The zinc finger proteins Mxr1p and repressor of phosphoenolpyruvate carboxykinase (ROP) have the same DNA binding specificity but regulate Methanol Metabolism antagonistically in pichia pastoris

The methanol-inducible alcohol oxidase I (AOXI) promoter of the methylotrophic yeast, Pichia pastoris, is used widely for the production of recombinant proteins. AOXI transcription is regulated by the zinc finger protein Mxr1p (methanol expression regulator 1). ROP (repressor of phosphoenolpyruvate carboxykinase, PEPCK) is a methanol- and biotin starvation-inducible zinc finger protein that acts as a negative regulator of PEPCK in P. pastoris cultured in biotin-deficient, glucose-ammonium medium. The function of ROP during methanol metabolism is not known. In this study, we demonstrate that ROP represses methanol-inducible expression of AOXI when P. pastoris is cultured in a nutrient-rich medium containing yeast extract, peptone, and methanol (YPM). Deletion of the gene encoding ROP results in enhanced expression of AOXI and growth promotion whereas overexpression of ROP results in repression of AOXI and growth retardation of P. pastoris cultured in YPM medium. Surprisingly, deletion or overexpression of ROP has no effect on AOXI gene expression and growth of P. pastoris cultured in a minimal medium containing yeast nitrogen base and methanol (YNBM). Subcellular localization studies indicate that ROP translocates from cytosol to nucleus of cells cultured in YPM but not YNBM. In vitro DNA binding studies indicate that AOXI promoter sequences containing 5' CYCCNY 3' motifs serve as binding sites for Mxr1p as well as ROP. Thus, Mxr1p and ROP exhibit the same DNA binding specificity but regulate methanol metabolism antagonistically in P. pastoris. This is the first report on the identification of a transcriptional repressor of methanol metabolism in any yeast species.

ROP Gadgets hiding techniques in Open Source Projects

Today there are many techniques that allows to exploit vulnerabilities of an application; there are also many techniques that are designed to stop these exploit attacks. This thesis wants to highlight how a specific type of attack, based on a technique called Return Oriented Programming (ROP), can be easily applied to binaries with particular characteristics. A new method that allows the injection of "useful" code in an Open Source projects without arousing suspicions is presented; this is possible because of the harmless aspects of the injected code. This useful code facilitate a ROP attack against an executable that contains vulnerable bugs. The injection process can be visualized in environment where an user can contribute with own code to a particular Open Source project. This thesis also highlights how current software protections are not correctly applied to Open Source project, thus enabling the proposed approach.

Cuidados de enfermería en neonatos con oxigenoterapia en la prevención de retinopatía (ROP)

El uso de oxígeno como forma de oxigenoterapia, es uno de los tratamientos que más se utilizan en las unidades de cuidado intensivo neonatal. El uso inadecuado y desmedido del mismo, puede llegar a traer graves consecuencias que afectan y atentan de manera silenciosa si no se mantiene un control periódico y una valoración constante a los recién nacidos bajo este tratamiento. Una de las principales patologías que se producen por causa del oxígeno, es la retinopatía del prematuro (ROP). Los objetivos de este estudio son: determinar el cumplimiento del rol del enfermero en la prevención de ROP con oxigenoterapia, indagar conocimientos de los enfermeros en cuidados de recién nacidos con oxigenoterapia, analizar las normas existentes, comparar las normas establecidas con las observaciones realizadas y caracterizar el rol de enfermería.

ROP-1, an RNA quality-control pathway component, affects Caenorhabditis elegans dauer formation

Caenorhabditis elegans dauer formation is an alternative larval developmental pathway that the worm can take when environmental conditions become detrimental. Animals can survive several months in this stress-resistant stage and can resume normal development when growth conditions improve. Although the worms integrate a variety of sensory information to commit to dauer formation, it is currently unknown whether they also monitor internal cellular damage. The Ro ribonucleoprotein complex, which was initially described as a human autoantigen, is composed of one major 60-kDa protein, Ro60, that binds to one of four small RNA molecules, designated Y RNAs. Ro60 has been shown to bind mutant 5S rRNA molecules in Xenopus oocytes, suggesting a role for Ro60 in 5S rRNA biogenesis. Analysis of ribosomes from a C. elegans rop-1(−) strain, which is null for the expression of Ro60, demonstrated that they contain a high percentage of mutant 5S rRNA molecules, thereby strengthening the notion of a link between the rop-1 gene product and 5S rRNA quality control. The Ro particle was recently shown to be involved in the resistance of Deinococcus radiodurans to UV irradiation, suggesting a role for the Ro complex in stress resistance. We have studied the role of rop-1 in dauer formation. We present genetic and biochemical evidence that rop-1 interacts with dauer-formation genes and is involved in the regulation of the worms' entry into the dauer stage. Furthermore, we find that the rop-1 gene product undergoes a proteolytic processing step that is regulated by the dauer formation pathway via an aspartic proteinase. These results suggest that the Ro particle may function in an RNA quality-control checkpoint for dauer formation.

Az európai uniós támogatások hatékonysága és hatásossága közötti különbség a ROP 1.2 intézkedésnél (The difference between the efficiency and effectiveness of using the EU-subsidies at Regional Operational Program)

A szerző cikkében megpróbál rávilágítani annak fontosságára, hogy az Európai Unióból érkező fejlesztési támogatások hasznosulása és a támogatási rendszer hatékonysága között jelentős különbség van. Ha nem tudjuk hatékonyan és hatásosan felhasználni az Unióból érkező pénzügyi eszközöket, akkor a cél: a kohézió, a konvergencia az Európai Unió régi tagállamainak fejlettségi szintjéhez, jólétéhez még nehezebben és lassabban érhető el. Az uniós támogatások hatékonysága alapvetően a rendelkezésre álló pénz lekötésének, lehívásának arányából, illetve a lekötött, lehívott és esetleg visszafizetett összeg arányából állapítható meg (kvantitatív megközelítés). A támogatások felhasználásának hatékonyságánál bonyolultabb, jóval összetettebb megközelítést igényel a hatásosság fogalma. A felhasználás hatásosságát a projektszinten a támogatás hatására megtermelt hozzáadott értékkel, programszinten a GDP hozzáadott növekedéssel lehet kifejezni. A cikk alapvetően a ROP 1.2 "Turisztikai fogadóképesség javítása" pályázati kiírás nyertes projektjeinél végzett kutatás eredményein keresztül kívánja szemléltetni a projektszintű vagy mikro hatásosság fontosságára (kvalitatív megközelítés). _________ The author tries to highlight the importance the difference between the efficiency and effectiveness of using the EU-subsidies. If Hungary cannot use the financial means of the EU efficiently and effectively, than the goal: cohesion and convergence to the level of the old, developed countries of the EU will be much harder and slowly. The efficiency of the EU-subsidies can be measured by the ratio of the amount of money obliged and the amount of money spent, and by the amount of money withdrawn by the Commission, which is actually lost (quantitative approach). The effectiveness of EU-subsidies needs a much more complicated and complex approach, than the efficiency. The effectiveness of usage on project level can be measured by the "added value" of the project, on program level by the added GDP growth or employment. The article is analysing basically the results of a survey made among the winner project of the application ROP 1.2 (Regional Operational Program 1.2) Improving the capacity for tourism (project level or micro effectiveness – qualitative approach).

Synthesis of 4-arm star poly(L-Lactide) oligomers using an in situ-generated calcium-based initiator

Using an in situ-generated calcium-based initiating species derived from pentaerythritol, the bulk synthesis of well-defined 4-arm star poly(L-lactide) oligomers has been studied in detail. The substitution of the traditional initiator, stannous octoate with calcium hydride allowed the synthesis of oligomers that had both low PDIs and a comparable number of polymeric arms (3.7 – 3.9) to oligomers of similar molecular weight. Investigations into the degree of control observed during the course of the polymerization found that the insolubility of pentaerythritol in molten L-lactide resulted in an uncontrolled polymerization only when the feed mole ratio of L-lactide to pentaerythritol was 13. At feed ratios of 40 and greater, a pseudo-living polymerization was observed. As part of this study, in situ FT-Raman spectroscopy was demonstrated to be a suitable method to monitor the kinetics of the ring-opening polymerization (ROP) of lactide. The advantages of using this technique rather than FT-IR-ATR and 1H NMR for monitoring L-lactide consumption during polymerization are discussed.

Policing of road safety in Oman: Perceptions and beliefs of traffic police officers

With increasing motorisation, road safety has become a major concern within Oman. Internationally, traffic policing plays a major role in improving road safety. Within Oman, the Royal Oman Police's (ROP) Directorate General of Traffic is responsible for policing traffic laws. Many common enforcement approaches originate from culturally different jurisdictions. The ROP is a relatively young policing force and may have different operational practices. Prior to applying practices from other jurisdictions it is important to understand the beliefs and expectations within the Directorate General of Traffic. Further, there is a need for individuals to understand their role and what is expected of them. Therefore, it is important to explore the agreement between levels of the ROP to determine how strategies and expectations transfer within the organisation. Interviews were conducted with 19 police officers from various levels of the ROP. A number of themes and findings emerged. Individuals at the upper level of the traffic police had a clear knowledge of the role of the ROP, believed that traffic police know what is expected of them, are well trained in their role and can have a very positive influence on road safety. These beliefs were less certain lower within the organisations with traffic officers having little knowledge of the role of the ROP or what was expected of them, felt undertrained, and believed their peers have little positive impact on road safety. There is a need to address barriers within the ROP in order to positively impact road safety.

Prevalence and characteristics of road traffic injuries among young drivers in Oman, 2009-2011

Objectives Studies from different parts of the world have indicated that the impact of road traffic incidents disproportionally affects young adults. Few known studies have been forthcoming from Arabian Gulf countries. Within Oman, a high proportion of the population is under the age of 20. Coupled with the drastic increase of motorization in recent years there is a need to understand the state of road safety among young people in Oman. The current research aimed to explore the prevalence and characteristics of road traffic injuries among young drivers aged 17-25 years. Methods Crash data from 2009-2011 was extracted from the Directorate General of Traffic, Royal Oman Police (ROP) database in Oman. The data was analyzed to explore the impact of road crashes on young people (17-25 years), the characteristics of young driver crashes and how these differ from older drivers and to identify key predictors of fatalities in young driver crashes. Results Overall, young people were over-represented in injuries and fatalities within the sample time period. While it is true that many young people in crashes were driving at the time, it was also evident that young people were often a victim in a crash caused by someone else. Thus, to reduce the impact of road crashes on young people, there is a need to generally address road safety within Oman. When young drivers were involved in crashes they were predominantly male. The types of crashes these drivers have can be broadly attributed to risk taking and inexperience. Speeding and night time driving were the key risk factors for fatalities. Conclusion The results highlight the need to address young driver safety in Oman. From these findings, the introduction of a graduated driver licensing system with night time driving restrictions could significantly improve young driver safety.

Catabolite repression of phosphoenolpyruvate carboxykinase by a zinc finger protein under biotin- and pyruvate carboxylase-deficient conditions in Pichia pastoris

We have identified a methanol- and biotin-starvation-inducible zinc finger protein named ROP [repressor of phosphoenolpyruvate carboxykinase (PEPCK)] in the methylotrophic yeast Pichia pastoris. When P. pastoris strain GS115 (wild-type, WT) is cultured in biotin-deficient, glucose-ammonium (Bio(-)) medium, growth is suppressed due to the inhibition of anaplerotic synthesis of oxaloacetate, catalysed by the biotin-dependent enzyme pyruvate carboxylase (PC). Deletion of ROP results in a strain (Delta ROP) that can grow under biotin-deficient conditions due to derepression of a biotin- and PC-independent pathway of anaplerotic synthesis of oxaloacetate. Northern analysis as well as microarray expression profiling of RNA isolated from WT and Delta ROP strains cultured in Bio(-) medium indicate that expression of the phosphoenolpyruvate carboxykinase gene (PEPCK) is induced in Delta ROP during biotin- or PC-deficiency even under glucose-abundant conditions. There is an excellent correlation between PEPCK expression and growth of Delta ROP in Bio(-) medium, suggesting that ROP-mediated regulation of PEPCK may have a crucial role in the biotin- and PC-independent growth of the Delta ROP strain. To our knowledge, ROP is the first example of a zinc finger transcription factor involved in the catabolite repression of PEPCK in yeast cells cultured under biotin- or PC-deficient and glucose-abundant conditions.

Trm1p, a Zn(II)(2)Cys(6)-type transcription factor, is essential for the transcriptional activation of genes of methanol utilization pathway, in Pichia pastoris

The zinc finger transcription factors Mxr1p and Rop are key regulators of methanol metabolism in the methylotrophic yeast, Pichia pastoris, while Trm1p and Trm2p regulate methanol metabolism in Candida boidinii. Here, we demonstrate that Trm1p is essential for the expression of genes of methanol utilization (mut) pathway in P. pastoris as well. Expression of AOXI and other genes of mut pathway is severely compromised in P. pastoris Delta Trm1 strain resulting in impaired growth on media containing methanol as the sole source of carbon. Trm1p localizes to the nucleus of cells cultured on glucose or methanol. The zinc finger domain of Mxr1p but not Trm1p binds to AOXI promoter sequences in vitro, indicating that these two positive regulators act by different mechanisms. We conclude that both Trm1p and Mxr1p are essential for the expression of genes of mut pathway in P. pastoris and the mechanism of transcriptional regulation of mut pathway may be similar in P. pastoris and C. boidinii. (C) 2014 Elsevier Inc. All rights reserved.

Biodegradable amphiphilic block copolymers bearing protected hydroxyl groups: Synthesis and characterization

A new biodegradable amphiphilic block copolymer, poly(ethylene glycol)-b-poly(L-factide-co-9-phenyl-2,4,8, 10-tetraoxaspiro[5,5]undecan-3-one) [PEG-b-P(LA-co-PTO)], was successfully prepared by ring-opening polymerization (ROP) Of L-lactide (LA) and functionalized carbonate monomer 9-phenyl-2,4,8,10-tetraozaspiro[5,5]undecan-3-one (PTO) in the presence of monohydroxyl poly(ethylene glycol) as macroinitiator using Sn(Oct)(2) as catalyst. NMR, FT-IR, and GPC studies confirmed the copolymer structure.