Structure and function of DsbA, a key bacterial oxidative folding catalyst

Since its discovery in 1991, the bacterial periplasmic oxidative folding catalyst DsbA has been the focus of intense research. Early studies addressed why it is so oxidizing and how it is maintained in its less stable oxidized state. The crystal structure of Escherichia coli DsbA (EcDsbA) revealed that the oxidizing periplasmic enzyme is a distant evolutionary cousin of the reducing cytoplasmic enzyme thioredoxin. Recent significant developments have deepened our understanding of DsbA function, mechanism, and interactions: the structure of the partner membrane protein EcDsbB, including its complex with EcDsbA, proved a landmark in the field. Studies of DsbA machineries from bacteria other than E. coli K-12 have highlighted dramatic differences from the model organism, including a striking divergence in redox parameters and surface features. Several DsbA structures have provided the first clues to its interaction with substrates, and finally, evidence for a central role of DsbA in bacterial virulence has been demonstrated in a range of organisms. Here, we review current knowledge on DsbA, a bacterial periplasmic protein that introduces disulfide bonds into diverse substrate proteins and which may one day be the target of a new class of anti-virulence drugs to treat bacterial infection. Antioxid. Redox Signal. 14, 1729–1760.

But how do we know they watched it? Adapting to the flipped classroom conundrum

Those who teach film and media need to use screen content to illustrate their subjects. For example, students want illustrations to accompany lectures on film or television genres. Our experience has been that student access to the film and television screen content underpinning a study of genres is not only desirable but is, in fact, crucial for effective teaching and learning outcomes. Not so long ago, a screening during or at the completion of a lecture was the expected method by which educators delivered screen content to illustrate their teaching. Even if student attendance fluctuated from week to week a quick head count confirmed that a certain number of students were physically present. It was assumed that this physical attendance encouraged students to reflect upon and contextualize the material post lecture. While simply attending a lecture will not translate into actual student learning, it does demonstrate a willingness by students to engage with the course content by making a commitment to attend a scheduled and recurring lecture and screening program. However, as flipped classroom models gain acceptance in educational institutions, this traditional lecture-screening model is giving way to online, off-site, and student-controlled mechanisms for screen content delivery and viewing. Nevertheless, care should be taken when assessing how online delivery translates into student engagement and learning. As Junco (2012) points out, “it’s not the technology that generates learning, but the ways in which the technology are used.” Discussed, debated, and embraced to varying degrees by educators, there remains no definitive model for the flipped classroom – although many models involve ‘flipping’ content and knowledge acquisition (including viewing films and television shows) from scheduled on-campus classes to online material viewed by students in advance of an on-campus lecture or class. The classroom or tutorial room then becomes a space to problem-solve, engage in collaborate learning, and advance and explain concepts. From an institutional perspective, the flipped classroom model could deliver an additional benefit beyond immediate pedagogical concerns. Tucker (2012) suggests through the flipped classroom model “all aspects of instruction can be rethought to best maximize the scarcest learning resource — time.” The narrative most often associated with this shift is that the move to online content delivery of lecture and cinematic / televisual material may also provide educators with more time to do other work such as engage in research, plan strategies to empower students. Experimentation with the flipped classroom model is playing out in various educational institutions. Yet several core concerns remain — one of these concerns is the crucial question of whether an online/digital flipped approach is more effective for student engagement and learning than the traditional lecture-screening mode for screen content delivery. Some urge caution in this regard, arguing that “new technology isn’t always supported by change management and professional development to ensure that digital isn’t just a goal within itself, but actually helps to transform education” (Fleming cited in Blain 2014). The most fundamental concern remains how do lecturers, instructors, and tutors know students have watched the films and television shows associated with a subject? The remainder of this discussion deals with these concerns, and possible solutions offered, through an analysis of the Film, Television and Screen Genres subject at the Queensland University of Technology (QUT) in Brisbane, Queensland.

Gene expression profiling reveals a downregulation in immune-associated genes in patients with AS

Objective: To identify differentially expressed genes in peripheral blood mononuclear cells (PBMCs) from patients with ankylosing spondylitis (AS) compared with healthy individuals. Methods: RNA was extracted from PBMCs collected from 18 patients with active disease and 18 gender-matched and age-matched controls. Expression profiles of these cells were determined using microarray. Candidate genes with differential expressions were confirmed in the same samples using quantitative reverse transcription-PCR (qRT-PCR). These genes were then validated in a different sample cohort of 35 patients with AS and 18 controls by qRT-PCR. Results: Microarray analysis identified 452 genes detected with 485 probes which were differentially expressed between patients with AS and controls. Underexpression of NR4A2, tumour necrosis factor AIP3 (TNFAIP3) and CD69 was confirmed. These genes were further validated in a different sample group in which the patients with AS had a wider range of disease activity. Predictive algorithms were also developed from the expression data using receiver-operating characteristic curves, which demonstrated that the three candidate genes have ∼80% power to predict AS according to their expression levels. Conclusions: The findings show differences in global gene expression patterns between patients with AS and controls, suggesting an immunosuppressive phenotype in the patients. Furthermore, downregulated expression of three immune-related genes was confirmed. These candidate genes were also shown to be strong predictive markers for AS.

Diagnosing adult Attention Deficit Hyperactivity Disorder: Are late onset and subthreshold ciagnoses valid?

Objective Diagnosing attention deficit hyperactivity disorder (ADHD) in adults is difficult when diagnosticians cannot establish an onset before the DSM-IV criterion of age 7 or if the number of symptoms recalled does not achieve DSM’s diagnosis threshold. Method The authors addressed the validity of DSM-IV’s age-at-onset and symptom threshold criteria by comparing four groups of adults: 127 subjects with full ADHD who met all DSM-IV criteria for childhood-onset ADHD, 79 subjects with late-onset ADHD who met all criteria except the age-at-onset criterion, 41 subjects with subthreshold ADHD who did not meet full symptom criteria for ADHD, and 123 subjects without ADHD who did not meet any criteria. The authors hypothesized that subjects with late-onset and subthreshold ADHD would show patterns of psychiatric comorbidity, functional impairment, and familial transmission similar to those seen in subjects with full ADHD. Result Subjects with late-onset and full ADHD had similar patterns of psychiatric comorbidity, functional impairment, and familial transmission. Most children with late onset of ADHD (83%) were younger than 12. Subthreshold ADHD was milder and showed a different pattern of familial transmission than the other forms of ADHD. Conclusions The data about the clinical features of probands and the pattern of transmission of ADHD among relatives found little evidence for the validity of subthreshold ADHD among such subjects, who reported a lifetime history of some symptoms that never met DSM-IV’s threshold for diagnosis. In contrast, the results suggested that late-onset adult ADHD is valid and that DSM-IV’s age-at-onset criterion is too stringent.

Applying a simulation model to the management of an infestation of Miconia calvescens in the wet tropics of Australia

A simulation model that combines biological, search and economic components is applied to the eradication of a Miconia calvescens infestation at El Arish in tropical Queensland, Australia. Information on the year M. calvescens was introduced to the site, the number of plants controlled and the timing of control, is used to show that currently there could be M. calvescens plants remaining undetected at the site, including some mature plants. Modelling results indicate that the eradication programme has had a significant impact on the population of M. calvescens, as shown by simulated results for uncontrolled and controlled populations. The model was also used to investigate the effect of changing search effort on the cost of and time to eradication. Control costs were found to be negligible over all levels of search effort tested. Importantly, results suggest eradication may be achieved within several decades, if resources are increased slightly from their current levels and if there is a long-term commitment to funding the eradication programme.

Eyeglasses of Moses Mendelssohn.

Digital Image

Letter to congregation in Jamaica asking assistance to build a synagogue

Digital image

Letter of Reverend Joseph J. Pinto to K. K. S. I. announcing his intention to return to Europe

Digital image

Letter of Joseph Jessurun Pinto to Trustees of K. K. S. I.

Digital image

Letter to Rodriques Pacheco and Daniel Gomez

Digital image

Charge-transfer interaction mediated organogels from bile acid appended anthracenes: rheological and microscopic studies

In this article we present dual-component charge-transfer interaction (CT) induced organogel formation with bile acid anthracene conjugates as donors and 2,4,7-trinitrofluorenone (TNF) as the acceptor. The use of TNF (1) as a versatile electron acceptor in the formation of gels is demonstrated through the formation of gels with different steroidal groups on the anthracene moiety in a variety of solvents ranging from aromatic hydrocarbons to long chain alcohols. Thermal stability and variable temperature fluorescence experiments were performed on these CT gels. Dynamic rheological experiments conducted on these gels suggest that these are viscoelastic soft materials and with the gel strength can be modulated by varying the donor/acceptor ratios.

Development of a New Generation of Vectors for Gene Expression, Gene Replacement, and Protein-Protein Interaction Studies in Mycobacteria

Escherichia coli-mycobacterium shuttle vectors are important tools for gene expression and gene replacement in mycobacteria. However, most of the currently available vectors are limited in their use because of the lack of extended multiple cloning sites (MCSs) and convenience of appending an epitope tag(s) to the cloned open reading frames (ORFs). Here we report a new series of vectors that allow for the constitutive and regulatable expression of proteins, appended with peptide tag sequences at their N and C termini, respectively. The applicability of these vectors is demonstrated by the constitutive and induced expression of the Mycobacterium tuberculosis pknK gene, coding for protein kinase K, a serine-threonine protein kinase. Furthermore, a suicide plasmid with expanded MCS for creating gene replacements, a plasmid for chromosomal integrations at the commonly used L5 attB site, and a hypoxia-responsive vector, for expression of a gene(s) under hypoxic conditions that mimic latency, have also been created. Additionally, we have created a vector for the coexpression of two proteins controlled by two independent promoters, with each protein being in fusion with a different tag. The shuttle vectors developed in the present study are excellent tools for the analysis of gene function in mycobacteria and are a valuable addition to the existing repertoire of vectors for mycobacterial research.

Orientation Selection and Microstructural Evolution in Directionally Solidified Tb0.3Dy0.7Fe1.95

Tb0.3Dy0.7Fe1.95 alloy was directionally solidified by using a modified Bridgman technique at a wide range of growth rates of 5 to 100 cm/h. The directionally grown samples exhibited plane front solidification morphology up to a growth rate of 90 cm/h. Typical island banding feature was observed closer to the chilled end, which eventually gave rise to irregular peritectic coupled growth (PCG). The PCG gained prominence with an increase in the growth rate. The texture study revealed formation of strong aOE (c) 311 > texture in a lower growth rate regime, aOE (c) 110 > and ``rotated aOE (c) 110 > aEuroe in an intermediate growth regime, and aOE (c) 112 > in a higher growth rate regime. In-depth analysis of the atomic configuration of a solid-liquid interface revealed that the growth texture is influenced by the kinetics of atomic attachment to the solid-liquid interface, which is intimately related to a planar packing fraction and an atomic stacking sequence of the interfacial plane. The mechanism proposed in this article is novel and will be useful in addressing the orientation selection mechanism of topologically closed packed intermetallic systems. The samples grown at a higher growth rate exhibit larger magnetostriction (lambda) and d lambda/dH owing to the absence of pro-peritectic (Tb,Dy)Fe-3 and formation of aOE (c) 112 > texture, which lies closer to the easy magnetization direction (EMD).