BACKGROUND: The needle exchange programme (NEP) started in Sweden in 1986 in Lund and shortly after in Malmo. The first NEP in Stockholm opened in spring 2013. The NEP is a service aimed at intravenous drug users (IDU) from 18 years old, with a goal of preventing the blood borne diseases, such as HIV, Hepatitis B (HBV), and Hepatitis C (HCV). With the on going HIV and Hepatitis epidemics, numerous countries around the world have adopted control strategies, such as the NEP to halt the spread of HIV, HBV, and HCV. The objective of this study was to examine if the needle exchange programme has decreased the incidence of HIV, HBV, and HCV in Sweden over a six-year period.  METHODS: Data for incidence and prevalence was extracted from the yearly reports of the Stockholm’s needle exchange programme from 2013 to 2018 and the yearly reports of the public health agency in Sweden from 2013 to 2018. The data was collected for Stockholm, and compared to Västra Götaland, and the whole of Sweden. RESULTS: The incidence of HIV was zero in 2013 and 2015 in the NEP. The incidence of HBV decreased to zero in 2013 in the NEP. There is an increased incidence of HCV in the NEP. CONCLUSION: The NEP has a protective effect through its combination of needle exchange, opiate substitute therapy, counselling, and vaccinations in reducing and stabilising incidences of the infections, in some instances to zero, as well as providing surveillance and treating infections.

Although the level of HBsAg is determined only qualitatively in routine clinical practice, recent data suggest that its quantification can assist or replace the viral load of HBV DNA in monitoring of HBV replication, which would be an easier and more economical alternative. Thus, the aim of this study was to correlate the levels of HBsAg with viral load of HBV DNA and other laboratory (HBeAg, ALT and AST) and histological (activity and fibrosis) findings in patients with chronic hepatitis B. A prospective, observational, cross-sectional study was performed on 128 patients with chronic hepatitis B, aged over 18 years, from the Hepatology Service of the University Hospital of Sergipe. Categorical variables were presented as frequencies and percentages with range of 95% where applicable. For the correlation analysis we used the Spearman test. It was considered that the correlations had statistical significance when ρ≤ 0.05. The overall correlation between HBV viral load and quantitative HBsAg was weak (ρ= 0.197, ρ= 0.026), and this same correlation was also weak and not statistically significant in HBeAg-positive patients (ρ= 0.233, ρ= 0.263). However, a strong correlation between HBsAg and HBV DNA >20,000 in HBeAgpositive patients was found. A regular correlation between HBsAg and HBV DNA in patients who were not on treatment (*ρ= 0394; ρ<0.001) was found and there was no significant correlation in patients receiving treatment (*ρ= -0.061; ρ= 0.673). No statistically significant association was observed between the levels of HBsAg and HBeAg, even when considering only the positive HBeAg (ρ: 0.121; ρ=0.565) and even not only the negative HBeAg (ρ =-0.067; ρ=0.501). The correlation between HBV DNA and HBeAg positive was fair (ρ =0.444; ρ=0.026). There was no statistical correlation between the levels of HBsAg and aminotransferases (ALT and AST). The distribution of HBsAg values did not differ between the degree of activity (ρ= 0.17) and fibrosis (ρ= 0.20). Conclusion: Our results show that, in general, the correlation between levels of HBsAg and HBV DNA exists, but it proved to be weak. Also, they suggest that HBsAg better reflects HBV DNA in the initial replicative phase of chronic hepatitis B, when patients present HBV reagent and high titers of HBeAg viral load. They also show that there is no association of HBsAg with aminotransferases or with the degree of activity and liver fibrosis. / Embora o nível de HBsAg seja determinado apenas qualitativamente na prática clínica rotineira, dados recentes sugerem que a sua quantificação pode auxiliar ou substituir a carga viral do VHB DNA no monitoramento da replicação do VHB, o que seria uma alternativa mais fácil e econômica. Desta forma, objetivou-se com este estudo, correlacionar os níveis de HBsAg com a carga viral do VHB DNA e com outros achados laboratoriais (HBeAg, ALT e AST) e histológicos (atividade e fibrose) em pacientes com hepatite B Crônica. Foi realizado um estudo observacional, prospectivo, transversal, com 128 pacientes portadores de hepatite B crônica, com idade igual ou superior a 18 anos, oriundos do Serviço de Hepatologia do Hospital Universitário de Sergipe. As variáveis categóricas foram apresentadas através de frequências e percentuais com intervalo de 95% quando pertinente. Para a análise das correlações utilizou-se o teste de Spearman. Considerou-se que as correlações possuíam significância estatística quando p ≤ 0,05. A correlação global entre a carga viral do VHB e o HBsAg quantitativo foi fraca (ρ=0,197; p=0,026); esta mesma correlação também foi fraca e sem significância estatística nos pacientes HBeAg positivos (ρ =0.233; p=0,263). Porém, foi encontrada uma forte correlação entre o HBsAg e o VHB DNA > 20.000, nos pacientes HBeAg positivos. Foi observada uma correlação regular entre o HBsAg e o VHB DNA nos pacientes que não estavam em tratamento (*ρ= 0394; p <0,001) e não existiu correlação significante nos pacientes em tratamento (*ρ= -0,061; p= 0,673). Não se observou associação estatisticamente significante entre os níveis de HBsAg e HBeAg, nem quando se considerou apenas os HBeAg positivos (ρ: 0,121; p=0,565) e nem apenas os HBeAg negativos (ρ =-0,067; p=0,501). A correlação entre o VHB DNA e o HBeAg positivo foi regular (ρ =0,444; p=0,026). Não foi verificada correlação estatística entre os níveis de HBsAg e as aminotransferases (ALT e AST). A distribuição dos valores de HBsAg não diferiu entre os graus de atividade (p=0,17) e fibrose (p=0,20). Conclusão: Os nossos resultados mostram que, de uma forma geral, a correlação entre os níveis de HBsAg e VHB DNA existe, mas é fraca. E sugerem que o HBsAg reflete melhor o VHB DNA na fase replicativa inicial da hepatite B crônica, quando os pacientes possuem HBeAg reagente e altos títulos de carga viral do VHB. Demonstram também que não existe associação do HBsAg com aminotransferases nem com o grau de atividade e fibrose hepática.

京都大学 / 新制・課程博士 / 博士(生命科学) / 甲第23341号 / 生博第459号 / 新制||生||61(附属図書館) / 京都大学大学院生命科学研究科高次生命科学専攻 / (主査)教授 朝長 啓造, 教授 野田 岳志, 教授 千坂 修 / 学位規則第4条第1項該当 / Doctor of Philosophy in Life Sciences / Kyoto University / DFAM

MSc (Microbiology) / Department of Microbiology / See the attached abstract below

Obwohl der Wildtyp des Hepatitis-B-Virus (HBV) nicht zytopathogen und die Pathogenese der Hepatitis B generell immunvermittelt ist, können in immunsupprimierten Nierentransplantatempfängern mit chronischer Hepatitis B schwere Leberschäden bis hin zu Leberzirrhose und Leberversagen entstehen. Die Entwicklung von Leberzirrhose in den Nierentransplantierten ist assoziiert mit der Akkumulation und Persistenz von komplexen HBV-Varianten mit Mutationen im Core-Promotor / X-Gen, Deletionen im Core (C)-Gen und teilweise zusätzlichen Deletionen im präS-Bereich. Dies lässt eine Rolle der Varianten in der speziellen Pathogenese bei Immunsupprimierten vermuten. In der vorliegenden Arbeit wurden funktionelle Analysen der komplexen Varianten im Vergleich zu Referenz-Wildtypgenomen und Wildtyp-ähnlichen Genomen der Patienten aus der frühen Infektionsphase durchgeführt, um Hinweise auf den potentiellen Beitrag der Varianten zur Pathogenese zu erlangen. Die Analysen erfolgten durch transiente Transfektion der humanen Hepatomazelllinie HuH7 mit repräsentativen HBV-Gesamtgenomen, die aus 2 Patienten während des Krankheitsverlaufs von einer asymptomatischen Infektion hin zur Leberzirrhose isoliert und kloniert worden waren. Trotz einiger Unterschiede im Detail wiesen die komplexen Varianten einen gemeinsamen, drastisch vom Wildtyp abweichenden Phänotyp auf. Dieser war gekennzeichnet durch eine veränderte Transkription mit reduzierten präC- und Oberflächen-mRNAs und verstärkter Expression der prägenomischen RNA, eine starke Reduktion des häufigsten Spleißprodukts der prägenomischen RNA, SP1, eine extrem reduzierte oder fehlende Expression und/oder Sekretion aller Oberflächenproteine und des HBeAg, ein verändertes intrazelluläres Verteilungsmuster des schwach exprimierten Core-Proteins und teilweise der Oberflächenproteine sowie eine erhöhte Replikation und Anreicherung gegenüber Wildtyp-HBV aufgrund einer verstärkten reversen Transkription der prägenomischen RNA. Dieser Phänotyp basierte zum Teil auf den Mutationen in Core-Promotor und C-Gen, wurde jedoch deutlich durch zusätzliche Mutationen in den übrigen Genomabschnitten beeinflusst. Die vielfältigen Veränderungen der Varianten unterstützen ihren vermuteten Beitrag zur Pathogenese. / Although wild-type hepatitis B virus is not cytopathogenic and the pathogenesis of hepatitis B is generally immune mediated, also immuno-suppressed patients, such as renal transplant recipients, with chronic hepatitis B may develop liver cirrhosis and end-stage liver disease. In renal transplant recipients, the development of liver cirrhosis is associated with the accumulation and persistence of complex HBV variants with mutations in core promoter / X gene, deletions in core (C) gene and sometimes additional deletions in the preS region. This suggests a role of these variants in the special pathogenesis in immuno-suppressed patients. In the present work, the complex variants were functionally analyzed in comparison to reference wild-type genomes and wild-type-like HBV genomes from the early asymptomatic phase of infection. For the analyses, representative cloned full-length HBV genomes isolated from 2 patients before and during liver cirrhosis were transiently transfected into the human hepatoma cell line HuH7. In spite of some variations, the complex variants showed a common phenotype, which was drastically altered compared to wild-type. It was characterized by reduced preC and surface mRNAs and increased expression of pregenomic RNA, by a strong reduction of the major spliced pregenomic RNA, SP1, by a partial or complete defect in expression and/or secretion of surface proteins and HBeAg, by an aberrant intracellular localization of the weakly expressed core protein and in some cases of the surface proteins, and by an enhanced replication and enrichment over wild-type HBV due to an enhanced reverse transcription of variant pregenomic RNA. The phenotypic alterations were often based on the mutations in core promoter and C gene but were considerably influenced by the additional mutations in other genomic regions. The multiple functional changes of the variants support their assumed contribution to pathogenesis.

Malgré l’existence d’un vaccin préventif efficace, l’infection chronique par le virus de l’hépatite B (HBV) demeure un problème majeur de santé publique. La persistance de l’infection par HBV étant clairement associée à des réponses immunitaires insuffisantes, l’immunothérapie par le vaccin à base d’ADN nu, visant à stimuler les réponses humorales et cellulaires, apparaît comme particulièrement pertinente pour la thérapie des hépatites B chroniques. Toutefois, l’efficacité thérapeutique d’une telle stratégie reste limitée chez l’homme, d’où la nécessité d’optimiser cette approche vaccinale pour une utilisation ultérieure en clinique. Ainsi, l’objectif général de ce travail de thèse était d’explorer, avec le modèle du DHBV (« Duck Hepatitis B Virus »), étroitement apparenté au HBV humain, si l’administration du vaccin à ADN par électroporation (EP) pouvait davantage améliorer son efficacité prophylactique et thérapeutique. Nous avons montré, dans un 1er temps chez des canards naïfs, que l’administration du vaccin à ADN par EP permet de potentialiser le pouvoir neutralisant et d’élargir le répertoire épitopique de la réponse humorale dirigée contre la protéine d’enveloppe du DHBV, même avec des doses d’ADN relativement faibles. Dans un 2ème temps, nous avons montré chez des animaux chroniquement infectés par le DHBV, que l’administration par EP du vaccin à ADN ciblant les protéines structurales du DHBV et le DuIFN-γ améliore considérablement l’efficacité thérapeutique du vaccin, notamment au regard de la séroconversion et de la clairance virale. Les résultats ainsi obtenus confirment l’intérêt majeur de cette approche vaccinale pour la thérapie des hépatites B chroniques / Despite the existence of an effective prophylactic vaccine, chronic hepatitis B virus (HBV) infection remains a major public health problem. Since persistence of HBV infection is mostly associated with insufficient immune responses, therefore DNA vaccination capable of activating both humoral and cellular immune responses appears as a pertinent strategy for chronic hepatitis B therapy. However, the efficacy of such therapeutic approach remains limited in humans. Improvement of DNA vaccine efficacy is therefore needed for future therapeutic applications in clinic. The main objective of this thesis was to investigate in the duck hepatitis B virus (DHBV) model, whether the protective and therapeutic efficacy of DNA vaccine can be enhanced using EP-based delivery system. Firstly, we showed in naïve ducks that EP-based delivery was able to improve the dose efficiency of DNA vaccine and to maintain a highly neutralizing, multi-specific B-cell response even with relatively low DNA doses, suggesting that it may be an effective approach for chronic hepatitis B therapy at clinically feasible DNA dose. Secondly, we showed in chronic DHBV-carriers that in vivo EP is able to dramatically enhance the therapeutic potency of DNA vaccine targeting hepadnaviral proteins. Indeed, this approach was able to consistently restore humoral immune response and to sustainably decrease and even clear viral infection. Thus, these data strongly support the use of this approach for chronic hepatitis B therapy in humans

No description available.

This thesis explores key aspects for synthesizing data across spatiotemporal scales relevant for water resources management in an Eastern Africa context. Specifically, the potential of large scale global precipitation datasets (GPDs) in data limited regions to overcome spatial and temporal data gaps is considered. The thesis also explores the potential to utilize limited and non-continuous streamflow and stream water chemistry observations to increase hydrological process understanding. The information gained is then used to build a coherent hydro-climatic framework for streamflow modelling. In this thesis, Kilombero Valley Drainage Basin (KVDB) in Tanzania is used as an example of a data limited region targeted for rapid development, intensification and expansion of agriculture. As such, it is representative for many regions across the Eastern Africa. With regards to the data synthesis, two satellite products, three reanalysis products and three interpolated products were evaluated based on their spatial and temporal precipitation patterns. Streamflow data from KVDB and eight subcatchments were then assessed for quality with regards to missing data. Furthermore, recession analysis was used to estimate catchment-scale characteristic drainage timescale. Results from these streamflow analyses, in conjunction with a hydrological tracer-based analysis, were then used for improved understanding of streamflow generation in the region. Finally, a coherent modelling framework using the HBV rainfall-runoff model was implemented and evaluated based on daily streamflow simulation. Despite the challenges of data limited regions and the often large uncertainty in results, this thesis demonstrates that improved process understanding could be obtained from limited streamflow records and a focused hydrochemical sampling when experimental design natural variability were leveraged to gain a large  signal to noise ratio. Combining results across all investigations rendered information useful for the conceptualization and implementation of the hydro-climatic modelling framework relevant in Kilombero Valley. For example, when synthesized into a coherent framework the GPDs could be downscaled and used for daily streamflow simulations at the catchment scale with moderate success. This is promising when considering the need for estimating impacts of potential future land use and climate change as well as agricultural intensification. / Denna avhandling utforskar aspekter på att syntetisera data med olika rumslig och temporal upplösning, vilket är centralt för vattenförvaltning i östra Afrika. Särskilt fokus ligger på att undersöka möjligheten till att använda globala nederbördsdataset för att fylla rumsliga och temporala luckor där data saknas. Avhandlingen undersökeräven möjligheten till att använda flödesdata med icke-kompletta tidsserier samt kemidata från vattendrag för att utöka kunskap-en om hydrologiska processer. Informationen används för att bygga upp ett integrerande ram-verk för hydro-klimatologisk modellering som exempelvis kan användas för att utforska ef-fekten av ett utökat och intensifierat jordburk på vattenresurser. I denna avhandling användes Kilomberodalens avrinningsområde (Tanzania) som exempel på ett databegränsat område där det pågår en intensiv utökning av jordbruksverksamhet. Detta område kan ses som representa-tivt för ett stort antal områden inom östra Afrika.Datasyntesen innefattade två nederbördsprodukter baserade på satellitdata, tre baserade på återanalysprodukter samt två baserade på interpolering av observervationsdata från regnmä-tare. Dessa åtta produkter utvärderades baserat på deras nederbördsmönster i rum och tid. Ut-över detta utvärderades vattenföringsdata från Kilomberodalens avrinningsområde samt åtta delavrinningsområden utifrån mängden saknad data i respektive tidsserie. Vidare användes resultaten från hydrologisk recessionsanalysför att uppskatta den karaktäristiska avrinningsti-den för avrinningsområden. Resultaten från recessionsanalysensamthydrologiskt spårämnes-försök användessedan för att utöka kunskapen om avrinningsbildning och vattenföring i om-rådet samt som stöd i valet av hydrologiskt modelleringsverktyg. Avslutningsvis användes HBV-avrinningsmodellen för att simulera daglig vattenföring. Trots utmaningen i att arbeta iett databegränsat område och de osäkerheter i resultat som detta tenderar att leda till visar resultaten att det var möjligt att använda begränsad vattenfö-ringsdata och vattenkemidata för att utöka den hydrologiska processförståelsen av området. Detta möjliggjordes genom ett experimentellt upplägg som utnyttjade till ett stort signal-till-brusförhållande under rådande förhållanden av naturlig variabilitet. Kombinerade resultat från alla genomförda studier kunde utnyttjas vid konceptualiseringen och implementeringen av ramverket för hydroklimatologisk modellering av Kilomberodalens avrinningsområde. Till exempel kunde de globala nederbördsdataseten användas för lokal modellering av flödesdata med viss framgång efter syntes och implementering i det integrerande ramverket för hydro-klimatologisk modellering. Detta är lovande med tanke på behovet av att undersöka vilken påverkan möjliga framtida förändringar i markanvändning, klimat samt jordbruk har på den lokala och regionala miljön. / <p>At the time of the doctoral defense, the following papers were unpublished and had a status as follows: Paper 3: Manuscript. Paper 4: Manuscript.</p>

Hepatitis B virus (HBV) infection is one of the major causes of chronic and cancerous liver disease. Elimination of HBV from chronically infected patients by recombinant interferon α (IFNα) monotherapy shows that the mechanisms of the innate immunity play an important role in suppressing viral infection. However, the mechanisms of recognition of the HBV genome and its escape from the mechanisms of natural immunity are still little known. One of the principal factors enabling the virus to escape from cellular restriction mechanisms is the HBx viral protein. HBx is a 154 amino acid pleiotropic multifunctional protein affecting transcription, signal transduction, cell cycle, protein degradation, apoptosis, and chromosomal stability in the host cell. Previous results from our laboratory have shown that activation of the MEK1/2-ERK signaling pathway in plasmacytoid dendritic cells leads to inhibition of IFNα production. The aim of my work was to determine whether HBx activates the MEK1/2-ERK pathway and thus inhibits IFN type I production also in hepatocytes. For this purpose, I monitored HBx production in the Huh7 hepatoma cell line by transfecting the bicistronic plasmid pHBx- IRES-EGFP and Western blotting. Using the same method, I monitored activation of the MEK1/2-ERK signaling pathway by ERK...

在此篇論文中，我們提出一個慢性Ｂ型肝炎病毒感染病程之數學模型。因為在病症間的轉移機率(Transition probability)是隨著患者的年齡變動，所以在過去的文獻中，已經有學者提出，在疾病轉移機率模型中，應加入國民生命表(Life table)，藉此讓機率模型更符合Ｂ型肝炎病患的生命歷程。但是過去的文獻中，學者並沒有利用加入國民生命表之後疾病模型做進一步的病程分析。在這篇論文當中，我們假設原始的疾病轉移模型是符合馬可夫鏈的性質，並且提出一種加入國民生命表的方法，賦予疾病有年齡相關特性之模型。根據文獻數據和類馬可夫機率性質，我們使用著名的Chapman-Kolmogorov公式計算Ｂ型肝炎的自然病程機率，並畫出病人的生存機率曲線(Survival curve)。文章最後將會藉由兩個例子來介紹此篇論文提出的模型。實驗數據結果證實，此模型不僅提供了一個更精確的方法去分析在病症與死亡間的轉移機率、平均餘命(Life expectancy)、以及在不同年齡的存活機率(Survival probability)，並且可以更進一步的分析且瞭解病情狀態之間的轉移狀況。 / In this thesis, we propose a new mathematical model extending the natural history of hepatitis B virus (HBV) prognosis progression on chronic HBV infection. Since the actual transition probabilities between symptoms are dependent of ages, it has been proposed that the life table should be accommodated to the HBV prognosis progression model so that it can more properly explain the disease progression of the HBV patients. But in the literature, no further disease analysis and applications of it with the life table are discussed. In this thesis, we assume that the original disease progression is described by a Markov model, and propose a new method to combine the HBV progression with the life table so that the proposed model integrates data from the life table and allows the accommodation of age-dependent properties of the target disease. With clinical data based on annual incidence rates, the entire model is Semi-Markov based in nature. Computation methods similar to the celebrated Chapman-Kolmogorov equation can be applied to study the associated probability of each likely trajectory with desired initial ages and health states under the scenarios of natural history and various treatment policies. This method provides a more accurate way to analyze the transitions between symptoms, such as the mean life expectancy or the survival probabilities at different ages. We will give examples to demonstrate the proposed method in this thesis. Numerical results show the proposed model not only provides a more accurate method to analyze the mean life expectancy, the survival probabilities at different ages, and the transition probabilities from symptoms to death but also helps us to understand the transitions between symptoms.