Host immune response against DENV and ZIKV infections.

Dengue is a major public health concern, affecting almost 400 million people worldwide, with about 70% of the global burden of disease in Asia. Despite revised clinical classifications of dengue infections by the World Health Organization, the wide spectrum of the manifestations of dengue illness continues to pose challenges in diagnosis and patient management for clinicians. When the Zika epidemic spread through the American continent and then later to Africa and Asia in 2015, researchers compared the characteristics of the Zika infection to Dengue, considering both these viruses were transmitted primarily through the same vector, the Aedes aegypti female mosquitoes. An important difference to note, however, was that the Zika epidemic diffused in a shorter time span compared to the persisting feature of Dengue infections, which is endemic in many Asian countries. As the pathogenesis of viral illnesses is affected by host immune responses, various immune modulators have been proposed as biomarkers to predict the risk of the disease progression to a severe form, at a much earlier stage of the illness. However, the findings for most biomarkers are highly discrepant between studies. Meanwhile, the cross-reactivity of CD8+ and CD4+ T cells response to Dengue and Zika viruses provide important clues for further development of potential treatments. This review discusses similarities between Dengue and Zika infections, comparing their disease transmissions and vectors involved, and both the innate and adaptive immune responses in these infections. Consideration of the genetic identity of both the Dengue and Zika flaviviruses as well as the cross-reactivity of relevant T cells along with the actions of CD4+ cytotoxic cells in these infections are also presented. Finally, a summary of the immune biomarkers that have been reported for dengue and Zika viral infections are discussed which may be useful indicators for future anti-viral targets or predictors for disease severity. Together, this information appraises the current understanding of both Zika and Dengue infections, providing insights for future vaccine design approaches against both viruses.

The Cytokines CXCL10 and CCL2 and the Kynurenine Metabolite Anthranilic Acid Accurately Predict Patients at Risk of Developing Dengue With Warning Signs.

BACKGROUND: The resolution or aggravation of dengue infection depends on the patient's immune response during the critical phase. Cytokines released by immune cells increase with the worsening severity of dengue infections. Cytokines activate the kynurenine pathway (KP) and the extent of KP activation then influences disease severity. METHODS: KP metabolites and cytokines in plasma samples of patients with dengue infection (dengue without warning signs [DWS-], dengue with warning signs [DWS+], or severe dengue) were analyzed. Cytokines (interferon gamma [IFN-É£], tumor necrosis factor, interleukin 6, CXCL10/interferon-inducile protein 10 [IP-10], interleukin 18 [IL-18], CCL2/monocyte chemoattractant protein-1 [MCP-1], and CCL4/macrophage inflammatory protein-1beta [MIP-1ß] were assessed by a Human Luminex Screening Assay, while KP metabolites (tryptophan, kynurenine, anthranilic acid [AA], picolinic acid, and quinolinic acid) were assessed by ultra-high-performance liquid chromatography and Gas Chromatography Mass Spectrophotometry [GCMS] assays. RESULTS: Patients with DWS+ had increased activation of the KP where kynurenine-tryptophan ratio, anthranilic acid, and picolinic acid were elevated. These patients also had higher levels of the cytokines IFN-É£, CXCL10, CCL4, and IL-18 than those with DWS-. Further receiver operating characteristic analysis identified 3 prognostic biomarker candidates, CXCL10, CCL2, and AA, which predicted patients with higher risks of developing DWS+ with an accuracy of 97%. CONCLUSIONS: The data suggest a unique biochemical signature in patients with DWS+. CXCL10 and CCL2 together with AA are potential prognostic biomarkers that discern patients with higher risk of developing DWS+ at earlier stages of infection.

Improved spectrophotometric human interferon-gamma bioassay.

Interferon gamma (IFNγ) is a cytokine involved in many anti-viral and immunoregulatory processes. One of the major mechanisms through which IFNγ exerts these effects is by inducing expression of indoleamine 2,3 dioxygenase-1 (IDO1), an enzyme that catalyses the first, rate-limiting step of the kynurenine pathway. In this pathway, tryptophan can be catabolised to many products, including picolinic acid and nicotinamide adenine dinucleotide. However, in endothelial cells, the pathway ends at the production of kynurenine. This is due to little or no expression of enzymes that metabolise kynurenine. Production of kynurenine has been used as an indicator of human IDO1 activity, and hence as an hIDO1 bioassay. Due to IFNγ's ability to induce IDO1 expression, kynurenine production can also be a measure of human IFNγ (hIFNγ) bioactivity. Previously, the levels of hIFNγ have been commonly determined by anti-viral assays, high performance liquid chromatography and ELISA. Apart from their technical complexity, these assays are costly and only the anti-viral assay measures bioactive IFNγ. Here, we report the development of an improved IFNγ spectrophotometric bioassay using a human brain endothelial cell line (HBEC 5i). The method is sensitive, easy to perform and cost efficient.

Integrating an open-source course management system (Moodle) into the teaching of a first-year medical physiology course: a case study.

Educators in medical schools around the world are presently experimenting with innovative ways of using web-based learning to supplement the existing teaching and learning process. We have recently used a popular open-source course management system (CMS) called the modular object-oriented dynamic learning environment (Moodle) to construct an online site (DPhysiol) to facilitate our face-to-face teaching of physiology to a group of first-year students in the Bachelor of Medicine and Bachelor of Surgery program. The integration of the Moodle site into our teaching was assessed using online log activity, student examination marks, and feedback from students. The freely available Moodle platform was simple to use, helped to effectively deliver course materials, and has features that allowed cooperative learning. Students who used the CMS throughout their academic year and commented favorably regarding its use as a complement to the face-to-face classroom sessions. The group of students used the CMS obtained significantly higher scores in the final examination compared with the previous class that did not use the CMS. In addition, there was a significant correlation between student participation and performance in online quizzes and their final examination marks. However, students' overall online usage of the CMS did not correlate with their examination marks. We recommend Moodle as a useful tool for physiology educators who are interested in integrating web-based learning into their existing teaching curriculum.