Home About us Contact
|
Home About us Contact | ||
|
|
||
Different Constructs (different + construct)
Selected AbstractsFocal electroporation in ovoDEVELOPMENTAL DYNAMICS, Issue 12 2009J. E. Simkin Abstract Gene expression fields in embryogenesis are spatially precise and often small, so experimental gene expression often requires similar spatial definition. For in ovo electroporation, typically a gene construct is injected into a natural body cavity in the embryo prior to electroporation. Limited control of the size and location of the electroporated field can be obtained by varying electrode placement and geometry, and by altering the miscibility and viscosity of the construct vehicle but it is difficult to tightly constrain electroporation to small regions. Electroporation of different constructs in close proximity has not been possible. We show that loading the construct into an agarose bead, which is then microsurgically implanted, allows for focal electroporation. Different constructs can be electroporated in close proximity by emplacing several agarose beads. This technique is simple, cheap, rapid, and requires no more specialised equipment than that required for conventional in ovo electroporation. Developmental Dynamics 238:3152,3155, 2009. © 2009 Wiley-Liss, Inc. [source] Evaluating the inter-respondent (consumer vs. staff) reliability and construct validity (SIS vs.JOURNAL OF INTELLECTUAL DISABILITY RESEARCH, Issue 4 2009Vineland) of the Supports Intensity Scale on a Dutch sample Abstract Background Despite various reliability studies on the Supports Intensity Scale (SIS), to date there has not been an evaluation of the reliability of client vs. staff judgments. Such determination is important, given the increasing consumer-driven approach to services. Additionally, there has not been an evaluation of the instrument's construct validity on a non-English speaking sample. This is important as the SIS is currently translated into 13 languages. Method Data were collected in two different samples, using the Dutch translation of the SIS and the Vineland-Z. Results There was a significant correlation between ratings of staff and consumers on the SIS; however, the relationship between the mean scores of consumer and staff responses indicated significant differences in staff and consumer scores. All correlations between the Vineland-Z domains and the SIS subscales were significant and negative, ranging from ,0.37 to ,0.89. Conclusions Analyses of the inter-respondent reliability suggest that one needs to consider the source of information regarding needed supports carefully. The significant negative correlations between SIS and Vineland-Z reflect that the SIS is measuring a different construct (needed support) than the Vineland-Z (adaptive behaviour). The results of the two studies provide additional support for the etic (universal) properties of the SIS, as both hypotheses were confirmed. In conclusion, SIS users are provided with a wealth of information that can be used for multiple purposes. [source] Focal electroporation in ovoDEVELOPMENTAL DYNAMICS, Issue 12 2009J. E. Simkin Abstract Gene expression fields in embryogenesis are spatially precise and often small, so experimental gene expression often requires similar spatial definition. For in ovo electroporation, typically a gene construct is injected into a natural body cavity in the embryo prior to electroporation. Limited control of the size and location of the electroporated field can be obtained by varying electrode placement and geometry, and by altering the miscibility and viscosity of the construct vehicle but it is difficult to tightly constrain electroporation to small regions. Electroporation of different constructs in close proximity has not been possible. We show that loading the construct into an agarose bead, which is then microsurgically implanted, allows for focal electroporation. Different constructs can be electroporated in close proximity by emplacing several agarose beads. This technique is simple, cheap, rapid, and requires no more specialised equipment than that required for conventional in ovo electroporation. Developmental Dynamics 238:3152,3155, 2009. © 2009 Wiley-Liss, Inc. [source] Quality of life and health-related quality of life of adolescents with cerebral palsyDEVELOPMENTAL MEDICINE & CHILD NEUROLOGY, Issue 7 2007Peter L Rosenbaum MD FRCP(C) This study assessed quality of life (QOL) and health-related quality of life (HRQOL) of 203 adolescents with cerebral palsy (111 males, 92 females; mean age 16y [SD 1y 9mo]). Participants were classified using the Gross Motor Function Classification System (GMFCS), as Level I (n=60), Level II (n=33), Level III (n=28), Level IV (n=50), or Level V (n=32). QOL was assessed by self (66.5%) or by proxy (33.5%) with the Quality of Life Instrument for People With Developmental Disabilities, which asks about the importance and satisfaction associated with the QOL domains of Being, Belonging, and Becoming; HRQOL was captured through proxy reports with the Health Utilities Index, Mark 3 (HUI3), which characterizes health in terms of eight attributes, each having five or six ordered levels of function. GMFCS level was not a source of variation for QOL domain scores but was significantly associated with the eight HRQOL attributes and overall HUI3 utility scores (p<0.05). Some QOL domain scores varied significantly by type of respondent (self vs proxy; p<0.05). Overall HUI3 utility values were significantly but weakly correlated with QOL Instrument scores for Being (r=0.37), Belonging (r=0.17), Becoming (r=0.20), and Overall QOL (r=0.28), and thus explain up to 14% of the variance (r2). These findings suggest that although QOL and HRQOL are somewhat related conceptually, they are different constructs and need to be considered as separate dimensions of the lives of people with functional limitations. [source] Mild cognitive impairment1, a review of prevalence, incidence and outcome according to current approachesACTA PSYCHIATRICA SCANDINAVICA, Issue 6 2002J. Bischkopf Objective:, Mild cognitive impairment is associated with an increased risk of developing dementia. However, agreement needs to be reached on clearly specified diagnostic criteria for mild cognitive impairment. The present paper critically reviews the different constructs of mild cognitive impairment on the basis of the available empirical evidence. Method:, All published papers on mild cognitive impairment during the last 15 years on Medline and other databases were reviewed. Results:, Age-specific prevalence and incidence rates according to the different constructs as well as the prognostic significance of the different constructs concerning the development of dementia are reported. Furthermore, a brief summary of recent research on possible risk factors for a negative course of mild cognitive impairment is provided. Conclusion:, As there is no construct to date that pools all efforts of defining mild cognitive impairment, the review provides suggestions for an agreement on constructive terminology and research practice. [source] Extra terminal residues have a profound effect on the folding and solubility of a Plasmodium falciparum sexual stage-specific protein over-expressed in Escherichia coliFEBS JOURNAL, Issue 21 2002Sushil Prasad Sati The presence of extra N- and C- terminal residues can play a major role in the stability, solubility and yield of recombinant proteins. Pfg27 is a 27K soluble protein that is essential for sexual development in Plasmodium falciparum. It was over-expressed using the pMAL-p2 vector as a fusion protein with the maltose binding protein. Six different constructs were made and each of the fusion proteins were expressed and purified. Our results show that the fusion proteins were labile and only partially soluble in five of the constructs resulting in very poor yields. Intriguingly, in the sixth construct, the yield of soluble fusion protein with an extended carboxyl terminus of 17 residues was several fold higher. Various constructs with either N-terminal or smaller C-terminal extensions failed to produce any soluble fusion protein. Furthermore, all five constructs produced Pfg27 that precipitated after protease cleavage from its fusion partner. The sixth construct, which produced soluble protein in high yields, also gave highly stable and soluble Pfg27 after cleavage of the fusion. These results indicate that extra amino acid residues at the termini of over-expressed proteins can have a significant effect on the folding of proteins expressed in E. coli. Our data suggest the potential for development of a novel methodology, which will entail construction of fusion proteins with maltose binding protein as a chaperone on the N-terminus and a C-terminal ,solubilization tag'. This system may allow large-scale production of those proteins that have a tendency to misfold during expression. [source] Not Enough Science or Not Enough Learning?HIGHER EDUCATION QUARTERLY, Issue 4 2008Exploring the Gaps between Leadership Theory, Practice This paper addresses the relationships between leadership theory, practice and development, drawing on both the higher education and wider leadership literature. It explores why challenges and problems exist within the contested field of leadership theory and why gaps remain between theory and practice after more than a century of research , and indeed, with increasing levels of research, scholarship and development in the last 25 years. After highlighting the importance of context for theory, practice and development, the first section of the paper examines a range of factors that contribute to theoretical ,contests' including different starting assumptions made by researchers, the different focus of studies, examination of different causal links to explain leadership, differences in values and cultural lenses and different constructs, terminology and perspectives. The second section examines the challenges faced by leadership practitioners, as individuals, and through exercising leadership as a collective responsibility in the context of changing operating environments within higher education institutions and across sectors and countries. The author highlights three areas where some re-thinking of the links between theory and practice are necessary , at the input stage, linking research findings and recruitment practices; in terms of outcomes, by researching links between leaders, leadership and performance; and in process terms, to examine more deeply complex and relational dynamic of leadership in action. The third section offers a number of specific suggestions as to how closer alignment between theory, practice and development can be achieved. The paper concludes by arguing for greater maturity (in research, practice and development) that acknowledges that leadership is played out in complex, dynamic and changing social systems. A stronger emphasis on ,leadership learning' should deliver both better science and better outcomes for leaders and led in higher education. [source] What do we know about quality of life in dementia?INTERNATIONAL JOURNAL OF GERIATRIC PSYCHIATRY, Issue 1 2009A review of the emerging evidence on the predictive, explanatory value of disease specific measures of health related quality of life in people with dementia Abstract Objective Given its complexity, there is growing consensus on the need to measure patient-rated broad outcomes like health-related quality of life (HRQL) as well as discrete functions like cognition and behaviour in dementia. This review brings together current data on the distribution, determinants and course of HRQL in dementia to investigate the predictive and explanatory value of measures of HRQL in people with dementia. Design A systematic review of papers in English published up to October 2007 to identify data on the use of disease-specific measures of HRQL in dementia. Results There are no clear or consistent associations between socio-demographic variables and HRQL. There is no convincing evidence that lower cognition or greater activity limitation is associated with lower HRQL. There is a strong suggestion that depression is consistently associated with decreased HRQL in dementia. However, the magnitude of the associations observed is moderate only and the proportion of variance explained is low suggesting that depression and HRQL are different constructs. We currently know almost nothing about the natural history of HRQL in dementia or what attributes or interventions promote or inhibit HRQL life for people with dementia. Conclusions While in other illnesses there may be simple association between HRQL and an easily measurable clinical variable, in dementia this is not so. There are now instruments available with which to measure disease-specific HRQL directly in clinical trials and other studies that can yield informative data. Copyright © 2008 John Wiley & Sons, Ltd. [source] Grade expectations: the effects of expectations on fairness and satisfaction perceptionsJOURNAL OF BEHAVIORAL DECISION MAKING, Issue 5 2003Bennett Cherry Abstract Previous research has documented the impact of self--derived expectations as reference points in the evaluation of outcomes (e.g. Ordóñez, Connolly, & Coughlan, 2000; van den Bos et al., 1998). In the present paper we extend these studies by investigating the effects of individuals' performance expectations on their subsequent evaluations of personally--relevant outcomes. In three separate studies, both in the laboratory and in the field, students' actual grade outcomes fell short, met, or exceeded grade expectations. From this information, the students evaluated their fairness and satisfaction with the actual grade outcome. The studies provide complementary results that distinguish fairness and satisfaction as different constructs based on the impact of expectations on evaluations of actual outcomes. Results demonstrate that expectations are important to perceptions of fairness and are less important to perceptions of satisfaction. Fairness judgments appear to be governed by an expectation matching proposition; whereby if the expectation is met, the outcome is fair. Whereas, satisfaction judgments are determined by the value of the actual outcome to the individual. Participants also evaluate the fairness of outcomes differently using hypothetical scenarios than they do when they experience actual outcomes in natural contexts. Copyright © 2003 John Wiley & Sons, Ltd. [source] Burning and cutting: Identifying the traits of individuals with an enduring propensity to tan and to undergo cosmetic surgeryJOURNAL OF CONSUMER BEHAVIOUR, Issue 5 2009John C. Mowen The research investigated the trait predictors of the propensity to use two beautification procedures that have the potential to harm health , the propensities to tan and to obtain cosmetic surgery. The results of a survey of 231 adult consumers revealed that a different combination of hierarchically arranged traits was predictive of the two procedures. These results suggest that different strategies are required in order to market or demarket tanning and plastic surgery. The research also made methodological contributions by revealing that two forms of vanity identified in the literature , physical view vanity and physical concern vanity , represent two different constructs rather than two dimensions of an underlying construct. Copyright © 2009 John Wiley & Sons, Ltd. [source] Effect of cell seeding concentration on the quality of tissue engineered constructs loaded with adult human articular chondrocytesJOURNAL OF TISSUE ENGINEERING AND REGENERATIVE MEDICINE, Issue 1 2008Sebastian Concaro Abstract Many aspects of the process of in vitro differentiation of chondrocytes in three-dimensional (3D) scaffolds need to be further investigated. Chitosan scaffolds were produced by freeze-drying 3% w/v 90% DDA chitosan gels. The effect of the cell seeding concentration was evaluated by culturing human adult chondrocytes in chitosan scaffolds After the first passage, cells were seeded into chitosan scaffolds with a diameter of 8 mm. The final cell seeding concentration per cm3 of chitosan scaffold was: Group A, 3 × 106; Group B, 6 × 106; Group C, 12 × 106; and Group D, 25 × 106 cells. After 14 and 28 days in 3D culture, the constructs were assesed for collagen, glucosaminoglycans and DNA content. The mechanical properties of the constructs were determined using a dynamic oscillatory shear test. The histological aspect of the constructs was evaluated using the Bern score. The collagen and GAG concentration increased, varying the cell seeding concentration. There was a significant increase in proteoglycan and hydroxyproline production between groups C and D. The sulphated GAG content increased significantly in the group D as compared to the other groups. The mechanical properties of the different constructs increased over time, from 9.6 G,/kPa at 14 days of 3D culture to 14.6 G,/kPa at 28 days under the same culture conditions. In this study we were able to determine that concentrations of 12,25 million cells/cm2 are needed to increase the matrix production and mechanical properties of human adult chondrocytes under static conditions. Copyright © 2008 John Wiley & Sons, Ltd. [source] Femoral intra-arterial injection: a tool to deliver and assess recombinant AAV constructs in rodents whole hind limbTHE JOURNAL OF GENE MEDICINE, Issue 6 2005Patrick Gonin Abstract With the aim of simplifying recombinant-adeno-associated virus (rAAV) delivery in muscle, a new femoral intra-arterial technique was designed and tested in rodents (rats and mice). Two serotypes, several promoters and transgenes (reporter or therapeutic) were tested using this administration route. The new route is both easy to perform and efficient. Its usefulness as a tool to assess gene delivery constructs in the muscle was established in the context of recombinant AAV serotypes 1 and 2, and with the ubiquitous CMV and two muscle-specific (C5-12 and CK6) promoters. Both serum monitoring of a secreted protein (murine alkaline phosphatase: muSEAP) and slide staining were used to compare the different constructs. Significantly different patterns of expression in kinetics of expression (muSEAP) and homogeneity of fiber transduction (staining) were evidenced with the different promoters tested, and compared with intra-muscular expression patterns. Detailed studies of differential transduction in leg and thigh muscles showed equivalent efficacy, except in rectus femoris, and to a lesser extent in soleus. In light of these results and prior data, intra-arterially mediated gene transfer mechanism is discussed. Copyright © 2005 John Wiley & Sons, Ltd. [source] Rational shape engineering of the filamentous protein , prefoldin through incremental gene truncationBIOPOLYMERS, Issue 6 2009Timothy A. Whitehead Abstract An enticing possibility in nanotechnology is to use proteins as templates for the positioning of molecules in regular patterns with nanometer precision over large surface areas. However, the ability to redesign protein quaternary structure to construct new shapes remains underdeveloped. In the present work, we have engineered the dimensions of a filamentous protein, the , prefoldin (, PFD) from the hyperthermophile Methanocaldococcus jannaschii, and have achieved controllable attachment of filaments in a specific orientation on a carbon surface. Four different constructs of , PFD were generated in which the coiled coils extending from the association domain are progressively truncated. Three of the truncation constructs form well-defined filaments with predictable dimensions according to transmission electron microscopy. Two of these constructs had 2D persistence lengths similar to that of , PFD at 300,740 nm. In contrast, the 2D persistence length of the shortest truncation mutant was 3500 nm, indicating that the filament adsorbs along a different axis than the other constructs with its two rows of coiled coils facing out from the surface. The elastic moduli of the filaments range from 0.7,2.1 GPa, similar to rigid plastics and within the lower limit for proteins whose primary intermolecular interaction is hydrogen bonding. These results demonstrate a versatile approach for controlling the overall dimensions and surface orientation of protein filaments, and expand the toolbox by which to tune two overall dimensions in protein space for the creation of templated materials over a wide variety of conditions. © 2009 Wiley Periodicals, Inc. Biopolymers 91: 496,503, 2009. This article was originally published online as an accepted preprint. The "Published Online" date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com [source] Cloning the bacterial bphC gene into Nicotiana tabacum to improve the efficiency of PCB phytoremediationBIOTECHNOLOGY & BIOENGINEERING, Issue 1 2009M. Novakova Abstract The aim of this work is to increase the efficiency of the biodegradation of polychlorinated biphenyls (PCBs) by the introduction of bacterial genes into the plant genome. For this purpose, we selected the bphC gene encoding 2,3-dihydroxybiphenyl-1,2-dioxygenase from Pseudomonas testosteroni B-356 to be cloned into tobacco plants. The dihydroxybiphenyldioxygenase enzyme is the third enzyme in the biphenyl degradation pathway, and its unique function is the cleavage of biphenyl. Three different constructs were designed and prepared in E. coli: the bphC gene being fused with the ,-glucuronidase (GUS) gene, with the luciferase (LUC) gene, and with histidine tail in three separate plant cloning vectors. The GUS and LUC genes were chosen because they can be used as markers for the easy detection of transgenic plants, while histidine tail better enables the isolation of protein expressed in plant tissue. The prepared vectors were then introduced into cells of Agrobacterium tumefaciens. The transient expression of the prepared genes was first studied in cells of Nicotiana tabacum. Once this ability had been established, model tobacco plants were transformed by agrobacterial infection with the bphC/GUS, bphC/LUC, and bphC/His genes. The transformed regenerants were selected on media using a selective antibiotic, and the presence of transgenes and mRNA was determined by PCR and RT-PCR. The expression of the fused proteins BphC/GUS and BphC/LUC was confirmed histochemically by analysis of the expression of their detection markers. Western blot analysis was performed to detect the presence of the BphC/His protein immunochemically using a mouse anti-His antibody. Growth and viability of transgenic plants in the presence of PCBs was compared with control plants. Biotechnol. Bioeng. 2009;102: 29,37. © 2008 Wiley Periodicals, Inc. [source] Are normal and complicated grief different constructs?CLINICAL PSYCHOLOGY AND PSYCHOTHERAPY (AN INTERNATIONAL JOURNAL OF THEORY & PRACTICE), Issue 6 2008A confirmatory factor analytic test Nowadays, much debate in the bereavement domain is directed towards the inclusion of Complicated Grief (CG) as a separate category in the Diagnostic and Statistical Manual of Mental Disorders. Important within this discussion is the conceptual relationship between CG reactions and normal grief (NG) reactions. This study aims at elucidating this relationship by using data from 456 bereaved young adults, aged 17 to 25 years. We examined the structural distinctiveness of CG and NG reactions, using two criteria sets. The first set ties in with previous research in bereaved adults on the distinctiveness of CG and NG and allows to test the replicability of earlier findings. The second set links up with the recently revised criteria for CG and permits to investigate whether earlier findings hold for the new criteria. For both sets, two models for NG and CG were compared using confirmatory factor analytic procedures. These analyses revealed that CG and NG reactions can be distinguished by their very nature, except for one CG reaction (viz. ,yearning'), that loaded on both factors.,Copyright © 2008 John Wiley & Sons, Ltd. [source] | |||||||||||||||||||