XB130 is a relatively unstudied novel signal adaptor protein. In the first phase of this study, an in silico search for proteins related to XB130 was conducted. Two other proteins (AFAP and AFAP1L1) were found to have a significant similarity to XB130 and were compared in detail. After an analysis of these three proteins, it was proposed that they are members of a novel protein family, termed the “AFAP family of signal adaptor proteins”. XB130 has previously been found to regulate cell cycle progression, death, and migration in lung epithelial cells. It was therefore hypothesized that XB130 is protective in acute lung injury (ALI) and important for facilitating repair after injury. XB130 was found to be differentially regulated in ALI depending on the initial insult. Engineering XB130 transgenic mice to further characterize the role of XB130 in lung injury/regeneration revealed that this protein could be essential for early embryo development.

Thesis (Ph. D.)--University of Virginia, 2008. / In title: [A] is subscript upper case A; [B] is subscript upper case B. Includes bibliographical references. Also available online through Digital Dissertations.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O objetivo do presente estudo foi investigar o envolvimento do estresse oxidativo na lesão pulmonar aguda (LPA) induzida por lipopolissacarídeo (LPS) e as repercussões inflamatórias, estruturais e funcionais, através de análises bioquímicas de estresse oxidativo, prova de função pulmonar, análise histológica e RT-PCR para citocinas e fatores de transcrição pró-inflamatórios. Na primeira etapa foram utilizados camundongos machos C57BL6 foram divididos em sete grupos: Grupo controle (CTR) (50 &#956;L de solução fisiológica) administrados via intratraqueal [it], LPS 6 horas (10 &#956;L de LPS) [it], LPS 12 horas (10 &#956;L de LPS) [it], LPS 24 horas (10 &#956;L de LPS) [i], LPS 48 horas (10 &#956;L de LPS). Para verificar que as alterações observadas eram estresse oxidativo dependentes camundongos machos C57BL6 foram pré-tratados com N-acetilcisteína (NAC) 1 hora antes do estímulo com LPS e sacrifícados 24 horas depois do estímulo com LPS. Os animais foram divididos da seguinte forma: Grupo LPS 24 horas (10 &#956;L) [it], grupo NAC 40 mg/kg (gavagem) + LPS (10 &#956;L) [it] e grupo NAC 100 mg/kg (gavagem) + LPS (10 &#956;L) [it]. O sistema antioxidante enzimático protegeu o pulmão do estresse oxidativo nas primeiras 12 horas. O estresse oxidativo foi caracterizado em 24 horas e em 48 horas observou-se falência do sistema antioxidante enzimático. Parâmetros de função pulmonar se mostraram alterados nos grupo estimulados com LPS principalmente no grupo LPS. A elastância (p<0,001), resistência de via aérea periférica (&#916;P2) (p<0,001), resistência de via aérea central (&#916;P1) (p<0,001) e resistência de via aérea total (&#916;Ptot) (p<0,001) se mostraram principlamente alteradas no grupo LPS 24 horas. O pré-tratamento com NAC impediu o aumento dos parâmetros de elastância (p<0,001), resistência de via aérea periférica (&#916;P2) (p<0,001) resistência de via aérea central (&#916;P1) (p<0,05) e resistência de via aérea total (&#916;Ptot) (p<0,001) comparado com o grupo LPS 24 horas. As alterações histológicas como espessamento de septo alveolar, influxo de células inflamatórias e hemorragia mostraram-se tempo dependentes. O pré-tratamento NAC impediu as alterações observadas nos grupo estimulados com LPS. Alterações inflamatórias foram observadas no grupo estimulado com LPS como IL-6 (p<0,001), iNOS (p<0,001), COX2 (p<0,05), TNF-&#945; (p<0,001) e NF&#954;B (p<0,001) quando comparados ao grupo controle. O pré-tratamento com NAC impediu o aumento desses parâmetros como IL-6 (p<0,001), iNOS (p<0,001), COX2 (p<0,05), TNF-&#945; (p<0,05) e NF&#954;B (p<0,001) quando comparados ao grupo LPS 24 horas. Nossos resultados sugerem que o estresse oxidativo desempenha um papel importante nas respostas inflamatórios, estruturais e funcionais no modelo de LPA induzido por LPS e que essas alterações são estresse oxidativo dependentes. / The aim of this study was to investigate the involvement of oxidative stress in acute lung injury (ALI) induced by lipopolysaccharide (LPS) and inflammatory effects, structural and functional, through biochemical analysis of oxidative stress, pulmonary function test, histological and RT-PCR for cytokines and transcription factors pro-inflammatory. In the first stage were used C57BL6 male mice were divided into seven groups: control group (CTR) (50 &#956;L saline) administered via intratracheal [it], LPS 6 hours (10 &#956;L of LPS) [it], LPS 12 hours (10 &#956;L of LPS) [it], LPS 24 hours (10 &#956;L of LPS) [it], LPS 48 hours (10 &#956;L of LPS). To confirm that the observed changes were dependent on oxidative stress in C57BL6 male mice were pretreated with N-acetylcysteine (NAC) 1 hours before stimulation with LPS and sacrificed 24 hours after stimulation with LPS. The animals were divided as follows: LPS group 24 hours (10 &#956;L) [it], NAC group 40 mg / kg (gavage) + LPS (10 &#956;L) [it] NAC group and 100 mg / kg (gavage) + LPS (10 &#956;L) [it]. The enzymatic antioxidant system protected the lungs against oxidative stress in the first 12 hours. Oxidative stress was characterized in 24 hours and 48 hours there was failure of the enzymatic antioxidant system. Pulmonary function parameters were shown in the altered group stimulated with LPS mainly in the LPS group. Elastance (p <0,001), peripheral airway resistance (&#916;P2) (p <0,001), central airway resistance (&#916;P1) (p <0,001) and total airway resistance (&#916;Ptot) (p <0,001) if especially in the group showed altered LPS 24 hours. Pretreatment with NAC prevented the increase in elastance parameters (p <0,001), peripheral airway resistance (&#916;P2) (p <0,001) central airway resistance (&#916;P1) (p <0,05) and resistance total airway (&#916;Ptot) (p <0,001) compared with the LPS group 24 hours. Histological changes such as thickening of alveolar septa, inflammatory cells and hemorrhage proved to be time dependent. The NAC pretreatment prevented the changes observed in the group stimulated with LPS. Inflammatory changes were observed in the group stimulated with LPS and IL-6 (p <0,001), iNOS (p <0,001), COX2 (p <0,05), TNF-&#945; (p <0,001) and NF&#954;B (p <0,001) compared with the control group. Pretreatment with NAC prevented the increase of these parameters as IL-6 (p <0,001), iNOS (p <0,001), COX2 (p <0,05), TNF-&#945; (p <0,05) and NF&#954;B ( p <0,001) when compared with LPS 24 hours. Our results suggest that oxidative stress plays an important role in inflammatory responses, structural and functional model of ALI induced by LPS and that these changes are dependent oxidative stress.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O objetivo do presente estudo foi investigar o envolvimento do estresse oxidativo na lesão pulmonar aguda (LPA) induzida por lipopolissacarídeo (LPS) e as repercussões inflamatórias, estruturais e funcionais, através de análises bioquímicas de estresse oxidativo, prova de função pulmonar, análise histológica e RT-PCR para citocinas e fatores de transcrição pró-inflamatórios. Na primeira etapa foram utilizados camundongos machos C57BL6 foram divididos em sete grupos: Grupo controle (CTR) (50 &#956;L de solução fisiológica) administrados via intratraqueal [it], LPS 6 horas (10 &#956;L de LPS) [it], LPS 12 horas (10 &#956;L de LPS) [it], LPS 24 horas (10 &#956;L de LPS) [i], LPS 48 horas (10 &#956;L de LPS). Para verificar que as alterações observadas eram estresse oxidativo dependentes camundongos machos C57BL6 foram pré-tratados com N-acetilcisteína (NAC) 1 hora antes do estímulo com LPS e sacrifícados 24 horas depois do estímulo com LPS. Os animais foram divididos da seguinte forma: Grupo LPS 24 horas (10 &#956;L) [it], grupo NAC 40 mg/kg (gavagem) + LPS (10 &#956;L) [it] e grupo NAC 100 mg/kg (gavagem) + LPS (10 &#956;L) [it]. O sistema antioxidante enzimático protegeu o pulmão do estresse oxidativo nas primeiras 12 horas. O estresse oxidativo foi caracterizado em 24 horas e em 48 horas observou-se falência do sistema antioxidante enzimático. Parâmetros de função pulmonar se mostraram alterados nos grupo estimulados com LPS principalmente no grupo LPS. A elastância (p<0,001), resistência de via aérea periférica (&#916;P2) (p<0,001), resistência de via aérea central (&#916;P1) (p<0,001) e resistência de via aérea total (&#916;Ptot) (p<0,001) se mostraram principlamente alteradas no grupo LPS 24 horas. O pré-tratamento com NAC impediu o aumento dos parâmetros de elastância (p<0,001), resistência de via aérea periférica (&#916;P2) (p<0,001) resistência de via aérea central (&#916;P1) (p<0,05) e resistência de via aérea total (&#916;Ptot) (p<0,001) comparado com o grupo LPS 24 horas. As alterações histológicas como espessamento de septo alveolar, influxo de células inflamatórias e hemorragia mostraram-se tempo dependentes. O pré-tratamento NAC impediu as alterações observadas nos grupo estimulados com LPS. Alterações inflamatórias foram observadas no grupo estimulado com LPS como IL-6 (p<0,001), iNOS (p<0,001), COX2 (p<0,05), TNF-&#945; (p<0,001) e NF&#954;B (p<0,001) quando comparados ao grupo controle. O pré-tratamento com NAC impediu o aumento desses parâmetros como IL-6 (p<0,001), iNOS (p<0,001), COX2 (p<0,05), TNF-&#945; (p<0,05) e NF&#954;B (p<0,001) quando comparados ao grupo LPS 24 horas. Nossos resultados sugerem que o estresse oxidativo desempenha um papel importante nas respostas inflamatórios, estruturais e funcionais no modelo de LPA induzido por LPS e que essas alterações são estresse oxidativo dependentes. / The aim of this study was to investigate the involvement of oxidative stress in acute lung injury (ALI) induced by lipopolysaccharide (LPS) and inflammatory effects, structural and functional, through biochemical analysis of oxidative stress, pulmonary function test, histological and RT-PCR for cytokines and transcription factors pro-inflammatory. In the first stage were used C57BL6 male mice were divided into seven groups: control group (CTR) (50 &#956;L saline) administered via intratracheal [it], LPS 6 hours (10 &#956;L of LPS) [it], LPS 12 hours (10 &#956;L of LPS) [it], LPS 24 hours (10 &#956;L of LPS) [it], LPS 48 hours (10 &#956;L of LPS). To confirm that the observed changes were dependent on oxidative stress in C57BL6 male mice were pretreated with N-acetylcysteine (NAC) 1 hours before stimulation with LPS and sacrificed 24 hours after stimulation with LPS. The animals were divided as follows: LPS group 24 hours (10 &#956;L) [it], NAC group 40 mg / kg (gavage) + LPS (10 &#956;L) [it] NAC group and 100 mg / kg (gavage) + LPS (10 &#956;L) [it]. The enzymatic antioxidant system protected the lungs against oxidative stress in the first 12 hours. Oxidative stress was characterized in 24 hours and 48 hours there was failure of the enzymatic antioxidant system. Pulmonary function parameters were shown in the altered group stimulated with LPS mainly in the LPS group. Elastance (p <0,001), peripheral airway resistance (&#916;P2) (p <0,001), central airway resistance (&#916;P1) (p <0,001) and total airway resistance (&#916;Ptot) (p <0,001) if especially in the group showed altered LPS 24 hours. Pretreatment with NAC prevented the increase in elastance parameters (p <0,001), peripheral airway resistance (&#916;P2) (p <0,001) central airway resistance (&#916;P1) (p <0,05) and resistance total airway (&#916;Ptot) (p <0,001) compared with the LPS group 24 hours. Histological changes such as thickening of alveolar septa, inflammatory cells and hemorrhage proved to be time dependent. The NAC pretreatment prevented the changes observed in the group stimulated with LPS. Inflammatory changes were observed in the group stimulated with LPS and IL-6 (p <0,001), iNOS (p <0,001), COX2 (p <0,05), TNF-&#945; (p <0,001) and NF&#954;B (p <0,001) compared with the control group. Pretreatment with NAC prevented the increase of these parameters as IL-6 (p <0,001), iNOS (p <0,001), COX2 (p <0,05), TNF-&#945; (p <0,05) and NF&#954;B ( p <0,001) when compared with LPS 24 hours. Our results suggest that oxidative stress plays an important role in inflammatory responses, structural and functional model of ALI induced by LPS and that these changes are dependent oxidative stress.

Inhaled drug delivery is currently the gold standard for treating many respiratory diseases. However, improved treatments are needed for lung diseases like Cystic Fibrosis (CF) and Acute Respiratory Distress Syndrome (ARDS), where mucus or fluid build-up in the lung limits ventilation and, thus, delivery of inhaled drugs. Delivery is most needed in the diseased or damaged regions of the lung, but if an area is not ventilated, inhaled drug will simply not reach it. To overcome this, this research proposes delivering drugs to the lungs within a perfluorocarbon (PFC) liquid. The lungs will be filled with a reverse emulsion containing a disperse phase of aqueous drugs within the bulk PFC and then ventilated. The PFC functions as both a respiratory medium, providing gas exchange, and as a delivery vehicle, providing a more uniform deposition of drugs. After treatment, the highly volatile PFCs are exhaled, returning the patient to normal respiration. This technique improves upon current therapies as follows. First, drugs are delivered directly to where they are needed, yielding higher concentrations in the lung and lower systemic concentrations. Second, PFCs are ideal for washing out lung exudate and mucus. The low surface tension and high density of PFC allows it to easily penetrate plugged or collapsed alveoli, detach infected mucus from the airway walls, and force these fluids to the top of the lungs where they can then be removed via suction. Mucus and exudate removal should allow drugs to penetrate previously plugged airways during emulsion delivery and subsequent treatment with inhaled therapies. Thus, drug delivery via emulsion would be used as a pre-treatment to enhance inhaled or systemic drug therapy. Third, PFC’s anti-inflammatory properties help return to normal lung function. This research examines two applications of this technology: delivery of antibiotics to combat respiratory infections (antibacterial perfluorocarbon ventilation, APV) or delivery of growth factors to enhance alveolar repair (perfluorocarbon emulsions for alveolar repair, PEAR). This work represents an in-depth analysis of the emulsions used during APV and PEAR. Initial efforts evaluated emulsion efficacy under in vitro setting that better simulated lung in vivo antibiotic delivery. The subsequent studies utilized an in vivo rat model of bacterial respiratory infection to validate the effects of emulsion on pharmacokinetics and to assess APVs potential treatment benefits. Lastly, in vitro methods of cellular response assessed the utility of delivering growth factors in PEAR. Significant advancements were made in optimizing the emulsion as a viable means of pulmonary drug delivery. Final efforts resulted in a promising emulsion formulation that overcame the quick transport of tobramycin away from the lung and successfully reduced pulmonary bacterial load in vivo. In vitro applications of PEAR showed the emulsions posed a significant barrier to the availability and, thus, the biological effect of lysophosphatidic acid growth factors. Further in vivo work is required to improve APV’s efficacy over conventional treatments and to determine PEAR’s feasibility and efficacy in promoting lung repair.

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第20250号 / 医博第4209号 / 新制||医||1020(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 伊達 洋至, 教授 戸口田 淳也, 教授 開 祐司 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

Enteral nutrition (EN) is often delayed in critically ill patients despite strong evidence to support that early enteral nutrition feeding is beneficial in this population. Adverse outcomes in critically ill patients in which nutrition is delayed include a longer length of stay and time on the ventilator, and a higher incidence of pneumonia and hospital mortality. The purpose of this literature review was to evaluate the current evidence regarding trophic enteral feeds in mechanically ventilated adult patients with acute respiratory distress syndrome (ARDS)/acute lung injury (ALI) and associated clinical outcomes. A retrospective literature review was performed to identify articles published on the topic of trophic feeds in mechanically ventilated adult patients with ALI/ARDS, with a focus on associated clinical outcomes. The studies included in this literature review indicated that the dose and timing of enteral nutrition in critically ill patients with ARDS/ALI had an effect on clinical outcomes. It is possible that additional variables such as the level of organ dysfunction and varying definitions for trophic enteral nutrition also influenced clinical outcomes. The United States (U.S.) and Canadian guidelines for nutrition supportrecommend either trophic or full EN for patients with ARDS/ALI on the basis that these two feeding strategies have similar patient outcomes over the first week of hospitalization. After reviewing the literature, we conclude that caution is warranted when following this recommendation. Regressions suggest full calorie enteral nutrition administered early in the course of critical illness significantly increased the odds of mortality, whereas full calorie enteral nutrition administered later reduced the odds of mortality.

No description available.

Une coagulopathie aigue endogène (EAC) est présente chez 25% des patients de traumatologie dès leur arrivée. Des résultats d’études récentes montrent que cette EAC est liée à l’activation de la voie de la protéine C (aPC). Quelques heures après, se développe un état pro-coagulant associant un niveau abaissé d’aPC et un taux plasmatique élevé de l’inhibiteur de l’activateur du plasminogene (PAI-1). Nous trouvons que l’incidence des pneumopathies associées à la ventilation est significativement augmentée chez ces patients sans toutefois connaître le rôle exact de ces anomalies de coagulation. Basé sur cette hypothèse central de susceptibilité augmentée a l’infection et plus particulièrement aux pneumopathie a P.aeruginosa (PA) le but de ce travail est d’identifier les mécanismes par lesquels PAI-1 et aPC peuvent moduler la perméabilité de la barrière alveolo capillaire et ceci a travers 3 objectifs spécifiques1 – Objectif 1 : déterminer les mécanismes par lequel PA augmente la perméabilité endothéliale. 2 – Objectif 2 : déterminer le rôle d’aPC dans la modulation des effets de PA sur l’œdème pulmonaire lésionnel.3 – Objectif 3 : déterminer le rôle de PAI-1 dans la modulation des effets de PA sur l’œdème pulmonaire lésionnel.En utilisant un inhibiteur spécifique des petites GTPases nous démontrons le rôle centrale joué par RhoA dans le développement de l’œdème pulmonaire induit par PA. PAI-1 et aPC sont impliquées dans le mécanisme lésionnel pulmonaire. aPC et l’inhibition de la voie du RhoA attenue le développement de l’œdème pulmonaire et diminue la dissémination systémique bactérienne. Cependant le blocage invivo de la voie de PAI-1 est associé à une surmortalité et à une augmentation de la charge bactérienne suggérant un rôle de PAI-1 dans l’activation de la réponse inflammatoire nécessaire a l’éradication de PA / A clinically significant acute endogenous coagulopathy (EAC) is present in 25% of major trauma patients upon arrival in the emergency department, before any fluid resuscitation. Results from recent clinical studies indicate that EAC is primarily caused by the activation of the anticoagulant protein C pathway. Several hours later, there is the development of a systemic procoagulant activity associated with low plasma levels of activated protein C (aPC) and an inhibition of the fibrinolysis caused by elevated plasma levels of plasminogen activator inhibitor 1 (PAI-1). We have found that the incidence of ventilator-associated pneumonia (VAP) is significantly increased in trauma patients with these coagulation abnormalities [6, 9]. However, whether these coagulation abnormalities play a mechanistic role in the increased susceptibility to nosocomial lung infection observed after severe posttraumatic hemorrhage is unknown. Thus, the central hypothesis is that the increased susceptibility to P. aeruginosa (PA) pneumonia following severe trauma with tissue hypoperfusion is mediated in part by these posttraumatic coagulation abnormalities within the airspaces of the lung. Specifically, in this work, we will identify through 3 specific aims the mechanisms by which PAI-1 and aPC modulate PA–mediated increase in alveolar-capillary barrier permeability.1 - Specific Aim 1: To determine the mechanisms by which PA increases lung endothelial permeability.2 - Specific Aim 2 : To determine the Role of aPC in modulating the effect of PA on the lung endothelial barrier function3 - Specific Aim 3 : To determine the Role of PAI-1 in modulating the effect of PA on the lung endothelial barrier functionIn the present work, we demonstrated the central role small GTPases RhoA plays in the increase of permeability induced by pseudomonas infection. PAI-1 and aPC are deeply involved in the control of early lung inflammation. aPC and inhibition of the RhoA pathway attenuates the development of pulmonary edema and decrease in the systemic dissemination of P. aeruginosa. However, in vivo disruption of PAI-1 signalling is associated with higher mortality at 24 h and significant increase in the bacterial burden suggesting that PAI-1 is required for the activation of the innate immune response necessary for the eradication of PA from the distal airspaces of the lung

A diabetes tipo 1 (DT1) está associada `a inflamação estéril (IE) e maior susceptibilidade a sepse. A sepse induz a síndrome da resposta inflamatória sistêmica (SIRS) e a inflamação pulmonar aguda (ALI). O leucotrieno (LT) B4 produzido condições inflamatórias induz a expressão de MyD88 em macrófagos (MA). Hipotetizamos que a DT1 induz a síntese de LTB4 promovendo a IE e isto contribui para SIRS, susceptibilidade a sepse e ALI. Os diabéticos apresentaram níveis elevados de LTB4 e IL-1b no soro e seu MA expressaram mais MyD88/STAT-1. A expressão de STAT-1 foi induzida por c-Jun de forma dependente de LTB4. O tratamento com insulina restaurou os níveis de LTB4 e STAT-1/MyD88 e a inibição de LTB4 restaurou os níveis de MyD88 e IL-1b. Na sepse, a inibição de 5LO prolongou a sobrevida dos diabéticos e diminuiu a SIRS menos IL-1b e IL-10 no soro e TNF-a e IL-1b na cavidade peritoneal. O pulmão dos diabéticos apresentaram ALI menos intensa que se correlacionou com um altos níveis de SOCS-1, baixos níveis de MyD88 e falha na ativação de NFkB nos macrófagos alveolares. / Type 1 diabetes (T1D) is associated with sterile inflammation (SI) and increased sepsis susceptibility. Sepsis induces Systemic Inflammatory Response Syndrome (SIRS) and Acute Lung Injury (ALI). Leukotriene (LT) B4 is produced in inflammatory conditions and induces MyD88 expression in macrophages (MA). We hypothesized that T1D induce LB4 that promotes SI contributing to SIRS, sepsis susceptibility and ALI. Diabetics presented higher levels of LTB4 and e IL-1b in the serum and MA expressed more MyD88/STAT-1. STAT-1 expression was induced by c-Jun on LTB4 dependent manner. Insulin treatment restored LTB4 and STAT-1/MyD88 levels and inhibition of LTB4 restored MyD88 and IL-1b levels. During sepsis, 5LO inhibition increased diabetics survival and inhibited SIRS- lower levels of IL-1b and IL-10 in the serum and TNF-a and IL-1b in the peritoneal cavity. Lungs from diabetics presented milder ALI that correlated with high levels of SOCS-1, low levels of MyD88 and impaired NFkB activation in alveolar macrophages.