In silico approaches in the identification of Cryptococcus neoformans chemoreceptors

Cryptococcus neoformans, a pathogenic, yeast-like fungus, typically forms biofilms on medical devices that prevents the efficient penetration of antifungal drugs. As quorum sensing (QS)-related chemoreceptors play a crucial role in the pathogenesis of Cryptococcus neoformans particularly in biofilm formation, studying them would enable a clearer understanding on the host-pathogen interaction. However, information regarding C. neoformans QS-related chemoreceptors such as the encoding DNA sequences, protein structures and the binding ligands remain unknown till today. Here, we provide information to facilitate researchers to comprehend the basic views on gene homology and its various applications in the identification of novel chemoreceptors in microorganism especially the novel QS-related chemoreceptors for future use.Keywords: Cryptococcus neoformans, homology-based transfer, quorum sensing, chemoreceptor

Cloning, expression and purification of squalene synthase from Candida tropicalis in Pichia pastoris

Squalene synthase (SS) is the key precursor and first committed enzyme of the sterol biosynthesis pathway. In a previous work, SS has been identified as one of the immunogenic proteins that could be a potential diagnostic candidate for the pathogenic fungus Candida tropicalis. In this study, SS from C. tropicalis was cloned and expressed as recombinant protein in Pichia pastoris to investigate its reactivity with serum antibodies. ERG9 gene that encodes for SS was amplified by PCR and cloned in-frame into pPICZB expression vector. The recombinant construct was then transformed into P. pastoris GS115 host strain. Expression of the recombinant protein was confirmed by SDS–PAGE and Western blot analysis using anti-His tag probe. Optimal protein production was achieved by cultivating the culture with 1.0% methanol for 72 h. The recombinant protein was purified to approximately 97% pure in a single step immobilized metal affinity chromatography with a yield of 70.3%. Besides, the purified protein exhibited specific reactivity with immune sera on Western blot. This is the first report on heterologous expression of antigenic SS from C. tropicalis in P. pastoris which can be exploited for large-scale production and further research. The results also suggested that the protein might be of great value as antigen candidate for serodiagnosis of Candida infection

Inhibition of hyphae formation and SIR2 expression in Candida albicans treated with fresh Allium sativum (garlic) extract

Aims: The aims of the present study were to determine whether Allium sativum (garlic) extract has any effect on the morphology transformation of Candida albicans, and to investigate whether it could alter the gene expression level of SIR2, a morphogenetic control gene and SAP4, a gene encoding secreted aspartyl proteinase. Methods and Results: Candida albicans cells were incubated with a range of concentrations of fresh garlic extract, and the morphology was monitored via light microscopy. Garlic extract treatment caused the transition of yeast form to hyphal form to be obviated. The expression of SIR2 was down-regulated from 1.2- to 2.5-fold with increasing concentration of the garlic extract, as determined from relative quantitative reverse transcription- polymerase chain reaction. There was no difference in the SAP4 expression in control vs treated cultures. Conclusions: Garlic and its bioactive components have the ability to suppress hyphae production and to affect the expression level of SIR2 gene. Significance and Impact of the Study: Hyphal production is an essential virulence determinant of C. albicans for invasive infections, therefore garlic and its constituents can be effective not only against colonizing C. albicans strains present in mucosal infections, but also virulent strains causing systemic or invasive candidiasis

Perspective on Marriage Among Young Chinese Women in Malaysia

Aims: This paper is based on the subjective views of young Chinese women in Malaysia. The percentage of 30 to 34-year-olds who had never been married doubled, rising from 6% to 12% for women and from 12% to 25% for men over three decades from the 1970s to the 2000s. The aim of this survey is to shed light on young Chinese Malaysian ladies’ perspectives and challenges related to marriage. Methods: Forty participants between the age of 25 to 40 participated in this study using a selective sampling and snowball method. The data was collected using an online questionnaire consisting of both qualitative and quantitative elements. Results: The findings demonstrated that 61% of participants felt it is not necessary to get married at a specific age. Most of them believed that marriage is an option in life and it does not symbolize the completeness of self. Among the more common reasons for getting married are companionship, the formation of a family that symbolized the continuity of a family line, and practical benefits such as legal protection and financial support. Contrary to most western studies in which love is a major factor in facilitating their decision to get married, Malaysian Chinese ladies cited companionship being a crucial reason. Most women hesitated to commit to a marriage mainly because of fear and failed to find a partner that meets their expectations (the right person). The fears include fear of uncertainties, fear of bad outcomes (a preference for monogamy), fear of unmet family expectations, and fear of commitment to potential unhealthy family dynamics especially those with previous traumatic experiences from original or other known families. They preferred to avoid involving in such vicious cycles, e.g., the strained relationship between two families or extended families. Having limited social circles and perceiving that marriage is not the top priority in present life had hindered the ladies from actively seeking a life partner for marriage. Participants also suggested ways to improve their situation if they were to get engaged in marriage. One of the possible practicable steps is to expand their social circle. This could be done by attending some love matching activities or with the use of social apps. An increase in personal willingness and readiness to grab the opportunity, and practice more self-love and self-care are other important steps. Approximately 12% of them were clueless about how to involve themselves in a relationship in the future. Discussion and Suggestions: Marriage is perceived as a not-the-only option and is obligatory for young Malaysian Chinese ladies in the age range of 30s. The authors opined that young Malaysian Chinese ladies are more concerned about quality of living rather than marriage, and preferred to be self-dependent, mainly due to the exponential growth of the economic and educational system, in addition to the advocacy for the women rights in recent years. This is contrary to the traditional Chinese culture of male superiority. The current study only allows us to have an overview of the group’s perspective and an in-depth interview shall be conducted for further understanding

A modified in situ RT-PCR method for localizing fungal-specific gene expression in Candida-infected mice renal cells

In situ Reverse Transcriptase PCR (in situ RT-PCR) can amplify mRNA and localize gene expression in cells. However, this method is not feasible in fungi as the thick fungal cell wall constitutes a barrier to this procedure. We developed a two step in situ RT-PCR procedure which enabled the detection and localization of Candida tropicalis mRNA expression in formalin-fixed, paraffin-embedded (FFPE) mouse kidney sections. This in situ hybridization study revealed the first direct evidence for deposition of Candida tropicalis secreted aspartic proteinase 2 (CtSAP2) in the tip of pseudohyphae and its involvement in acute systemic candidiasis. We conclude that in situ RT-PCR can be successfully applied to FFPE tissues and will offer new perspectives in studying gene expression in Candida species

Knowledge transfer of eLearning objects: Lessons learned from an intercontinental capacity building project

Background Effective knowledge transfer of eLearning objects can hasten the adoption and dissemination of technology in teaching and learning. However, challenges exist which hinder inter-organisational knowledge transfer, particularly across continents. The ACoRD project aimed to transfer knowledge on digital learning development from UK/EU (provider) to Malaysian (receiver) higher education institutions (HEIs). This study explores the challenges encountered during the knowledge transfer process and lessons learned. Methods This is a qualitative study involving both the knowledge providers and receivers in focus group discussions (n = 25). Four focus group discussions were conducted in the early (n = 2) and mid-phase (n = 2) of the project by trained qualitative researchers using a topic guide designed to explore experiences and activities representing knowledge transfer in multi-institutional and multi-cultural settings. The interviews were audio-recorded, transcribed verbatim, and checked. The transcripts were analysed using thematic analysis. Results Five main themes emerged from this qualitative study: mismatched expectations between providers and receivers; acquiring new knowledge beyond the professional "comfort zone"; challenges in cascading newly acquired knowledge to colleagues and management; individual and organisational cultural differences; and disruption of knowledge transfer during the COVID-19 pandemic. Conclusion This study highlights the need to create a conducive platform to facilitate continuous, timely and bi-directional needs assessment and feedback; this should be done in the early phase of the knowledge transfer process. The challenges and strategies identified in this study could guide more effective knowledge transfer between organisations and countries.publishedVersio

Prioritising topics for developing e-learning resources in healthcare curricula: A comparison between students and educators using a modified Delphi survey

Background Engaging students in the e-learning development process enhances the effective implementation of e-learning, however, students’ priority on the topics for e-learning may differ from that of the educators. This study aims to compare the differences between the students and their educators in prioritising the topics in three healthcare curricula for reusable e-learning object (RLO) development. Method A modified Delphi study was conducted among students and educators from University Malaya (UM), Universiti Putra Malaysia (UPM) and Taylor’s University (TU) on three undergraduate programmes. In Round 1, participants were asked to select the topics from the respective syllabi to be developed into RLOs. Priority ranking was determined by using frequencies and proportions. The first quartile of the prioritised topics was included in Round 2 survey, which the participants were asked to rate the level of priority of each topic using a 5-point Likert scale. The mean score of the topics was compared between students and educators. Result A total of 43 educators and 377 students participated in this study. For UM and TU Pharmacy, there was a mismatch in the prioritised topics between the students and educators. For UPM, both the educators and students have prioritised the same topics in both rounds. To harmonise the prioritisation of topics between students and educators for UM and TU Pharmacy, the topics with a higher mean score by both the students and educators were prioritised. Conclusion The mismatch in prioritised topics between students and educators uncovered factors that might influence the prioritisation process. This study highlighted the importance of conducting needs assessment at the beginning of eLearning resources development.publishedVersio

Prioritising topics for developing e-learning resources in healthcare curricula: A comparison between students and educators using a modified Delphi survey

Background Engaging students in the e-learning development process enhances the effective implementation of e-learning, however, students’ priority on the topics for e-learning may differ from that of the educators. This study aims to compare the differences between the students and their educators in prioritising the topics in three healthcare curricula for reusable e-learning object (RLO) development. Method A modified Delphi study was conducted among students and educators from University Malaya (UM), Universiti Putra Malaysia (UPM) and Taylor’s University (TU) on three undergraduate programmes. In Round 1, participants were asked to select the topics from the respective syllabi to be developed into RLOs. Priority ranking was determined by using frequencies and proportions. The first quartile of the prioritised topics was included in Round 2 survey, which the participants were asked to rate the level of priority of each topic using a 5-point Likert scale. The mean score of the topics was compared between students and educators. Result A total of 43 educators and 377 students participated in this study. For UM and TU Pharmacy, there was a mismatch in the prioritised topics between the students and educators. For UPM, both the educators and students have prioritised the same topics in both rounds. To harmonise the prioritisation of topics between students and educators for UM and TU Pharmacy, the topics with a higher mean score by both the students and educators were prioritised. Conclusion The mismatch in prioritised topics between students and educators uncovered factors that might influence the prioritisation process. This study highlighted the importance of conducting needs assessment at the beginning of eLearning resources development

25th annual computational neuroscience meeting: CNS-2016

The same neuron may play different functional roles in the neural circuits to which it belongs. For example, neurons in the Tritonia pedal ganglia may participate in variable phases of the swim motor rhythms [1]. While such neuronal functional variability is likely to play a major role the delivery of the functionality of neural systems, it is difficult to study it in most nervous systems. We work on the pyloric rhythm network of the crustacean stomatogastric ganglion (STG) [2]. Typically network models of the STG treat neurons of the same functional type as a single model neuron (e.g. PD neurons), assuming the same conductance parameters for these neurons and implying their synchronous firing [3, 4]. However, simultaneous recording of PD neurons shows differences between the timings of spikes of these neurons. This may indicate functional variability of these neurons. Here we modelled separately the two PD neurons of the STG in a multi-neuron model of the pyloric network. Our neuron models comply with known correlations between conductance parameters of ionic currents. Our results reproduce the experimental finding of increasing spike time distance between spikes originating from the two model PD neurons during their synchronised burst phase. The PD neuron with the larger calcium conductance generates its spikes before the other PD neuron. Larger potassium conductance values in the follower neuron imply longer delays between spikes, see Fig. 17.Neuromodulators change the conductance parameters of neurons and maintain the ratios of these parameters [5]. Our results show that such changes may shift the individual contribution of two PD neurons to the PD-phase of the pyloric rhythm altering their functionality within this rhythm. Our work paves the way towards an accessible experimental and computational framework for the analysis of the mechanisms and impact of functional variability of neurons within the neural circuits to which they belong

EXTENSION OF TIME & LIQUIDATED ASCERTAINED DAMAGES IN CONSTRUCTION CONTRACTS

Bachelor'sBACHELOR OF SCIENCE (BUILDING