Transitions to K–12 Education Systems: Experiences from Five Case Countries

Currently more than 140 countries offer, or are in transition to, what has become the international norm for pre-tertiary education, namely a kindergarten through grade 12 (K–12) school education system—kindergarten because of the preponderance of research asserting the long-term learning and social benefits of school readiness programs; and 12 years of primary and secondary schooling due to the time needed to acquire the knowledge and skills sets necessary for 21st century university education, postsecondary training, or decent1 work. This desk study2 conveys the experiences of four countries and one province in preparing and implementing a transition to a K–12 school education system: Mongolia, Ontario (Canada), the Philippines, Poland, and Turkey. Looking at K–12 transition in countries and systems that vary as broadly as this set enables common threads to stand out and divergent options to be noted.

Embedding Indigenous knowledges: An Australian case study of urban and remote teaching practicum

In this chapter we propose that there are certain conditions that enable the agency of pre-service teachers to enact curriculum decision-making within their pedagogical relationships with their supervising teachers as they endeavour to embed Indigenous knowledges (IK) during the teaching practicum. The case study, underpinned by decolonising methodologies, centred upon pre-service teacher preparation at one Australian university, where we investigated how role modelling in urban and remote schools occurred in the learning and teaching relationships between pre-service teachers on practicum and their supervising teachers. This chapter draws from an Office of Learning and Teaching (OLT) sponsored project at one Australian university; a full report on this project has been documented (see McLaughlin, Whatman and Nielsen, 2014). We commence with a discussion of decolonising and critical pedagogical spaces as the conceptual framework for the embedding Indigenous knowledges and perspectives in curricula and pedagogy. Our focus then shifts to a contextual overview of the development of Indigenous Knowledges (IK) in Australian school and university curriculum, providing a standpoint from which to consider the unfolding case study.

Inquiry learning: A pedagogical and curriculum framework for information literacy

This chapter presents an inquiry learning framework that can be used as a pathway for the development of information literacy in both K-12 and higher education. Inquiry learning is advocated as an authentic and active approach that draws upon students’ natural curiosity. The pedagogical and curriculum framework incorporates three major elements: questioning frameworks, information literacy and an iterative research cycle. Models and strategies for the elements of the framework are presented and discussed. The chapter ends with an acknowledgement of the challenges associated with implementing inquiry learning.

Process makes perfect: High-performance design creates healthy environments for high-performance learning

Considering the staggering benefits of high-performance schools, it seems an obvious choice to “go green.” High-performance schools offer an exceptionally cost-effective means to enhance student learning, using on average 33 percent less energy than conventionally designed schools, and provide substantial health gains, including reduced respiratory problems and absenteeism. According to the 2006 study, Greening America's Schools, Costs and Benefits, co-sponsored by the American Institute of Architects (AIA) and Capital E, a green building consulting firm, high-performance lighting is a key element of healthy learning environments, contributing to improved test scores, reduced off-task behavior, and higher achievement among students. Few argue this point more convincingly than architect Heinz Rudolf, of Portland-Oregon-based Boora Architects, who has designed sustainable schools for more than 80 school districts in Oregon, Washington, Colorado, and Wyoming, and has pioneered the high-performance school movement. Boora's recently completed project, the Baker Prairie Middle School in Canby, Oregon is one of the most sustainable K-12 facilities in the state, and illustrates Rudolf's progressive and research-intensive approach to school design.

sRNAscanner: A Computational Tool for Intergenic Small RNA Detection in Bacterial Genomes

Background:Bacterial non-coding small RNAs (sRNAs) have attracted considerable attention due to their ubiquitous nature and contribution to numerous cellular processes including survival, adaptation and pathogenesis. Existing computational approaches for identifying bacterial sRNAs demonstrate varying levels of success and there remains considerable room for improvement. Methodology/Principal Findings: Here we have proposed a transcriptional signal-based computational method to identify intergenic sRNA transcriptional units (TUs) in completely sequenced bacterial genomes. Our sRNAscanner tool uses position weight matrices derived from experimentally defined E. coli K-12 MG1655 sRNA promoter and rho-independent terminator signals to identify intergenic sRNA TUs through sliding window based genome scans. Analysis of genomes representative of twelve species suggested that sRNAscanner demonstrated equivalent sensitivity to sRNAPredict2, the best performing bioinformatics tool available presently. However, each algorithm yielded substantial numbers of known and uncharacterized hits that were unique to one or the other tool only. sRNAscanner identified 118 novel putative intergenic sRNA genes in Salmonella enterica Typhimurium LT2, none of which were flagged by sRNAPredict2. Candidate sRNA locations were compared with available deep sequencing libraries derived from Hfq-co-immunoprecipitated RNA purified from a second Typhimurium strain (Sittka et al. (2008) PLoS Genetics 4: e1000163). Sixteen potential novel sRNAs computationally predicted and detected in deep sequencing libraries were selected for experimental validation by Northern analysis using total RNA isolated from bacteria grown under eleven different growth conditions. RNA bands of expected sizes were detected in Northern blots for six of the examined candidates. Furthermore, the 5'-ends of these six Northern-supported sRNA candidates were successfully mapped using 5'-RACE analysis. Conclusions/Significance: We have developed, computationally examined and experimentally validated the sRNAscanner algorithm. Data derived from this study has successfully identified six novel S. Typhimurium sRNA genes. In addition, the computational specificity analysis we have undertaken suggests that similar to 40% of sRNAscanner hits with high cumulative sum of scores represent genuine, undiscovered sRNA genes. Collectively, these data strongly support the utility of sRNAscanner and offer a glimpse of its potential to reveal large numbers of sRNA genes that have to date defied identification. sRNAscanner is available from: http://bicmku.in:8081/sRNAscanner or http://cluster.physics.iisc.ernet.in/sRNAscanner/.

Under-representation of intrinsic terminators across bacterial genomic islands: Rho as a principal regulator of xenogenic DNA expression

Two transcription termination mechanisms - intrinsic and Rho-dependent - have evolved in bacteria. The Rho factor occurs in most bacterial lineages, and has been hypothesized to play a global regulatory role. Genome-wide studies using microarray, 2D-gel electrophoresis and ChIP-chip provided evidence that Rho serves to silence transcription from horizontally acquired genes and prophages in Escherichia coli K-12, implicating the factor to be a part of the ``cellular immune mechanism'' protecting against deleterious phages and aberrant gene expression from acquired xenogenic DNA. We have investigated this model by adopting an alternate in silico approach and have extended the study to other species. Our analysis shows that several genomic islands across diverse phyla have under-representation of intrinsic terminators, similar to that experimentally observed in E. coli K-12. This implies that Rho-dependent termination is the predominant process operational in these islands and that silencing of foreign DNA is a conserved function of Rho. From the present analysis, it is evident that horizontally acquired islands have lost intrinsic terminators to facilitate Rho-dependent termination. These results underscore the importance of Rho as a conserved, genome-wide sentinel that regulates potentially toxic xenogenic DNA. (C) 2012 Elsevier B.V. All rights reserved.

Functional Analysis of the Genes Encoding Diaminopropionate Ammonia Lyase in Escherichia coli and Salmonella enterica Serovar Typhimurium

Diaminopropionate ammonia lyase (DAPAL) is a pyridoxal-5'phosphate (PLP)-dependent enzyme that catalyzes the conversion of diaminopropionate (DAP) to pyruvate and ammonia and plays an important role in cell metabolism. We have investigated the role of the ygeX gene of Escherichia coli K-12 and its ortholog, STM1002, in Salmonella enterica serovar Typhimurium LT2, presumed to encode DAPAL, in the growth kinetics of the bacteria. While Salmonella Typhimurium LT2 could grow on DL-DAP as a sole carbon source, the wild-type E. coli K-12 strain exhibited only marginal growth on DL-DAP, suggesting that DAPAL is functional in S. Typhimurium. The expression of ygeX in E. coli was low as detected by reverse transcriptase PCR (RT-PCR), consistent with the poor growth of E. coli on DL-DAP. Strains of S. Typhimurium and E. coli with STM1002 and ygeX, respectively, deleted showed loss of growth on DL-DAP, confirming that STM1002 (ygeX) is the locus encoding DAPAL. Interestingly, the presence of DL-DAP caused a growth inhibition of the wild-type E. coli strain as well as the knockout strains of S. Typhimurium and E. coli in minimal glucose/glycerol medium. Inhibition by DL-DAP was rescued by transforming the strains with plasmids containing the STM1002 (ygeX) gene encoding DAPAL or supplementing the medium with Casamino Acids. Growth restoration studies using media lacking specific amino acid supplements suggested that growth inhibition by DL-DAP in the absence of DAPAL is associated with auxotrophy related to the inhibition of the enzymes involved in the biosynthetic pathways of pyruvate and aspartate and the amino acids derived from them.

Síntese, caracterização e avaliação da capacidade bactericida de copolímeros à base de 2-vinilpiridina funcionalizados

Neste trabalho, dois copolímeros à base de 2-vinilpiridina (2Vpy), estireno (Sty) e divinilbenzeno (DVB) foram sintetizados empregando a técnica de polimerização em suspensão aquosa via radical livre. Os copolímeros com diferentes características morfológicas foram preparados variando a composição da mistura diluente, constituída por tolueno e n-heptano, solventes solvatantes e não solvatantes para as cadeias poliméricas. A caracterização estrutural desses materiais foi feita através de medidas de área específica, volume de poros, diâmetro médio de poros e densidade aparente. Além disso, esses materiais foram avaliados por microscopia ótica e eletrônica de varredura, por espectroscopia na região do infravermelho (FTIR), análise termogravimétrica (TGA) e análise elementar. Os copolímeros Sty-DVB-2Vpy foram modificados quimicamente através de reação de quaternização das unidades de 2Vpy usando dois reagentes: iodeto de metila e acrilonitrila. A quaternização dos copolímeros Sty-DVB-2Vpy foi confirmada através de espectroscopia de infravermelho, pelo aparecimento das bandas de absorção características do íon peridíneo e do grupo nitrila e também pela análise do comportamento térmico. Os produtos dessas reações foram submetidos à avaliação da capacidade bactericida através do método de contagem em placas contra suspensão de Escherichia coli. O copolímero do tipo gel quaternizado com iodeto de metila apresentou a maior ação bactericida registrada, com eficiência até a concentração de 104 células/mL. Contudo, a maioria dos materiais quaternizados não apresentou ação biocida significativa. Com o objetivo de maximizar a atividade bactericida dos materiais preparados, os copolímeros quaternizados e não quaternizados foram impregnados com iodo por meio de duas metodologias: em solução e em fase vapor. O teor de iodo incorporado foi quantificado por análise gravimétrica. Foi possível observar que os copolímeros quaternizados e impregnados com iodo se mostraram mais eficientes como agentes bactericidas que os copolímeros não funcionalizados impregnados com iodo. De uma forma geral, foi possível perceber que a ação bactericida dos polímeros é fruto da associação entre as suas características de porosidade, o grau de quaternização alcançado e o teor de iodo incorporado. Para efeito de comparação foram feitos também ensaios bactericidas com uma resina comercial à base de Sty-DVB com grupo amônio quaternário, VP OC 1950. Os testes mostraram que a resina comercial não possui atividade bactericida. A impregnação de iodo a essa resina comercial forneceu um material com ação biocida semelhante à do copolímero do tipo gel, quaternizado com iodeto de metila e impregnado com iodo

退火对ZrO2薄膜微结构及激光损伤阈值的影响

利用电子束蒸发方法在Yb:YAG晶体和熔融石英衬底上沉积单层ZrO2薄膜,分别在673 K和1 073 K的温度下经过12 h退火以后,通过X射线衍射(XRD)分析了薄膜晶相,计算了薄膜的晶粒尺寸;利用表面热透镜技术获得了薄膜的吸收;测量了退火后薄膜的激光损伤阈值.实验结果表明:两种衬底上的薄膜结构受到退火温度和衬底表面结构的影响,高温退火有利于单斜相的形成,含单斜相的ZrO2薄膜具有较高的激光损伤阈值,而由于衬底的吸收,Yb:YAG晶体上薄膜的损伤阈值远小于石英衬底上薄膜的损伤阈值.

Ta2O5膜在532, 800 和 1064 nm波长的n-on-1体系下的激光损伤阈值研究

Ta2O5膜采用传统的电子束蒸发方法制备，并在氧气中673 K的条件下进行了退火12 h处理。首先在1-on-1体系下对Ta2O5膜进行了532和1064 nm波长下的激光损伤阈值（LIDT） 研究，然后在n-on-1体系下分别对532，800和1064 nm三种波长下的激光损伤阈值进行了研究。结果表明在n-on-1体系下Ta2O5膜在532和1064 nm波长下的阈值均高于1-on-1体系下的阈值。此外，在n-on-1体系下，薄膜的阈值随波长增加而增大。同时发现，在氧气中进行退火对Ta2O5膜的光学性能

基于错配修复系统的基因突变检测生物芯片研究

许多人类疾病和微生物抗药性的产生都是由基因组中单个碱基的替换、插入或缺失等基因突变引起的。因此，迫切需要发展快速、高通量基因突变检测方法来实现对基因疾病和细菌抗药性的早期诊断。本研究针对匕述需求发展了纂于DN八错配修复系统的墓因突变检测生物芯片方法。根据DNA错配修复MtltS蛋白结构与功能上的高度保守性，通过PCR从E.coli K-12基因组中扩一增出DNA错配修复基因，甩石（2.56kb）。通过基因水平的分子操纵，构建了Trx-His6-MutS（THM）、Trx-His6-Linker peptide-Muts（THLM）、Trx-His6-GFP-Linker peptide-MutS（THGLM）和Trx-His6-Linker peptide-Strep-tagll-Linker peptide-MutS（THLSLM）的融合基因并在大肠杆菌中进行了IPTG诱导表达。SDS-PAGE分析表明均有一与预期分子量相应的诱导表达条带出现，其表达量占菌体蛋白的30％左右，且以可溶形式存在。融合蛋白中Trx和His6亲和肤能增加表达蛋白的可溶性及便于蛋白的纯化。连接肤的加入增大了融合蛋白各个成分之间的距离，减少空间位阻，使各个蛋白能够较大程度地保持其原有的生物活性。MLltS融合蛋白的生物活性鉴定结果表明：它们既能识别、结合含有错配碱基的DNA双链，又保留了其它融合成分的生物活性。利用融合蛋白THLSLM中的Strep-tagII与Streptavidin相互作用的天然特性，使融合蛋白THLSLM在StrePtavidin修饰过的芯片基质上自动布阵沉积，制作成蛋白质芯片来识别、结合样品中含有错配或未配刘碱基的DNA双链。THGLM、THLM-Cy3和THLSLM能够使MutS蛋白显示不同的标记信号，通过它们识别并结合固定在DNA芯片基质上的基因片段来发展基因突变检测DNA芯片方法。利用基于MutS的蛋白质芯片和DNA芯片方法对含有不同错配类型、不同长度的DNA片段和错配序列背景对错配结合的影响做了深入研究，证明了MutS介导的基因突变检测生物芯片方法的可行性。基于MutS蛋白的鳌因突变检测生物芯片方法借用了生物系统本身的DNA错配修复（Mismatch Repair，MMR）机制。DNA错配修复过程是许多修复蛋白之间的相互作用共同完成的，其中蛋白MutS、MutL和MutH在肠道细菌例如大肠杆菌的甲基定向错配修复中起决定作用。这些修复蛋白的相关研究也引起了越来越多学者的关注，但对于MutL蛋白的体外生物功能一直存在争议，从而限制了该蛋白的应用研究。本研究利用基因的体外拼接技术构建了融合蛋白Trx-Hi56-Linker peptide-MutL（THLL）、Trx-His6-GFP-Linker peptide-MutL（THGLL）和Trx-His6-Linker peptide-Strep-tagII-Linker peptide-MutL（THLSLL）。非变性凝胶电泳鉴定MutL融合蛋白体外生物功能结果表明：THLL、THGLL和THLsLL都能增加融合蛋白Trx-His6-Linker peptide-MutS（THLM）与含有错配碱基DNA双链的结合，但受ATP浓度变化的影响很大。通过融合蛋白THGLL中绿色荧光蛋白（Green Fluorescent Protein，GFP）的荧光信号或THLSLL中Strep-tagII的特性并利用酶学反应来指示该蛋白的存在，发展了体外研究DNA错配修复蛋白MtuS和MutL之间相互作用的简便方法。本研究以构建的MutS融合蛋白为分子识别元件发展了基因突变检测生物芯片并利用构建的MutL融合蛋白发展了体外研究DNA错配修复蛋白MLuS和MutL之间相互作用的简便方法。建立的融合分子系统方法也为研究其它的蛋白质或生物大分子之间的相互作用提供了一个技术平台。此外，本研究构建的融合蛋白THGLL及其 DNA错配修复蛋白与GFP的融合构想还可用来进行DNA错配修复基因产物的表达与基因突变频率和人类肿瘤恶性程度的相关性研究。

Effects of solution viscosity on heterogeneous electron transfer across a liquid/liquid interface

Scanning electrochemical microscopy (SECM) is employed to investigate the effect of solution viscosity on the rate constants of electron transfer (ET) reaction between potassium ferricyanide in water and 7,7,8,8-tetracyanoquinodimethane (TCNQ) in 1,2-dichloroethane. Either tetrabutylammonium (TBA(+)) or ClO4- is chosen as the common ion in both phases to control the interfacial potential drop. The rate constant of heterogeneous ET reaction between TCNQ and ferrocyanide produced in-situ, k(12), is evaluated by SECM and is inversely proportional to the viscosity of the aqueous solution and directly proportional to the diffusion coefficient of K4Fe(CN)(6) in water when the concentration of TCNQ in the DCE phase is in excess. The k(12) dependence on viscosity is explained in terms of the longitudinal relaxation time of the solution. The rate constant of the heterogeneous ET reaction between TCNQ and ferricyanide, k(21), is also obtained by SECM and these results cannot be explained by the same manner.

Photoluminescent multilayer films based on polyoxometalates

Ultrathin multilayer films consisting of the polyoxotungstoeuropate cluster K-12[EuP5W30O110] (EuP5W30) and poly( allylamine hydrochloride) (PAH) have been prepared by the layer-by-layer self-assembly method. The (EuP5W30 /PAH) multilayer films have been characterized by small-angle X-ray reflectivity measurements, X-ray photoelectron spectra, and atomic force microscopy (AFM). From the AFM images, the thickness of the {PEI/PSS/PAH(EuW30/PAH)} multilayer film was estimated to be 6.5 nm, corresponding to an average thickness of ca. 1.1 nm for a EuW30/PAH layer pair. The photoluminescent behavior of the film at room temperature was investigated to show the characteristic Eu3+ emission pattern of D-5(0)-->F-7(J). The fluorescence behavior of the multilayer film is essentially identical to that of H-n[EuP5W30O110]((12-n)-) in a concentrated aqueous solution, except for the relative intensities and peak bandwidths. The occurence of photoluminescent activity confirms the potential for creating luminescent multilayers with polyoxometalates (see ref. 23).

Antisense Gene Silencing and Bacteriophages as Novel Disinfection Processes for Engineered Systems

The growth and proliferation of invasive bacteria in engineered systems is an ongoing problem. While there are a variety of physical and chemical processes to remove and inactivate bacterial pathogens, there are many situations in which these tools are no longer effective or appropriate for the treatment of a microbial target. For example, certain strains of bacteria are becoming resistant to commonly used disinfectants, such as chlorine and UV. Additionally, the overuse of antibiotics has contributed to the spread of antibiotic resistance, and there is concern that wastewater treatment processes are contributing to the spread of antibiotic resistant bacteria.

Due to the continually evolving nature of bacteria, it is difficult to develop methods for universal bacterial control in a wide range of engineered systems, as many of our treatment processes are static in nature. Still, invasive bacteria are present in many natural and engineered systems, where the application of broad acting disinfectants is impractical, because their use may inhibit the original desired bioprocesses. Therefore, to better control the growth of treatment resistant bacteria and to address limitations with the current disinfection processes, novel tools that are both specific and adaptable need to be developed and characterized.

In this dissertation, two possible biological disinfection processes were investigated for use in controlling invasive bacteria in engineered systems. First, antisense gene silencing, which is the specific use of oligonucleotides to silence gene expression, was investigated. This work was followed by the investigation of bacteriophages (phages), which are viruses that are specific to bacteria, in engineered systems.


For the antisense gene silencing work, a computational approach was used to quantify the number of off-targets and to determine the effects of off-targets in prokaryotic organisms. For the organisms of Escherichia coli K-12 MG1655 and Mycobacterium tuberculosis H37Rv the mean number of off-targets was found to be 15.0 + 13.2 and 38.2 + 61.4, respectively, which results in a reduction of greater than 90% of the effective oligonucleotide concentration. It was also demonstrated that there was a high variability in the number of off-targets over the length of a gene, but that on average, there was no general gene location that could be targeted to reduce off-targets. Therefore, this analysis needs to be performed for each gene in question. It was also demonstrated that the thermodynamic binding energy between the oligonucleotide and the mRNA accounted for 83% of the variation in the silencing efficiency, compared to the number of off-targets, which explained 43% of the variance of the silencing efficiency. This suggests that optimizing thermodynamic parameters must be prioritized over minimizing the number of off-targets. In conclusion for the antisense work, these results suggest that off-target hybrids can account for a greater than 90% reduction in the concentration of the silencing oligonucleotides, and that the effective concentration can be increased through the rational design of silencing targets by minimizing off-target hybrids.

Regarding the work with phages, the disinfection rates of bacteria in the presence of phages was determined. The disinfection rates of E. coli K12 MG1655 in the presence of coliphage Ec2 ranged up to 2 h-1, and were dependent on both the initial phage and bacterial concentrations. Increasing initial phage concentrations resulted in increasing disinfection rates, and generally, increasing initial bacterial concentrations resulted in increasing disinfection rates. However, disinfection rates were found to plateau at higher bacterial and phage concentrations. A multiple linear regression model was used to predict the disinfection rates as a function of the initial phage and bacterial concentrations, and this model was able to explain 93% of the variance in the disinfection rates. The disinfection rates were also modeled with a particle aggregation model. The results from these model simulations suggested that at lower phage and bacterial concentrations there are not enough collisions to support active disinfection rates, which therefore, limits the conditions and systems where phage based bacterial disinfection is possible. Additionally, the particle aggregation model over predicted the disinfection rates at higher phage and bacterial concentrations of 108 PFU/mL and 108 CFU/mL, suggesting other interactions were occurring at these higher concentrations. Overall, this work highlights the need for the development of alternative models to more accurately describe the dynamics of this system at a variety of phage and bacterial concentrations. Finally, the minimum required hydraulic residence time was calculated for a continuous stirred-tank reactor and a plug flow reactor (PFR) as a function of both the initial phage and bacterial concentrations, which suggested that phage treatment in a PFR is theoretically possible.

In addition to determining disinfection rates, the long-term bacterial growth inhibition potential was determined for a variety of phages with both Gram-negative and Gram-positive bacteria. It was determined, that on average, phages can be used to inhibit bacterial growth for up to 24 h, and that this effect was concentration dependent for various phages at specific time points. Additionally, it was found that a phage cocktail was no more effective at inhibiting bacterial growth over the long-term than the best performing phage in isolation.

Finally, for an industrial application, the use of phages to inhibit invasive Lactobacilli in ethanol fermentations was investigated. It was demonstrated that phage 8014-B2 can achieve a greater than 3-log inactivation of Lactobacillus plantarum during a 48 h fermentation. Additionally, it was shown that phages can be used to protect final product yields and maintain yeast viability. Through modeling the fermentation system with differential equations it was determined that there was a 10 h window in the beginning of the fermentation run, where the addition of phages can be used to protect final product yields, and after 20 h no additional benefit of the phage addition was observed.

In conclusion, this dissertation improved the current methods for designing antisense gene silencing targets for prokaryotic organisms, and characterized phages from an engineering perspective. First, the current design strategy for antisense targets in prokaryotic organisms was improved through the development of an algorithm that minimized the number of off-targets. For the phage work, a framework was developed to predict the disinfection rates in terms of the initial phage and bacterial concentrations. In addition, the long-term bacterial growth inhibition potential of multiple phages was determined for several bacteria. In regard to the phage application, phages were shown to protect both final product yields and yeast concentrations during fermentation. Taken together, this work suggests that the rational design of phage treatment is possible and further work is needed to expand on this foundation.