The critical function of the kidney is to regulate the body’s extracellular fluid volume to maintain homeostasis. When insults to the kidney occur, as in the case of kidney ischemia, the function of the kidney to filter metabolic wastes and reabsorb essential solutes is compromised, leading to a variety of clinical manifestations. Current metrics of kidney function are measured by the rise of a single analyte, the serum creatinine, which implies injury of the kidney tubule and its epithelial cells and is encapsulated by the term Acute Kidney Injury (AKI). Yet, creatinine does not specify the etiology, the cell type, or the molecular pathways that are affected by the acute decreases in kidney excretory function. During my thesis work, I hypothesized that there is a pathogenetic heterogeneity of kidney injury and a specificity of location, timing, and molecular mechanisms, unique to each of these three injury models: kidney ischemia, volume depletion, and urinary tract infection. Using genetic mouse models, RNA-sequencing, and a range of molecular biology techniques, I have found (1) kidney ischemia activates inflammatory responses, signal transduction pathways, and epithelial repair and reprogramming, that are not activated in volume depletion, (2) which in contrast, is a transient metabolic condition, inducing genes regulating energy metabolism that were reversible upon rehydration. Lastly, (3) I have found that urinary tract infection, particularly one that invades the kidney, involves a novel heme transport system in the collecting duct of the kidney, that may contribute to nutritional defenses against bacterial pathogens. Each of these findings is explored in specific aims and experiments, which I detail here in my thesis.

Translocator Protein 18 kDa (TSPO) is a protein that is expressed at low levels in the brain, but upon brain injury or inflammation, increases its expression in the areas of the brain specific to injury. In this way, TSPO can be used as a biomarker of brain inflammation and injury. TSPO is primarily expressed in two cell types, microglia and astrocytes, and is used as a marker of reactive gliosis in various brain pathologies. Currently, there is a paucity of knowledge on the function(s) of TSPO in glial cells. Recent studies using conditional and global TSPO knockout mice have questioned the role of TSPO in translocating cholesterol across the outer mitochondrial membrane as the first step in steroidogenesis. In the brain, microglia and astrocytes exhibit distinct spatial and temporal patterns of TSPO upregulation. These differential patterns are not well characterized across disease models and in particular, are poorly characterized in the early stages of disease, prior to behavioral and clinical disease manifestations. Importantly, these distinct patterns of TSPO upregulation may indicate different functions of TSPO in microglia and astrocytes. We examined TSPO levels in a neurodegenerative transgenic mouse model of Sandhoff disease (SD) and longitudinally compared TSPO levels to behavioral manifestations of disease and other neuropathological endpoints (neurodegeneration, reactive gliosis, ganglioside accumulation). This study confirmed TSPO upregulation prior to neurodegeneration in a brain region-dependent and disease course-dependent way. In brain regions with increased TSPO levels, there was a differential pattern of glial cell activation with astrocytes being activated earlier than microglia during the progression of disease. Immunofluorescent confocal imaging confirmed that TSPO colocalizes with both microglia and astrocyte markers, but the glial source of the TSPO response differs by brain region and age in SD mice. We next wanted to gain insight into the function of TSPO in microglia. We previously demonstrated that TSPO ligands (TSPO-L) (1-100 nM) induced intracellular ROS production which was abrogated by NADPH oxidase (NOX2) inhibitors, thereby indicating an association between TSPO and NOX2. To further elucidate the relationship between TSPO and NOX, we determined the source of ROS production resulting from microglia exposure to TSPO-L. Intracellular and extracellular ROS production was inhibited by NOX inhibitors, but not by a mitochondria permeability transition pore inhibitor, indicating that the source of ROS production is from NOX and not from mitochondria. These findings were confirmed using the mitochondria specific ROS probe MitoSOX. To further explore the TSPO-NOX2 association, we used 3 molecular approaches to examine protein-protein interactions under unstimulated or stimulated conditions (100 ng/mL lipopolysaccharide (LPS) for 18 hours) in primary microglia. 1) Co-immunoprecipitation (co-IP) revealed that the NOX2 subunits, gp91phox (gp91) and p22phox (p22), co-IP with TSPO supporting a protein-protein interaction. TSPO’s association with gp91 and p22 decreased with activation, but TSPO’s association with VDAC, a mitochondrial protein, remained constant. These findings suggest that microglia activation changes the dynamics of the TSPO-NOX2 interaction. 2) Confocal imaging and colocalization analysis of TSPO/gp91 or TSPO/p22 immunofluorescence confirmed that TSPO colocalizes with both NOX subunits. Under stimulated conditions, TSPO associated with gp91 and TSPO associated with p22, exhibit significantly decreased colocalization with VDAC suggesting a movement from the mitochondria to other cellular compartments. 3) Duolink Proximity Ligation Assay confirmed that TSPO interacts with p22, gp91 and VDAC. Our results suggest a novel TSPO-gp91-p22 interaction with VDAC in primary microglia that is disrupted by microglia activation and may be involved with redox homeostasis with significant implications for a new understanding of TSPO glial cell biology. In summary, the present studies have strengthened the use of TSPO as a preclinical biomarker, confirmed its specific spatiotemporal upregulation in two cell types and have provided a new potential function of TSPO in microglia that has the possibility to revolutionize the TSPO field and to inform neurotoxicity assessments and neurological disease treatments.

Injuries to the peripheral nervous system rank among the most common causes of chronic neuropathic pain. Afflicting millions of people for months or even years, symptoms of this condition have proven difficult to treat clinically. A thorough understanding of the pathophysiological changes induced by such nerve lesions is essential to the development of more efficient therapeutic options. Peripheral nerve injury induces a robust and tightly regulated innate immune response in the dorsal horn of the spinal cord. The precise molecular mechanisms regulating the spatiotemporal dynamics and functional impact of the response remain incompletely understood. Preclinical evidence suggests mitigating this immune response can have a significant therapeutic benefit in the treatment of neuropathic pain, however these findings have yet to be clinically validated. To elucidate the mechanisms regulating the spinal immune response, we used a mouse model of partial sciatic nerve injury exclusively in male adult (2-3-month-old) mice. The spared nerve injury (SNI) model employed throughout our studies induces robust, persistent neuropathic pain-like behavior. We established a time course for the spinal immune response to SNI and used mRNA extracted from the ipsilateral dorsal horn of lumbar spinal cord segments L4 and L5 to analyze changes in the transcriptome at the peak of the immune reaction 7 days after nerve lesion. We discovered upregulation of multiple elements of the triggering receptor expressed on myeloid cells 2 (Trem2) pathway. Trem2 is considered a regulator of toll-like receptor signaling in innate immune cells. It also promotes microglia-mediated phagocytosis in the central nervous system. Recent work from our lab has established neuronal apoptosis in the ipsilateral dorsal horn after SNI as an essential mechanism leading to the development of chronic neuropathic pain-like behavior. We used TUNEL staining of L4 spinal cord sections to compare the clearance of apoptotic cell profiles in Trem2-/- mice to wild-type littermates and discovered a key role for Trem2 in the clearance of apoptotic cells after SNI. We further used genetic deletion of Trem2 as well as administration of a Trem2 agonist in C57Bl/6 mice to assess the impact of Trem2 signaling on both the spinal immune response and neuropathic pain-like behavior after SNI. Neither removal nor augmentation of Trem2 signaling significantly affected the development of neuropathic pain-like behavior. Utilizing flow cytometry, we also evaluated the cellular composition of the spinal immune response. We found no evidence that monocytes from the peripheral circulation invade the spinal cord after SNI, as has been previously suggested. These findings were corroborated by immunohistochemical analysis of spinal cord sections from transgenic mice that express distinct fluorescent proteins in their monocyte and microglia cell populations. To better understand the different mechanisms modulating the spinal immune response, we further examined several transcriptionally regulated signaling pathways. We achieved the greatest reduction of mechanical allodynia in nerve-lesioned mice treated with a P2x4r antagonist. Surprisingly, the removal of fractalkine (Cx3cl1) signaling, another prominent chemokine signaling pathway in microglia, had no significant impact on either the spinal immune response or mechanical allodynia after SNI. Reducing the number of spinal microglia by blocking Csf1r activation did not prevent the development of mechanical allodynia after SNI either. Our findings reveal a more nuanced concept of microglial activation after nerve injury. The impact on neuropathic pain-like behavior and phagocytosis appear to be regulated by pathways that differ from those controlling immune cell recruitment and global activation. These findings provide a greater understanding of the complex mechanisms governing microglial function and offer new insight into molecular targets essential to the development of more efficient treatment options for neuropathic pain.

Sun Yehuan. / "September 2004." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (p. 213-223). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. Ann Arbor, MI : ProQuest Information and Learning Company, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.

Cerebral haemodynamic status defined as cerebral vasoreactivity to carbon dioxide and pressure autoregulatory response, have been shown to be affected after traumatic brain injury (TBI) and correlate with the neurological condition and clinical outcome. Therefore, it is important to have a reliable method to determine the cerebral haemodynamic status in brain-injured patients. Blood flow velocity (BFV) measurement by transcranial Doppler ultrasonography (TCD) has been shown to give accurate indication of changes in cerebral blood flow (CBF). Transient hyperaemic response (THR) test with TCD measurement to assess the BFV response of middle cerebral artery to a brief compression of the ipsilateral carotid artery, provides a simple method for repeated assessment of the cerebrovascular autoregulatory reserve in brain injured patients. However, the test has not been validated systematically against classical assessment tests using TCD and gold standard CBF measurement. / The aims of this thesis are (1) to validate the non-invasive TCD and its haemodynamic tests with a more involved gold standard CBF measurement using stable xenon-enhanced computerized tomography. (2) To correlate the cerebral haemodynamic abnormalities with the patterns of neurochemical disturbance detected by intracerebral microdialysis. (3) To investigate the possibility to reverse or minimized the cerebral haemodynamic abnormalities and metabolic derangement by treatment. (Abstract shortened by UMI.) / The goal of intensive care management for TBI is to provide them with a favourable physiological and metabolic environment for recovery of the injured-compromised cells. The development of clinical intracerebral microdialysis has enabled documentation of the metabolic derangement that provides more understanding of the mechanism of brain damage. Continuous measurement of both neurochemical and physiological parameters including CPP defined as mean arterial blood pressure (ABP) minus intracranial pressure, BFV and CBF, enables study of the relationship between metabolic events and physiologic changes. Clinical management of patients with TBI has emphasized on maintaining an optimal cerebral perfusion pressure (CPP). This critical CPP can then be defined by TCD, CBF as well as the metabolic measurements. / Ng Chi Ping. / "June 2005." / Adviser: Wai-sang Poon. / Source: Dissertation Abstracts International, Volume: 67-01, Section: B, page: 0122. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2005 / Includes bibliographical references (p. 147-154). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese. / School code: 1307.

O trauma desponta como mais uma doença a que os idosos podem estar vulneráveis. Além disso, tendo em vista o aumento desta faixa etária, é possível que, em breve, a realidade do trauma nesta população também cresça. Dessa forma, é fundamental que os profissionais dos serviços de saúde conheçam as alterações que ocorrem com o processo de envelhecimento e as características específicas do trauma, com a finalidade de melhor assistir esta população. Assim, os objetivos deste estudo foram: identificar o perfil sociodemográfico de idosos, vítimas de trauma; caracterizar as doenças preexistentes e os medicamentos em uso; descrever as características do trauma e sua evolução; verificar a existência de associação entre variáveis sociodemográficas, doenças preexistentes, características e evolução do trauma; verificar a existência de correlação entre dias internados em CTI e ISS. Trata-se de um estudo não experimental, retrospectivo e exploratório. Realizado a partir da análise de dados de natureza secundária contidos em um banco de dados do Núcleo Hospitalar de Epidemiologia do Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto da Universidade de São Paulo, referentes às notificações dos pacientes, vítimas de trauma, atendidos na Unidade de Emergência, deste hospital, no período de 2008 a 2010. Dessa forma, a coleta dos dados seguiu as informações contidas no referido banco, além da busca nos prontuários médicos para identificação de doenças preexistentes, do uso de medicamentos em domicílio e das complicações clínicas após o trauma. Foram estudados 131 idosos, vítimas de trauma, média de idade 69,9 anos (s=7,7); 73,3% eram homens; 55,1%, casados; 54,7%, aposentados; 65,6% possuíam doenças preexistentes, sendo 38,9% hipertensão arterial sistêmica e 19,8% etilismo, média de doenças 2,3 (s=1,4); 48,9% tomavam medicação em domicílio, média 3,2 medicamentos (s=2,3). Quanto às características do trauma, para 31,3%, o mecanismo de trauma foi queda e para 28,2%, pedestre; 83,2% por trauma contuso; 59,5% possuíam lesão em cabeça/pescoço, 45,8% em extremidades e ossos da pelve, média 1,8 (s=1,0); 44,3% obtiveram ISS entre 9 e 15 (trauma moderado) e 30,5% ISS de 25 ou mais (trauma muito grave); 80,2% apresentaram TRISS entre 51% ou mais (francas condições de se evitar o óbito). Com relação à evolução do trauma, 30,5% internaram em CTI, média de 4,2 dias; 62,6% desenvolveram complicações clínicas, sendo 43,5% infecciosas e 30,5% cardiovasculares; 46,1% foram submetidos à cirurgia ortopédica; 66,4% sobreviveram ao trauma, 47,3% receberam alta hospitalar com limitações moderadas e 33,6% faleceram, sendo 36,4% por traumatismo cranioencefálico e 22,7% por sepse. Houve associação entre mecanismo do trauma e doença preexistente (p=0,01) e associação entre mecanismo do trauma e sexo (p=0,03); a presença de doenças aumentou em 3,10 a chance para desenvolver complicações em relação aos que não apresentavam doenças (p=0,02); para os internados em CTI, a chance de ter complicações aumenta em 28,2 (p<0,01); conforme aumenta o índice de gravidade do trauma, maiores são as chances de complicações, odds = 3,07 entre ISS 16 e 24 (grave) e odds = 6,50 com ISS 25+ (muito grave) em relação ao ISS 9 a 15 (moderado); para idosos com complicações, a chance de morte aumenta em 5,56, quando comparados com aqueles que não apresentaram (p<0,01); para idosos com TRISS <50% (sobrevida inesperada), a chance de óbito foi de 10,13 em relação àqueles com TRISS >=50% (morte evitável) (p<0,01); a correlação entre os dias de internação no CTI e os escores do ISS foi fraca e positiva (r=0,18), indicando que quanto maior o número de dias de internação no CTI maiores são os índices de gravidade do trauma (p=0,03). O conhecimento das características e da evolução do trauma pode possibilitar aos profissionais de saúde o planejamento de medidas preventivas, além de viabilizar melhor atendimento aos idosos na atenção intra-hospitalar e após a alta, com vistas a melhorar a qualidade de vida. / Trauma emerges as another condition to which elderly individuals are vulnerable. Considering the increase in this population, trauma events are also likely to increase among older individuals. Hence, it is essential that health care providers are aware of changes that may occur with the aging process and the specific characteristics of trauma aiming to better care for this population. This study identifies the sociodemographic profile of elderly trauma victims; characterizes pre-existent diseases and used medications; describes the characteristics of trauma and its progression; verifies potential correlation between days hospitalized in ICU and Injury Severity Score (ISS). This non-experimental, retrospective and exploratory study was based on secondary data collected from a database of the Hospital Epidemiology Center at the Hospital das Clinicas, Medical School, University of São Paulo at Ribeirão Preto concerning reports of elderly trauma victims cared for in the hospital\'s emergency department from 2008 to 2010. Data collection was based on information contained in the database and search on medical charts to identify pre-existent diseases, medication used at home, and clinical complications after the trauma. A total of 131 elderly trauma victims participated in the study: 69.9 years old in average (sd=7.7); 73.3% men; 55.1% married; 54.7% retired; 65.6% with pre-existent diseases: 38.9% systemic arterial hypertension, and 19.8% alcoholism; average of diseases 2.3 (sd=1.4); 48.9% took medication at home, average of 3.2 medications (sd=2.3). In relation to the characteristics of trauma: 31.3% was caused by falls and 28.2% pedestrian; 83.2% was contusion trauma; 59.5% had head and neck injury; 45.8% had limbs and pelvic bones affected, average 1.8 (sd=1.0); 44.3% obtained ISS between 9 and 15 (moderate trauma) and 30.5% ISS was 25 or above (very severe trauma); 80.2% presented Trauma and Injury Severity Score (TRISS) between 51% or above (real conditions to avoid death). In relation to trauma progression, 30.5% was hospitalized in ICU, 4.2 days in average; 62.6% developed clinical complications: 43.5% infections and 30.5% cardiovascular; 46.1% was submitted to orthopedic surgery; 66.4% survived, 47.3% was discharged with moderate impairment and 33.6% died: 36.4% due to brain injury and 22.7% due to sepsis. An association between the mechanism of trauma and pre-existent diseases was found (p=0.01) as well as association between mechanism of trauma and gender (p=0.03). Pre-existent diseases increased 3.10 times the chance of complications comparing to those with no pre-existent diseases (p=0.02). The chances of complications increased 28.2 times for those hospitalized in ICU (p<0.01); the higher the index of trauma severity, the greater the chances of complications, odds = 3.07 between ISS 16 to 24 (severe) and odds = 6.50 with ISS 25+ (very severe) in relation to ISS 9 to 15 (moderate). The chances of dying increased 5.56 times for those with complications compared to those with no complications (p<0.01); chances of death was 10.13 times higher for individuals with TRISS <50% (unexpected survival) in relation to those with TRISS >=50% (evitable death) (p<0.01). Correlation between duration of hospitalization in ICU and ISS scores was weak and positive (r=0.18) indicating that the longer the hospitalization in ICU, the higher the trauma severity index (p=0.03). Knowledge concerning the trauma characteristics and progression can enable health care providers to plan preventive measures and provide better care to elderly individuals both at the hospital and after discharge aiming to improve their quality of life.

O objetivo do estudo foi realizar a adaptação cultural do STAR Skin Tear Classification System, para a língua portuguesa no Brasil e testar a validade de conteúdo e a confiabilidade inter-observadores da versão adaptada. O estudo é do tipo metodológico com abordagem quantitativa. A adaptação cultural foi desenvolvida em três fases: tradução, avaliação por comitê de juízes e retro-tradução. Foram testadas duas propriedades de medida: validade de conteúdo e confiabilidade inter-observadores. Para as análises estatísticas foi utilizado o índice kappa ponderado. A versão adaptada para o português obteve um nível regular de concordância (kw = 0,286), embora estatisticamente significativo (p = 0,000), quando de sua aplicação por enfermeiros em fotografias de lesão por fricção. Quando de sua aplicação na prática clínica, a versão adaptada em português obteve um nível moderado e estatisticamente significativo de concordância (kw = 0,596; p < 0,001). O estudo sobre o processo de adaptação cultural e validação das propriedades de medida do STAR Skin Tear Classification System possibilitou atestar a validade de conteúdo e a confiabilidade inter-observadores da versão adaptada para uso na língua portuguesa do Brasil / This study aims to perform a cultural adaptation of the STAR Skin Tear Classification System, to the Portuguese language spoken in Brazil and to test its contents validity and the reliability in the translated version. This is a methodological type of study with a quantitative approach. The cultural adaptation was developed in three stages: translation, evaluation by a judges committee and back translation. Two measures were tested: validity and reliability. The statistical analysis used the weighted kappa index. The adapted version had a regular concordance level (kw = 0,286), although statistically significant (p < 0,000) when tested by nurses in skin tears photographs. When tested in clinical practice, the adapted version achieved a moderate and statistically significant concordance level (kw = 0,596; p < 0,001). The study about the cultural adaptation process and validation of the measurements properties of the STAR Skin Tear Classification System confirmed the adapted Brazilian Portuguese version content validity and reliability

Thesis (Master of Emergency Medical Care)--Cape Peninsula University of Technology, 2018. / Physical injury is a major cause of premature death and disability worldwide (WHO, 2015). Mortality statistics for South Africa indicate that approximately half of all injury-related deaths were intentionally inflicted, often as a result of sharp-force injuries (Donson 2009). Cape Town is reputed to be a violent city (Nicol et al., 2014). Pre-hospital emergency care providers are often the first medical contact for injured patients. Previously, it was understood that high volume crystalloid administration would improve survival and was standardised in the management of shock (Santry & Alam 2010). However, over-administration of crystalloid fluid can cause patient harm by potentially worsening injuries and can be detrimental to a patients survival. Current evidence supports the practice of lower volume crystalloid intravenous fluid administration- permissive hypotension. Little is known about pre-hospital emergency care providers intravenous fluid management practices for penetrating injury. Injury surveillance data for victims of penetrating injury is also scarce with the majority of current data taken from mortality sources. Surveilling pre-hospital cases may yield opportunities for prevention from premature mortality and morbidity. The aim of this study is to undertake surveillance of penetrating injury and related intravenous fluid resuscitation in the pre-hospital emergency care environment. A prospective observational descriptive survey was conducted in the Cape Metropole1. Over three consecutive months, emergency care providers documented parameters related to mechanism of injury, scene vital signs, hospital vital signs, intravenous fluid resuscitation and basic patient demographic information for patients with penetrating injury. A predetermined inclusion and exclusion criteria was used to sample patients.

Injury now surpasses disease as the leading global cause of premature death and disability, claiming over 5.8 millions lives each year. However, unlike disease, which has been subjected to a rigorous epidemiologic approach, the field of injury prevention and control has been a relative newcomer to scientific investigation. With the distribution of injury now well described (i.e. ???who???, ???what???, ???where??? and ???when???), the underlying hypothesis is that progress in understanding ???how??? and ???why??? lies in classifying injury occurrences aetiologically. The advancement of a means of classifying injury aetiology has so far been inhibited by two related limitations: 1. Structural limitation: The absence of a cohesive and validated aetiological taxonomy for injury, and; 2. Methodological limitation: The need to manually classify large numbers of injury cases to determine aetiological patterns. This work is directed at overcoming these impediments to injury research. An aetiological taxonomy for injury was developed consistent with epidemiologic principles, along with clear conventions and a defined three-tier hierarchical structure. Validation testing revealed that the taxonomy could be applied with a high degree of accuracy (coder/gold standard agreement was 92.5-95.0%), and with high inter- and intra- coder reliability (93.0-96.3% and 93.5-96.3%). Practical application demonstrated the emergence of strong aetiological patterns which provided insight into causative sequences leading to injury, and led to the identification of effective control measures to reduce injury frequency and severity. However, limitations related to the inefficient and error-prone manual classification process (i.e. average 4.75 minute/case processing time and 5.0-7.5% error rate), revealed the need for an automated approach. To overcome these limitations, a knowledge acquisition (KA) software tool was developed, tested and applied, based on an expertsystems technique known as ripple down rules (RDR). It was found that the KA system was able acquire tacit knowledge from a human expert and apply learned rules to efficiently and accurately classify large numbers of injury cases. Ultimately, coding error rates dropped to 3.1%, which, along with an average 2.50 minute processing time, compared favourably with results from manual classification. As such, the developed taxonomy and KA tool offer significant advantages to injury researchers who have a need to deduce useful patterns from injury data and test hypotheses regarding causation and prevention.

"April 2002" Bibliography: leaves 190-202. Examines the suitability of the [alpha] 2 adrenoreceptor agonist xylazine for providing safe and effective analgesia in 2 settings: for post-surgical pain in adult sheep used for biomedical research, and for routine husbandry procedures applied to lambs on farms, such as mulesing, tail-docking and castration. Concludes in setting 1 that intramuscular administration of xylazine was simple to perform yet was characterized by a rapid peak analgesic effect with a reasonable duration of action and minimal deleterious effects on cardiac output, blood pressure or arterial blood gases. In setting 2 the anti-nociceptive effects in lambs are of a similar magnitude and duration to those in adult sheep when the dose was scaled for body weight. A ranking of the relative painfulness of husbandry procedures was developed and used to assess the efficacy of intramuscular xylazine.