京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第18504号 / 医博第3924号 / 新制||医||1005(附属図書館) / 31390 / 京都大学大学院医学研究科医学専攻 / (主査)教授 妻木 範行, 教授 開 祐司, 教授 中辻 憲夫 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

This thesis deals with filtering selected DDoS attacks on denial of the service. The the toretical part deals with the problems of general mechanisms used for DDoS attacks, defense mechanisms and mechanisms of detection and filtration. The practical part deals with the filtering of attacks using the iptables and IPS Suricata firewall on the Linux operating system in an experimental workplace using a network traffic generator to verify its functionality and performance, including the statistical processing of output data from filter tools using the Elasticsearch database.

The detection and intrusion prevention systems could be realized as independent hardware or set in the software form on to the host. The primary purpose of these protective elements is the undesirable activity detection such as integrity intrusion of the files, invalid attempts while connecting to the remote service or acquisition of the local network data. The systems react to the event on the basis of the action that is defined by internal rules. We can include the caution sending or communication blocking among possible counteractions. The base principals of the detection and intrusion prevention systems are described in the dissertation. Various types of captured data analyses and processes of the inhere rules creation and further more caution formats are mentioned in the dissertation. There are also considered the alternatives of their location including advantages of selected situations. There is described the installation and setting up of particular elements of the realized network and security systems. In order to the verification of functionality and factor of the protection providing there was realized several selected types of attacks.

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第21657号 / 医博第4463号 / 新制||医||1035(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 戸口田 淳也, 教授 松田 秀一, 教授 安達 泰治 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

In comparison to the southern pine beetle (SPB), (Dendroctonus frontalis Zimmermann), less is known about the three species of Ips bark beetles; the six-spined engraver, Ips calligraphus (Germar); the eastern five-spined engraver, Ips grandicollis (Eichhoff); and the small southern pine engraver, Ips avulses (Eichhoff). Ips commonly enter a stand following the first commercial thinning, feeding on slash and stressed individuals. However, the factors that influence Ips severity are poorly understood. Therefore, this project was designed to study the seasonal effects of first commercial thinning operations on Ips activity in north Mississippi loblolly pine stands. Treatments represented the most commonly used thinning practices in this area (i.e., a fifth row harvest with select, a third row harvest with select, and control). Treatments were duplicated three times per site and two sites were utilized to compare Ips movement to harvesting season.

京都大学 / 新制・課程博士 / 博士(医学) / 甲第23765号 / 医博第4811号 / 新制||医||1056(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 辻川 明孝, 教授 寺田 智祐, 教授 濵﨑 洋子 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

Bakgrund: Sveriges befolkning blir äldre och den allmänna pensionen sjunker. I en sådan situation blir ett privat pensionssparande allt viktigare. Trots detta har Regeringen valt att slopa avdragsrätten i produkten individuellt pensionssparande (IPS). Efterslopandet av avdragsrätten i IPS är det fortfarande kunder som inte har ändrat sitt beteende. En problematik uppstår kring hur dessa kunder ska hanteras och vilka marknadsmässiga mekanismer som styr deras beteende. Syfte: Inom ramen för marknadsföring är syftet att undersöka kundbeteendet när ett incitament försvinner och tidigare förutsättningar förändras. Genomförande: Studien utgår från en kvalitativ ansats med fokus på intervjuer. Sakkunniga inom området, rådgivare och privatpersoner bidrar till empirins kvalité. För att identifiera privatpersoner med en relation till IPS utfördes en förstudie. Utifrån den insamlade empirin utfördes en empiristyrd tematisk analysmetod för att urskilja likheter och skillnader, vilket genererat en djup diskussion och slutsats i ämnet. Slutsats: Okunskap samt informationsspridning är viktiga beståndsdelar för hur kunder beter sig vid sparande. Borttagandet av incitament leder till en negativ inställning tillvarumärket IPS som således kan orsaka svårigheter vid implementering av en ny produkt för privat pensionssparande. Det uppdagas ett utbudsunderskott och vidare krävs det att regering, banker och finansiella institut tar hänsyn till kringliggande faktorer, inklusive kundbeteendet, för att fylla detta tomrum. / Background: The population of Sweden is getting older and the national pension decreases. In that situation a private pension becomes important. Despite this, the government decided to abolish the deductibility of IPS. After the abolition of the deductibility of the IPS there still are customers who have not changed their savings behavior. A problem arises as to how these customers should be handled. Purpose: Within the boundaries of marketing the purpose is to explore the customer behavior when an incentive disappears and conditions held before, changes. Completion: The study is based on a qualitative approach, focusing on interviews. Experts in the area, counselors and individuals contribute to the quality of the empiric. To identify individuals with a relation to IPS, the authors first conducted a feasibility study. Based on the collected empirical data, an empirical driven thematic analysis is followed to discern and categorize different similarities and differences. Which, in the end resulted in a deep reflected discussion and conclusion regarding the subject. Conclusion: Regarding savings, ignorance and the dissemination of information are key elements in customer behavior. Eliminating the incentive also adds negativity to the brand IPS that could cause difficulties in the implementation of a potential new product. It has been revealed that there is a supply deficit and further, it requires that the government, banks and financial institutions needs to analyze these surrounding elements, and the customer behavior which occur in the study, into consideration in order to fulfill the customer demand.

La dystrophie Facio-Scapulo-Humérale, ou FSHD, est une maladie d'hérédité autosomique dominante, caractérisée par un affaiblissement des muscles de la face, et progressant de façon asymétrique au reste du corps. Des analyses de liaison ont montré que 95% des cas sont associés à la délétion de D4Z4, macrosatellite polymorphe de 3,3Kb, répété en tandem dans la région subtélomérique du bras long du chromosome 4, mais aucun gène candidat n'a clairement été identifié, et l'existence de patients sans contraction (FSHD2), suggère des mécanismes complexes dans la survenue de la maladie. Parmi ceux-ci, l'implication d'altérations épigénétique a été suggérée. En effet, des changements de marques épigénétiques chez les patients, tels que l'hypométhylation de l'ADN et la diminution de la triméthylation des résidus H3K9, indiquent que des modifications de l'organisation chromatinienne et des altérations épigénétiques interviendraient dans la FSHD. De plus la localisation particulière du locus 4q35 dans le noyau pourrait contribuer à la régulation de ce locus. La disposition et la localisation des gènes n'est pas aléatoire au sein du noyau, c'est un processus dynamique impliqué dans la régulation des gènes au cours de la prolifération cellulaire et la différenciation. L'objectif nos travaux a donc été de comprendre la conformation tridimensionnelle du locus 4q35 chez les malades et les individus non atteints, pour déterminer de potentielles interactions entre séquences, proches ou distantes, impliquées dans la survenue de la FSHD. / Facio-Scapulo-Humeral-Dystrophy (FSHD) is an autosomal dominant, neuromuscular disorder, characterized by facial muscular weakness with an asymmetric progression to the rest of the body. Linkage analyses have shown that 95% of cases are associated with D4Z4 deletion, a 3.3 kb tandemly repeated sequence located in the subtelomeric region of the long arm of chromosome 4, 4q35. However, no candidate gene has been clearly identified, and families with typical FSHD phenotype, but no linkage to 4q35 (FSHD2) suggest complex epigenetic mechanisms. The deletion of a certain number of this repetitive DNA sequence associated to changes in the epigenetic marks across the D4Z4 array such as DNA hypomethylation or decrease in H3K9 trimethylation also indicates that FSHD involves changes in chromatin organization and epigenetic alterations. Among these changes, the positioning of the 4q35 region within the nuclear space might contribute to the regulation of the disease-associated locus. Indeed, DNA sequences are not randomly distributed within the nuclear space and gene localization is a dynamic process that contributes to the control of gene expression during cell proliferation and differentiation. The aim of our study was to compare chromatin conformation of the 4q35 locus in patients and unaffected individuals, and to identify interactions between sequences, in close proximity or distant to the D4Z4 array in FSHD pathogenesis. Therefore, we have developed an immuno-FISH technique in 3D, in different cellular models, in order to study the tridimensional distribution of DNA sequences within the nuclear volume.

A geração de células-tronco pluripotentes induzidas (iPS) a partir de células somáticas demonstrou que células adultas de mamíferos podem ser reprogramadas a um estágio de pluripotência através da inserção de fatores de transcrição embrionários. Esta descoberta tem levantado questões fundamentais sobre os mecanismos, que através destes fatores de transcrição, influenciam epigeneticamente as células e seus potenciais de diferenciação após a reprogramação e um normal desenvolvimento. Componentes lipídicos e lipoprotéicos afetam vários aspectos no comportamento celular durante sua manutenção e diferenciação, podendo afetar diretamente fatores essenciais em processos de reprogramação celular, manutenção da pluripotência e perfil epigenético das células. Nesse sentido, esta tese propôs o estudo da composição lipídica com diferentes abordagens entre células iPS, células-tronco embrionárias (H1) e células fibroblastos (BJ). Foram produzidas três linhagens de células pluripotentes induzidas no modelo humano que foram caracterizadas quanto 1a sua pluripotência e utilizadas, juntamente às linhagens H1 e BJ como modelos para o estudo da composição lipídica proposto. Foram identificadas e estudadas um total de 44 espécies lipídicas das classes PC, PE, PI, SM e PS, e discutidas frente a reprogramação celular e manutenção da pluripotência. Foi identificado um padrão de composição fosfolipídica distinta entre células pluripotentes e não pluripotentes, e especulamos que a presença dessas espécies parecem ter um envolvimento fundamental para a manutenção da pluripotência. Este padrão, mostrou pela análise de componente principal, que durante o processo de reprogramação, alterações na composição lipídica ocorrem de forma com que a pluripotência surge durante a reprogramação, evidenciando alterações lipídicas particulares do estádio da pluripotência, sugerindo uma ligação entre estas alterações na composição lipídica com as alterações metabólicas da própria reprogramação celular. O estudo da quantificação de fosfolipídios entre linhagens celulares pluripotentes e não pluripotentes evidenciaram que existe uma diferença fosfolipídica entre estas linhagens, observamos que as linhagens iPS e H1, do ponto de vista das classes observadas e os fosfolipídios quantificados, são similares entre si e diferentes de células não pluripotentes. É evidente que estas moléculas lipídicas, individualmente, não são capazes de modular processos como a reprogramação celular, entretanto, é de extrema importância o entendimento das mesmas dentro da reprogramação celular e manutenção da pluripotência. Nossos dados sugerem que a composição lipídica de células pluripotentes tem importante papel para o desenvolvimento e evolução do processo de reprogramação celular e o entendimento da manutenção da pluripotência / The generation of induced pluripotent stem cells (iPS) from adult somatic cells has shown that mammalian cells can be reprogrammed to a pluripotent state by the insertion of embryonic transcription factors. This finding has raised questions about the fundamental mechanisms through which these transcription factors epigenetically influence cells, their potential of differentiation after reprogramming and normal development. Lipid and lipoprotein components affect numerous aspects of cell behavior during its maintenance and differentiation, which can directly affect main factors in cell reprogramming processes, maintenance of pluripotency and epigenetic profile of the cells. Thus, this thesis proposed to study, with different approaches, the lipid composition of iPS cells, embryonic stem cells (H1) and fibroblast cells (BJ). Three induced pluripotent cell lines were produced in the human model. They were characterized regarding their pluripotency and used along with H1 and BJ cell lines, as models for the proposed lipid composition study. A total of 44 species of lipid from the classes PC, PE, PI, PS and SM have been identified, studied and discussed regarding cellular reprogramming and maintenance of pluripotency. A different phospholipid composition pattern was observed between pluripotent and non-pluripotent cells, and it is speculated that the presence of these species appears to have a major involvement on the maintenance of pluripotency. This array showed, by the principal component analysis, that during the reprogramming process changes in the lipid composition occur, so that pluripotency takes place during reprogramming, highlighting lipid changes particular of the pluripotency state, suggesting a connection between these changes in lipid composition and the metabolic changes of cell reprogramming. The study of the quantitation of phospholipids from pluripotent and non-pluripotent cell lines indicated a phospholipid difference between these cell lines when considering the observed classes and quantified phospholipid. It was eminent that iPS lines and H1 are similar and differ from non-pluripotent cells. It is clear that these lipid molecules are not individually capable of modulating processes such as cell reprogramming, however, it is extremely important to understand them within cellular reprogramming and maintenance of pluripotency. Our data suggests that the lipid composition of pluripotent cells has important role in the development and evolution of cellular reprogramming process and the understanding the maintenance of pluripotency

A inserção de fatores de transcrição conhecidos em células somáticas é capaz de reprograma-las levando a um estágio de pluripotência, gerando células-tronco pluripotentes induzidas (iPS). A utilização destas células na medicina regenerativa tem grande potencial tanto na medicina humana quanto na veterinária, e a influência da origem da célula somática utilizada na geração de iPS é discutida atualmente. Mamíferos domésticos, como por exemplo o equino, são bons modelos para o estudo para medicina humana e veterinária, com destaque para terapia celular utilizando células mesenquimais em lesões ortopédicas e articulares. Tendo como hipótese que células equinas com diferentes origens apresentam potenciais de indução à pluripotência e capacidade de diferenciação in vitro variáveis; esta proposta tem como objetivo gerar células iPS equinas obtidas através da transdução viral dos fatores de transcrição OSKM humanos ou murinos em fibroblastos (eFibro) adultos e células mesenquimais derivadas de tecido adiposo (eAdMSC), medula óssea (eMO) e tecido de cordão umbilical (etCU). Foram isoladas e cultivadas 4 linhagens de células do tecido de cordão umbilical, 3 linhagens de fibroblastos equinos, 3 linhagens de células mesenquimais de tecido adiposo equinas e 2 linhagens de células de medula óssea equina. Essas células tiveram seu tempo de duplicação celular calculado em horas (eMO 61,3±15; etCU 53,8±5; eFIBRO 27±2; eAdMSC 18,2±4,5) e foram caracterizadas por diferenciação condrogênica, osteogênica e audiogênica e por imunofenotipagem. A partir destas células foram produzidas 85 linhagens iPS equinas (eiPS- 30 linhagens derivadas de fibroblasto, 33 linhagens derivadas de células de tecido adiposo, 21 linhagens derivadas de células de tecido de cordão umbilical com os fatores humanos e 1 linhagem de derivada de células de tecido adiposo com os fatores murinos). Testes de detecção de fosfatase alcalina e OCT4 foram realizados para células eiPS obtidas de cada linhagem e as células eiPS derivadas de tecido adiposo e fibroblastos foram positivas para detecção de NANOG, TRA1-60, TRA1-81 e as células eiPS derivadas de eAdMSC foram positivas para detecção de SSEA-1. As células eiPS derivadas de cada tipo celular geraram corpos embrióides, que posteriormente foram dissociados para teste de diferenciação espontânea in vitro. Os resultados mostram que células equinas podem ser mais facilmente reprogramadas com fatores humanos quando comparadas com os fatores murinos (P<0.05). Enquanto os fatores murinos produziram apenas uma linhagem de células iPS derivada de eAdMSC, os fatores de reprogramação humanos foram capazes de produzir variadas linhagens de células iPS, sendo a formação de colônias mais eficiente para células derivadas de eAdMSC (322, P<0.01) do que para células derivadas de eFibro (65) e etCU (58), que não diferiram (P=0.95), e não foi possível produzir células eiPS a partir de eMO. Usando o sistema de indução à pluripotência padronizado em nosso laboratório, a utilização de fatores humanos na reprogramação direta gera uma maior eficiência na produção de células iPS equinas quando comparados com fatores murinos. No nosso conhecimento, este é o primeiro relato de células eiPS produzidas unicamente dependentes de bFGF, sem necessidade de adição de LIF no meio de cultivo. / The insertion of known transcription factors into somatic cells is capable of reprogramming them into a pluripotency stage, generating induced pluripotent stem cells (iPS). The influence of the origin of the somatic cell used in the generation of iPS is currently discussed, and the use of these cells in regenerative medicine has great potential in both human and veterinary medicine. Domestic mammals, such as the equine are great models for the study of human and veterinary medicine, highlighting cell therapy using mesenchymal cells in orthopedic and articular injuries. Hypothesizing that equine cells derived from different tissues show variable pluripotency induction potentials and in vitro differentiation capacity, depending on the source of derivation; this proposal aims to generate equine iPS cells obtained by viral transduction of human or murine OSKM transcription factors in adult fibroblasts (eFibro) and mesenchymal cells derived from adipose tissue (eAdMSC), bone marrow(eMO), umbilical cord tissue (etCU). Four umbilical cord tissue cell lines, three equine fibroblast cells lines, three equine adipose tissue mesenchymal cell lines and 2 equine bone marrow cell lines were isolated and cultured. These cells had their doubling time calculated in hours (eMO 61.3 ± 15, etCU 53.8 ± 5, eFIBRO 27 ± 2, and ADMSC 18.2 ± 4.5) and were characterized by chondrogenic, osteogenic and adipogenic and by immunophenotyping. From these cells, 85 equine iPS (eiPS) cell lines were produced (30 fibroblast-derived cell lines, 33 cell lines derived from adipose tissue cells, 21 cell lines derived from umbilical cord tissue cells, with human factors and 1 cell line derived from eAdMSC using murine factors). Alkaline phosphatase and OCT4 detection tests were performed on eiPS cells obtained from each cell line and eAdMSC and eFibro derived iPS cells were positive for the detection of NANOG,TRA1-60, TRA1-81 and eiPS derived from eAdMSC were positive for SSEA-1 detection. Embryonic bodies, which were later dissociated for spontaneous differentiation test in vitro were derived from each cell type. Results show that equine cells can be more easily reprogrammed with human factors when compared to murine factors (P<0.05). While murine factors produced only one eiPS cell line derived from eAdMSC, human reprogramming factors were able to produce multiple eiPS cell lines, and colony formation was more efficient for cells derived from eAdMSC (322 P <0.01) than for cells derived from eFibro (65) and etCU (58), which did not differ (P = 0.95) and It has not yet been possible to produce iPS cells from the eMO cell lines. Using the induction system standardized in our laboratory, the use of human factors in direct reprogramming results in greater efficiency in the production of equine iPS cells when compared to murine factors. To our knowledge, this is the first report of eiPS cells produced solely dependent on bFGF, without the need for addition of LIF in the culture medium.