Philosophiae Doctor - PhD / Background: Oropharyngeal candidiasis, caused by the fungus Candida, is the most common opportunistic infection affecting the quality of life of immunocompromised patients. Fluconazole is widely used as the first line of treatment for fungal infections. However, the inappropriate and misguided use of the drug has led to the evolvement of fluconazole-resistant Candida organisms. This arising resistance resulted in the urgent need for the development of new antimicrobial drugs. The aim of the present study was to investigate the antifungal action of K21, a novel antimicrobial quarternary ammonium compound, on fluconazole-resistant Candida species.

Multiresistens hos gramnegativa bakterier är ett stort problem i stora delar av världen. Avsaknaden av nya preparat har resulterat i återintroduktion av ett tidigare stoppat antibiotikum, kolistin, men dessvärre kantas resistensbestämningen av flertalet problem. Kolistin är en positivt laddad molekyl som binder till lipid A i yttermembranet, vilket destabiliseras och resulterar i celldöd. Flertalet resistensmekanismer mot kolistin finns beskrivna och majoriteten resulterar i förändrad lipid A-struktur, vilket leder till försämrad kolistininbindning. Syftet med examensarbetet var att utvärdera fyra potentiella screeningmetoder för att upptäcka kolistinresistens och jämföra resultaten mot buljongspädning som referensmetod. De fyra screeningmetoderna var buljongeluering och diskdiffusion samt de kommersiellt tillgängliga screeningtesterna Superpolymyxin och Rapid polymyxin NP. Totalt analyserades 57 helgenomsekvenserade Enterobacterales, varav 28 Escherichia coli och 29 Klebsiella pneumoniae, där aktuella resistensmekanismer var kända. Samtliga isolat analyserades en gång per metod och utöver detta analyserades även tre kontrollstammar. Vid buljongspädning erhölls värde för minsta inhiberande koncentration (MIC) kolistin, vilket användes för klassificering som känslig (MIC ≤ 2 mg/l) eller resistent (MIC > 2 mg/l). De fyra screeningtesterna var designade för att upptäcka resistenta isolat och innehöll 2–4 mg/l kolistin. Resultaten från screeningtesterna var kvalitativa och känslighetskategoriserade isolaten. Sensitivitet och specificitet beräknades för metoderna. Högst sensitivitet (100 %) noterades för Superpolymyxin och Rapid polymyxin NP, och lägst specificitet (76 %) för buljongeluering. Med hänsyn till resultat, prevalens, hållbarhets-, tids- och kostnadseffektivitet drogs slutsatsen att Rapid polymyxin NP är bästa alternativet som screeningmetod av utvärderade metoder. / Infections due to multidrug resistant Gram-negative bacteria are a major problem in large parts of the world. Due to lack of treatment options, a previously banned antibiotic, colistin, has been reintroduced. Unfortunately, susceptibility testing of colistin is problematic and currently the only recommended method is broth microdilution (BMD). Colistin is a positively charged molecule that binds to lipid A at the negatively charged outer cell membrane, which leads to cell disruption. Several mutations causing colistin resistance have been identified, most of them cause alterations of lipid A, resulting in impaired colistin outer cell membrane interaction. The aim of this study was to evaluate four potential screening methods for colistin resistance and compare the results with BMD (reference method). The four screening methods were broth disk elution, disk diffusion and the commercially available tests Superpolymyxin and Rapid polymyxin NP. A collection of 57 whole genome sequenced Enterobacterales consisting of 28 Escherichia coli and 29 Klebsiella pneumoniae were included in the study. Reference results were obtained by BMD, where the estimated minimum inhibitory concentration (MIC) values were used for classification of the isolates as sensitive (MIC ≤ 2 mg/l) or resistant (MIC > 2 mg/l). Sensitivity and specificity were calculated for the methods. The highest sensitivity (100 %) was noted for Superpolymyxin and Rapid polymyxin NP, and the lowest specificity (76 %) for broth disk elution. Considering the results, prevalence, durability, time and cost efficiency, it was concluded that Rapid polymyxin NP is the best alternative as a screening method of evaluated methods.

Glaze damage to canopy trees, gap size and seedling regeneration was surveyed relating to site topographic features after a major glaze storm at Fishburn Forest in the central Appalachian mountain region. The survey sites were stratified according to aspect and landform, and sample plots were randomly assigned to each topographic category. The canopy trees in the plots were tallied and their damage was visually classified. The results demonstrated that different species had different susceptibilities to glaze damage and susceptibility of species varied as topography changed. Virginia pine was the most susceptible species. In general, trees had the greatest damage on steep slopes and eastern aspects, and least damage on toe-slopes. Some species varied greatly in their susceptibility to glaze damage across the topographic categories, such as red maple, while some species varied little, like blackgum. Basal area was reduced 6% and varied due to species and topography. Where glaze damage was severe, gaps in the forest canopy were created. Gap sizes and the effects of gap size and topography on seedling regeneration within gaps caused by glaze damage were assessed. Seedlings were tallied by species and height in microplots within each sample plot and in the nearest gap, where gap size was measured. Gap size varied from 14,000 m2 to less than 20 m2. Average gap size was 255 m2. Canopy tree damage, forest type and topography were the most important factors affecting gap size. Gaps tended to be larger on steep backslopes. Common seedling species in both gaps and understory were sassafras, serviceberry and red maple. Disturbance affected both seedling height and density, as did topography. However gap size had effect only on seedling height. The future forest in this area is unlikely to change significantly, as species composition of seedlings was largely unaffected by the glaze storm damage. / Master of Science

Background: Previous reports have suggested that there may be gene×gender interaction for bipolar disorder (BD)-associated genes/loci at 22q11-13. This study aimed to investigate the associations of SEZ6L genetic variants with bipolar disorder I (BD-I) and to examine gender-specific genetic associations. Methods: 605 BD-I Caucasian cases and 1034 controls were selected from the publicly available data of the Whole Genome Association Study of BD. To increase power, an additional 362 Caucasian controls were added to this study from the Genome-Wide Association Study of Schizophrenia. In total, 605 BD-I cases and 1396 controls (934 males and 1067 females) were available for genetic association analysis of 118 SNPs within the SEZ6L gene using PLINK software. Results: 16 SNPs showed significant gene x gender interactions influencing BD-I (P<0.01). In addition, significant differences in the distribution of the alleles for these 16 SNPs were observed between the female BD-I patients and healthy controls (P<0.015) but no significant associations were found for the male sample (P>0.05). The SNP rs4822691 showed the strongest association with BD-I in the female sample (P=2.18×10-4) and the strongest gene×gender interaction in influencing BD-I (P=9.16×10 -5). Limitations: The findings of this study need to be replicated in independent samples. Conclusions: This is the first demonstration that genetic variants in the SEZ6L gene are associated with BD-I in female patients and provides additional compelling evidence for genetic variation at 22q11-13 that influences BD-I risk. The present findings highlight the gene x gender interactions modifying BD-I susceptibility.

Background: Previous reports have suggested that there may be gene×gender interaction for bipolar disorder (BD)-associated genes/loci at 22q11-13. This study aimed to investigate the associations of SEZ6L genetic variants with bipolar disorder I (BD-I) and to examine gender-specific genetic associations. Methods: 605 BD-I Caucasian cases and 1034 controls were selected from the publicly available data of the Whole Genome Association Study of BD. To increase power, an additional 362 Caucasian controls were added to this study from the Genome-Wide Association Study of Schizophrenia. In total, 605 BD-I cases and 1396 controls (934 males and 1067 females) were available for genetic association analysis of 118 SNPs within the SEZ6L gene using PLINK software. Results: 16 SNPs showed significant gene x gender interactions influencing BD-I (P<0.01). In addition, significant differences in the distribution of the alleles for these 16 SNPs were observed between the female BD-I patients and healthy controls (P<0.015) but no significant associations were found for the male sample (P>0.05). The SNP rs4822691 showed the strongest association with BD-I in the female sample (P=2.18×10-4) and the strongest gene×gender interaction in influencing BD-I (P=9.16×10 -5). Limitations: The findings of this study need to be replicated in independent samples. Conclusions: This is the first demonstration that genetic variants in the SEZ6L gene are associated with BD-I in female patients and provides additional compelling evidence for genetic variation at 22q11-13 that influences BD-I risk. The present findings highlight the gene x gender interactions modifying BD-I susceptibility.

Objectives: To compare and contrast the geographic and demographic distribution of bro β-lactamase and antibiotic MIC50/90 for 1440 global Moraxella catarrhalis isolates obtained from children and adults between 2001 and 2002. Methods: One thousand four hundred and forty M. catarrhalis isolates originating from seven world regions were investigated. The isolates were recovered from 411 children <5 years of age and 1029 adults >20 years of age. PCR-restriction fragment length polymorphism (RFLP) was performed to determine bro prevalence and to distinguish between bro types. MIC values of 12 different antibiotics were determined using the CLSI (formerly NCCLS) broth microdilution method. Results: Of the 1440 isolates, 1313 (91%) possessed the bro-1 gene and 64 (4%) possessed the bro-2 gene. Additionally, the prevalence of bro positivity between the child and adult age groups was significantly different (P<0.0001), though bro-1 and bro-2 prevalences within age groups were not significantly different. Consistently higher β-lactam MICs were observed for M. catarrhalis isolates originating in the Far East. Significant correlations in MICs were observed for several antibiotic combinations, including all five β-lactams with each other, and among the two quinolones. Conclusions: The worldwide prevalence of bro gene carriage in clinical isolates of M. catarrhalis is now approaching 95%, with children significantly more likely to harbour bro-positive isolates than adults. Further, statistically significant differences in the distribution of β-lactam MICs were observed between different world regions, particularly with respect to the Far East.

High incidences of slope movement are observed throughout Cuyahoga River watershed in northeast Ohio, USA. The major type of slope failure involves rotational movement in steep stream walls where erosion of the banks creates over-steepened slopes. The occurrence of landslides in the area depends on a complex interaction of natural as well as human induced factors, including: rock and soil strength, slope geometry, permeability, precipitation, presence of old landslides, proximity to streams and flood-prone areas, land use patterns, excavation of lower slopes and/or increasing the load on upper slopes, alteration of surface and subsurface drainage. These factors were used to evaluate the landslide-induced hazard in Cuyahoga River watershed using logistic regression analysis, and a landslide susceptibility map was produced in ArcGIS. The map classified land into four categories of landslide susceptibility: low, moderate, high, and very high. The susceptibility map was validated using known landslide locations within the watershed area. The landslide susceptibility map produced by the logistic regression model can be efficiently used to monitor potential landslide-related problems, and, in turn, can help to reduce hazards associated with landslides.

Background. Severe periodontal disease is highly prevalent worldwide, affecting 20% of the population between the ages of 35 and 44 years. The etiological epidemiology in Peru is scarce, even though some studies describe a prevalence of 48.5% of periodontal disease in the general population. Periodontitis is one of the most prevalent oral diseases associated with site-specific changes in the oral microbiota and it has been associated with a socioeconomic state. This study aimed to determine the etiology and resistance profile of bacteria identified in a group of Peruvian patients with periodontal disease. Methods. Six subgingival plaque samples were collected from eight patients with severe periodontitis. Bacterial identification was carried out by an initial culture, PCR amplification, and subsequently DNA sequencing. We evaluated the antibiotic susceptibility by the disk diffusion method. Results. Variable diversity in oral microbiota was identified in each one of the eight patients. The bacterial genus most frequently found was Streptococcus spp. (15/48, 31.3%) followed by Rothia spp. (11/48, 22.9%), Actinomyces spp. (9/48, 18.8%), and Eikenella spp. (4/48, 8.3%). The most common species found was Rothia dentocariosa (8/48, 16.7%). The antimicrobial susceptibility assay varied according to the species tested; however, among all the isolates evaluated, Actinomyces naeslundii was resistant to penicillin and tetracycline; Eikenella corrodens was resistant to dicloxacillin; and Rothia dentocariosa was resistant to amoxicillin + clavulanic acid and metronidazole but also susceptible to trimethoprim-sulfamethoxazole. Conclusions. The most prevalent periodontal bacterium found in this study was Rothia dentocariosa. Specific antimicrobial therapy is required to improve the treatment outcomes of patients with periodontal disease and avoid antibiotic resistance. / Revisión por pares

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第18647号 / 医博第3946号 / 新制||医||1006(附属図書館) / 31561 / 京都大学大学院医学研究科医学専攻 / (主査)教授 福山 秀直, 教授 平岡 眞寛, 教授 宮本 享 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DGAM

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第21298号 / 医博第4387号 / 新制||医||1030(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 中川 一路, 教授 木原 正博, 教授 小池 薫 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM