Upregulated mRNA expression of desaturase and elongase, two enzymes involved in highly unsaturated fatty acids biosynthesis pathways during follicle maturation in zebrafish

<p>Abstract</p> <p>Background</p> <p>Although unsaturated fatty acids such as eicosapentaenoic acid (EPA, C20:5n-3), docosahexaenoic acid (DHA, C22:6n-3) and arachidonic acid (ARA, C20:4n-6), collectively known as the highly unsaturated fatty acids (HUFA), play pivotal roles in vertebrate reproduction, very little is known about their synthesis in the ovary. The zebrafish (Danio rerio) display capability to synthesize all three HUFA via pathways involving desaturation and elongation of two precursors, the linoleic acid (LA, C18:2n-6) and linolenic acid (LNA, C18:3n-3). As a prerequisite to gain full understanding on the importance and regulation of ovarian HUFA synthesis, we described here the mRNA expression pattern of two enzymes; desaturase (fadsd6) and elongase (elovl5), involved in HUFA biosynthesis pathway, in different zebrafish ovarian follicle stages. Concurrently, the fatty acid profile of each follicle stage was also analyzed.</p> <p>Methods</p> <p>mRNA levels of fadsd6 and elovl5 in different ovarian follicle stages were determined by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) assays. For analysis of the ovarian follicular fatty acid composition, gas chromatography was used.</p> <p>Results</p> <p>Our results have shown that desaturase displayed significant upregulation in expression during the oocyte maturation stage. Expression of elongase was significantly highest in pre-vitellogenic follicles, followed by maturation stage. Fatty acid composition analysis of different ovarian follicle stages also showed that ARA level was significantly highest in pre-vitellogenic and matured follicles. DHA level was highest in both late vitellogenic and maturation stage.</p> <p>Conclusion</p> <p>Collectively, our findings seem to suggest the existence of a HUFA synthesis system, which could be responsible for the synthesis of HUFA to promote oocyte maturation and possibly ovulation processes. The many advantages of zebrafish as model system to understand folliculogenesis will be useful platform to further elucidate the regulatory and mechanism aspects of ovarian HUFA synthesis.</p

Electrical impedance tomography with fuzzy logic classification in lung image reconstruction

Electrical Impedance Tomography (EIT) estimates the electrical impedance distribution within a medium and produces cross-sectional images of an admittivity distribution inside an electrically conducting object. EIT in biomedicine application was first applied in lung due to it being large organs that allow large conductivity changes and is a promising technique since it allows continuous monitoring of the ventilation distribution. This study aims to explore the potential EIT technique in medical applications, with strategies to enhance the image reconstruction of the lung images. Performance of the enhanced image reconstruction is analyzed through simulation on the thorax Finite Element Model (FEM) based on a thorax CT image generated using NETGEN Mesher. To integrate and simulate EIT image of the thorax model, data are obtained from the forward and inverse model. Graz consensus Reconstruction algorithm for EIT (GREIT) technique is then applied as the consensus linear reconstruction algorithm for lung EIT images. Subsequently, the involvement of 3D imaging opens the opportunity to explore more electrode placement strategies for enhancement in image reconstruction. Performance of the reconstructed images based on electrode numbers and placement strategies are analyzed using the five figures of merits and classified into poor, average and good using Fuzzy Logic (FL). From the analysis, planar-offset configuration with 16-electrodes outperforms all others while planar configuration with 16-electrodes followed closely

Gut microbiota in early life and its influence on health and disease: a position paper by the Malaysian Working Group on Gastrointestinal Health

The role of gut microbiota in early life and its impact on gut health and subsequent diseases remain unclear. There is a lack of research and awareness in this area, especially in the Asia-Pacific region, including Malaysia. This paper reports the position of a Malaysian Working Group on some key issues surrounding gut microbiota in early life and its role in gut health and diseases, as well as experts’ stand on probiotics and prebiotics. The group reached a consensus that certain factors, including elective caesarean; premature deliveries; complementary feeding; use of antibiotics, prebiotics and/or probiotics; and exposure to the external environmental, have an impact on gut microbiota in early life. However, as evidence is lacking, especially from the Asia-Pacific region, further studies are needed to understand how gut microbiota in early life affects subsequent diseases, including allergy, inflammatory bowel disease, obesity and infantile colic. Lastly, although beneficial in acute diarrhoeal disease and probably allergic eczema, probiotics (and/or prebiotics) should be used cautiously in other gut dysbiotic conditions until more data are available

Mindful gratitude journaling: psychological distress, quality of life and suffering in advanced cancer: a randomised controlled trial

Context Numerous studies have shown that gratitude can reduce stress and improve quality of life. Objective Our study aimed to examine the effect of mindful gratitude journaling on suffering, psychological distress and quality of life of patients with advanced cancer. Methods We conducted a parallel-group, blinded, randomised controlled trial at the University of Malaya Medical Centre, Malaysia. Ninety-two adult patients with advanced cancer, and an overall suffering score ≥4/10 based on the Suffering Pictogram were recruited and randomly assigned to either a mindful gratitude journaling group (N=49) or a routine journaling group (N=43). Results After 1 week, there were significant reductions in the overall suffering score from the baseline in both the intervention group (mean difference in overall suffering score=−2.0, 95% CI=−2.7 to −1.4, t=−6.125, p=0.000) and the control group (mean difference in overall suffering score=−1.6, 95% CI=−2.3 to −0.8, t=−4.106, p=0.037). There were also significant improvements in the total Hospital Anxiety and Depression Scale score (mean difference=−3.4, 95% CI=−5.3 to −1.5, t=−3.525, p=0.000) and the total Functional Assessment of Chronic Illness Therapy-Spiritual Well-Being score (mean difference=7.3, 95% CI=1.5 to 13.1, t=2.460, p=0.014) in the intervention group after 7 days, but not in the control group. Conclusion The results provide evidence that 7 days of mindful gratitude journaling could positively affect the state of suffering, psychological distress and quality of life of patients with advanced cancer

Characterisation of protein-truncating and missense variants in PALB2 in 15 768 women from Malaysia and Singapore

Funder: National University Cancer Institute, Singapore; FundRef: http://dx.doi.org/10.13039/501100011105Background: Rare protein-truncating variants (PTVs) in partner and localiser of BRCA2 (PALB2) confer increased risk to breast cancer, but relatively few studies have reported the prevalence in South-East Asian populations. Here, we describe the prevalence of rare variants in PALB2 in a population-based study of 7840 breast cancer cases and 7928 healthy Chinese, Malay and Indian women from Malaysia and Singapore, and describe the functional impact of germline missense variants identified in this population. Methods: Mutation testing was performed on germline DNA (n=15 768) using targeted sequencing panels. The functional impact of missense variants was tested in mouse embryonic stem cell based functional assays. Results: PTVs in PALB2 were found in 0.73% of breast cancer patients and 0.14% of healthy individuals (OR=5.44; 95% CI 2.85 to 10.39, p<0.0001). In contrast, rare missense variants in PALB2 were not associated with increased risk of breast cancer. Whereas PTVs were associated with later stage of presentation and higher-grade tumours, no significant association was observed with missense variants in PALB2. However, two novel rare missense variants (p.L1027R and p.G1043V) produced unstable proteins and resulted in a decrease in homologous recombination-mediated repair of DNA double-strand breaks. Conclusion: Despite genetic and lifestyle differences between Asian and other populations, the population prevalence of PALB2 PTVs and associated relative risk of breast cancer, are similar to those reported in European populations

Robust estimation of bacterial cell count from optical density

Optical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing E. coli. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals &lt;1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data

1999 study on the profile of individual investors in Singapore.

This report investigates the profile of individual investors in Singapore resulting in a survey conducted in late 1999. Its fundamental objective is to investigate how the economic crisis has affected the investment decisions of investors in Singapore. Statistical analysis has been used to asses the impact of the crisis on investors' decisions across several of their demographics

Viable short-term directed energy weapon naval solutions: a systems analysis of current prototypes

With conventional weapons nearing their peak capability, the need to identify alternative war fighting solutions suggests a look at Directed Energy Weapons (DEWs). The goal is to change the means by which warfare is conducted to improve operational efficiencies and overall effectiveness. The Naval Postgraduate School Systems Engineering and Analysis (SEA-19B) Capstone project team examined how existing directed energy technologies can provide performance across multiple warfare area domains and mission subsets for the U.S. Navy. The aim was to identify and characterize the capability gaps with conventional weapons systems, produce a coherent vision of naval missions that incorporate DEWs, and generate a roadmap for a DEW fleet. By conducting a thorough Analysis of Alternatives based on system performance, integration, schedule, and cost, the project team identified that the Tactical Laser System (with a laser beam power of 10 kW) provided the best overall capability to defend surface combatants, although none of the analyzed DEWs have the capability to replace a current conventional weapon. The Active Denial System (microwave) provided a niche capability in the Anti-Terrorism/Force Protection mission set.http://archive.org/details/viableshorttermd1094534734Approved for public release; distribution is unlimited

Additional file 1: Figure S1. of Membrane feeding of dengue patient’s blood as a substitute for direct skin feeding in studying Aedes-dengue virus interaction

Patient’s arm inside a glove-box during feeding experiments. (DOCX 25 kb

Structural determinants for peptide-bond formation by asparaginyl ligases

Asparaginyl endopeptidases (AEPs) are cysteine proteases which break Asx (Asn/Asp)-Xaa bonds in acidic conditions. Despite sharing a conserved overall structure with AEPs, certain plant enzymes such as butelase 1 act as a peptide asparaginyl ligase (PAL) and catalyze Asx-Xaa bond formation in near-neutral conditions. PALs also serve as macrocyclases in the biosynthesis of cyclic peptides. Here, we address the question of how a PAL can function as a ligase rather than a protease. Based on sequence homology of butelase 1, we identified AEPs and PALs from the cyclic peptide-producing plants Viola yedoensis (Vy) and Viola canadensis (Vc) of the Violaceae family. Using a crystal structure of a PAL obtained at 2.4-Å resolution coupled to mutagenesis studies, we discovered ligase-activity determinants flanking the S1 site, namely LAD1 and LAD2 located around the S2 and S1' sites, respectively, which modulate ligase activity by controlling the accessibility of water or amine nucleophile to the S-ester intermediate. Recombinantly expressed VyPAL1-3, predicted to be PALs, were confirmed to be ligases by functional studies. In addition, mutagenesis studies on VyPAL1-3, VyAEP1, and VcAEP supported our prediction that LAD1 and LAD2 are important for ligase activity. In particular, mutagenesis targeting LAD2 selectively enhanced the ligase activity of VyPAL3 and converted the protease VcAEP into a ligase. The definition of structural determinants required for ligation activity of the asparaginyl ligases presented here will facilitate genomic identification of PALs and engineering of AEPs into PALs.Ministry of Education (MOE)Accepted versionThis research was supported by Academic Research Grant Tier 3 (MOE2016-T3-1-003) from the Singapore Ministry of Education (MOE) to the J.P.T., J.L., and C.-F.L. laboratories, and NMRC Grants CBRG/0028/2014 and NRF2016NRF-CRP001-063