Designing Primers with a Plant Signal Peptide to Enhance the Expression of GBA1 in Transgenic Soybean Plants

تقدم النباتات المعدلة وراثيا مزايا لإنتاج بروتينات مركبة حاوية على جزيئات سكر المانوزفي طرف سلاسل الغليكان، لذلك يتم اختيار النباتات كمصدر للمنتجات الصيدلانيه بالاضافه الى تطوير انظمة تعبير بديله لانتاج إنزيمات الليزوزوميه المركبة. في الدراسة الحالية التتابع الجيني الطبيعي لجزيئة الديوكسي رايبوز ثلاثي الفوسفات المتممه والتي تشفر لانتاج الانزيم البشري الغلوكوسيربيروزيداز حورت لتعزيز التعبير الجيني لهذا الانزيم داخل نبات فول الصويا،حيث ان اشارة البدا لجين GBA تم استبدالها باشارة البدا لجين انزيم اندوجينيز لنبات Arabidopsis thaliana وذلك لتسهيل انتقال البروتين المركب الى الشبكه الاندوبلازميه كما تم اضافه التتابع الخاص باشاره جين الجيتينيز لنبات التبغ والمسؤل عن خزن البروتين المركب داخل حويصلات الخزن وذلك بتصميم بادئات خاصه لتضخيم الجين البشري GBA حاويه على كلا من اشاره النقل الى الشبكه الاندوبلازميه واشاره النقل الى الحويصلات الخازنه. استخدمت تقنية PCR لتضخيم Hu-GBA من البلازمد المركب PGEM-GBA بعدها تم كلونة الجين المضخم داخل ناقل التعبير النباتي المسمى Agrobacterium tumefaciens سلالةLBA4404 والتي بدورها استخدمت لتحويل فلقات نبات فول الصويا، حيث أن الفلقه وبعمر 5ايام وبارتفاع0.5 سم قطعت واستخدمت كنموذج للاستزراع وبعد اصابة الفلقات بالبكتريا المحوله واعادة استنباتها استخدم مضاد الهايكرومايسين بي في وسط الاختبار لاختيار الفلقات المحورة وراثيا. استخدمت تقنيهPCR لتحديد وجود GBA في جينومك النبات المحور وكانت نتيجة التضخيم حزمه بحجم 1587زوج قاعدي، كما تم الكشف عن التعبير الجيني للجين في نباتات فول الصويا المحور بواسطة الريال تايم مقارنة مع Hu-GBA القياسيTransgenic plants offer advantages for the manufacture of recombinant proteins with terminalmannose residues on their glycan chains. So plants are chosen as source of pharmaceutical products and forthe development of alternative expression systems to produce recombinant lysosomal enzymes. In thepresent study the sequence of the natural cDNA encoding for the human lysosomal enzymeglucocerebrosidase (GCD) was modified to enhance its expression in soybean plants. The glucocerebrosidasegene signal peptide was substituted with that signal peptide for the Arabidopsis thaliana basic endochitinasegene to support the co-translational translocation into the endoplasmic reticulum (ER), and the storagevacuole. So, targeting signal from tobacco chitinase A, to facilitate GCD trafficking from the ER to thestorage vacuole, appropriate primers were designed containing both an ER and vacuolar targeting signals,(VTS). Those primers were used for PCR amplification of the human GBA gene (Hu-GBA) gene fromconstructed PGEM-GBA plasmid which was cloned in the plant expression vector pCAMBIA1304. Theresulted construct was transported in Agrobacterium tumefaciens strain LBA4404 and was used fortransformation of cotyledon explants. After 5-day of seedling, cotyledons were cut and used as explants.After infection and co-cultivation, hygromycin B was added in selection media as a selective agent for thetransformants cotyledons. The presence of the Hu-GBA transgene in the genomes of transgenic plants wasdetermined by polymerase chain reaction PCR as a band of size1587 bp. The GBA mRNA expression inmodified soybean was detected by qRT-PCR compared with control GBA mRNA

Soseske učenja: Vseživljenjsko učenje, skupnost in trajnostni razvoj v učečem se mestu Cork

Cork Learning City is piloting a programme called Learning Neighbourhoods. While it is an evolving model, it has potential for animating a culture of learning that is community based and with the prospect of giving ordinary citizens a chance to contribute both to broader issues at city level as well as to significant global debates of the day. The Learning Neighbourhood concept is essentially about collaboration and coalition building, extending networks and prioritising learning (horizontally and vertically) among and between generations and the positive leverage of all and every resource to enhance the process. It is a community building concept articulated through learning. Learning is not something abstract and removed from the practicalities of living. Learning encapsulates the four types of capital, human, social, identity, and cultural; it also extends to capacity building, resilience, and the challenges of the sustainable development goals (SDGs). This paper contextualises the theory and practice informing Learning Neighbourhoods.Učeče se mesto Cork poskusno izvaja program, imenovan Soseske učenja. Čeprav je še v razvojni fazi, ima program zmožnost za spodbujanje kulture učenja, ki temelji na skupnosti in omogoča ljudem, da sodelujejo pri reševanju pomembnih vprašanj na mestni in globalni ravni. Zamisel soseske učenja temelji na sodelovanju in združevanju, na širjenju omrežij in dajanju prednosti učenju (horizontalno in vertikalno) tako znotraj posamezne generacije kot med njimi ter na učinkoviti uporabi prav vseh virov pri krepitvi tega procesa. Na takšen način se z učenjem gradi skupnost. To pa ni nekaj abstraktnega ali nekaj, kar bi bilo ločeno od vsakdanjega življenja. Učenje obsega štiri vrste kapitala, človeškega, družbenega, identitetnega in kulturnega, nanaša pa se tudi na področja krepitve zmogljivosti in odpornosti ter soočanja z izzivi, ki nam jih prinašajo cilji trajnostnega razvoja. Članek predstavlja teorijo in prakso, na podlagi katerih je oblikovan koncept sosesk učenja

Judicial review under the Irish Constitution: More American than Commonwealth

The Irish legal system emerged from the British system. Yet the Irish constitution diverges from the Commonwealth model and more closely resembles the American constitution in both substance and application. This means that while the Irish parliamentary system is very much based on the Westminster model, the legal system, while remaining within the Common Law family, has important distinctions. One of the key areas of divergence from the Commonwealth model is in judicial review. This has important ramifications for the recognition and elaboration of human rights under the constitution. This article explores some of the historical reasons for the shift from the Commonwealth model and the nature and consequence of some of those differences. It also contrasts certain aspects of the Irish judicial review process with those pertaining in the United States; in particular it highlights some anti-democratic tendencies in the Irish system

Aesthetic Experience and Place Configurations in the Poetry of Zakia Malollah

يقدّم هذا البحث دراسة جديدة في موضوعها؛ إذ يعالج التجربة الجمالية في شعر زكيّة مال الله، من خلال تعاملها مع المكان وتشكيلها إياه، بوصفه وسيلة للتعبير عن تجربة جمالية. وهنا يظهر المكان على أنه مشبّه به، أو استعارة، أو كناية، أو رمزٌ، أو معادلٌ موضوعيٌّ لحالة، إضافة إلى ظهوره بوصفه مكانًا أيضًا، تتم معالجته بتشكيل مكاني. وقد جرى تفحّص هذه الوسيلة المكانية للتعبير من خلال تجارب جمالية عدة، تشمل جوانب الحياة والكون والمجتمع، منها تجربة المكان نفسه، والبحث عن الذات، والحب، والوضع الاجتماعي للمرأة، وحب الإله. وقد تبيّن ظهور المكان في التجربة الجمالية عند زكيّة مال الله ظهورًا واضحًا، وموظَّفًا بتقنية فنية عالية، وتأكّد من خلال البحث ارتباط المكان بعمق التجربة الجمالية عند الشاعرة، وهو ما قاد إلى ظهوره الفني في تشكيلات جمالية متنوعة.This study deals with the aesthetic experience in the poetry of Zakia Malollah, through its focus on place formation as a means for articulating an aesthetic experience. Place is created as a metaphor, a symbol, or an objective reckoning of a condition, in addition to its being a spatial formation. Place as a spatial medium of expression has been examined through several aesthetic experiences, covering aspects of life, the universe, and society, including the experience of the place itself, the search for self, love, the social status of women, and the love of God. Place in the aesthetic experience stands out in Zakia Malollah’s work, and it is technically well-employed. Research avers that place is related to the deep aesthetic experience of the poet, which in turn has resulted in its artistic appearance in a variety of aesthetic formations

International pricing and distribution of therapeutic pharmaceuticals – An ethical minefield

This paper seeks to identify and consider ethical issues relating to the international pricing of pharmaceuticals, drawing especially on liberal rights theories. It suggests why and how some of these issues might be resolved. It examines and critiques arguments presented by major pharmaceutical manufacturers. It addresses a range of ancillary issues like current pricing policies, R&D, intellectual property rights, rights to profits, the public good and regulation. It proposes a potential model for moving forward on the pricing of pharmaceuticals, with a view to increasing access to essential drugs

Limiting key drilling parameters to control mud losses in Basra\u27s oil fields, Iraq

Wells in Basra\u27s oil fields are highly susceptible to lost circulation problems when drilling through the Dammam, Hartha, and Shuaiba formations. Drilling engineers are challenged to select the optimum value for key drilling parameters such as ECD, Yp, MW, WOB, ROP, and RPM to mitigate mud losses in these formations. Many of these drilling parameters are inter-related and the overall impact of changing key parameters require extensive drilling experience and study. A multi-regression analysis was performed on mud loss event data for more than 300 wells drilled in Basra\u27s oil fields. From this analysis, a model was developed to predict the total mud losses. Mud loss was found to be significantly affected by MW, ECD, and ROP. Models for ECD and ROP were also developed from the multi-regression analysis. In the same vein, proactive approaches are made prior entering the Dammam, Hartha, and Shuiaba formations to prevent or mitigate the occurrence of the lost circulation. Key drilling parameters are estimated to use during drilling through these formations. In case preventive measures didn\u27t work, corrective actions are determined for each type of the mud losses to provide effective remedies, minimize NPT, and reduce cost. This dissertation will contribute toward a better understanding of how the different factors could affect the lost circulation which is believed to have a significant impact on the mitigation mud losses. Therefore, optimized preventive and corrective solutions will be designed for combating lost circulation --Abstract, page iv

A comprehensive study of lost circulation with application in the South Rumaila field, Iraq

“Drilling mud losses and problems associated with lost circulation while drilling account for a major expense in drilling oil and gas wells. By industry estimates, more than 2 billion USD is spent annually to combat and mitigate this problem (Ali et al., 2015). The South Rumaila field in Iraq is one of the largest oilfields in the world. Wells drilled in this field are highly susceptible to lost circulation problems when drilling through the Dammam, Hartha and Shuaiba formations. Lost circulation events range from seepage losses to complete loss of the borehole and are a critical issue in field development. This thesis describes a study of the lost circulation events in more 50 wells drilled in the Rumaila field. Lost circulation events were extracted from daily drilling reports. Key drilling parameters (e.g. RPM, RPM, bit type) and mud properties (e.g. density, yield point, gel strength) at the time of each event were recorded along with the lost circulation remedies attempted, and the outcome of those remedies. These data were analyzed to determine ranges for the key drilling parameters and mud properties that have the greatest chance of mitigating lost circulation in each of the three formations. Practical field information from a range of sources were reviewed and summarized to develop an integrated methodology and flowchart for handling lost circulation events in the South Rumaila Field. Best field procedures for avoiding or minimizing lost circulation events in the South Rumaila field were identified and were provided as tabled procedures, or as additional data in the appendices of this thesis. This study provides a unique compilation of information regarding traditional approaches and the latest approaches of lost circulation control. The thesis attempts to provide useful guidelines or references for both situations”--Abstract, page iii

Combined Effects of Frequency Quantization and Additive Input Noise in a First-order Digital PLL

AbstractA recent work by Gardner [Gardner, F.M., Frequency granularity in digital phase-locked loops, IEEE Trans. Commun., 44 (1996), 749758] on the subject of digital phase-locked loops (DPLLs) investigated, via simulation, the characteristics of the phase-jitter caused by frequency quantization in the numerically-controlled oscillator. Further works by Feely, Teplinsky et al [Feely, O., Rogers, A., and Teplinsky, A., Phase-jitter dynamics of digital phaselocked loops, IEEE Trans. Circuits and Systems, Part I: Fundamental Theory and Applications, 46 (1999), 545–558], [Feely, O., and Teplinsky, A., Phase-jitter dynamics of digital phase-locked loops: Part II, IEEE Trans. Circuits and Systems, Part I: Fundamental Theory and Applications., 47 (2000), 458–473] used the theory of nonlinear dynamics to provide a complete analytical explanation of this phase-jitter.This paper examines in detail the case where the input signal is embedded in additive noise, a scenario earlier investigated by Gardner where no satisfactory method of characterising the phase-jitter was found. Here, further numerical results for the 1-D DPLL are presented and it is shown analytically how the DPLL noise-jitter dynamics may be approximated by a noisy circle rotation map for reasonable levels of additive noise. The noise in this case is unique and highly nonlinear in nature and thus not amenable to traditional analysis. By considering the the probability flow over time, a time-dependent difference-delay equation is derived for the probability density function (PDF) of the phase-jitter. It is shown that this PDF reaches a steady-state and that this state is described by a non-local equation. The solutions of this equation are investigated, both numerically and analytically, and used to explain the interaction between the additive and quantization noise that was previously not understood