Development of a new genome engineering technology, CRISPR-PCDup, to create segmental aneuploid and exploration of synthetic lethality in yeast

博士（工学）崇城大

Can randomness alone tune the fractal dimension?

We present a generalized stochastic Cantor set by means of a simple {\it cut and delete process} and discuss the self-similar properties of the arising geometric structure. To increase the flexibility of the model, two free parameters, $m$ and $b$, are introduced which tune the relative strength of the two processes and the degree of randomness respectively. In doing so, we have identified a new set with a wide spectrum of subsets produced by tuning either $m$ or $b$. Measuring the size of the resulting set in terms of fractal dimension, we show that the fractal dimension increases with increasing order and reaches its maximum value when the randomness is completely ceased.Comment: 6 pages 2-column RevTeX, Two figures (presented in the APCTP International Symposium on Slow Dynamical Processes in Nature, Nov. 2001, Seoul, Korea

Solution Blow Spinning (SBS): A Promising Spinning System for Submicron/Nanofibre Production

Submicron/nanofibres possess great potential for application in different areas because of their amazingly high surface area-to-weight ratio. The demand for fabrication of such fibres on a huge scale is increasing with the fast improvement of nanotechnology. Traditionally, nanofibre fabrication methods have intrinsic faults, limiting their application in industry. Solution blow spinning (SBS) is a viable option for producing adaptable and conformable submicron/nanofibre mats on a variety of surfaces. The technique can be employed to produce submicron/ nanofibres with only a simple commercial airbrush, a concentrated polymer solution, and a compressed gas source. It depends on the high velocity of decompressed air that allows the rapid stretching and evaporation of the solvent from a polymeric solution jet at the outlet of the concentric nozzles system. Along with recent advancements, the importance and drawbacks of the solution blow spinning system in comparison to other methods, such as electrospinning and melt blowing, are briefly discussed. Furthermore, the mechanisms of co-axial SBS spinning and micro SBS spinning system for submicron/nanofibre fabrication are also described. Drawbacks and research challenges of SBS are also addressed in this paper

Proliferation and Differentiation of Intestinal Caco-2 Cells Are Maintained in Culture with Human Platelet Lysate Instead of Fetal Calf Serum

Cell lines are widely used as in vitro model systems and substitute for animal experiments. The frequently used Caco-2 cell line is considered to reflect characteristics of differentiated intestinal epithelium. However, the need to culture the cells with fetal calf serum (FCS) induces a high variability, risk of contamination and is ethically disputed. We tested the culture of Caco-2 cells with human platelet lysate (PL) instead of FCS. We compared cell viability and differentiation by measuring ATP levels, gene and protein expression of specific markers in total cell extracts, brush border membrane vesicles (BBM) and lipid rafts (LR). Cell viability was slightly enhanced in cells grown with PL compared to FCS. The cells differentiated to an intestinal phenotype like the cells cultured in FCS, as indicated by the similar gene expression levels of hexose and protein transport proteins and the structural protein VILLIN. BBM showed a comparable distribution of the intestinal hydrolases, indicating a maintained cell membrane polarity. The distribution of the marker protein FLOTILLIN-2 in LR was also similar. We conclude that PL is an exquisite and suitable replacement for FCS in the culture of Caco-2 cells that can eliminate many disadvantages incurred due to the use of FCS

Essays in Economic and Political Choice

This thesis is a collection of essays. Each essay is a whole chapter. I have prepared the following separate abstracts for each chapter. Chapter 1 Chance Constrained Dynamic Programming Model of Water Reservoir with Joint Products This chapter presents two models for reservoir management. Model l is a dynamic programming formulation which only allows exporting and importing of water to correct excesses or deficiencies caused by the optimizing decisions. It shows that if the one-period profit is a function of the water releases, water stock, and the physical capacity of the reservoir, then optimal decisions regarding water releases and capacity exist and are unique both for the N-period and the infinite period problems. Moreover, the model shows that if the profit function is separable and linear in the releases, then the optimal decision rule is linear and the long-run distribution for the stock of water exists and is a simple form. The model in the Appendix of this chapter is a deterministic equivalent chance constraint formulation in which an approximation for the long-run distribution of the water stock is determined. Chapters 2 and 3 On the Retrospective Effect of Economic Conditions in Congressional Elections: A Simultaneous Equation Model Approach In these chapters a simultaneous equation model is employed to investigate the relative effects of: 1) economic conditions, 2) incumbency, and 3) recognition of the presidential party's candidate on the dual decisions of the individual to participate and vote in congressional elections. My finding is decidedly negative regarding the effect of economic conditions on both turnout and voting for the presidential party. I have, however, established the relative effects of both incumbency and recognition. Chapter 4 The Decline of Competition in Congressional Elections: Mayhew May Still Be Right Several theories have been advanced to explain the reduction in the number of competitive congressional districts during the past decade. Among these is Mayhew's theory, which attributes the reduction to the increasing control of campaign resources by incumbents. Ferejohn presents evidence which casts doubt on Mayhew's thesis. In this chapter, Ferejohn's evidence is examined within the framework of a simultaneous equation model. I conclude that Mayhew's thesis, although bloodied by Ferejohn's attack, is still very much alive. Chapter 5 Leftist Ideological Shifts in Arab Contemporary Politics: A Spatial Theory Approach This chapter presents a formal model of some aspects of leftist ideological shifts in contemporary Arab politics. In particular it focuses on the effects of information costs and the cost of ideological vagueness on the competitive parties of the left. A spatial model is used to examine analytically an observation originally advanced by the Baath Arab Socialist party. This observation states that both the cost of vagueness and the inability to meet the high information costs inherent to an articulate ideology may have been the factors which caused the adoption of Marxism Leninism by some leftist groups. Certain reasonable assumptions generated results consistent with the Baath observation. Chapter 6 The Effect of a Random Planning Horizon on Production and Investment for Petroleum Reservoir -- A Note on Kuller's and Cumming's Model There have been several formulations of models for crude oil production which tried to identify the elements of user cost and show their effect on production and investment decisions. In this chapter, previous results are extended by incorporating the uncertainty regarding the date of arrival of the backstop technology in the model. This uncertainty adds a new element to the user cost identified previously and is shown to affect the production and investment decisions.</p

Visokotlačna tekućinska kromatografija s detektorom raspršenja svjetla u uparenom uzorku za određivanje količine glavnih razreda fosfolipida u magarećem serumu

An HPLC method with an evaporative laser scattering detector was used to quantify major phospholipids fractions in donkeys’ serum. Blood samples were collected bimonthly for a whole year from 20 donkeys (10 male and 10 female) from the Sudanese breed kept at the premises of the Central Veterinary Research Laboratory (CVRL), Soba. The method used made the excellent separation possible of phosphatidylglycerine (PG), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylcholine (PC), and sphingomyelin (SM). in 27 minutes, including the regeneration of the column. The SM resulted in two peaks (saturated and unsaturated fatty acids), as described by other researchers. The method was unable to separate phosphatidylserine (PS), which appeared in one peak with phosphatidylinositol. There is a signififi cant difference in the level of PG, PE, PI and SM between females and males.The method fully discussed in this study and the obtained values of major phospholipids can be used for health control and diagnosis of diseases.Visokotlačna tekućinska kromatografija s detektorom raspršenja svjetla u uparenom uzorku rabljena je za određivanje količine glavnih frakcija fosfolipida u magarećem serumu. Uzorci krvi bili su uzeti od 20 životinja (10 magaraca i 10 magarica sudanske pasmine) držanih u prostorima Središnjega veterinarskoga istraživačkoga laboratorija, Soba. Rabljena metoda omogućila je izvrsno odvajanje fosfatidilglicerola (FG), fosfatidiletanolamina (FE), fosfatidilinozitola (FI), fosfatidilkolina i sfifi ngomijelina (SM) za 27 minuta uključujući i regeneraciju kolone. Sfingomijelin je pokazivao dva vrška (zasićene i nezasićene masne kiseline) kao što je opisano od drugih autora. Rabljenom metodom nije se moglo odvojiti fosfatidilserin od fosfatidilinozitola. Ustanovljena je signifikantna razlika u razini FG, FE, FI i SM u mužjaka i ženki. U radu se iscrpno raspravlja o metodi, a dobivene vrijednosti glavnih fosfolipida mogu se upotrijebiti za kontrolu zdravlja i dijagnosticiranje bolesti

Context Aware Mobile Application Architecture (CAMAA) for Health Care systems : standardization and abstraction of context aware layers

Context awareness was introduced recently in several fields in quotidian human activities. Among context aware applications, health care systems are the most important ones. Such applications, in order to perceive the context, rely on sensors which maybe physical or virtual. However, these applications lack of standardization in handling the context and the perceived sensors data. In this work, we propose a formal context aware application architecture model to deal with the context taking into account the scalability and interoperability as key features towards an abstraction of the context relatively to end user applications. As a proof of concept, we present also a case study and simulation explaining the operational aspect of this architecture in health care systems

Trafficking pathways of Cx49-GFP in living mammalian cells

In the present study we examined the trafficking pathways of connexin49 (Cx49) fused to green fluorescent protein (GFP) in polar and non-polar cell lines. The Cx49 gene was isolated from ovine lens by RT-PCR. Cx49 cDNA was fused to GFP and the hybrid cDNA was transfected into several cell lines. After transfection of Cx49-GFP cDNA into HeLa cells, it was shown using the double whole-cell patch-clamp technique that the expressed fusion protein was still able to form conducting gap junction channels. Synthesis, assembly, and turnover of the Cx49-GFP hybrid protein were investigated using a pulse-chase protocol. A major 78-kDa protein band corresponding to Cx49-GFP could be detected with a turnover of 16-20 h and a half-life time of 10 h. The trafficking pathways of Cx49-GFP were monitored by confocal laser microscopy. Fusion proteins were localized in subcellular compartments, including the endoplasmic reticulum (ER), the ER-Golgi intermediate compartment, the Golgi apparatus, and the trans-Golgi network, as well as vesicles traveling towards the plasma membrane. Time-dependent sequential localization of Cx49-GFP in the ER and then the Golgi apparatus supports the notion of a slow turnover of Cx49-GFP compared to other connexins analyzed so far. Gap junction plaques resembling the usual punctuate distribution pattern could be demonstrated for COS-1 and MDCK cells. Basolateral distribution of Cx49-GFP was observed in polar MDCK cells, indicating specific sorting behavior of Cx49 in polarized cells. Together, this report describes the first characterization of biosynthesis and trafficking of lens Cx49.Fritz Thyssen-Stiftun

Synergistic Antibacterial Effects of Polyphenolic Compounds from Olive Mill Wastewater

Polyphenols or phenolic compounds are groups of secondary metabolites widely distributed in plants and found in olive mill wastewater (OMW). Phenolic compounds as well as OMW extracts were evaluated in vitro for their antimicrobial activity against Gram-positive (Streptococcus pyogenes and Staphylococcus aureus) and Gram-negative bacteria (Escherichia coli and Klebsiella pneumoniae). Most of the tested phenols were not effective against the four bacterial strains when tested as single compounds at concentrations of up to 1000 μg mL−1. Hydroxytyrosol at 400 μg mL−1 caused complete growth inhibition of the four strains. Gallic acid was effective at 200, and 400 μg mL−1 against S. aureus, and S. pyogenes, respectively, but not against the gram negative bacteria. An OMW fraction called AntiSolvent was obtained after the addition of ethanol to the crude OMW. HPLC analysis of AntiSolvent fraction revealed that this fraction contains mainly hydroxytyrosol (10.3%), verbascoside (7.4%), and tyrosol (2.6%). The combinations of AntiSolvent/gallic acid were tested using the low minimal inhibitory concentrations which revealed that 50/100–100/100 μg mL−1 caused complete growth inhibition of the four strains. These results suggest that OMW specific fractions augmented with natural phenolic ingredients may be utilized as a source of bioactive compounds to control pathogenic bacteria

Ex vivo biochemical analysis of CFTR in human rectal biopsies

AbstractThis report describes the first biosynthetic analysis of the cystic fibrosis transmembrane conductance regulator (CFTR) in freshly excised human rectal biopsies. Expression of functional CFTR was assessed by intestinal current measurement (ICM) prior to biosynthetic studies. Several structural features of CFTR are found to be comparable to those established in CFTR-expressing cell lines. Interestingly, maturation of CFTR increases substantially in tissue incubated at 26 °C. Our data provide a solid basis for future studies on the characterisation of CFTR in pathological cases