Mapping sites of interaction of p47-phox and flavocytochrome b with random-sequence peptide phage display libraries.

During assembly of the phagocyte NADPH oxidase, cytosolic p47-phox translocates to the plasma membrane and binds to flavocytochrome b, and binding domains for p47-phox have been identified on the C-terminal tails of both flavocytochrome b subunits. In the present report, we further examine the interaction of these two oxidase components by using random-sequence peptide phage display library analysis. Screening p47-phox with the peptide libraries identified five potential sites of interaction with flavocytochrome b, including three previously reported regions of interaction and two additional regions of interaction of p47-phox with gp91-phox and p22-phox. The additional sites were mapped to a domain on the first predicted cytosolic loop of gp91-phox encompassing residues S86TRVRRQL93 and to a domain near the cytosolic C-terminal tail of gp91-phox encompassing residues F450EWFADLL457. The mapping also confirmed a previously reported binding domain on gp91-phox (E554SGPRGVHFIF564) and putative Src homology 3 domain binding sites on p22-phox (P156PRPP160 and G177GPPGGP183). To demonstrate that the additional regions identified were biologically significant, peptides mimicking the gp91-phox sequences F77LRGSSACCSTRVRRQL93 and E451WFADLLQLLESQ463 were synthesized and assayed for their ability to inhibit NADPH oxidase activity. These peptides had EC50 values of 1 microM and 230 microM, respectively, and inhibited activation when added prior to assembly but did not affect activity of the preassembled oxidase. Our data demonstrate the usefulness of phage display library analysis for the identification of biologically relevant sites of protein-protein interaction and show that the binding of p47-phox to flavocytochrome b involves multiple binding sites along the C-terminal tails of both gp91- and p22-phox and other regions of gp91-phox nearer to the N terminus.

Investigating and Understanding Student Learning Outcomes in an Online and Face-to-Face Graduate Level Legal Administration Course: An Embedded Mixed Methods Design

Online education is a new teaching and learning medium with few current guidelines for faculty, administrators or students. Its rapid growth over the last decade has challenged academic institutions to keep up with the demand, while also providing a quality education. Our understanding of the factors that determine quality and effective online learning experiences that lead to student learning outcomes is still evolving. There is a lack of consensus on the effectiveness of online versus face-to-face education in the current research. The U.S. Department of Education conducted a meta-analysis in 2009 and concluded that student-learning outcomes in online courses were equal to and, often times, better than face-to-face traditional courses. Subsequent research has found contradictory findings, and further inquiry is necessary. The purpose of this embedded mixed methods design research study is to further our understanding of the factors that create quality and successful educational outcomes in an online course. To achieve this, the first phase of this study measured and compared learning outcomes in an online and in class graduate-level legal administration course. The second phase of the study entailed interviews with those students in both the online and face-to-face sections to understand their perspectives on the factors contributing to learning outcomes. Six themes emerged from the qualitative findings: convenience, higher order thinking, discussions, professor engagement, professor and student interaction, and face-to-face interaction. Findings from this study indicate the factors students perceive as contributing to learning outcomes in an online course are consistent among all students and are supported in the existing literature. Higher order thinking, however, emerged as a stronger theme than indicated in the current research, and the face-to-face nature of the traditional classroom may be more an issue of familiarity than a factor contributing to learning outcomes. As education continues to reach new heights and developments in technology advance, the factors found to contribute to student learning outcomes will be refined and enhanced. These developments will continue to transform the ways in which we deliver and receive knowledge in both traditional and online classrooms. While there is a growing body of research on online education, the field’s evolution has unsettled earlier findings and posed new areas to investigate.

Interaction of peptides, peptidomimetics and other drug candidates with the DI/tripeptide transport system in intestinal epithelial cells, using the in vitro caco-2 cell culture system

An uptake system was developed using Caco-2 cell monolayers and the dipeptide, glycyl-[3H]L-proline, as a probe compound. Glycyl-[3H]L-proline uptake was via the di-/tripeptide transport system (DTS) and, exhibited concentration-, pH- and temperature-dependency. Dipeptides inhibited uptake of the probe, and the design of the system allowed competitors to be ranked against one another with respect to affinity for the transporter. The structural features required to ensure or increase interaction with the DTS were defined by studying the effect of a series of glycyl-L-proline and angiotensin-converting enzyme (ACE)-inhibitor (SQ-29852) analogues on the uptake of the probe. The SQ-29852 structure was divided into six domains (A-F) and competitors were grouped into series depending on structural variations within specific regions. Domain A was found to prefer a hydrophobic function, such as a phenyl group, and was intolerant to positive charges and H+ -acceptors and donors. SQ-29852 analogues were more tolerant of substitutions in the C domain, compared to glycyl-L-proline analogues, suggesting that interactions along the length of the SQ-29852 molecule may override the effects of substitutions in the C domain. SQ-29852 analogues showed a preference for a positive function, such as an amine group in this region, but dipeptide structures favoured an uncharged substitution. Lipophilic substituents in domain D increased affinity of SQ-29852 analogues with the DTS. A similar effect was observed for ACE-NEP inhibitor analogues. Domain E, corresponding to the carboxyl group was found to be tolerant of esterification for SQ-29852 analogues but not for dipeptides. Structural features which may increase interaction for one series of compounds, may not have the same effect for another series, indicating that the presence of multiple recognition sites on a molecule may override the deleterious effect of anyone change. Modifying current, poorly absorbed peptidomimetic structures to fit the proposed hypothetical model may improve oral bioavailability by increasing affinity for the DTS. The stereochemical preference of the transporter was explored using four series of compounds (SQ-29852, lysylproline, alanylproline and alanylalanine enantiomers). The L, L stereochemistry was the preferred conformation for all four series, agreeing with previous studies. However, D, D enantiomers were shown in some cases to be substrates for the DTS, although exhibiting a lower affinity than their L, L counterparts. All the ACE-inhibitors and β-lactam antibiotics investigated, produced a degree of inhibition of the probe, and thus show some affinity for the DTS. This contrasts with previous reports that found several ACE inhibitors to be absorbed via a passive process, thus suggesting that compounds are capable of binding to the transporter site and inhibiting the probe without being translocated into the cell. This was also shown to be the case for oligodeoxynucleotide conjugated to a lipophilic group (vitamin E), and highlights the possibility that other orally administered drug candidates may exert non-specific effects on the DTS and possibly have a nutritional impact. Molecular modelling of selected ACE-NEP inhibitors revealed that the three carbonyl functions can be oriented in a similar direction, and this conformation was found to exist in a local energy-minimised state, indicating that the carbonyls may possibly be involved in hydrogen-bond formation with the binding site of the DTS.

The interaction of sodium chlorite with phospholipids and glutathione:a comparison of effects in vitro, in mammalian and in microbial cells

In this study the interaction of the preservative sodium chlorite with unsaturated lipids and glutathione was investigated, in comparison with peroxides, sodium hypochlorite, and benzalkonium chloride. The aim was to determine whether the action of sodium chlorite could involve membrane lipid damage or antioxidant depletion, and how this related to toxicity in both mammalian and microbial cells. The treatment of phospholipids with chlorite yielded low levels of hydroperoxides, but sodium chlorite oxidized the thiol-containing antioxidant glutathione to its disulfide form very readily in vitro, with a 1:4 oxidant:GSH stoichiometry. In cultured cells, sodium chlorite also caused a substantial depletion of intracellular glutathione, whereas lipid oxidation was not very prominent. Sodium chlorite had a lower toxicity to ocular mammalian cells than benzalkonium chloride, which could be responsible for the different effects of long-term application in the eye. The fungal cells, which were most resistant to sodium chlorite, maintained higher percentage levels of intracellular glutathione during treatment than the mammalian cells. The results show that sodium chlorite can cause oxidative stress in cells, and suggest that cell damage is more likely to be due to interaction with thiol compounds than with cell membrane lipids. The study also provides important information about the differential resistance of ocular cells and microbes to various preservatives and oxidants.

A paradigm shift:alternative interaction techniques for use with mobile & wearable devices

Desktop user interface design originates from the fact that users are stationary and can devote all of their visual resource to the application with which they are interacting. In contrast, users of mobile and wearable devices are typically in motion whilst using their device which means that they cannot devote all or any of their visual resource to interaction with the mobile application -- it must remain with the primary task, often for safety reasons. Additionally, such devices have limited screen real estate and traditional input and output capabilities are generally restricted. Consequently, if we are to develop effective applications for use on mobile or wearable technology, we must embrace a paradigm shift with respect to the interaction techniques we employ for communication with such devices.This paper discusses why it is necessary to embrace a paradigm shift in terms of interaction techniques for mobile technology and presents two novel multimodal interaction techniques which are effective alternatives to traditional, visual-centric interface designs on mobile devices as empirical examples of the potential to achieve this shift.

An analysis of Haitian parents' perceptions of their children with disabilities

Parental involvement is an integral part of the educational system in the U.S. Yet, parents from non-mainstream racial/ethnic backgrounds have not fully grasped the nature of parental involvement expectations in the educational process and how these expectations may impact student achievement. The purpose of this study was to identify Haitian parents’ perceptions of their children with disabilities and the education these children were receiving. Several authors have conducted studies on parents of children with disabilities to better gain an understanding of the level of their involvement with their children’s education, their perceptions of the children, and their views on the school system (Harry, 1992a, 1992b). In this study, Haitian parents of children with disabilities were interviewed using an interview protocol. Through these interviews, this study explored 10 Haitian parents’ perceptions of their child with a disability, the education the child was receiving, their interaction with the school system, and how the disability had affected their relationship with their child and their involvement with the school. Findings of the present study revealed that these Haitian parents seldom disagreed with school personnel and did not seem to fully grasp the different methods available to address their concerns as parents of children with disabilities nor the role they were expected to play in the process. The majority did not have basic literacy skills in Creole or English. The parents in this study were overwhelmed by school written communication. Additionally, this study discovered that parents’ perceptions were guided by two core concepts: coping mechanisms and locus of control. Parents with an internal locus of control, who tended to be more educated, focused inward to find solutions to problems encountered. Those with an external locus of control relied on outside influences to resolve their problems. Parental involvement was strongly influenced by their values, beliefs, customs, and conceptual knowledge about disability; all closely aligned with culture and acculturation. Overall, these parents’ perceptions greatly influenced their thoughts and behaviors when they realized that their children with disabilities might fall short of their immigrant dreams of success they held for these children.

An Analysis of Haitian Parents' Perceptions of Their Children With Disabilities

Parental involvement is an integral part of the educational system in the U.S. Yet, parents from non-mainstream racial/ethnic backgrounds have not fully grasped the nature of parental involvement expectations in the educational process and how these expectations may impact student achievement. The purpose of this study was to identify Haitian parents’ perceptions of their children with disabilities and the education these children were receiving. Several authors have conducted studies on parents of children with disabilities to better gain an understanding of the level of their involvement with their children’s education, their perceptions of the children, and their views on the school system (Harry, 1992a, 1992b). In this study, Haitian parents of children with disabilities were interviewed using an interview protocol. Through these interviews, this study explored 10 Haitian parents’ perceptions of their child with a disability, the education the child was receiving, their interaction with the school system, and how the disability had affected their relationship with their child and their involvement with the school. Findings of the present study revealed that these Haitian parents seldom disagreed with school personnel and did not seem to fully grasp the different methods available to address their concerns as parents of children with disabilities nor the role they were expected to play in the process. The majority did not have basic literacy skills in Creole or English. The parents in this study were overwhelmed by school written communication. Additionally, this study discovered that parents’ perceptions were guided by two core concepts: coping mechanisms and locus of control. Parents with an internal locus of control, who tended to be more educated, focused inward to find solutions to problems encountered. Those with an external locus of control relied on outside influences to resolve their problems. Parental involvement was strongly influenced by their values, beliefs, customs, and conceptual knowledge about disability; all closely aligned with culture and acculturation. Overall, these parents’ perceptions greatly influenced their thoughts and behaviors when they realized that their children with disabilities might fall short of their immigrant dreams of success they held for these children.

School participation of students with autism spectrum disorders

This paper addresses the participation of students with autism spectrum disorders (ASD) in main- stream schools. There are different benefits for ASD students to be educated in an inclusive environment (Gena, 2006; Whitaker, 2004). They challenge the school community by presenting difficulties in essential domains for school activi- ties (Chamberlain, Kasari and Rotheram-Fuller, 2006; Eman and Farrell, 2009; Humphrey and Symes, 2010). Thus, these are students with increased difficulties participating in inclusive environments, reinforcing the need of an ade- quate inclusion process (Gena, 2006; Hall and McGregor, 2000; Hestenes and Carroll, 2000). We characterised this students ’ participation with a questionnaire to the students from mainstream classes in which ASD students were included, a questionnaire applied to each class teacher/head teacher and an interview to four of the school educational assistants. The location of the ASD student in mainstream classroom was also ana- lysed, trying to understand if it influences the quality of ASD students ’ participation, hypothesis- ing that there is an influence. Results showed a good perception of the students with ASD and their behaviour, low frequency of behaviours involving interaction with these students, good feelings about their presence at the school/class and an overall acceptance of them in the peer groups of typical development students. Results are mostly consistent across the different infor- mation sources. We found a significant effect of the location on the quality of participation. Results are mainly consistent with the literature reviewed and enlighten the need to keep making progress on inclusion practices related to ASD students in mainstream schools.

Role of the Tryptophan Residues in the Specific interaction of the Sea Anemone Stichodacty la helianthus’s Actinoporin Sticholysin II with Biological Membranes

Actinoporins are pore-forming toxins from sea anemones. Upon interaction with sphingomyelin-containing bilayers, they become integral oligomeric membrane structures that form a pore. Sticholysin II from Stichodactyla helianthus contains five tryptophans located at strategic positions; its role has now been studied using different mutants. Results show that W43 and W115 play a eterminant role in maintaining the high thermostability of the protein, while W146 provides specific interactions for protomer−protomer assembly. W110 and W114 sustain the hydrophobic effect, which is one of the major driving forces for membrane binding in the presence of Chol. However, in its absence, additional interactions with sphingomyelin are required. These conclusions were confirmed with two sphingomyelin analogues, one of which had impaired hydrogen bonding properties. The results obtained support actinoporins’ Trp residues playing a major role in membrane recognition and binding, but their residues have an only minor influence on the diffusion and oligomerization steps needed to assemble a functional pore.

Experience of problem-based learning (PBL) in virtual space : a phenomenographical study

This thesis reports the outcomes of an investigation into students’ experience of Problem-based learning (PBL) in virtual space. PBL is increasingly being used in many fields including engineering education. At the same time many engineering education providers are turning to online distance education. Unfortunately there is a dearth of research into what constitutes an effective learning experience for adult learners who undertake PBL instruction through online distance education. Research was therefore focussed on discovering the qualitatively different ways that students experience PBL in virtual space. Data was collected in an electronic environment from a course, which adopted the PBL strategy and was delivered entirely in virtual space. Students in this course were asked to respond to open-ended questions designed to elicit their learning experience in the course. Data was analysed using the phenomenographical approach. This interpretative research method concentrated on mapping the qualitative differences in students’ interpretations of their experience in the course. Five qualitatively different ways of experiencing were discovered: Conception 1: ‘A necessary evil for program progression’; Conception 2: ‘Developing skills to understand, evaluate, and solve technical Engineering and Surveying problems’; Conception 3: ‘Developing skills to work effectively in teams in virtual space’; Conception 4: ‘A unique approach to learning how to learn’; Conception 5: ‘Enhancing personal growth’. Each conception reveals variation in how students attend to learning by PBL in virtual space. Results indicate that the design of students’ online learning experience was responsible for making students aware of deeper ways of experiencing PBL in virtual space. Results also suggest that the quality and quantity of interaction with the team facilitator may have a significant impact on the student experience in virtual PBL courses. The outcomes imply pedagogical strategies can be devised for shifting students’ focus as they engage in the virtual PBL experience to effectively manage the student learning experience and thereby ensure that they gain maximum benefit. The results from this research hold important ramifications for graduates with respect to their ease of transition into professional work as well as their later professional competence in terms of problem solving, ability to transfer basic knowledge to real-life engineering scenarios, ability to adapt to changes and apply knowledge in unusual situations, ability to think critically and creatively, and a commitment to continuous life-long learning and self-improvement.

Investigating Familiarity in Older Adults to Facilitate Intuitive Interaction

This paper discusses how intuitive interaction is a possible way to increase the efficiency and effectiveness of interaction with older adults. It provides insights into existing research on intuitive interaction, and the role of prior experience and familiarity in intuition. An experiment is discussed which investigates differences in familiarity between younger and older adults. A comprehensive coding system has been developed to help analyse the data collected. This research is currently in progress.

Proton-transfer versus nontransfer in compounds of the diazo-dye precursor 4-(phenyldiazenyl) aniline (aniline yellow) with strong organic acids: the 5-sulfosalicylate and the dichroic benzenesulfonate salts, and the 1:2 adduct with 3,5-dinitrobenzoic

The structures of two 1:1 proton-transfer red-black dye compounds formed by reaction of aniline yellow [4-(phenyldiazenyl)aniline] with 5-sulfosalicylic acid and benzenesulfonic acid, and a 1:2 nontransfer adduct compound with 3,5-dinitrobenzoic acid have been determined at either 130 or 200 K. The compounds are 2-(4-aminophenyl)-1-phenylhydrazin-1-ium 3-carboxy-4-hydroxybenzenesulfonate methanol solvate, C12H12N3+.C7H5O6S-.CH3OH (I), 2-(4-aminophenyl)-1-hydrazin-1-ium 4-(phenydiazinyl)anilinium bis(benzenesulfonate), 2C12H12N3+.2C6H5O3S-, (II) and 4-(phenyldiazenyl)aniline-3,5-dinitrobenzoic acid (1/2) C12H11N3.2C~7~H~4~N~2~O~6~, (III). In compound (I) the diaxenyl rather than the aniline group of aniline yellow is protonated and this group subsequently akes part in a primary hydrogen-bonding interaction with a sulfonate O-atom acceptor, producing overall a three-dimensional framework structure. A feature of the hydrogen bonding in (I) is a peripheral edge-on cation-anion association involving aromatic C--H...O hydrogen bonds, giving a conjoint R1/2(6)R1/2(7)R2/1(4)motif. In the dichroic crystals of (II), one of the two aniline yellow species in the asymmetric unit is diazenyl-group protonated while in the other the aniline group is protonated. Both of these groups form hydrogen bonds with sulfonate O-atom acceptors and thee, together with other associations give a one-dimensional chain structure. In compound (III), rather than proton-transfer, there is a preferential formation of a classic R2/2(8) cyclic head-to-head hydrogen-bonded carboxylic acid homodimer between the two 3,5-dinitrobenzoic acid molecules, which in association with the aniline yellow molecule that is disordered across a crystallographic inversion centre, result in an overall two-dimensional ribbon structure. This work has shown the correlation between structure and observed colour in crystalline aniline yellow compounds, illustrated graphically in the dichroic benzenesulfonate compound.

St. John’s wort and Kava in treating major depressive disorder with comorbid anxiety : a randomised double-blind placebo-controlled pilot trial

Objective - We report the first randomised controlled trial (RCT) using a combination of St. John’s wort (SJW) and Kava for the treatment of major depressive disorder (MDD) with comorbid anxiety. Methods - Twenty-eight adults with MDD and co-occurring anxiety were recruited for a double-blind RCT. After a placebo run-in of 2 weeks, the trial had a crossover design testing SJW and Kava against placebo over two controlled phases, each of 4 weeks. The primary analyses used intention-to-treat and completer analyses. Results - On both intention-to-treat ( p¼0.047) and completer analyses ( p¼0.003), SJW and Kava gave a significantly greater reduction in self-reported depression on the Beck Depression Inventory (BDI-II) over placebo in the first controlled phase. However, in the crossover phase, a replication of those effects in the delayed medication group did not occur. Nor were there significant effects on anxiety or quality of life. Conclusion - There was some evidence of antidepressant effects using SJW and Kava in a small sample with comorbid anxiety. Possible explanations for the absence of anxiolysis may include a potential interaction with SJW, the presence of depression, or an inadequate dose of Kava.

A pilot test on the differences between domestic and international students’ perceptions towards online learning in a Malaysian institution (Part I)

Teaching to an international audience online can be significantly different as compared to a traditional classroom setting. In a traditional classroom setting, the students are usually removed from their own cultural context and required to operate in the lecturer’s context. International students coming to Malaysia to study are implicitly expected to, and often do, become familiar with the Malaysian culture and style of education. The use of educational technologies as a blended strategy in higher education programs offers challenges and opportunities for all students but this may be different for international students who come from varied backgrounds. With an increasingly competitive global demand for higher education, Malaysian institutions strive to be the hub of educational excellence and a preferred option for international students in coping with the challenges of studying abroad in a different culture. This research will evaluate how undergraduate students perceive their online learning experiences in a Malaysian university. The OLES (Online Learning Environment Survey) will be used to explore the international and domestic students’ perception on e-learning and the findings of the first six OLES scales varying from (Computer Usage, Teacher Support, Student Interaction & Collaboration, Personal Relevance, Authentic Learning, and Student Autonomy) will be reported in this research. An in-depth study will be conducted to compare and contrast the challenges of international students with domestic students. Major difficulties encountered and how these students actually cope with e-learning, as well as the strategies and tools used to overcome the challenges will be investigated.

Thai students' engagement with cooperative learning in mathematics classrooms

Over the past decade, Thai schools have been encouraged by the Thai Ministry of Education to introduce more student-centred pedagogies such as cooperative learning into their classrooms (Carter, 2006). However, prior research has indicated that the implementation of cooperative learning into Thai schools has been confounded by cultural traditions endemic within Thai schools (Deveney, 2005). The purpose of the study was to investigate how 32 Grade 3 and 32 Grade 4 students enrolled in a Thai school engaged with cooperative learning in mathematics classrooms after they had been taught cooperative learning strategies and skills. These strategies and skills were derived from a conceptual framework that was the outcome of an analysis and synthesis of social learning, behaviourist and socio-cognitive theories found in the research literature. The intervention began with a two week program during which the students were introduced to and engaged in practicing a set of cooperative learning strategies and skills (3 times a week). Then during the next four weeks (3 times a week), these cooperative learning strategies and skills were applied in the contexts of two units of mathematics lessons. A survey of student attitudes with respect to their engagement in cooperative learning was conducted at the conclusion of the six-week intervention. The results from the analysis of the survey data were triangulated with the results derived from the analysis of data from classroom observations and teacher interviews. The analysis of data identified four complementary processes that need to be considered by Thai teachers attempting to implement cooperative learning into their mathematics classrooms. The paper concludes with a set of criteria derived from the results of the study to guide Thai teachers intending to implement cooperative learning strategies and skills in their classrooms.