Understanding the role that Quality of Work Life of food and beverage employees plays in perceived service delivery and productivity

3The purpose of this article is to determine the role that Quality ofWork Life plays in the perceived service delivery and productivityof food and beverage employees in Potchefstroom, South Africa.The questionnaires were distributed at selected food and beverageestablishments, and a total of 224 questionnaires were included in thestatistical analysis. The data analysis consisted of a demographic profile,a factor analysis and a structural equation model. The results indicatedthat job, creativity and aesthetics, actualisation, organisational supportand employee commitment attributes each exert an influence on theperceived service delivery and productivity of employees. This impliesthat food and beverage managers should try to improve the workingconditions of employees as well as provide an appropriate level ofrecognition to hardworking employees. Management should considerthe recommendations that are made in terms of the job satisfactionlevels of food and beverage service employees, as they are able topositively influence organisational performance and success.Key words: employee management, food and beverage sector, structural equation model,Quality of Work Lif

The role and attributes of entrepreneurs at South Africa’s largest arts festival.

The Klein Karoo National Arts Festival (KKNK) in Oudtshoorn, South Africa, is the largest arts festival in South Africa. The purpose of this research was to determine the attributes and role of the entrepreneurs at the Klein Karoo National Arts Festival. This was done by means of a questionnaire survey (N=249). After data capturing was completed, two factor analyses were conducted. The first factor analysis revealed six factors (entrepreneurial attributes), namely organisational skills, resourcefulness, self-edification, explorative, acquired skill and drive, of which resourcefulness had the highest mean value. The second factor analysis identified the role of entrepreneurs at KKNK and revealed three primary roles, namely festival promotion, product promotion and income generation, of which product promotion had the highest mean value. This is the first time that the roles of entrepreneurs at festivals were investigated in South AfricaEl Festival de las Artes Nacional Klein Karoo (KKNK) en Oudtshoorn, Sudáfrica, es el mayor festival de las artes en el sur de África. El propósito de esta investigación fue determinar las características y el papel de los empresarios en éste. Lo cual se hizo por medio de una encuesta (N = 249). Después de la captura de datos, se realizaron dos análisis. El análisis factorial reveló por primera vez seis factores (atributos empresariales): capacidad de organización, capacidad de auto-edificación, de exploración, adquirió la habilidad y la unidad, de la que el ingenio tuvo el valor más alto. El análisis del segundo factor identificado como el papel de los empresarios en el KKNK reveló tres papeles principales, a saber, la promoción de festivales, promoción de productos y generación de ingresos, de los cuales tenían la promoción de los productos de mayor valor. Esta es la primera vez que se ha investigado el papel de los empresarios en los festivales de Sudáfrica

A paradigm shift in the provision of recreation in the North West Province of South Africa.

Recreation professionals in the 21st century are faced with one of the most challenging tasks, to be innovative in the provision of recreation services to communities. The modern day requires recreation professionals to anticipate and effect changes rather than to wait and react to change as a result of the demand for recreation services from the communities. The purpose of this study was to investigate the possibility of utilising new ways of delivering recreation services to communities by local governments in the North West Province in South Africa. A mixed-methods research design using a questionnaire and focus groups was used on 20 targeted personnel who are responsible for managing recreation at local government level in the Province. Frequencies and phi coefficients were calculated for the quantitative data, while for qualitative data patterns were established thematically. The results show that 75% of the local governments are located in rural settings whilst 25% are found in urban areas with limited resources. Most respondents (70%) indicated that they do not provide programmes for human services that go beyond traditional recreation activities. A large proportion (65%) of local governments indicated that they do not use staff resources to coach citizens until they can provide their own leadership. About 35% of local governments indicated that they do not justify budgets in terms of social need and programme results. The results indicate that local governments do not adapt to new ways of responding to community needs with regard to recreation. The study recommends that if a new approach to recreation is accepted (emerging paradigm) not only can communities help to implement programmes that should satisfy their needs, but service providers will be able to afford and incorporate emerging activities. Local governments should be at the forefront in designing and implementing effective recreation programmes in consultation with communities, and also liaising with the relevant departments in provincial government

Deriving four functional anti-HIV siRNAs from a single Pol III-generated transcript comprising two adjacent long hairpin RNA precursors

Several different approaches exist to generate expressed RNA interference (RNAi) precursors for multiple target inhibition, a strategy referred to as combinatorial (co)RNAi. One such approach makes use of RNA Pol III-expressed long hairpin RNAs (lhRNAs), which are processed by Dicer to generate multiple unique short interfering siRNA effectors. However, because of inefficient intracellular Dicer processing, lhRNA duplexes have been limited to generating two independent effective siRNA species. In this study, we describe a novel strategy whereby four separate anti-HIV siRNAs were generated from a single RNA Pol III-expressed transcript. Two optimized lhRNAs, each comprising two active anti-HIV siRNAs, were placed in tandem to form a double long hairpin (dlhRNA) expression cassette, which encodes four unique and effective siRNA sequences. Processing of the 3′ position lhRNA was more variable but effective multiple processing was possible by manipulating the order of the siRNA-encoding sequences. Importantly, unlike shRNAs, Pol III-expressed dlhRNAs did not compete with endogenous and exogenous microRNAs to disrupt the RNAi pathway. The versatility of expressed lhRNAs is greatly expanded and we provide a mechanism for generating transcripts with modular lhRNAs motifs that contribute to improved coRNAi

The biogenesis and characterization of mammalian microRNAs of mirtron origin

Mirtrons, short hairpin pre-microRNA (miRNA) mimics directly produced by intronic splicing, have recently been identified and experimentally confirmed in invertebrates. While there is evidence to suggest several mammalian miRNAs have mirtron origins, this has yet to be experimentally demonstrated. Here, we characterize the biogenesis of mammalian mirtrons by ectopic expression of splicing-dependent mirtron precursors. The putative mirtrons hsa-miR-877, hsa-miR-1226 and mmu-miR-1224 were designed as introns within eGFP. Correct splicing and function of these sequences as introns was shown through eGFP fluorescence and RT–PCR, while all mirtrons suppressed perfectly complementary luciferase reporter targets to levels similar to that of corresponding independently expressed pre-miRNA controls. Splicing-deficient mutants and disruption of key steps in miRNA biogenesis demonstrated that mirtron-mediated gene knockdown was splicing-dependent, Drosha-independent and had variable dependence on RNAi pathway elements following pre-miRNA formation. The silencing effect of hsa-miR-877 was further demonstrated to be mediated by the generation of short anti-sense RNA species expressed with low abundance. Finally, the mammalian mirtron hsa-miR-877 was shown to reduce mRNA levels of an endogenous transcript containing hsa-miR-877 target sites in neuronal SH-SY5Y cells. This work confirms the mirtron origins of three mammalian miRNAs and suggests that they are a functional class of splicing-dependent miRNAs which are physiologically active

A direct comparison of strategies for combinatorial RNA interference

<p>Abstract</p> <p>Background</p> <p>Combinatorial RNA interference (co-RNAi) is a valuable tool for highly effective gene suppression of single and multiple-genes targets, and can be used to prevent the escape of mutation-prone transcripts. There are currently three main approaches used to achieve co-RNAi in animal cells; multiple promoter/shRNA cassettes, long hairpin RNAs (lhRNA) and miRNA-embedded shRNAs, however, the relative effectiveness of each is not known. The current study directly compares the ability of each co-RNAi method to deliver pre-validated siRNA molecules to the same gene targets.</p> <p>Results</p> <p>Double-shRNA expression vectors were generated for each co-RNAi platform and their ability to suppress both single and double-gene reporter targets were compared. The most reliable and effective gene silencing was achieved from the multiple promoter/shRNA approach, as this method induced additive suppression of single-gene targets and equally effective knockdown of double-gene targets. Although both lhRNA and microRNA-embedded strategies provided efficient gene knockdown, suppression levels were inconsistent and activity varied greatly for different siRNAs tested. Furthermore, it appeared that not only the position of siRNAs within these multi-shRNA constructs impacted upon silencing activity, but also local properties of each individual molecule. In addition, it was also found that the insertion of up to five promoter/shRNA cassettes into a single construct did not negatively affect the efficacy of each individual shRNA.</p> <p>Conclusions</p> <p>By directly comparing the ability of shRNAs delivered from different co-RNA platforms to initiate knockdown of the same gene targets, we found that multiple U6/shRNA cassettes offered the most reliable and predictable suppression of both single and multiple-gene targets. These results highlight some important strengths and pitfalls of the currently used methods for multiple shRNA delivery, and provide valuable insights for the design and application of reliable co-RNAi.</p

Silencing of Parkinson's disease-associated genes with artificial mirtron mimics of miR-1224

Mirtrons are a recently described category of microRNA (miRNA) relying on splicing rather than processing by the microprocessor complex to generate pre-miRNA precursors of the RNA interference (RNAi) pathway. Their discovery and subsequent verification provides important information about a distinct class of miRNA and inherent advantages that could be exploited to silence genes of interest. These include micro-processor-independent biogenesis, pol-II-dependent transcription, accurate species generation and the delivery of multiple artificial mirtrons as introns within a single host transcript. Here we determined the sequence motifs required for correct processing of the mmu-miR-1224 mirtron and incorporated these into artificial mirtrons targeting Parkinson's disease-associated LRRK2 and α-synuclein genes. By incorporating these rules associated with processing and splicing, artificial mirtrons could be designed and made to silence complementary targets either at the mRNA or protein level. We further demonstrate with a LRRK2 targeting artificial mirtron that neuronal-specific silencing can be directed under the control of the human synapsin promoter. Finally, multiple mirtrons were co-delivered within a single host transcript, an eGFP reporter, to allow simultaneous targeting of two or more targets in a combinatorial approach. Thus, the unique characteristics of artificial mirtrons make this an attractive approach for future RNAi applications

The Efficacy of Generating Three Independent Anti-HIV-1 siRNAs from a Single U6 RNA Pol III-Expressed Long Hairpin RNA

RNA Interference (RNAi) effectors have been used to inhibit rogue RNAs in mammalian cells. However, rapidly evolving sequences such as the human immunodeficiency virus type 1 (HIV-1) require multiple targeting approaches to prevent the emergence of escape variants. Expressed long hairpin RNAs (lhRNAs) have recently been used as a strategy to produce multiple short interfering RNAs (siRNAs) targeted to highly variant sequences. We aimed to characterize the ability of expressed lhRNAs to generate independent siRNAs that silence three non-contiguous HIV-1 sites by designing lhRNAs comprising different combinations of siRNA-encoding sequences. All lhRNAs were capable of silencing individual target sequences. However, silencing efficiency together with concentrations of individual lhRNA-derived siRNAs diminished from the stem base (first position) towards the loop side of the hairpin. Silencing efficacy against HIV-1 was primarily mediated by siRNA sequences located at the base of the stem. Improvements could be made to first and second position siRNAs by adjusting spacing arrangements at their junction, but silencing of third position siRNAs remained largely ineffective. Although lhRNAs offer advantages for combinatorial RNAi, we show that good silencing efficacy across the span of the lhRNA duplex is difficult to achieve with sequences that encode more than two adjacent independent siRNAs