The field of systems biology is often held back by difficulties in obtaining comprehensive, highquality, quantitative data sets. In this paper, we undertook an interlaboratory effort to generate such a data set for a very large number of cellular components in the yeast Saccharomyces cerevisiae, a widely used model organism that is also used in the production of fuels, chemicals, food ingredients and pharmaceuticals. With the current focus on biofuels and sustainability, there is much interest in harnessing this species as a general cell factory. In this study, we characterized two yeast strains, under two standard growth conditions. We ensured the high quality of the experimental data by evaluating a wide range of sampling and analytical techniques. Here we show significant differences in the maximum specific growth rate and biomass yield between the two strains. on the basis of the integrated analysis of the highthroughput data, we hypothesize that differences in phenotype are due to differences in protein metabolism.

We here present simple and rapid methods for fast screening of yeast lipids in Saccharomyces cerevisiae. First we introduced a microwave-assisted technique for fast lipid extraction that allows the extraction of lipids within 10 min. The new method enhances extraction rate by 27 times, while maintaining product yields comparable to conventional methods (n = 14, P > 0.05). The recovery (n = 3) from spiking of synthetic standards were 92 ± 6% for cholesterol, 95 ± 4% for triacylglycerol, and 92 ± 4% for free fatty acids. Additionally, the new extraction method combines cell disruption and extraction in one step, and the approach, therefore, not only greatly simplifies sample handling but also reduces analysis time and minimizes sample loss during sample preparation. Second, we developed a chromatographic separation that allowed separation of neutral and polar lipids from the extracted samples within a single run. The separation was performed based on a three gradient solvent system combined with hydrophilic interaction liquid chromatography-HPLC followed by detection using a charged aerosol detector. The method was shown to be highly reproducible in terms of retention time of the analytes (intraday; 0.002-0.034% RSD; n = 10, interday; 0.04-1.35% RSD; n = 5) and peak area (intraday; 0.63-6% RSD; n = 10, interday; 4-12% RSD; n = 5).

Nutrient sensing and coordination of metabolic pathways are crucial functions for living cells. A combined analysis of the yeast transcriptome, phosphoproteome and metabolome is used to investigate the interactions between the Snf1 and TORC1 pathways under nutrient-limited conditions.

We present a fast and accurate method for preparation of fatty acid methyl esters (FAMEs) using microwave-assisted derivatization of fatty acids present in yeast samples. The esterification of free/bound fatty acids to FAMEs was completed within 5 min, which is 24 times faster than with conventional heating methods. The developed method was validated in two ways: (1) through comparison with a conventional method (hot plate) and (2) through validation with the standard reference material (SRM) 3275-2 omega-3 and omega-6 fatty acids in fish oil (from the Nation Institute of Standards and Technology, USA). There were no significant differences (P>0.05) in yields of FAMEs with both validations. By performing a simple modification of closed-vessel microwave heating, it was possible to carry out the esterification in Pyrex glass tubes kept inside the closed vessel. Hereby, we are able to increase the number of sample preparations to several hundred samples per day as the time for preparation of reused vessels was eliminated. Pretreated cell disruption steps are not required, since the direct FAME preparation provides equally quantitative results. The new microwave-assisted derivatization method facilitates the preparation of FAMEs directly from yeast cells, but the method is likely to also be applicable for other biological samples.

Several reports have shown antimicrobial and anticancer activities of mucous glycoproteins extracted from the giant African snail Achatina fulica. Anticancer properties of the snail mucous peptides remain incompletely revealed. The aim of this study was to predict anticancer peptides from A. fulica mucus. Two of HPLC-separated mucous fractions (F2 and F5) showed in vitro cytotoxicity against the breast cancer cell line (MCF-7) and normal epithelium cell line (Vero). According to the mass spectrometric analysis, 404 and 424 peptides from the F2 and F5 fractions were identified. Our comprehensive bioinformatics workflow predicted 16 putative cationic and amphipathic anticancer peptides with diverse structures from these two peptidome data. These peptides would be promising molecules for new anti-breast cancer drug development.

Bioluminescence, which living organisms such as fireflies emit light, has been studied extensively for over half a century. This intriguing reaction, having its origins in nature where glowing insects can signal things such as attraction or defense, is now widely used in biotechnology with applications of bioluminescence and chemiluminescence. Luciferase, a key enzyme in this reaction, has been well characterized; however, the enzymes involved in the biosynthetic pathway of its substrate, luciferin, remains unsolved at present. To elucidate the luciferin metabolism, we performed a de novo transcriptome analysis using larvae of the firefly species, Luciola aquatilis. Here, a comparative analysis is performed with the model coleopteran insect Tribolium casteneum to elucidate the metabolic pathways in L. aquatilis. Based on a template luciferin biosynthetic pathway, combined with a range of protein and pathway databases, and various prediction tools for functional annotation, the candidate genes, enzymes, and biochemical reactions involved in luciferin metabolism are proposed for L. aquatilis. The candidate gene expression is validated in the adult L. aquatilis using reverse transcription PCR (RT-PCR). This study provides useful information on the bio-production of luciferin in the firefly and will benefit to future applications of the valuable firefly bioluminescence system.

IntroductionDental and oral diseases are major concerns in Thailand, affecting the country's health system. Everyone, at any age, is at risk of losing teeth. Tooth loss can lead to many health problems, including difficulty chewing, which increases the risk of poor nutrition. In addition, tooth loss leaves a large gap, leading to plaque accumulation that can cause periodontal disease. Moreover, the remaining teeth start to shift in an attempt to fill in the gap, leading to misalignment of teeth. Losing teeth may also affect personality, including a loss of confidence that may affect work or social activities. Restoration, with dentures or dental implants, is needed to replace the missing teeth.Current research has focused on dental and bone implants capable of promoting cell differentiation Xavier Acasigua et al., 2014;Naddeo et al., 2015). Cells isolated from tooth pulp contain odontoblasts, fibroblasts, immune cells, and undifferentiated mesenchymal cells called dental pulp stem cells (DPSCs). These findings may lead to restoration using dental pulp cells (DPCs), together with dental implants, to replace lost teeth. Various research groups have proposed the possibility of producing bioactive dental implants by adding natural products, such as natural hydroxyapatite/ zircon (Karamian et al., 2014), plant products (Varoni et al., 2012), or herbal extracts (Wang et al., 2012), to dental materials.Snails have been used in medicine since ancient times. Until the late 19th century, the synthetic peptide ziconotide (SNXIII), which is found in the venom of Conus magus, was used as a painkiller following approval by the United States Food and Drug Administration (Bonnemain, 2005). According to reports, snails contain several medically useful substances, including mytimacinlike antimicrobial peptides, which are exclusively found in the mucus of giant African snails (Zhong et al., 2013). In addition, lectin, which acts as an antimicrobial, is also present in the mucus of giant African snails (Ito et al., 2011). High-performance liquid chromatography was used to determine the amount of allantoin and glycolic Abstract: Dental pulp tissue contains stem cells that can be isolated and used for regenerative medicine in tooth restoration or autologous transplantation. The aim of this study was to observe the mineralization and gene expression in dental pulp cells (DPCs) following treatment with snail mucus. Snail mucus was collected from adult Achatina fulica and processed as a dry powder by the freeze-drying technique. The mucus powder was dissolved in a culture medium at a concentration of 15 µg/mL. DPCs were prepared by the outgrowth technique and cultured in a 6-well plate at a density of 5 × 10 4 cells per well. A mucus-supplemented medium (15 µg/mL) was added to each well. Cell culture was maintained for 3 weeks. The results of Alizarin Red S staining indicated that the DPCs cultured in a medium supplemented with snail mucus showed a higher number of mineralized nodules as compared with the control group cultured in a norm...

BackgroundIn the yeast Saccharomyces cerevisiae, genes containing UASINO sequences are regulated by the Ino2/Ino4 and Opi1 transcription factors, and this regulation controls lipid biosynthesis. The expression level of INO2 and INO4 genes (INO-level) at different nutrient limited conditions might lead to various responses in yeast lipid metabolism.MethodsIn this study, we undertook a global study on how INO-levels (transcription level of INO2 and INO4) affect lipid metabolism in yeast and we also studied the effects of single and double deletions of the two INO-genes (deficient effect). Using 2 types of nutrient limitations (carbon and nitrogen) in chemostat cultures operated at a fixed specific growth rate of 0.1 h-1 and strains having different INO-level, we were able to see the effect on expression level of the genes involved in lipid biosynthesis and the fluxes towards the different lipid components. Through combined measurements of the transcriptome, metabolome, and lipidome it was possible to obtain a large dataset that could be used to identify how the INO-level controls lipid metabolism and also establish correlations between the different components.ResultsIn this study, we undertook a global study on how INO-levels (transcription level of INO2 and INO4) affect lipid metabolism in yeast and we also studied the effects of single and double deletions of the two INO-genes (deficient effect). Using 2 types of nutrient limitations (carbon and nitrogen) in chemostat cultures operated at a fixed specific growth rate of 0.1 h-1 and strains having different INO-level, we were able to see the effect on expression level of the genes involved in lipid biosynthesis and the fluxes towards the different lipid components. Through combined measurements of the transcriptome, metabolome, and lipidome it was possible to obtain a large dataset that could be used to identify how the INO-level controls lipid metabolism and also establish correlations between the different components.ConclusionsOur analysis showed the strength of using a combination of transcriptome and lipidome analysis to illustrate the effect of INO-levels on phospholipid metabolism and based on our analysis we established a global regulatory map.