Genetic and physical mapping of DNA replication origins in Haloferax volcanii

The halophilic archaeon Haloferax volcanii has a multireplicon genome, consisting of a main chromosome, three secondary chromosomes, and a plasmid. Genes for the initiator protein Cdc6/Orc1, which are commonly located adjacent to archaeal origins of DNA replication, are found on all replicons except plasmid pHV2. However, prediction of DNA replication origins in H. volcanii is complicated by the fact that this species has no less than 14 cdc6/orc1 genes. We have used a combination of genetic, biochemical, and bioinformatic approaches to map DNA replication origins in H. volcanii. Five autonomously replicating sequences were found adjacent to cdc6/orc1 genes and replication initiation point mapping was used to confirm that these sequences function as bidirectional DNA replication origins in vivo. Pulsed field gel analyses revealed that cdc6/orc1-associated replication origins are distributed not only on the main chromosome (2.9 Mb) but also on pHV1 (86 kb), pHV3 (442 kb), and pHV4 (690 kb) replicons. Gene inactivation studies indicate that linkage of the initiator gene to the origin is not required for replication initiation, and genetic tests with autonomously replicating plasmids suggest that the origin located on pHV1 and pHV4 may be dominant to the principal chromosomal origin. The replication origins we have identified appear to show a functional hierarchy or differential usage, which might reflect the different replication requirements of their respective chromosomes. We propose that duplication of H. volcanii replication origins was a prerequisite for the multireplicon structure of this genome, and that this might provide a means for chromosome-specific replication control under certain growth conditions. Our observations also suggest that H. volcanii is an ideal organism for studying how replication of four replicons is regulated in the context of the archaeal cell cycle. © 2007 Norais et al

Managing Learning Experiences In An AACSB Environment: Beyond The Classroom

The study explores the development and management of a rich learning environment that extends the traditional classroom to include significant co-curricular programs. Learning enrichment is guided by the individual mission of the business school, accreditation agency (AACSB), and in our case, the Jesuit mission. That central framework provides a student centric focus to achieve our mission as well as our specific Assurance of Learning objectives. Key concepts discussed include identifying management models/approaches, how to measure the richness of the learning experience, maintaining Assurance of Learning standards, as well as a variety of implementation issues

Predictors of breastfeeding duration among women in Kuwait: results of a prospective cohort study

The purposes of this paper are to report the prevalence of breastfeeding to six months among women in Kuwait and to determine the factors that are associated with the duration of breastfeeding. A cohort of 373 women recruited from maternity wards in four hospitals in Kuwait city were followed from birth to 26 weeks postpartum. The association of any and full breastfeeding duration and predictor variables were explored using multivariate Cox’s proportional hazards models. At six months, 39% of all infants were receiving some breast milk and only 2% of infants had been fully breastfed to 26 weeks. Women born in other Arab countries were less likely to discontinue breastfeeding than women born in Kuwait. Other factors positively associated with breastfeeding duration were level of maternal education, higher parity, infant being demand fed in hospital and a preference for breastfeeding on the part of the infant’s father and maternal grandmother. The introduction of a pacifier before four weeks of age and the mother intending to return to work by six months were negatively associated with duration. These findings present a number of opportunities for prolonging breastfeeding duration in Kuwait

Software for Autonomous Spacecraft Maneuvers

The AutoCon computer programs facilitate and accelerate the planning and execution of orbital control maneuvers of spacecraft while analyzing and resolving mission constraints. AutoCon-F is executed aboard spacecraft, enabling the spacecraft to plan and execute maneuvers autonomously; AutoCon-G is designed for use on the ground. The AutoCon programs utilize advanced techniques of artificial intelligence, including those of fuzzy logic and natural-language scripting, to resolve multiple conflicting constraints and automatically plan maneuvers. These programs can be used to satisfy requirements for missions that involve orbits around the Earth, the Moon, or any planet, and are especially useful for missions in which there are requirements for frequent maneuvers and for resolution of complex conflicting constraints. During operations, the software targets new trajectories, places and sizes maneuvers, and controls spacecraft burns. AutoCon-G provides a userfriendly graphical interface, and can be used effectively by an analyst with minimal training. AutoCon-F reduces latency and supports multiple-spacecraft and formation-flying missions. The AutoCon architecture supports distributive processing, which can be critical for formation- control missions. AutoCon is completely object-oriented and can easily be enhanced by adding new objects and events. AutoCon-F was flight demonstrated onboard GSFC's EO-1 spacecraft flying in formation with Landsat-7

Practicing Collective Biography

Collective biography uses researchers\u27 written memories about a set of experiences as texts for collective analysis. As a feminist approach to research, collective biography draws centrally on the idea that significant memories are critical in the constitution of the self, and maintains that in analyzing memories collectively, researchers can begin to tap into wider social processes and structures. Though rarely used in geography, collective biography could be useful in data collection and analysis for geographers. In this paper, we provide a brief history and description of collective biography. We situate collective biography in relation to life writing methods. We then identify a set of attributes that mark collective biography as a distinct research approach. In closing, we reflect on our experiences working with collective biography

Protein detection using hydrogel-based molecularly imprinted polymers integrated with dual polarisation interferometry

A polyacrylamide-based molecularly imprinted polymer (MIP) was prepared for bovine haemoglobin (BHb). A 3 mg/ml solution of BHb was injected over a dual polarisation interferometer (DPI) sensor to form a physisorbed layer typically of 3.5 ± 0.5 nm thickness. Onto the pre-adsorbed protein layer, MIP and NIP (non-imprinted polymer) were separately injected to monitor the interaction of BHb MIP or NIP particles under different loading conditions with the pre-adsorbed protein layer. In the case of NIP flowing of the protein layer, there was negligible surface stripping of the pre-adsorbed protein. When a protein-eluted sample of MIP particles was flowed over a pre-adsorbed protein layer on the sensor chip, the sensor detected significant decreases in both layer thickness and mass, suggestive that protein was being selectively bound to MIP after being stripped-off from the sensor surface. We also integrated thin-film MIPS for BHb and BSA onto the DPI sensor surface and were able to show that whereas BHb bound selectively and strongly to the BHb MIP thin film (resulting in a sustained increase in thickness and mass), the BHb protein only demonstrated transient and reversible binding on the BSA MIP. MIPs were also tested after biofouling with plasma or serum at various dilutions. We found that serum at 1/100 dilution allowed the MIP to still function selectively. This is the first demonstration of MIPs being integrated with DPI in the development of synthetic receptor-based optical protein sensors. © 2012 Elsevier B.V. All rights reserved

TGFβ-induced osteogenic potential of human amniotic fluid stem cells via CD73-generated adenosine production

The human amniotic fluid stem cell (hAFSC) population consists of two morphologically distinct subtypes, spindle-shaped and round-shaped cells (SS-hAFSCs and RS-hAFSCs). Whilst SS-hAFSCs are routinely expanded in mesenchymal-type (MT) conditions, we previously showed that they acquire broader differentiation potential when cultured under embryonic-type (ET) conditions. However, the effects of culture conditions on RS-hAFSCs have not been determined. Here, we show that culturing RS-hAFSCs under ET conditions confers faster proliferation and enhances the efficiency of osteogenic differentiation of the cells. We show that this occurs via TGFβ-induced activation of CD73 and the associated increase in the generation of extracellular adenosine. Our data demonstrate that culture conditions are decisive for the expansion of hAFSCs and that TGFβ present in ET conditions causes the phenotype of RS-hAFSCs to revert to an earlier state of stemness. Cultivating RS-hAFSCs in ET conditions with TGFβ may therefore increase their therapeutic potential for clinical applications

Generation of High Dose Inhalable Effervescent Dispersions against Pseudomonas aeruginosa Biofilms

Purpose: Novel particle engineering approach was used in this study to generate high dose inhalable effervescent particles with synergistic effects against Pseudomonas aeruginosa biofilms. Methods: Spray dried co-amorphous salt of ciprofloxacin (CFX) and tartaric acid (TA) was prepared and coated with external layer of sodium bicarbonate and silica coated silver nanobeads. Design of experiments (DOE) was used to optimize physicochemical properties of particles for enhanced lung deposition. Results: Generated particles were co-amorphous CFX/TA showing that CFX lost its zwitterionic form and exhibiting distinct properties to CFX/HCl as assessed by FTIR and thermal analysis. Particles exhibited mass mean aerodynamic diameter (MMAD) of 3.3 μm, emitted dose of 78% and fine particle dose of 85%. Particles were further evaluated via antimicrobial assessment of minimum inhibitory concentrations (MIC) and minimum biofilm eradication concentration (MBEC). MIC and MBEC results showed that the hybrid particles were around 3–5 times more effective when compared to CFX signifying that synergistic effect was achieved. Diffusing wave spectroscopy results showed that the silver containing particles had a disruptive effect on rheological properties as opposed to silver free particles. Conclusions: Overall, these results showed the potential to use particle engineering to generate particles that are highly disruptive of bacterial biofilms