Humans evolved in a world of giant creatures. Current evidence suggests that most ice age megafauna went extinct around the transition to our current Holocene epoch. The ecological reverberations associated with the loss of over 65% of Earth’s largest terrestrial animals transformed ecosystems and human lifeways forever thereafter. However, there is still substantial debate as to the cause of this mass extinction. Evidence variously supports climate change and anthropogenic factors as primary drivers in the restructuring of the terrestrial biosphere. Much of the ongoing debate is driven by the insufficient resolution accessible via macro-remains. To help fill in the gaps in our understandings of the Pleistocene-Holocene transition, I utilized the growing power of sedimentary ancient DNA (sedaDNA) to reconstruct shifting signals of plants and animals in central Yukon. To date, sedaDNA has typically been analyzed by amplifying small, taxonomically informative regions. However, this approach is not ideally suited to the degraded characteristics of sedaDNA and ignores most of the potential data. Means of isolating sedaDNA have also suffered from the use of overly aggressive purification techniques resulting in substantial loss. To address these limitations, I first experimentally developed a novel means of releasing and isolating sedaDNA. Secondly, I developed a novel environmental bait-set designed to simultaneously capture DNA informative of macro-scale ecosystems. When combined, we identify a substantial improvement in the quantity and breadth of biomolecules recovered. These optimizations facilitated the unexpected discovery of horse and mammoth surviving thousands of years after their supposed extirpation. I followed up these results by extracting DNA from multiple permafrost cores where we confirm the late survival signal and identify a far more complex and high-resolution dataset beyond those identifiable by complementary methods. I was also able to reconstruct mitochondrial genomes from multiple megafauna simultaneously solely from sediment, demonstrating the information potential of sedaDNA. / Dissertation / Doctor of Philosophy (PhD) / A new addition to the rapidly growing field of palaeogenetics is environmental DNA (eDNA) with its immense wealth of biomolecules preserved over millennia outside of biological tissues. Organisms are constantly shedding cells, and while most of this DNA is metabolized or otherwise degraded, some small fraction is preserved through sedimentary mineral-binding. I experimentally developed new ancient eDNA methods for recovery, isolation, and analysis to maximize our access to these biomolecules and demonstrate that this novel approach outperforms alternative protocols. Thereafter, I used these methods to extract DNA from ice age permafrost samples dating between 30,000–6,000 years before present. These data demonstrate the power of ancient eDNA for reconstructing ecosystem change through time, as well as identifying evidence for the Holocene survival of caballine horse and woolly mammoth in continental North America. This late persistence of Pleistocene fauna has implications for understanding the human ecological and climatological factors involved in the Late Pleistocene mass extinction event. This effort is paralleled with megafaunal mitogenomic assembly and phylogenetics solely from sediment. This thesis demonstrates that environmental DNA can significantly augment macro-scale buried records in palaeoecology.

本研究は中国農村部における貧困問題、特に2000年代以降の南西部少数民族地域の貧困と経済発展、および政府の貧困削減政策について研究するものである。世界銀行や中国国家統計局などの公的機関が公開したデータ、および研究対象地域で収集された農家家計や地域経済発展に関するデータを用いて、農村貧困の実態、政府による「精準扶貧」政策の実施と効果、少数民族地域貧困地域の生活と貧困、および自力で貧困状況を改善した地域の事例について分析した。 / This study sheds a light on the poverty issues in rural China from the 2000s, with a specific focus on the poverty and economic development in ethnic areas in south-western China, and the poverty alleviation policies implemented by the government. Utilizing the data published by organizations such as World Bank and National Bureau of Statistics of China, and the data collected from the research area, the following research questions are discussed: the actual conditions of poverty in rural China; the implementation and the effect of the governmental poverty alleviation policy, named Targeted Poverty Relief Strategy (jingzhun fupin); the livelihood and poverty of ethnic minorities living in deprived area; and the case study of a region which has lifted themselves out of poverty. / 博士(現代アジア研究) / Doctor of Philosophy in Contemporary Asian Studies / 同志社大学 / Doshisha University

Диссертация посвящена вопросам социального маркетинга – Social Media Marketing (SMM). Рассматривается статистика продвижения товаров и услуг через работу в социальных сетях. Предметом анализа выступает инструментарий SMM, статистика активных пользователей и капитализация компаний. Результаты исследования могут быть применены в сфере организации SMM-продвижения бизнес-процессов и подготовки студентов в области онлайн-маркетинга. / The article is devoted to the issues of network marketing – Social Media Marketing (SMM). We Сonsider the statistics of the promotion of goods and services through the work in social networks. The subject of the analysis is the SMM toolkit and its system application. The results of the study can be applied in the organization of SMM-support business processes and prepare students in the field of online marketing.

Les lipides « switch » - bascules - appartiennent à la famille des matériaux sensibles à un stimulus. Quand ces lipides bascules sont incorporés aux nanoparticules lipidiques (LNP), ils permettent la délivrance contrôlée grâce à un changement de conformation activé par une baisse de pH. Des expériences précédentes avaient démontré que les LNP bascules ont transfecté le petits ARN interférents (siRNA) in vitro et in vivo, silençant la protéine fluorescente verte (GFP) et la protéine hépatique Facteur VII, respectivement. La double administration de micro ARN (miRNA) et d'agent anticancéreux melphalan a également été réalisée par les LNP bascule sur un modèle de rétinoblastome murin. Ces résultats prometteurs nous ont encouragé à élargir les applications de LNP bascules en tant que vecteur de siRNA. De plus, le mécanisme par lequel les LNP bascules induisent la déstabilisation de la membrane et la libération de matériaux encapsulé au milleu acide reste obscur. La compréhension de ce mécanisme est cruciale pour cerner les avantages et les limites des LNP bascules, pour proposer des futures applications et pour prévenir leur toxicité. Dans ce mémoire, nous avons comme objectif d’évaluer le potentiel des LNP bascules pour le traitement du cancer. Nous avons évalué les LNP bascules comme vecteur de livraison du siRNA ciblant l'une des protéines cancéreuses les plus spécifiques découvertes à ce jour, la survivine. En parallèle, nous avons étudié le comportement biophysique des membranes contenant des lipides bascules dans des vésicules de taille micromètrique. Dans la première étude, nous avons démontré que les LNP bascules ont permis le silençage de la survivine dans une gamme de lignées cellulaires cancéreuses (poumon, cervical, ovaire, sein, côlon, rétinoblastome). Dans les cellules du rétinoblastome humain (Y79), nous avons examiné plusieurs agents cytotoxiques utilisés en clinique quant à leur synergie avec le silençage de la survivine: melphalan, topotécan, téniposide et carboplatine. Le prétraitement avec les LNP chargées de siRNA-survivine a amélioré de manière synergique la cytotoxicité du carboplatine et du melphalan mais dans une moindre mesure celle du topotécan et du téniposide. Cet effet était spécifique aux cellules cancéreuses car les cellules saines (ARPE.19) n'exprimaient pas de survivine. L'inhibition de la survivine par silençage de siRNA s'est révélée plus spécifique et moins dommageable pour les cellules saines (ARPE.19) que le YM155, un inhibiteur moléculaire de la survivine. Dans la deuxième étude, nous avons observé par microscopie confocale que les lipides bascules induisaient rapidement le stress, la fission et une courbure positive dans les membranes des vésicules unilamellaires géantes lorsqu'elles étaient exposées à des conditions acides. La dynamique de la membrane a été confirmée par des expériences de diffusion dynamique de la lumière (DLS) et de fuite de calcéine. Ces phénomènes ont également été observés lorsque des lipides bascules ont été incorporés dans une membrane hybride polymère/lipide, fournissant des propriétés sensibles au pH aux vésicules hybrides. À notre connaissance, c'est la première fois qu'une vésicule hybride sensible au pH est reportée. Nos résultats corroborent l'applicabilité des LNP bascules en tant qu'agents de vectorisation des siRNA pour le traitement du cancer grâce au silençage de la survivine, en particulier comme adjuvant à la chimiothérapie. L'investigation biophysique a révélé que les lipides bascules agissent sur la fluidité de la membrane, en particulier à pH acide. Cette sélectivité en pH garantit leur biocompatibilité à pH neutre ainsi que la libération efficace et rapide de leur cargo à pH acide. La compatibilité avec les vésicules hybrides polymère/lipide ouvre de nouvelles applications au niveau de vésicules biomimétiques et l'administration de médicaments. / Cationic switchable lipids belong to the class of stimuli-responsive materials. When incorporated in lipid nanoparticles (LNP), switchable LNP promote pH-triggered delivery of payload based on a molecular switch mechanism. Previous studies have demonstrated that switchable LNP successfully delivered small interferring RNA (siRNA) in vitro and in vivo, promoting the silencing of a reporter Green Fluorescencen Protein (GFP) protein and liver-produced factor VII, respectively. Dual delivery of micro RNA (miRNA) and anticancer agent melphalan was also achieved through switchable LNP in a retinoblastoma rat model. These promising results encouraged us to enlarge the applications of switchable LNP as siRNA carrier. Moreover, the mechanism whereby switchable LNP mediate acid-triggered membrane destabilization and, thus, payload release remains elusive. Understanding this mechanism is crucial to draw the advantages and limitations of switchable LNP, and to tailor their future applications and prevent their potential toxicity. In this dissertation, we aimed to further understand the potential of switchable LNP for cancer treatment. We assessed switchable LNP as a siRNA delivery carrier by targeting one of the most specific cancer protein discovered to date, survivin. Meanwhile, we investigated the biophysical behavior of switchable-lipid containing membranes in micron-sized vesicles. In the first study, we demonstrated that switchable LNP efficiently silenced survivin in a range of cancer cell line models (lung, cervical, ovary, breast, colon, retinoblastoma). In retinoblastoma (RB) cells (Y79), several clinically used cytotoxic agents were screened for their synergy with survivin silencing: melphalan, topotecan, Teniposide, and carboplatin. Pretreatment with LNP loaded with siRNA targeted against survivin synergistically enhanced the cytotoxicity of carboplatin and melphalan but in lesser extent topotecan and teniposide. This effect was specific to cancer cells since healthy cells (ARPE.19) did not express survivin. Survivin inhibition through siRNA silencing revealed more specific and less damageable for healthy cells (ARPE.19) than a molecular approach, such as YM155. In the second study, we observed by confocal microscopy that switchable lipids rapidly induced stress, fission, and positive curvature in giant unilamellar vesicles’ membranes when submitted to acidic conditions. The membrane dynamics was confirmed by dynamic light scattering and calcein leakage experiments. Remarkably, these phenomena were also observed when switchable lipids were embedded into a hybrid polymer/lipid membrane, providing pH-sensitive properties to hybrid vesicles. To the best of our knowledge, this is the first time a pH-sensitive hybrid vesicle is reported. Our findings corroborate with the applicability of switchable LNP as siRNA delivery agents for cancer treatment through survivin silencing, especially as an adjuvant to chemotherapy. The biophysical investigation revealed that the switchable lipids act on the membrane fluidity, specifically at acidic pH. This pH selectivity guarantees their biocompatibility at neutral pH as well as its efficient and quick release of their cargo at acidic pH. Their compatibility with hybrid polymer/lipid vesicles opens new applications in biomimetic vesicles and drug delivery.

At low doses, relevant to nuclear incidents and accidental releases of radioactivity, the detriment of radiation extends beyond direct effects. This thesis investigates genomic instability, a subclass of non-targeted effects where damage and lethality is transmitted vertically and expressed in the progeny of cells many generations after initial radiation exposure. Through a series of experiments using clonogenic assay of human and fish cell culture, studies described in this thesis describe lethal mutations, hyper radiosensitivity and increased radioresistance – processes involving repair mechanisms that dictate survival in cells exposed to low doses. Further study investigates the difference in the relative biological effect of alpha particle radiation compared to what is expected at high doses. Results demonstrate increased radioresistance in a human cell line while also revealing increased lethality in a fish cell line confirming the need for consideration of dose-dependence as well as variance in behaviors of different cell lines and species. It is hoped the conclusions of this thesis will inspire the creation of protocols with greater attention to the indirect consequences of exposure to radiation at doses relevant to nuclear incidents and accidents. / Thesis / Master of Science (MSc)

Graft-versus-host disease (GvHD) of the skin is a severe allo-immune reaction and complication following allogeneic stem cell transplantation. Over the past years, intensive pre-clinical research has led to an improved understanding of the pathophysiology of acute and to a lesser extend chronic GvHD. This has translated into the approval of several new agents for the treatment of both forms of GvHD. This review summarizes the most recent advances in underlying pathomechanisms, clinical trials and newly approved agents for GvHD, with a special focus on skin involvement.

No / Beta amyloid plays a main role in the pathophysiology of Alzheimer's disease by inducing oxidative stress in the brain. Curcumin, a natural antioxidant, is known to inhibit beta amyloid and beta amyloid induced oxidative stress. However, low bioavailability and photodegradation are the major concerns for the use of curcumin. In the present study, we have formulated apolipoprotein E3 mediated poly(butyl) cyanoacrylate nanoparticles containing curcumin (ApoE3-C-PBCA) to provide photostability and enhanced cell uptake of curcumin by targeting. Prepared nanoparticles were characterized for particle size, zeta potential, entrapment efficiency and in vitro drug release. The entrapment of curcumin inside the nanoparticles was confirmed by X-ray diffraction analysis. Physicochemical characterization confirmed the suitability of the method of preparation. The photostability of curcumin was increased significantly in nanoparticles compared to plain curcumin. In vitro cell culture study showed enhanced therapeutic efficacy of ApoE3-C-PBCA against beta amyloid induced cytotoxicity in SH-SY5Y neuroblastoma cells compared to plain curcumin solution. Beta amyloid is known to induce apoptosis in neuronal cells, therefore antiapoptotic activity of curcumin was studied using flow cytometry assays. From all the experiments, it was found that the activity of curcumin was enhanced with ApoE3-C-PBCA compared to plain curcumin solution suggesting enhanced cell uptake and a sustained drug release effect. The synergistic effect of ApoE3 and curcumin was also studied, since ApoE3 also possesses both antioxidant and antiamyloidogenic activity. It was found that ApoE3 did indeed have activity against beta amyloid induced cytotoxicity along with curcumin. Hence, ApoE3-C-PBCA offers great advantage in the treatment of beta amyloid induced cytotoxicity in Alzheimer's disease.

Biological invasions provide a unique opportunity to study the mechanisms that regulate community composition and ecosystem function. Invasive species that are also ecosystem engineers can substantially alter physical features in an environment, and this can lead to cascading effects on the biological community. Aquatic-terrestrial interface ecosystems are excellent systems to study the interactions among invasive ecosystem engineers, physical features, and biological communities, because interactions among vegetation, sediment, and fluids within biophysical feedbacks create and modify distinct physical features. Further, these systems provide important ecosystem services including coastal protection afforded by their natural features. In this dissertation, I investigate the interactions and feedbacks among sand-binding beach grass species (a native, Elymus mollis (Trin.), and two non-natives, Ammophila arenaria (L.) Link and A. breviligulata Fernald), sediment supply, and dune shape along the U.S. Pacific Northwest coast. Dunes dominated by A. arenaria tend to be taller and narrower compared to the shorter, wider dunes dominated by A. breviligulata. These patterns suggest an ecological control on dune shape, and thus, coastal vulnerability to overtopping waves. I investigate the causes and consequences of these patterns with experiments, field observations, and modeling. Specifically, I investigate the relative roles of vegetation and sediment supply in shaping coastal dunes over inter-annual and multi-decadal time scales (Chapter 2), characterize a biophysical feedback between beach grass species growth habit and sediment supply (Chapter 3), uncover the mechanisms leading to beach grass coexistence and whether A. breviligulata can invade and dominate new sections of coastline (Chapter 4), and examine the non-target effects resulting from management actions that remove Ammophila for the recovery of the threatened Western Snowy plover (Charadrius alexandrinus nivosus) (Chapter 5). I found that vegetation and sediment supply play important roles in dune shape changes across inter-annual and multi-decadal time scales (Chapter 2). I determined that a biophysical feedback between the beach grass growth habits and sediment supply results in species-specific differences in sand capture ability, and thus, is a likely explanation for differences in dune shape (Chapter 3). I found that all three beach grass species can coexist across different sediment deposition rates, and that this coexistence is largely mediated by positive direct and indirect species interactions. I further determined that A. breviligulata is capable of invading and dominating the beach grass community in regions where it is currently absent (Chapter 4). Combined, these findings indicate that A. breviligulata is an inferior dune building species as compared to A. arenaria, and suggest that in combination with sediment supply gradients, these species differences ultimately lead to differences in dune shape. Potential further invasions of A. breviligulata into southern regions of the Pacific Northwest may diminish the coastal protection ability of dunes currently dominated by A. arenaria, but this effect could be moderated by the predicted near co-dominance of A. arenaria in these lower sediment supply conditions. Finally, I found that the techniques used to remove Ammophila for plover recovery have unintended consequences for the native and endemic dune plant communities, and disrupt the natural disturbance regime of shifting sand. A whole-ecosystem restoration focus would be an improvement over the target-species approach, as it would promote the return of the natural disturbance regime, which in turn, would help recover the native biological community. The findings from this dissertation research provide a robust knowledge base that can guide further investigations of biological and physical changes to the coastal dunes, can help improve the management of dune ecosystem services and the restoration of native communities, and can help anticipate the impacts of future beach grass invasions and climate change induced changes to the coast. / Graduation date: 2012 / Access restricted to the OSU Community at author's request from Sept. 22, 2011 - March 22, 2012

Background & Aims: Despite advances in surgical care and chemotherapeutic regimens, the five-year survival rate for Stage IV Hepatoblastoma (HB), the predominant pediatric liver tumor, remains at 27%. YAP1 and β-Catenin co-activation occurs in 80% of children’s HB; however, a lack of conditional genetic models precludes exploration of tumor maintenance and therapeutic targets. Thus, the clinical need for a targeted therapy remains unmet. Given the predominance of YAP1 and β-catenin activation in children’s tumors, I sought to evaluate YAP1 as a therapeutic target in HB. Approach & Results: Herein, I engineered the first conditional murine model of HB using hydrodynamic injection to deliver transposon plasmids encoding inducible YAP1S127A, constitutive β-CateninDelN90, and a luciferase reporter to murine liver. Tumor regression was evaluated using in vivo bioluminescent imaging, and tumor landscape characterized using RNA sequencing, ATAC sequencing and DNA foot-printing. Here I show that YAP1 withdrawal in mice mediates >90% tumor regression with survival for 230+ days. Mechanistically, YAP1 withdrawal promotes apoptosis in a subset of tumor cells and in remaining cells induces a cell fate switch driving therapeutic differentiation of HB tumors into Ki-67 negative “hbHep cells.” hbHep cells have hepatocyte-like morphology and partially restored mature hepatocyte gene expression. YAP1 withdrawal drives formation of hbHeps by modulating liver differentiation transcription factor (TF) occupancy. Indeed, tumor-derived hbHeps, consistent with their reprogrammed transcriptional landscape, regain partial hepatocyte function and can rescue liver damage in mice. Conclusions: YAP1 withdrawal, without modulation of oncogenic β-Catenin, significantly regresses hepatoblastoma, providing the first in vivo data to support YAP1 as a therapeutic target for HB. Modulating YAP1 expression alone is sufficient to drive long-term regression in hepatoblastoma because it promotes cell death in a subset of tumor cells and modulates transcription factor occupancy to reverse the fate of residual tumor cells to mimic functional hepatocytes.

The paper deals with the use of unmanned aircraft of the American national security policy. It examines the history of unmanned aviation, its military use, and cost- efficiency. It then examines the main obstacles and problems with their use in national security that this technology meets and will meet in the future after a higher level of autonomy is developed. These problems involve legal issues, international and domestic American law, the issue of civilian casualties, the role of the media, and public opinion. The final chapter focuses on the problems of technical, strategic and operational issues. In this section the research paper comes to the first conclusion which claims that the introduction of more autonomous systems to war will radically change its structure and, consequently, standard procedures and strategies. Case studies are included to illustrate how successful the drone strategy is applied in the five countries where the United States leads a war on terror. The research using the theory of the revolution in military affairs concludes that these UAVs pose the greatest challenge in history and it goes well beyond military matters. UAVs in national security affect the understanding of the basic principles of war in relation to the concepts of warrior ethos and just war.