Background: Focal segmental glomerulosclerosis (FSGS) is a component of childhood nephrotic syndrome occurring in 10%-20% of all cases. Over time, 25%-50% of children with FSGS develop kidney failure disease. We followed a cohort of children with FSGS in order to delineate the risk profile of progression to kidney failure (KF). Methods: We evaluated patient data collected from 1977 to 2002 at a regional mid-Atlantic nephrology center in the United States. KF was defined primarily for those patients whose serum creatinine (SCr) value doubled compared with the SCr value from a previous visit. Patients who received dialysis or a kidney transplant were also defined as having KF. We analyzed patient data for those who had at least two visits with SCr values recorded. Various baseline characteristics of patients who had developed KF and those with no kidney failure (NKF) were compared. Hazard ratios and correlation were used to further investigate potential risk factors of the kidney failure. We also compared the inverse SCr trend for KF and NKF patients using weighted linear regression. Results: Thirty-four of 43 FSGS patients had adequate follow-up data. About 60% of the patients developed KF over the study period. The average age of the KF patients at diagnosis of FSGS was 9 years, and that of NKF patients 12 years (P=0.05). FSGS patients with KF had a significantly higher mean diastolic blood pressure (DBP) at baseline, compared to those with NKF (P<0.0001). Other baseline characteristics including race, body mass index (BMI), systolic blood pressure, total cholesterol, urinary protein/creatinine ratio and calculated glomerular filtration rate (cGFR) were not significantly different. Baseline DBP was a significant risk factor in progression to KF (HR: 1.03; 95%CI: 1.01-1.06). Inverse SCr values were significantly decreased over time in KF patients (P=0.01). Conclusions: The data of this study indicate that children diagnosed with FSGS who are younger than 10 years and have elevated baseline DBP are more likely to develop kidney failure. The non-significant hazard ratios for other baseline characteristics including gender, race, and BMI are not instrumental risk factors. These results may help understand what may affect progression towards kidney failure in children with FSGS.

The focal adhesion kinase (FAK) and the protein tyrosine kinase 2-beta (PYK2) are two closely related non-receptor tyrosine kinases that link cell adhesion, migration and proliferation, and thus also promote cancer cell invasiveness. FAK and PYK2 have the same domain structure (comprising the FERM, kinase and FAT domains) and possess several overlapping functions, however their cellular roles can be different or even opposing. In particular, PYK2 can be activated by calcium, and has important functions in the brain and neurodegenerative disease. The molecular basis for calcium-based activation of PYK2 is unclear and controversial. In this work we combined biophysical and structural methods to determine the molecular basis for calcium-sensing in PYK2. For this, we investigated the least-studied region of these kinases, namely the long linker (KFL) region between the kinase and FAT domains. This linker is only ~20% conserved between FAK and PYK2, and, therefore, is a prime candidate for causing their differential properties. We find that the linker harbors a helical segment, which is conserved in both FAK and PYK2, and contributes to their dimerization (an important step in their activation). Helix-flanking regions differ between both proteins, and we show that these of PYK2 create a non-canonical dimeric binding site for calcium-bound calmodulin. Calmodulin-binding is synergistic with linker dimerization in PYK2, explaining how calcium influx can be translated into activation of PYK2. Collectively, our work clarifies the capacities for FAK and PYK2 to receive, process and transduce cellular signals, and may provide new opportunities for targeted therapeutic intervention.

京都大学 / 0048 / 新制・課程博士 / 博士(工学) / 甲第18657号 / 工博第3966号 / 新制||工||1610(附属図書館) / 31571 / 京都大学大学院工学研究科高分子化学専攻 / (主査)教授 岩田 博夫, 教授 木村 俊作, 教授 秋吉 一成 / 学位規則第4条第1項該当 / Doctor of Philosophy (Engineering) / Kyoto University / DFAM

Nephrotic syndrome (NS) is the second leading cause of chronic kidney disease (CKD) presenting under the age of 30. NS presents in children with edema and severe proteinuria, caused by the effacement of podocyte foot processes within the glomerular filtration barrier. Patients with steroid-resistant NS (SRNS) frequently develop end-stage renal disease (ESRD). Additionally, renal biopsies from these patients often reveal focal segmental glomerulosclerosis (FSGS). Pathogenic mutations in known monogenic disease genes have been found in 11-45% children with FSGS/SRNS. Notably, most Mendelian etiologies exhibit recessive inheritance, while dominant vertical inheritance with incomplete penetrance is observed in the remainder. The role of de novo variants (DNVs) in NS necessitates further investigation. Tripartite motif containing 8, TRIM8, is an E3 ubiquitin ligase. De novo TRIM8 variants were previously implicated in a syndromic disease consisting of neurodevelopmental delay, epilepsy, cerebral atrophy, and nephrotic syndrome. In this study, we recapitulate the patient-specific mutations in inducible overexpression cell lines and in CRISPR/Cas9-generated mouse models. N-terminal MYC or GFP-tagged TRIM8 inducible cell lines were generated and characterized using the pInducer21 system. Western blot and immunofluorescence data show that MYC- and GFP-TRIM8 were induced by doxycycline in immortalized podocyte cell lines. Candidate interactors for TRIM8 from the literature and stratified using kidney single cell mRNA sequencing expression were cloned into mammalian expression vectors. Finally, a Trim8 knockout allele (c. 56_162del; p.H20Qfs*124 and c.367_463+304delins46) was generated and bred to yield an allelic series of wildtype, heterozygous and homozygous animals. These mice exhibited normal survival and did not demonstrate proteinuria through three to four months of life. Overall, further studies are ongoing with regards to the continued monitoring of proteinuria and kidney dysfunction, as well as the potential interactor cloning and cell line characterization. / 2025-02-06T00:00:00Z

Most current augmented reality (AR) displays present content at a fixed focal demand. At the same time, real-world stimuli can occur at a variety of focal distances. To integrate information, users need to switch eye focus between virtual and real-world information continuously. Previously, Gabbard, Mehra, and Swan (2018) examined these issues, using a text-based visual search task on a monocular AR display. This thesis replicated and extended the previous experiment by including a new experimental variable stereopsis (stereo, mono) and fully crossing the variables of context switching and focal distance switching, using AR haploscope. The results from the monocular condition indicate successful replication, which is consistent with the hypothesis that the findings are a general property of AR. The outcome of the stereo condition supports the same adverse effects of context switching and focal distance switching. Further, participants have better performance and less eye fatigue in the stereo condition compared to the monocular condition.

No description available.

No description available.

京都大学 / 新制・課程博士 / 博士(医学) / 甲第23800号 / 医博第4846号 / 新制||医||1058(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 高橋 淳, 教授 髙折 晃史, 教授 辻川 明孝 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

Since its conception in 2005, irreversible electroporation (IRE), a non-thermal tumor ablation modality, was investigated for safety and efficacy in clinical applications concerning different organs. IRE utilizes high voltage (~3kV), short duration (~100us) pulses to create transient nanoscale defects in the plasma membrane to cause cell death due to irreversible defects, osmotic imbalances and ATP loss. More recently, high-frequency irreversible electroporation (H-FIRE), which employs narrow bipolar pulses (~0.5-10us) delivered in bursts (on time ~100us), was invented to provide benefits such as the mitigation of intense muscle contractions associated with IRE-based treatments. Furthermore, H-FIRE exhibits the potential to improve lesion predictability in homogeneous and heterogeneous tissue masses. Therapeutic IRE and H-FIRE utilize source and sink electrodes inserted into or around the tumor to deliver the treatment. Prediction of the ablation size, for a set of parameters, can be achieved by the use of pre-treatment planning algorithms that calculate the induced electric field distribution in the target tissue. An electric field above a certain threshold induces cell death and parameters are tuned to ensure complete tumor coverage while sparing the nearby healthy tissue. IRE studies have shown that the underlying field is influenced by the increase in tissue conductivity due to enhanced membrane permeability, and treatment outcome can be improved when this nonlinearity is accounted for in numerical models. Since IRE pulses far exceed the time constant of the cell (~1us), the tissue response can be treated as essentially DC a static approximation can be used to predict the field distribution. Alternately, as H-FIRE pulses are on the order of the time constant of the membrane, the tissue response can no longer be treated as DC. The complexity of the H-FIRE-induced field distribution is further enhanced due to the dispersion and non-linearity in biological tissue impedance during treatment. In this dissertation, we have studied the electromagnetic fields induced in tissue during H-FIRE using several experimental and modeling techniques. In addition, we have characterized the nonlinearity and dispersion in tissue impedance during H-FIRE treatments and proposed simpler methods to predict the field distribution to enable easier translation to the clinic. / Ph. D.

Irreversible electroporation (IRE) is a non-thermal ablation technique that allows for eradication of unresectable tumors in a minimally invasive procedure. While IRE will preferentially kill larger cells over smaller ones, it does not discriminate between cells with larger and small nuclei. Given that one of the hallmarks of cancer cell morphology is larger, more abundant nuclei, our team set out to explore the possibility of preferentially targeting this physical and geometrical characteristic. / Master of Science