Recovery of intracellular materials by electroporation

Electroporation is commonly used to transfer genetic materials such as DNA into the cell by exposing the cell to a high electric field that momentarily permeabilizes the cell membrane. This premise was utilized to use electroporation as a means of releasing intracellular proteins. A procedure has been developed for the release of intracellular proteins through the use of electroporation. This procedure is a breakthrough. In general, industrial scale techniques to recover proteins from microorganisms are inefficient (Clarkson, 1993). A benefit of this electroporation-based mechanism would be the reduction of steps in the downstream processing required to purify the desired product. Yeast cells in suspensions were subjected to high voltage electric pulses by using an E.coli Pulser from BIO-RAD. The pulses permeabilized the cell and allowed the release of intracellular materials. Research has shown that the amount of released materials increased with the number of times the cells were subjected to pulses. In addition, experiments show that even more material can be released by repulsing the cells. Other parameters to affect the release of intracellular materials were optical density, growth stage, and the storage of the cell suspensions. The protein assay was found to be an effective means to determine the released intracellular product concentration

THE EFFECTS OF ATOMOXETINE ON COGNITIVE PERFORMACE AND NEUROPLASTICITY AFTER TRAUMATIC BRAIN INJURY

Catecholaminergic neurotransmission is regionally altered following injury, and drugs aimed at these systems offer promising avenues for post-TBI pharmacotherapies. Atomoxetine is a selective norepinephrine transporter (NET) inhibitor currently indicated for treatment of attention-deficit hyperactivity disorder (ADHD). The studies in this dissertation were designed to test the efficacy of atomoxetine for treating cognitive deficits following experimental TBI and the potential mechanism for any beneficial effect. The first part of the study focused on behavioral recovery following atomoxetine treatment. Several important questions of dose, therapeutic window, and duration of treatment were addressed in these studies. Sprague-Dawley rats were subjected to lateral fluid-percussion injury (L-FPI) of moderate severity (2.08 atm +/- .05). Four experiments were performed. In the first study, atomoxetine (.3 mg/kg, 1mg/kg, 3 mg/kg, or 9 mg/kg) or vehicle was administered daily on post injury days (PID) 1-15. Cognitive assessment was performed using the Morris water maze on PID 11-15. L-FPI resulted in significant cognitive impairment when compared to Sham-Injury. Treatment with lower doses of atomoxetine (.3mg/kg, 1mg/kg, and 3mg/kg) significantly attenuated the cognitive deficits in injured animals. Treatment with the higher dosage (9mg/kg) of atomoxetine resulted in animals that were not significantly different than injured-vehicle treated animals. The optimal response was achieved using 1 mg/kg atomoxetine. In the second study, treatment with atomoxetine (1mg/kg) or vehicle was delayed for 11 days post-injury. Rats were administered atomoxetine daily for 15 days and cognitive assessment was performed on PID 25-29. In this study, treatment with atomoxetine (1 mg/kg) did not result in improved cognitive performance. In the next study atomoxetine was given daily on PID 1-7 and then treatment was terminated. The animals were tested in the MWM on PID 11-15. We found that atomoxetine treatment for 7 days post-injury provides an enhancement of cognitive deficits that is not significantly different from sham animals. We then investigated whether a single treatment of atomoxetine 24 h after brain injury could influence behavioral outcome on days 11-15. From this study, we found a single dose of atomoxetine is not as effective as chronic treatment. Finally, we investigated changes in the protein expression of brain-derived neurotrophic factor, growth-associated protein-43, and synaptophysin on day 7 PID to investigate what effect atomoxetine may have on brain plasticity and regeneration. We found that atomoxetine can enhance both GAP-43 and BDNF, but not synaptophysin at this time point. In conclusion, this is the first study to show that low doses of atomoxetine initiated early after experimental traumatic brain injury results in improved cognition. Furthermore, we show that enhancement of catecholamines via atomoxetine treatment during periods of postinjury-induced plasticity can provide long-term functional and structural benefits

How can technology make this work? Preservice teachers, off-campus learning and digital portfolios

Utilising appropriate Information Communication Technologies (ICT) as instructional tools in teacher education can be a challenging yet worthwhile endeavour. This paper reports the difficulties and benefits of a recent inter-university project requiring preservice primary teachers to construct professional digital portfolios using the support of ICT. Challenges with regard to communication and learning were numerous as 34 preservice teachers (PSTs) from three universities in NSW (situated in country towns, and in Sydney) worked together as a collaborative learning community. Meeting regularly face-to-face during the 12-month process was not always possible and so ICT resources were employed to facilitate instruction and communication. An action research methodology generated a data set of surveys, observational field notes, and student assessment artefacts. We share the findings in this paper so that others may learn from our experiences in using free access software and commonplace hardware when constructing digital portfolios with students who are some distance from campus

A new genus of Protasteridae (Ophiuridea) from the Lower Devonian Bokkeveld Group of South Africa

Gamiroaster tempestatis, a new genus and species of Palaeozoic ophiuroid, is described from four specimens identified in the Lower DevonianVoorstehoek Formation (Ceres Subgroup, Bokkeveld Group) of SouthAfrica. This ophiuroid belongs the family Protaseridae, a Middle–Late Ordovician taxon that continued into the late Palaeozoic. This new ophiuroid forms part of a much wider fauna of the Malvinokaffric Realm, a biogeographical termused to denote the cool- to cold-water, high-latitude endemic, benthic marine, Devonian faunas of southwestern Gondwana, which also includes the invertebrate fossil assemblages of the Argentine Precordillera and the Fox Bay Formation of the Falkland Islands. The specimens were collected from an obrution deposit excavated on Karbonaatjies farm, ~145 km northeast of Cape Town in theWestern Cape. The excavated rock samples contain >700 articulated specimens of Gamiroaster tempestatis that are closely associated with two types of less common mitrate stylophorans. Silicone casts and high-resolution three-dimensional digital models obtained via micro-CT scanning of these mould fossils provided detailed morphological proxies for this taxonomic description.DST-NRF Centre of Excellence, Palaeosciences (CoE-Pal)JNC201

Reassortant Pandemic (H1N1) 2009 Virus in Pigs, United Kingdom

Surveillance for influenza virus in pigs in the United Kingdom during spring 2010 detected a novel reassortant influenza virus. This virus had genes encoding internal proteins from pandemic (H1N1) 2009 virus and hemagglutinin and neuraminidase genes from swine influenza virus (H1N2). Our results demonstrate processes contributing to influenza virus heterogeneity

The sustainability of knowledge brokerage of the mental health improvement outcomes framework in Scotland:a follow-up analysis

This is a follow-up study to Reid et al (2017) which considered the barriers and facilitators of getting knowledge into policy when using a knowledge brokering approach. The previous study analysed the use of strategies to reduce barriers to the use of evidence in mental health strategy planning in Scotland using outcome frameworks. The main facilitators highlighted were the importance of local champions, cooperation within partnership networks, and national-level support. The barriers were local implementation cultures, local time pressures, perceived complexities of the framework, and timeliness of the framework. The present article details the results of a follow-up qualitative evaluation of the sustainability of the mental health improvement outcomes framework with local planners. There is a dearth of literature which focuses on the sustainability of outcome frameworks and the findings of this study suggest that the barriers highlighted by Reid et al (2017) remain acute issues. However, there are further aspects for learning for knowledge brokers themselves in terms of national and local relations and the wider challenges and opportunities relating to network governance and policy reform agendas.</jats:p

Whose depression relates to discrepancies? Testing relations between informant characteristics and informant discrepancies from both informants' perspectives

This study examined whether mothers’ and children’s depressive symptoms were each uniquely related to mother– child rating discrepancies on a multidimensional dyadic construct: domains associated with parental monitoring (i.e., Child Disclosure, Parental Knowledge, and Parental Solicitation). Participants included a community sample of 335 mother/female-caregiver and child dyads (182 girls, 153 boys; 9–16 years old). Children’s depressive symptoms were consistently related to each of the three domains of mother– child discrepancies. Mothers’ depressive symptoms were related to perceived discrepancies in two domains (Child Disclosure and Parental Knowledge). Furthermore, these relations could not be accounted for by other informant characteristics (maternal stress, child age, child gender, child ethnicity). Findings provide important empirical support for theory suggesting that both informants’ perspectives meaningfully contribute to their discrepancies in perceived behavior. Consideration of both informants’ perspectives leads to valuable information as to whether any particular characteristic is an important correlate of discrepancies.This work was supported, in part, by National Institute of Mental Health Grant F31 MH67540 (awarded to Andres De Los Reyes). It was also supported by National Institute on Drug Abuse Grant K01 DA015442 01A1 of the National Institutes of Health (awarded to Wendy Kliewer)

The Longitudinal Consistency of Mother–Child Reporting Discrepancies of Parental Monitoring and Their Ability to Predict Child Delinquent Behaviors Two Years Later

Correspondence regarding this manuscript should be addressed to Andres De Los Reyes, Comprehensive Assessment and Intervention Program, Department of Psychology, University of Maryland at College Park, Biology/Psychology Building, Room 3123H, College Park, MD 20742; 301-405-7049 (office); 301-314-9566 (fax); E-mail: [email protected] study examined the longitudinal consistency of mother–child reporting discrepancies of parental monitoring and whether these discrepancies predict children’s delinquent behaviors 2 years later. Participants included 335 mother/female-caregiver and child (46% boys,[90% African American; age range 9–16 years [M = 12.11, SD = 1.60]) dyads living in moderate-to-high violence areas. Mother–child discrepancies were internally consistent within multiple assessment points and across measures through a 2-year follow-up assessment. Further, mothers who at baseline consistently reported higher levels of parental monitoring relative to their child had children who reported greater levels of delinquent behaviors 2 years later, relative to mother–child dyads that did not evidence consistent discrepancies. This finding could not be accounted for by baseline levels of the child’s delinquency, maternal and child emotional distress, or child demographic characteristics. This finding was not replicated when relying on the individual reports of parental monitoring to predict child delinquency, suggesting that mother–child reporting discrepancies provided information distinct from the absolute frequency of reports. Findings suggest that mother–child discrepancies in reports of parental monitoring can be employed as new individual differences measurements in developmental psychopathology research.This work was supported, in part, by National Institutes of Health, National Institute on Drug Abuse Grants K01 DA015442 01A1 and R21DA020086-02 (awarded to Wendy Kliewer)

Characterising standard genetic parts and establishing common principles for engineering legume and cereal roots

Plant synthetic biology and cereal engineering depends on the controlled expression of transgenes of interest. Most engineering in plant species to date has relied heavily on the use of a few, well-established constitutive promoters to achieve high levels of expression; however, the levels of transgene expression can also be influenced by the use of codon optimisation, intron-mediated enhancement and varying terminator sequences. Most of these alternative approaches for regulating transgene expression have only been tested in small-scale experiments, typically testing a single gene of interest. It is therefore difficult to interpret the relative importance of these approaches and to design engineering strategies that are likely to succeed in different plant species, particularly if engineering multi-genic traits where the expression of each transgene needs to be precisely regulated. Here we present data on the characterisation of 46 promoters and 10 terminators in Medicago truncatula, Lotus japonicus, Nicotiana benthamiana and Hordeum vulgare, as well as the effects of codon optimisation and intron-mediated enhancement on the expression of two transgenes in H. vulgare. We have identified a core set of promoters and terminators of relevance to researchers engineering novel traits in plant roots. In addition, we have shown that combining codon optimisation and intron-mediated enhancement increases transgene expression and protein levels in barley. Based on our study, we recommend a core set of promoters and terminators for broad use, and also propose a general set of principles and guidelines for those engineering cereal species