京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第18880号 / 医博第3991号 / 新制||医||1008(附属図書館) / 31831 / 京都大学大学院医学研究科医学専攻 / (主査)教授 山下 潤, 教授 篠原 隆司, 教授 斎藤 通紀 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

This diploma thesis is focused on business plan of a freshly established company. The company's core business is pharmacy service. Theory part describes important features to create business plan. Analytics part uses various analysis methods of external and internal environment. For this purposes, PEST and Porter's analysis have been used. Purpose part brings exact solutions for creating a business plan for ILIPHARM, s.r.o. company.

Acute lower respiratory tract infections are a leading cause of morbidity and mortality world-wide. Streptococcus pneumoniae (pneumococcus) is the most common bacterial cause of community-acquired pneumonia. Recovery from pneumococcal pneumonia results in the formation of resident memory CD4+ T cells, which act on lung epithelial cells to accelerate immune responses. Alveolar macrophages (AMs) are tissue-resident macrophages localized in the air spaces, where they orchestrate the lung anti-microbial responses. We hypothesized that recovery from pneumococcal pneumonia results in remodeling of the pool of alveolar macrophages, which act in concordance with other immune cells to protect the lungs from future infections. Although AM numbers were unchanged in experienced lungs, their surface phenotype showed significant changes, most prominently an increased MHC-II and a decreased SiglecF. This experienced AM phenotype was regionally-localized and long-lasting. Experienced AMs also exhibited extensive remodeling on the metabolomics and transcriptional level. Experienced AMs demonstrated significant increases in phosphocreatine and its metabolite precursors. The transcriptional analyses also revealed extensive changes. At baseline, experienced AMs exhibited a significant reduction in cell cycle activity and mRNA processing compared to naïve mice. During acute pneumonia, experienced AMs exhibited significant increases in immune signaling and energy metabolism. Moreover, transcriptional data also revealed strong but imperfect enrichment of a signature previously associated with IFN𝛾 signaling and marrow-derived AMs. IFN𝛾 gain and loss of functions experiments corroborated transcriptional data and revealed an essential role for IFN𝛾 in directly driving the AM MHC-II remodeling. Several immune cells produced IFN𝛾, with neutrophils being the most prominent source after the 1st pneumococcal challenge but other cells predominating after the 2nd pneumococcal challenge. CD4+ T cell depletion studies demonstrated that AMs' experienced phenotype was independent of CD4+ T cells. In contrast to naïve mice, lineage-tracing studies demonstrated that marrow-derived AMs predominately constitute the experienced AM pool. Upon experience, both embryonic AMs and marrow-derived AMs demonstrated similar remodeling for both SiglecF and MHC-II on their surfaces. While all AM similarly remodeled independent of their origin, marrow-derived AMs in experienced lungs displayed some differences from their embryonic counterparts, being less phagocytic. In conclusion, recovery from pneumococcal pneumonia remodels the pool of alveolar macrophages to acquire adaptive characteristics. This remodeling involves a combination of recruitment of new cells and trained immunity via IFN𝛾 signaling.

Eukaryotes organize their genomes by wrapping DNA around positively charged proteins called histones to form a structure known as chromatin. This structure is ideal for keeping the genome safe from damage, but also becomes an obstacle for the transcriptional machinery to access information stored in the DNA. To facilitate a balance between storage and accessibility, eukaryotes utilize a family of enzymes known as ATP-dependent chromatin remodelers to directly manipulate chromatin structure. The diverse activities of these chromatin remodeling enzymes range from simply sliding nucleosomes to reveal transcription start sites, to editing the composition of a nucleosome by exchanging canonical histones for histone variants. Chromatin remodeling enzymes recognize features of the nucleosome that activate their ATPase domains and enable proper remodeling function. One nuclear epitope that has been extensively studied is the nucleosomal acidic patch. This negatively charged region on the face of the nucleosome has been shown to be essential for remodeling enzymes like Chd1, ISWI, and INO80C. The chromatin remodeler SWR1C edits nucleosomes by removing the canonical histone H2A from nucleosomes and exchanges it for the histone variant H2A.Z, but the role of the acidic patch in this process has not been investigated. In this work, I showed that SWR1C has normal binding affinity to acidic patch mutant nucleosomes and retains ATPase stimulation but can no longer exchange dimers on this substrate. This work also identified a novel arginine anchor on the essential SWR1C subunit, Swc5, that binds specifically to the nucleosomal acidic patch. The data in this work suggest a mechanism where SWR1C engages nucleosomes and uses the Swc5 subunit to recognize the nucleosomal acidic patch to couple ATPase activity to histone dimer exchange.

Cardiac remodeling involves changes in heart shape, size, structure, and function after injury to the myocardium. The proinflammatory adaptor protein myeloid differentiation protein 88 (MyD88) contributes to cardiac remodeling. To investigate whether excessive MyD88 levels initiate spontaneous cardiac remodeling at the whole-organism level, we generated a transgenic MyD88 mouse model with a cardiac-specific promoter. MyD88 mice (male, 20–30 g, n=~80) were born at the expected Mendelian ratio and demonstrated similar morphology of the heart and cardiomyocytes with that of wild-type controls. Although heart weight was unaffected, cardiac contractility of MyD88 hearts was mildly reduced, as shown by echocardiographic examination, compared with wild-type controls. Moreover, the cardiac dysfunction phenotype was associated with elevation of ANF and BNP expression. Collectively, our data provide novel evidence of the critical role of balanced MyD88 signaling in maintaining physiological function in the adult heart.

Background: Stroke volume (SV) and aortic valve area calculations require the left ventricular (LV) outflow tract (LVOT) or aortic annular area calculations that involve squaring the respective diameters. Area calculation errors became evident with transcatheter aortic valve replacement where areas were underestimated due to an elliptical annulus. We hypothesized that LVOT and annular shape are more elliptical in patients with greater relative LV wall thickness (RWT) leading to underestimation of SV index using 2D Doppler echocardiography. Methods: We studied 203 consecutive patients referred to an outpatient noninvasive laboratory for Doppler echocardiograms which included acceptable 3-dimensional images. 3-dimensional assessment of the LVOT at 3–5 mm from the valve insertion, at the site of valve insertion, and at the sinus of Valsalva (SOV) was performed with assessment of the minor axis (MN), major axis (MJ), and areas at mid-systole. SV index was calculated from LVOT and annular diameters obtained from 2-dimensional echo and from 3-dimensional LVOT areas. Results: An inverse relation of RWT with MN/MJ at mid-systole for the LVOT (r = 0.5812, P < 0.0001) and annulus (r = 0.6865, P < 0.0001) was noted. LVOT and annulus areas were similar among groups at mid-systole. SV index calculated from 2D LVOT dimensions was significantly smaller than using 3D LVOT areas (35.6 ± 8.9 vs 53.6 ± 16.1 mL, P < 0.0001). Conclusion: There is an inverse relation between MN/MJ and RWT at the LVOT and aortic annulus despite the LVOT and annular areas being similar across most geometries resulting in SV index underestimation calculated using LVOT diameters vs 3D LVOT areas.

Objective: Midlife obesity is a risk factor for dementia development. Obesity has also been linked to hyperaldosteronism, and this can be modeled in rats by high fat (HF) feeding from weaning. Aldosterone, or activation of the mineralocorticoid receptor (MR) causes cerebrovascular injury in lean hypertensive rats. We hypothesized that rats fed a HF diet would show inward middle cerebral artery (MCA) remodeling that could be prevented by MR antagonism. We further proposed that the cerebral artery remodeling would be associated with white mater injury. Methods: Three-week-old male Sprague-Dawley rats were fed a HF diet ± the MR antagonist canrenoic acid (Canr) for 17 weeks. Control rats received normal chow (control NC). MCA structure was assessed by pressure myography. Results: The MCAs from HF fed rats had smaller lumens and thicker walls when compared to arteries from control NC rats; Canr prevented the MCA remodeling associated with HF feeding. HF feeding increased the mRNA expression of markers of cell proliferation and vascular inflammation in cerebral arteries and Canr treatment prevented this. White mater injury was increased in the rats fed the HF diet and this was reduced by Canr treatment. The expression of doublecortin, a marker of new and immature neurons was reduced in HF fed rats, and MR antagonism normalized this. Conclusions: These data suggest that HF feeding leads to MR dependent remodeling of the MCA and this is associated with markers of dementia development.

Thesis (M. Arch.)--Massachusetts Institute of Technology, Dept. of Architecture; and, (M.S.)--Massachusetts Institute of Technology, Dept. of Civil Engineering, 1981. / MICROFICHE COPY AVAILABLE IN ARCHIVES AND ROTCH. / Includes bibliographical references (leaves 84-86). / Because of the decreasing use of private automobiles in city centers and because of usual development pressures, some urban parking garages will become available for replacement or recycling. The choice between replacement or recycling of an abandoned garage is based on cost, but many other factors influence this decision. The suitability of a garage for recycling can often be determined by the consideration of three simple indicators: the type of garage, the horizontal depth of the building, and the typical floor-to-ceiling height. Following the determination of basic suitability, several architectural and structural issues must be considered in order to identify potential problems in the intended recycling and in order to discover practical solutions to these problems. The determination of suitability and the consideration of architectural and structural problems are discussed generally, and are demonstrated in the study of the West Garage. / by Michael Johannes Paul. / M.S. / M.Arch.

Indiana University-Purdue University Indianapolis (IUPUI) / Insulin-stimulated glucose uptake into skeletal muscle cells requires translocation of the glucose transporter-4 (GLUT4) from the cell interior to the plasma membrane. Insulin-stimulated GLUT4 vesicle translocation is dysregulated in Type 2 diabetes (T2D). The Group I p21–activated kinase (PAK1) is a required element in insulin-stimulated GLUT4 vesicle translocation in mouse skeletal muscle in vivo, although its placement and function(s) in the canonical insulin signaling cascade in skeletal muscle cells, remain undetermined. Therefore, the objective of my project is to determine the molecular mechanism(s) underlying the requirement for PAK1 in the process of insulin-stimulated GLUT4 vesicle translocation and subsequent glucose uptake by skeletal muscle cells. Toward this, my studies demonstrate that the pharmacological inhibition of PAK1 activation blunts insulin-stimulated GLUT4 translocation and subsequent glucose uptake into L6-GLUT4myc skeletal myotubes. Inhibition of PAK1 activation also ablates insulin-stimulated F-actin cytoskeletal remodeling, a process known to be required for mobilizing GLUT4 vesicles to the plasma membrane. Consistent with this mechanism, PAK1 activation was also required for the activation of cofilin, another protein implicated in F-actin remodeling. Interestingly, my studies reveal a novel molecular mechanism involving PAK1 signaling to p41-ARC, a regulatory subunit of the cytoskeletal Arp2/3 complex, and its interactions with another cytoskeletal factor, N-WASP, to elicit the insulin-stimulated F-actin remodeling in skeletal muscle cells. Pharmacological inactivation of N-WASP fully abrogated insulin-stimulated GLUT4 vesicle translocation to the cell surface, coordinate with blunted F-actin remodeling. Furthermore, my studies revealed new insulin-induced interactions amongst N WASP, actin, p41-ARC and PAK1; inactivation of PAK1 signaling blocked these dynamic interactions. Taken together, the above studies demonstrate the significance of PAK1 and its downstream signaling to F-actin remodeling in insulin-stimulated GLUT4 vesicle translocation and glucose uptake, revealing new signaling elements that may prove to be promising targets for future therapeutic design.

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第20606号 / 医博第4255号 / 新制||医||1023(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 羽賀 博典, 教授 篠原 隆司, 教授 近藤 玄 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM