel to this, quantum optical devices have also seen rapid developments. Notably, the need for new quantum communications and quantum imaging devices has recently risen. The thesis outlines advancements in both of these areas and, in many ways, bridges gaps between them. It discusses the development of optics that compress free space, the design of holographic optical elements, and the generation of entangled photon states in thin-film sources. First, we describe an optic designed to miniaturize free space, termed the spaceplate. Spaceplates achieve the propagation of light for a distance greater than their thickness.Therefore, they compress optical space, reducing the required distance between optical elements in an imaging system. In this thesis, we describe a spaceplate based on conventional optics in a 4-f arrangement, mimicking the transfer function of free space in a thinner system - we term this device a three-lens spaceplate. It is broadband, polarization-independent, and achieves meter-scale space compression. We experimentally measure compression ratios up to 15.6, replacing up to 4.4 meters of free space, three orders of magnitude greater than previous spaceplates. We demonstrate that three-lens spaceplates reduce the length of a full-color imaging system, albeit with reductions in resolution and contrast. We also present theoretical limits on the numerical aperture and the compression ratio. Our design presents a simple, accessible, cost-effective method for optically compressing large volumes of space. Second, we discuss the design of holographic optical elements. Holograms are extraordinarily versatile optics. They have many applications, including interferometry, spectrometry, data storage, optical filtration, and sensing. We can design various optical elements such as filters, lenses, beam splitters, and solar concentrators by tailoring the phase response of a hologram. In this thesis, we describe the nature and function of holograms, and we experimentally characterize holography in lithium niobate and photopolymers. Using this characterization, we assess the limitations of different holographic analysis methods. Further, we describe novel holographic optical elements like the holographic spaceplate - a holographic optic element whose phase response mimics free space. Lastly, this thesis outlines the production of entangled photon pairs, or biphotons, via non-phase-matched spontaneous parametric down-conversion in micrometer- and nanometer-scale devices. By producing biphotons in micrometer-scale crystals rather than in bulk, as is done traditionally, we are allowed to ignore phase matching. These devices produce broadband emission in both angle and frequency not seen in phase-matched bulk sources. We measure entangled biphotons produced via spontaneous parametric down-conversion in gallium arsenide (111) and lithium niobate - both undoped and iron-doped. Lastly, we outline and present initial experiments towards a holographic spontaneous parametric down-conversion optic that combines photon production and mode sorting - an optic of cascaded miniaturization.

Interactive handheld electronic displays use hydrogenated amorphous silicon (a-Si:H) thin film transistor (TFT) as a backplane and a Touch Screen Panel (TSP) on top as an input device. The low mobility and instability of a-Si:H TFT threshold voltage are major two issues for driving constant current as required for Active Matrix Organic Light Emitting Ddiode (AMOLED) displays. Low mobility is compensated by increasing transistor width or resorting to more expensive material TFTs. On the other hand, the ever increasing threshold voltage shift degrades the drain current under electrical operation causing OLED display to dim. Mutual capacitive TSP, the current cell phone standard, requires two layers of metals and a dielectric to be put in front of the display, further dimming the device and adding to visual noise due to sun reflection, not to mention increased integration cost and decreased yield. This thesis focuses on the aforementioned technological hurdles of a handheld electronic display by proposing a dual-gate TFT used as an OLED current driving TFT and a novel phase response readout scheme that can be applied to a one metal track TSP. Our dual-gate TFT has shown on average 20% increase in drive current over a single gate TFT fabricated in the same batch, attributed to the aid of a top channel to the convention bottom channel TFT. Furthermore the dual gate TFT shows three times the Poole-Frenkel current than the single gate TFT attributed to the increase in gate to drain overlap. The dual-gate TFT shows a 50% improvement in threshold voltage shift over a single gate TFT at room temperature, but only ~8% improvement under 75ºC. This is an important observation as it shows an accelerated threshold voltage shift in the dual-gate. This difference in the rate of threshold voltage change under varying temperature is attributed to the difference in interface states, supporting Libsch and Kanicki’s multi-level temperature dependant dielectric trapping model. The phase response TSP readout scheme requires IC only on one side of the display. Cadence Spectre simulation results showed that both touch occurrence and touch position can be obtained using only one metal layer.

Interactive handheld electronic displays use hydrogenated amorphous silicon (a-Si:H) thin film transistor (TFT) as a backplane and a Touch Screen Panel (TSP) on top as an input device. The low mobility and instability of a-Si:H TFT threshold voltage are major two issues for driving constant current as required for Active Matrix Organic Light Emitting Ddiode (AMOLED) displays. Low mobility is compensated by increasing transistor width or resorting to more expensive material TFTs. On the other hand, the ever increasing threshold voltage shift degrades the drain current under electrical operation causing OLED display to dim. Mutual capacitive TSP, the current cell phone standard, requires two layers of metals and a dielectric to be put in front of the display, further dimming the device and adding to visual noise due to sun reflection, not to mention increased integration cost and decreased yield. This thesis focuses on the aforementioned technological hurdles of a handheld electronic display by proposing a dual-gate TFT used as an OLED current driving TFT and a novel phase response readout scheme that can be applied to a one metal track TSP. Our dual-gate TFT has shown on average 20% increase in drive current over a single gate TFT fabricated in the same batch, attributed to the aid of a top channel to the convention bottom channel TFT. Furthermore the dual gate TFT shows three times the Poole-Frenkel current than the single gate TFT attributed to the increase in gate to drain overlap. The dual-gate TFT shows a 50% improvement in threshold voltage shift over a single gate TFT at room temperature, but only ~8% improvement under 75ºC. This is an important observation as it shows an accelerated threshold voltage shift in the dual-gate. This difference in the rate of threshold voltage change under varying temperature is attributed to the difference in interface states, supporting Libsch and Kanicki’s multi-level temperature dependant dielectric trapping model. The phase response TSP readout scheme requires IC only on one side of the display. Cadence Spectre simulation results showed that both touch occurrence and touch position can be obtained using only one metal layer.

No description available.

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No. of bitstreams: 1 girardi_am_dr_jabo.pdf: 19183057 bytes, checksum: cf7a9853bcc87aef37b3ca91a2afceea (MD5) Previous issue date: 2016-01-26 / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Este estudo avaliou o uso da termografia infravermelha e do exame radiológico digital como ferramentas de diagnóstico precoce da laminite em ovinos com acidose ruminal subaguda induzida experimentalmente e os efeitos da ingestão prolongada de alta proporção de alimento concentrado sobre as variáveis clínicas e laboratoriais. Foram utilizadas sete ovelhas adultas, com cânulas ruminais permanentes, que não tiveram prévio contato com alimento concentrado. Para a indução da acidose ruminal, incluiu-se ao volumoso, diariamente, 10% de alimento concentrado até atingir 80%, porcentagem mantida até completarem 19 semanas. A observação dos sinais de diarreia deu-se do quarto ao 22º dia e de laminite do quinto ao 24º dia. O fluido ruminal foi predominantemente líquido, de odor ácido e coloração amarelada. Observou-se elevação inicial nos valores da frequência cardíaca, tempo de sedimentação e flotação no fluido ruminal, plaquetas, neutrófilos segmentados, AST, FA, GGT, cálcio ionizado, magnésio, glicose, ureia, triglicérides e bilirrubinas indireta e total. No início do experimento, notou-se redução da frequência respiratória, linfocitopenia, monocitopenia, hipocalcemia, hipofosfatemia, hiponatremia, hipoproteinemia e hipocolesterolemia. A frequência de movimentos ruminais, o tempo de redução do azul de metileno, os níveis sanguíneos de creatinina, bilirrubina direta, lactato e CK foram reduzidos, enquanto as médias de temperatura, peso e escore corporal, eosinófilos, albumina, cloretos e potássio no sangue aumentaram durante todo o período de observação. Redução do pH ruminal ocorreu nos primeiros dias, a despeito de sua manutenção acima de 5,5 durante as 19 semanas de observação. Mecanismos compensatórios respiratórios e metabólicos, principalmente a acidificação da urina, mantiveram o pH sanguíneo dentro dos limites fisiológicos considerados para a espécie ovina. Dezesseis frações proteicas séricas foram separadas pela eletroforese, dentre elas oito proteínas identificadas pelos seus pesos moleculares, de funções desconhecidas. A maioria das proteínas de fase aguda e as imunoglobulinas identificadas no fracionamento eletroforético variaram durante o período de observação, sugerindo reação inflamatória. Nenhuma alteração macroscópica foi detectada à necropsia. A radiologia digital não permitiu a identificação de alterações podais decorrentes de laminite durante o período de avaliação. A termografia infravermelha detectou aumentos na temperatura da parede do casco mesmo sem a manifestação clínica de dor, indicando que esta técnica pode ser uma ferramenta útil para detectar laminite em ovinos numa fase precoce de evolução da doença. / This study evaluated the use of infrared termography and digital radiologic examination as early diagnostic tools for laminitis in sheep with experimentally induced subacute ruminal acidosis, and the effects of long-term ingestion of a high concentrate diet on clinical and laboratory variables. Seven adult ewes with permanent rumen cannula were used, which did not have any previous ingestion of concentrate feed. For the induction of ruminal acidosis, it was added to the roughage, 10% of concentrate feed until reach 80%, percentage held to the end of the 19-week experiment. Diarrhea signs were observed from fourth to 22nd day and laminitis from fifth to 24th day. Ruminal fluid was predominantly liquid, sour-smelling and yellowish. Initial increases of heart rate, sedimentation and flotation time, platelets, segmented neutrophils, AST, ALP, GGT, ionized calcium, magnesium, glucose, urea, triglycerides, indirect and total bilirubin were noted. At the beginning of the experiment, there was reduction of respiratory rate, lymphocytopenia, monocytopenia, hypocalcemia, hypophosphatemia, hyponatremia, hypoproteinemia, and hypocholesterolemia. Ruminal movement rate, methylene blue reduction time, creatinine, direct bilirubin, lactate and CK blood levels decreased, while body temperature, weight and score, eosinophils, albumin, chloride and potassium blood levels increase throughout the period. Ruminal pH reduction occurred within the first days, despite its maintenance above 5.5 during the 19 weeks. Respiratory and metabolic mechanisms, mainly urine acidification, remained the blood pH between physiologic limits for sheep. Sixteen protein fractions were separated by electrophoresis, among them eight proteins of unknown functions, identified by their molecular weights. Most acute phase proteins and the immunoglobulins identified in the electrophoretic fractionation varied throughout the observation period, suggesting inflammation. No macroscopic alteration was detected at necropsy. The digital radiology did not allow the identification of podal changes resulting from laminitis during the trial period. Infrared thermography has detected increases in the maximum temperature of the hoof wall, even without the clinical manifestation of pain, indicating that this technique can be an useful tool to detect laminitis in sheep at an early stage of the disease. / FAPESP: 2012/09111-7

京都大学 / 新制・課程博士 / 博士(医学) / 甲第24800号 / 医博第4992号 / 新制||医||1067(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 伊藤 貴浩, 教授 波多野 悦朗, 教授 妹尾 浩 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

C-reactive protein (CRP) is an acute phase protein produced by hepatocytes whose serum concentration increases in inflammatory conditions including cardiovascular complications. Statins that are used in the treatment of cardiovascular diseases to reduce cholesterol also lower serum CRP levels. In human hepatoma Hep3B cells, CRP is induced in response to cytokines IL-6 and IL-1β. The objective of the study was to determine the mechanism of regulation of CRP gene expression in Hep3B cells in response to cytokines and to determine the effect of statins on CRP expression. Key findings of our research were: 1. IL-1β-activated NF-κB p50/p65 acted synergistically with IL-6-activated C/EBPβ in inducing CRP transactivation through the proximal CRP promoter. 2. A NF-κB site was localized in the proximal CRP promoter centered at position -69 overlapping the known OCT-1/HNF-1/HNF-3 sites. 3. The synergy between IL-6 and IL-1β in inducing CRP gene expression was partially mediated through the NF-κB site. 4. In the absence of C/EBPβ, a complex containing C/EBPζ and RBP-Jκ was formed at the C/EBP-p50-site. 5. Overexpressed C/EBPζ repressed both (IL-6+IL-1β)-induced and C/EBPβ-induced CRP expression. 6. OCT-1 repressed (IL-6+IL-1β)-induced CRP transactivation through the proximal CRP promoter. 7. Statins reduce cytokine-induced CRP gene expression at the transcriptional level. These findings led us to conclude that: 1. CRP transcription is determined by the relative levels of various transcription factors such as C/EBPβ, C/EBPζ, NF-κB and OCT-1 and their interaction with the proximal CRP promoter. 2. Inhibition of CRP transcription by statins is not due to an anti-inflammatory effect but due to the direct effect on CRP gene expression.