Potential for Cell-Transplant Therapy with Human Neuronal Precursors to Treat Neuropathic Pain in Models of PNS and CNS Injury: Comparison of hNT2.17 and hNT2.19 Cell Lines

Effective treatment of sensory neuropathies in peripheral neuropathies and spinal cord injury (SCI) is one of the most difficult problems in modern clinical practice. Cell therapy to release antinociceptive agents near the injured spinal cord is a logical next step in the development of treatment modalities. But few clinical trials, especially for chronic pain, have tested the potential of transplant of cells to treat chronic pain. Cell lines derived from the human neuronal NT2 cell line parentage, the hNT2.17 and hNT2.19 lines, which synthesize and release the neurotransmitters gamma-aminobutyric acid (GABA) and serotonin (5HT), respectively, have been used to evaluate the potential of cell-based release of antinociceptive agents near the lumbar dorsal (horn) spinal sensory cell centers to relieve neuropathic pain after PNS (partial nerve and diabetes-related injury) and CNS (spinal cord injury) damage in rat models. Both cell lines transplants potently and permanently reverse behavioral hypersensitivity without inducing tumors or other complications after grafting. Functioning as cellular minipumps for antinociception, human neuronal precursors, like these NT2-derived cell lines, would likely provide a useful adjuvant or replacement for current pharmacological treatments for neuropathic pain.

The role of teleost grazers in a relatively pristine seagrass ecosystem

Trophic downgrading of ecosystems necessitates a functional understanding of trophic cascades. Identifying the presence of cascades, and the mechanisms through which they occur, is particularly important for seagrass meadows, which are among the most threatened ecosystems on Earth. Shark Bay, Western Australia provides a model system to investigate the potential importance of top-down effects in a relatively pristine seagrass ecosystem. The role of megagrazers in the Shark Bay system has been previously investigated, but the role of macrograzers (i.e., teleosts), and their importance relative to megagrazers, remains unknown. The objective of my dissertation was to elucidate the importance of teleost macrograzers in transmitting top-down effects in seagrass ecosystems. Seagrasses and macroalgae were the main food of the abundant teleost Pelates octolineatus, but stable isotopic values suggested that algae may contribute a larger portion of assimilated food than suggested by gut contents. Pelates octolineatus is at risk from numerous predators, with pied cormorants (Phalacrocorax varius) taking the majority of tethered P. octolineatus. Using a combination of fish trapping and unbaited underwater video surveillance, I found that the relative abundance of P. octolineatus was greater in interior areas of seagrass banks during the cold season, and that the mean length of P. octolineatus was greater in these areas compared to along edges of banks. Finally, I used seagrass transplants and exclosure experiments to determine the relative effect of megagrazers and macrograzers on the establishment and persistence of three species of seagrasses in interior microhabitats. Teleost grazing had the largest impact on seagrass species with the highest nutrient content, and these impacts were primarily observed during the warm season. My findings are consistent with predictions of a behaviorally-mediated trophic cascade initiated by tiger sharks (Galeocerdo cuvier) and transmitted through herbivorous fishes and their predators.

Transplantes de córneas no estado do Rio Grande do Norte: aspectos epidemiológicos e clínicos

This study aimed to characterize in a clinical and epidemiological way the patients who are on a waiting list for transplantation and the patients transplanted with corneal tissue in a corneal transplants reference service in the state of Rio Grande do Norte. It is an epidemiological study of a quantitative approach, with cross-sectional, descriptive and analytical cut including all patients on the waiting list for transplantation (population A) and the patients already transplanted with a corneal tissue (population B) in a reference service. In population A, there was a census conducted of patients on the waiting list for corneal transplantation (n=62 patients). In population B, the sample was non-probabilistic and corresponded to all corneal transplants performed in the service in the period from 2010 to 2014 (n=258). This study is approved by the Research Ethics Committee of the Federal University of Rio Grande do Norte, in Opinion 876 177 and CAAE 37533014.8.0000.5537. Data were collected in full in the period from January to April 2015, by two instruments built to systematize the necessary data collection. After being coded and tabulated, data were analyzed using the Statistical Package for Social Sciences software, version 20.0. The definition of variables and their distribution patterns were presented as frequencies and measures of central tendency while, for multivariate analysis, the effect of magnitude measures were applied (prevalence ratio) and measures of association (chi-square test or Fisher's exact test) for a 0.05 significance level. The results are shown in two scientific articles coming from the field survey data. It was found that the epidemiological profile of patients on the waiting list (n=62) showed a prevalence of individuals aged over 50 years old, female (54.84%) and residents of the middle region of East Rio Grande do Norte (66.13%). The clinical profile of patients with corneal transplantation (n=258) was characterized by being male (51.16%) with an average age of 49.33 years old and 57.75% were coming from East Rio Grande do Norte. The average time on the waiting list was 172.63 days in elective transplants and 9.03 days in urgent transplants. Keratoconus was the main indicator condition to perform the transplant. For patients on the waiting list, the variable “type of disorder of the cornea” showed statistically significant association with gender and age. For patients with corneal transplants, the variable "type of disorder of the cornea" was associated with the variables gender, age, previous surgery, failure of previous graft, classification of the eye and glaucoma. By characterizing the clinical and epidemiological profile of corneal transplants, it is possible to question the reality, pointing about the care that should be offered and develop targeted interventions to collective and individual needs intrinsic to patients who need this surgery as a treatment option.

Global Japanization? The transnational transformation of the labour process

Global Japaniziation? Brings together research from North America, Japan, Europe and Latin America to analyse the influence of Japanese manufacturing investment and Japanese working practices across the global economy. The editors present original case studies of work reorganization and workers' experiences within both Japanese companies and those of their competitors in diverse sectors and national settings. These studies provide a wide-ranging critique of conventional accounts of Japanese models of management and production, and their implications for employees. They offer new evidence and fresh perspectives on the role of "transplants" in disseminating manufacturing innovations, and on the responses of non-Japanese firm in reorganizing production operations and industrial relations.

Facteurs influençant le succès de la restauration des populations de carex faux-lupulina

Ce mémoire visait à déterminer les pratiques et les facteurs favorables au succès des réintroductions du carex faux-lupulina (Carex lupuliformis), une espèce vivace menacée poussant dans divers types de milieux humides ouverts du nord-est de l’Amérique du Nord. Pour répondre à cet objectif, des expériences de germination en serres ont été réalisées, et la survie, la vigueur et le microhabitat de chaque plant de carex faux-lupulina connu au Québec (sauvage ou réintroduit) ont été mesurés. Nos résultats montrent que le carex faux-lupulina est une espèce facile à propager ex situ. Une stratification froide (4°C) des semences dans du sable humide pendant un mois induit des taux de germination élevés. Nos résultats montrent également que des pucerons exotiques, ainsi qu’une transplantation dans des habitats inadaptés aux besoins de l’espèce en termes d’humidité du sol et de lumière disponible, ont causé la mort prématurée d’un grand nombre de transplants. Notre programme de réintroduction a permis d’établir une nouvelle population de l’espèce au Québec, et a potentiellement permis de sextupler la taille des populations de la province. Globalement, cette étude a permis de mieux comprendre l’écologie du carex faux-lupulina, et ainsi de faciliter la restauration de ses populations.

Seawater carbonate chemistry and biological processes duirng experiments with bryozoan Myriapora truncata, 2010

There are serious concerns that ocean acidification will combine with the effects of global warming to cause major shifts in marine ecosystems, but there is a lack of field data on the combined ecological effects of these changes due to the difficulty of creating large-scale, long-term exposures to elevated CO2 and temperature. Here we report the first coastal transplant experiment designed to investigate the effects of naturally acidified seawater on the rates of net calcification and dissolution of the branched calcitic bryozoan Myriapora truncata (Pallas, 1766). Colonies were transplanted to normal (pH 8.1), high (mean pH 7.66, minimum value 7.33) and extremely high CO2 conditions (mean pH 7.43, minimum value 6.83) at gas vents off Ischia Island (Tyrrhenian Sea, Italy). The net calcification rates of live colonies and the dissolution rates of dead colonies were estimated by weighing after 45 days (May-June 2008) and after 128 days (July-October) to examine the hypothesis that high CO2 levels affect bryozoan growth and survival differently during moderate and warm water conditions. In the first observation period, seawater temperatures ranged from 19 to 24 °C; dead M. truncata colonies dissolved at high CO2 levels (pH 7.66), whereas live specimens maintained the same net calcification rate as those growing at normal pH. In extremely high CO2 conditions (mean pH 7.43), the live bryozoans calcified significantly less than those at normal pH. Therefore, established colonies of M. truncata seem well able to withstand the levels of ocean acidification predicted in the next 200 years, possibly because the soft tissues protect the skeleton from an external decrease in pH. However, during the second period of observation a prolonged period of high seawater temperatures (25-28 °C) halted calcification both in controls and at high CO2, and all transplants died when high temperatures were combined with extremely high CO2 levels. Clearly, attempts to predict the future response of organisms to ocean acidification need to consider the effects of concurrent changes such as the Mediterranean trend for increased summer temperatures in surface waters. Although M. truncata was resilient to short-term exposure to high levels of ocean acidification at normal temperatures, our field transplants showed that its ability to calcify at higher temperatures was compromised, adding it to the growing list of species now potentially threatened by global warming.

Exploring several nonconventional interactions of ice-binding proteins

Traditionally, ice-binding proteins (IBPs), also known as antifreeze proteins (AFPs), have been defined by two universal activities: ice recrystallization inhibition and thermal hysteresis. However, there remains the possibility IBPs have other complementary functions given the diversity found within this protein group. This thesis explores some of these in both natural and applied settings, in the hopes of furthering our understanding of this remarkable group of proteins. Plant IBPs could function as part of a defensive strategy against ice nucleators produced by certain pathogens. To assess this hypothesis, recombinant IBPs from perennial ryegrass and purple false brome were combined with the ice nucleation protein (INP) from the plant pathogen, Pseudomonas syringae. Strikingly, the plant proteins depressed the freezing point of the bacterial INP, while a fish AFP could not, nor did the INPs have any effect on IBP activity. Thus, the interaction between these two different proteins suggests a role in plant defensive strategies against pathogenic bacteria as another IBP function. In addition, the potential use of hyperactive insect IBPs in organ preservation was investigated. Current kidney preservation techniques involve storing the organ at 4 °C for a maximum of 24 h prior to transplantation. Extending this “safe” time would have profound effects on renal transplants, however, ischemic injury is prevalent when storage periods are prolonged. Experiments described here allowed subzero preservation for 72 h with the addition of a beetle IBP to CryoStasis® solution. Kidneys stored using the traditional technique for 24 h and the method developed here for 72 h showed similar levels of biomarker enzymes, underscoring the potential utility of insect IBPs for future transplant purposes. Finally, IBP function in the freeze-tolerant gall fly, Eurosta solidaginis, was examined. Larvae representing the mid-autumn stage displayed ice-binding activity, suggesting an IBP is being expressed, possibly as a protective measure against freezing damage when fall temperatures can unpredictably drop. IBP activity was also observed in the larvae’s host plant, Solidago spp. Mass spectrometry analysis of ice-affinity purified plant extracts provided three candidate pathogenesis-related proteins that could be responsible for the detected activity, further demonstrating additional functions of IBPs.

Découverte de nouveaux marqueurs pharmacogénomiques de la maladie du greffon contre l'hôte en transplantation de cellules souches hématopoïétiques

La transplantation de cellules souches hématopoïétiques (CSH) constitue une avenue thérapeutique potentiellement curative pour plusieurs cancers hématologiques comme la leucémie. L’utilisation d’une thérapie immunosuppressive pour prévenir la maladie du greffon contre l’hôte (GvHD) est un déterminant majeur du succès de la greffe. Malgré tout, cette complication survient chez 25 à 50% des transplantés et est une cause majeure de mortalité. L’optimisation du régime d’immunosuppression est un facteur facilement modifiable qui pourrait améliorer le pronostic des patients. Particulièrement, les polymorphismes du génome du donneur ou du receveur dans les voies pharmacogénomiques des immunosuppresseurs pourraient influencer l’exposition et l’action de ces médicaments, de même que le pronostic du patient. Le profilage de 20 pharmacogènes prioritaires chez des paires de donneurs-receveurs en greffe de CSH a permis d’identifier des variations génétiques liées au risque de la GvHD aiguë. Principalement, le statut génétique du receveur pour les protéines ABCC1 et ABCC2, impliquées dans le transport du méthotrexate (MTX), ainsi que des cibles moléculaires de ce médicament (ATIC et MTHFR) ont été associées au risque de GvHD aiguë. Similairement, le NFATc1, codant pour une cible moléculaire de la cyclosporine, augmentait lui aussi le risque de la maladie. Les porteurs de deux génotypes à risque et plus étaient particulièrement prédisposés à développer cette complication. Par surcroît, le statut génétique du donneur influençait également le pronostic du receveur après la greffe. Entre autres, des allèles protecteurs ont été identifiés dans les voies liées au transport (SLC19A1) et à l’action du MTX (DHFR). Inversement, NFATc2 a été associé à une augmentation du risque de GvHD aiguë. Afin de mieux comprendre les associations observées entre ces marqueurs génétiques et le risque de GvHD aiguë, une étude prospective innovante est en cours chez des greffés de CSH. Cette étude permettra d’étudier comment la génétique du patient ou du donneur peut influencer la pharmacocinétique et la pharmacodynamie des immunosuppresseurs, de même que leurs liens avec la GvHD aiguë. Ces paramètres sont quantifiés grâce à des approches analytiques que nous avons mises au point afin de répondre aux besoins spécifiques et uniques de cette étude. Les approches proposées dans cette thèse sont complémentaires aux méthodes classiques de suivi des immunosuppresseurs et pourraient aider à optimiser la pharmacothérapie du patient. Une meilleure identification des patients à haut risque de GvHD aiguë avant la greffe, basée sur des marqueurs pharmacogénomiques identitaires, pourrait guider le choix de la prophylaxie immunosuppressive, et ainsi améliorer l’issue clinique de la greffe.

Regenerative Cues of Myeloid Cells on Pancreatic Epithelium

Thesis (Ph.D.)--University of Washington, 2016-07

Rôle des nutriments dans l’enracinement et le rendement en fruits chez la chicouté (Rubus chamaemorus)

La chicouté (Rubus chamaemorus L.) pousse naturellement dans les tourbières ombrotrophes. La culture de la chicouté dans les tourbières en fin d’exploitation serait très intéressante afin de maintenir des activités économiques sur ces sites ainsi que d’améliorer la disponibilité de ce petit fruit pour une future commercialisation. L’implantation de cette culture fait toutefois face à certains problèmes tels la faible survie des boutures au cours de la première année et un rendement fruitier très variable. Des essais de fertilisation et d’application d’auxine ont été réalisés pour augmenter la production de racines sur les boutures de rhizome au moment de la plantation afin de réduire leur mortalité. La fertilisation a permis d’augmenter la longueur des racines, mais seulement à la fin de la saison. Les fertilisants ont également stimulé la croissance des plants. Par contre, les concentrations d’auxine utilisées ont entraîné une très forte mortalité des boutures de chicouté. Aucun de ces traitements n’a permis d’augmenter la survie des boutures lors de la plantation. Afin de mieux comprendre les limitations nutritives liées aux faibles rendements fruitiers, nous avons utilisé une analyse compositionnelle (CND) nous permettant d’identifier les débalancements nutritifs. Cette analyse a montré que les parcelles moins productives sont caractérisées par une concentration foliaire plus élevée en manganèse, fer, soufre et cuivre. Les résultats de ce projet de maîtrise vont permettre d’améliorer la régie de fertilisation lors de la plantation de la chicouté en tourbière résiduelle, mais d’autres recherches doivent être menées afin de réduire la mortalité des boutures lors de la plantation.

Unusual case of abdominal pain following liver transplant: causality or casualty?

This is an unusual case of chronic abdominal pain following two liver transplants with at least three potential causes: traumatic neuroma, intussusception of the small bowel of the Roux loop and biliary cast. Surgical removal of the latter two factors led to resolution of the pain. The management of the clinical case is discussed.