Unveiling the components of the signaling pathway between endosomes and the phagocytic uptake machinery in Dictyostelium discoideum

The soil amoebae Dictyostelium discoideum take up particles from their environment in order to obtain nutrition. The particle transits through the cell within a phagosome that fuses with organelles of different molecular compositions, undergoing a gradual degradation by different sets of hydrolytic enzymes. Griffiths’ concept of “phagosome individuality” predicts signaling from phagosomes into the cytoplasm, which might regulate many aspects of cell physiology. The finding that Dictyostelium cells depleted of the lysozyme AlyA or over-expressing the esterase Gp70 exhibit increased uptake of food particles, led to the postulation of a signaling cascade between endocytic compartments and the cytoskeletal uptake machinery at the plasma membrane. Assuming that Gp70 acts downstream of AlyA, gene-expression profiling of both mutants revealed different and overlapping sets of misregulated genes that might participate in this signaling cascade. Based on these results, we analyzed the effects of the artificial misregulation of six candidate genes by over-expression or negative genetic interference, in order to reconstruct at least part of the signaling pathway. SSB420 and SSL793 were chosen as candidates for the first signaling step, as they were up-regulated in AlyA-null cells and remained unaltered in the Gp70 over-expressing cells. The over-expression of SSB420 enhanced phagocytosis and raised the expression levels of Gp70, supporting its involvement in the signaling pathway between AlyA and Gp70 as a positive regulator of phagocytosis. However, this was not the case of cells over-expressing SSL793, as this mutation had no effects on phagocytosis. For the signaling downstream of Gp70, we studied four commonly misregulated genes in AlyA-depleted and Gp70 over-expressing cells. The expression levels of SLB350, SSB389 and TipD were lower in both mutants and therefore these were assumed as possible candidates for the negative regulation of phagocytosis. Cells depleted of SLB350 exhibited an increased phagocytic activity and no effect on Gp70 expression, proving its participation in the signaling pathway downstream of Gp70. Unlike SLB350, the disruption of the genes coding for SSB389 and TipD had no effects on particle uptake, excluding them from the pathway. The fourth candidate was Yipf1, the only gene that was commonly up-regulated in both mutants. Yet, the artificial over-expression of this protein had no effects on phagocytosis, so this candidate is also not included in the signaling pathway. Furthermore, localizing the products of the candidate genes within the cell helped unveiling several cellular organelles that receive signals from the phagosome and transduce them towards the uptake machinery.

Identifying the component structure of job satisfaction by principal components analysis among extension officers in North West Province, South Africa

The component structure of a 34-item scale measuring different aspects of job satisfaction was investigated among extension officers in North West Province, South Africa. A simple random sampling technique was used to select 40 extension officers from which data were collected. A structured questionnaire consisting of 34 job satisfaction and 10 personal characteristic items was administered to the extension officers. Items on job satisfaction were measured at interval level and analyzedwith Principal ComponentAnalysis. Most of the respondents (82.5%) weremales, between 40 to 45 years, 85% were married and 87.5% had a diploma as their educational qualification. Furthermore, 54% of the households size between 4 to 6 persons, whereas 75% were Christians. The majority of the extension officers lived in their job area (82.5), while 80% covered at least 3 communities and 3 farmer groups. In terms of number of farmers covered, only 40% of the extension officers covered more than 500 farmers and 45% travelled more than 40 km to reach their farmers. From the job satisfaction items 9 components were extracted to show areas for job satisfaction among extension officers. These were in-service training, research policies, communicating recommended practices, financial support for self and family, quality of technical help, opportunity to advance education, management and control of operations, rewarding system and sanctions. The results have several implications for motivating extension officers for high job performance especially with large number of clients and small number of extension agents.

Identification of Problem-based learning components and their effects on graduates' competencies

This study investigated the relationship between higher education and the requirement of the world of work with an emphasis on the effect of problem-based learning (PBL) on graduates' competencies. The implementation of full PBL method is costly (Albanese & Mitchell, 1993; Berkson, 1993; Finucane, Shannon, & McGrath, 2009). However, the implementation of PBL in a less than curriculum-wide mode is more achievable in a broader context (Albanese, 2000). This means higher education institutions implement only a few PBL components in the curriculum. Or a teacher implements a few PBL components at the courses level. For this kind of implementation there is a need to identify PBL components and their effects on particular educational outputs (Hmelo-Silver, 2004; Newman, 2003). So far, however there has been little research about this topic. The main aims of this study were: (1) to identify each of PBL components which were manifested in the development of a valid and reliable PBL implementation questionnaire and (2) to determine the effect of each identified PBL component to specific graduates' competencies. The analysis was based on quantitative data collected in the survey of medicine graduates of Gadjah Mada University, Indonesia. A total of 225 graduates responded to the survey. The result of confirmatory factor analysis (CFA) showed that all individual constructs of PBL and graduates' competencies had acceptable GOFs (Goodness-of-fit). Additionally, the values of the factor loadings (standardize loading estimates), the AVEs (average variance extracted), CRs (construct reliability), and ASVs (average shared squared variance) showed the proof of convergent and discriminant validity. All values indicated valid and reliable measurements. The investigation of the effects of PBL showed that each PBL component had specific effects on graduates' competencies. Interpersonal competencies were affected by Student-centred learning (β = .137; p < .05) and Small group components (β = .078; p < .05). Problem as stimulus affected Leadership (β = .182; p < .01). Real-world problems affected Personal and organisational competencies (β = .140; p < .01) and Interpersonal competencies (β = .114; p < .05). Teacher as facilitator affected Leadership (β = 142; p < .05). Self-directed learning affected Field-related competencies (β = .080; p < .05). These results can help higher education institution and educator to have informed choice about the implementation of PBL components. With this information higher education institutions and educators could fulfil their educational goals and in the same time meet their limited resources. This study seeks to improve prior studies' research method in four major ways: (1) by indentifying PBL components based on theory and empirical data; (2) by using latent variables in the structural equation modelling instead of using a variable as a proxy of a construct; (3) by using CFA to validate the latent structure of the measurement, thus providing better evidence of validity; and (4) by using graduate survey data which is suitable for analysing PBL effects in the frame work of the relationship between higher education and the world of work.

Optimization of the Mechanical and Optical Properties of Tunable Optical Sensor Arrays (TOSA) for a Nanospectrometer in the Visible and Near Infrared Spectral Range

Tunable Optical Sensor Arrays (TOSA) based on Fabry-Pérot (FP) filters, for high quality spectroscopic applications in the visible and near infrared spectral range are investigated within this work. The optical performance of the FP filters is improved by using ion beam sputtered niobium pentoxide (Nb2O5) and silicon dioxide (SiO2) Distributed Bragg Reflectors (DBRs) as mirrors. Due to their high refractive index contrast, only a few alternating pairs of Nb2O5 and SiO2 films can achieve DBRs with high reflectivity in a wide spectral range, while ion beam sputter deposition (IBSD) is utilized due to its ability to produce films with high optical purity. However, IBSD films are highly stressed; resulting in stress induced mirror curvature and suspension bending in the free standing filter suspensions of the MEMS (Micro-Electro-Mechanical Systems) FP filters. Stress induced mirror curvature results in filter transmission line degradation, while suspension bending results in high required filter tuning voltages. Moreover, stress induced suspension bending results in higher order mode filter operation which in turn degrades the optical resolution of the filter. Therefore, the deposition process is optimized to achieve both near zero absorption and low residual stress. High energy ion bombardment during film deposition is utilized to reduce the film density, and hence the film compressive stress. Utilizing this technique, the compressive stress of Nb2O5 is reduced by ~43%, while that for SiO2 is reduced by ~40%. Filters fabricated with stress reduced films show curvatures as low as 100 nm for 70 μm mirrors. To reduce the stress induced bending in the free standing filter suspensions, a stress optimized multi-layer suspension design is presented; with a tensile stressed metal sandwiched between two compressively stressed films. The stress in Physical Vapor Deposited (PVD) metals is therefore characterized for use as filter top-electrode and stress compensating layer. Surface micromachining is used to fabricate tunable FP filters in the visible spectral range using the above mentioned design. The upward bending of the suspensions is reduced from several micrometers to less than 100 nm and 250 nm for two different suspension layer combinations. Mechanical tuning of up to 188 nm is obtained by applying 40 V of actuation voltage. Alternatively, a filter line with transmission of 65.5%, Full Width at Half Maximum (FWHM) of 10.5 nm and a stopband of 170 nm (at an output wavelength of 594 nm) is achieved. Numerical model simulations are also performed to study the validity of the stress optimized suspension design for the near infrared spectral range, wherein membrane displacement and suspension deformation due to material residual stress is studied. Two bandpass filter designs based on quarter-wave and non-quarter-wave layers are presented as integral components of the TOSA. With a filter passband of 135 nm and a broad stopband of over 650 nm, high average filter transmission of 88% is achieved inside the passband, while maximum filter transmission of less than 1.6% outside the passband is achieved.