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1.
Sci Rep ; 14(1): 9855, 2024 04 29.
Article in English | MEDLINE | ID: mdl-38684759

ABSTRACT

Gut microbiome dysbiosis contributes to the pathophysiology of both gestational diabetes mellitus (GDM) and its associated adverse outcomes in the woman and offspring. Even though GDM prevalence, complications, and outcomes vary among different ethnic groups, limited information is available about the influence of ethnicity on gut microbiome dysbiosis in pregnancies complicated by GDM. This pilot prospective cohort study examined the impact of ethnicity on gut dysbiosis in GDM among three Asian ethnic groups (Chinese, Malay, Indian) living in Singapore, and investigated the potential modulatory roles of diet and lifestyle modifications on gut microbiome post-GDM diagnosis. Women with GDM (n = 53) and without GDM (n = 16) were recruited. Fecal samples were collected at 24-28- and 36-40-weeks' gestation and analyzed by targeted 16S rRNA gene-based amplicon sequencing. Permutational multivariate analysis of variance (PERMANOVA) analysis was performed to evaluate differences between groups. Differentially abundant taxa were identified by DeSeq2 based analysis. Functional prediction was performed using the phylogenetic investigation of communities by reconstruction of unobserved states (PICRUSt2). Among women with GDM, gut microbiome from different ethnicities harbored common microbial features. However, among those without GDM, there was contrasting microbiome composition between ethnic groups. Microbial members such as Collinsella, Blautia, Ruminococcus, Ruminococcus gnavus, Ruminococcus torques, and Eubacterium hallii groups were differentially enriched (p < 0.05) in women with GDM compared to those without. Among women with GDM, no differences in alpha- and beta- diversity were observed when comparing 24-28 weeks' samples with 36-40 weeks' samples, a period covering intense dietary and lifestyle modification, suggesting an inability to modulate gut microbiota through classic GDM management. Women with GDM have a distinct gut microbiome profile which harbours common features across different Asian ethnic groups, consistent with the notion that specific microbes are involved in the pathogenesis of insulin resistance, pro-inflammatory conditions, and other metabolic dysregulation known to be present in GDM.


Subject(s)
Diabetes, Gestational , Dysbiosis , Gastrointestinal Microbiome , Humans , Female , Pregnancy , Diabetes, Gestational/microbiology , Dysbiosis/microbiology , Pilot Projects , Adult , Singapore/epidemiology , Prospective Studies , Asian People , RNA, Ribosomal, 16S/genetics , Diet , Ethnicity , Feces/microbiology , Bacteria/classification , Bacteria/genetics , Bacteria/isolation & purification
2.
NPJ Biofilms Microbiomes ; 10(1): 8, 2024 Jan 23.
Article in English | MEDLINE | ID: mdl-38253600

ABSTRACT

Fires in tropical peatlands extend to depth, transforming them from carbon sinks into methane sources and severely limit forest recovery. Peat microbiomes influence carbon transformations and forest recovery, yet our understanding of microbiome shifts post-fire is currently limited. Our previous study highlighted altered relationships between the peat surface, water table, aboveground vegetation, and methane flux after fire in a tropical peatland. Here, we link these changes to post-fire shifts in peat microbiome composition and assembly processes across depth. We report kingdom-specific and depth-dependent shifts in alpha diversity post-fire, with large differences at deeper depths. Conversely, we found shifts in microbiome composition across all depths. Compositional shifts extended to functional groups involved in methane turnover, with methanogens enriched and methanotrophs depleted at mid and deeper depths. Finally, we show that community shifts at deeper depths result from homogeneous selection associated with post-fire changes in hydrology and aboveground vegetation. Collectively, our findings provide a biological basis for previously reported methane fluxes after fire and offer new insights into depth-dependent shifts in microbiome assembly processes, which ultimately underlie ecosystem function predictability and ecosystem recovery.


Subject(s)
Carbon , Microbiota , Methane , Soil
3.
Nat Chem Biol ; 20(4): 473-483, 2024 Apr.
Article in English | MEDLINE | ID: mdl-37957272

ABSTRACT

The rhizosphere is a niche surrounding plant roots, where soluble and volatile molecules mediate signaling between plants and the associated microbiota. The preferred lifestyle of soil microorganisms is in the form of biofilms. However, less is known about whether root volatile organic compounds (rVOCs) can influence soil biofilms beyond the 2-10 mm rhizosphere zone influenced by root exudates. We report that rVOCs shift the microbiome composition and growth dynamics of complex soil biofilms. This signaling is evolutionarily conserved from ferns to higher plants. Methyl jasmonate (MeJA) is a bioactive signal of rVOCs that rapidly triggers both biofilm and microbiome changes. In contrast to the planktonic community, the resulting biofilm community provides ecological benefits to the host from a distance via growth enhancement. Thus, a volatile host defense signal, MeJA, is co-opted for assembling host-beneficial biofilms in the soil microbiota and extending the sphere of host influence in the rhizosphere.


Subject(s)
Acetates , Cyclopentanes , Microbiota , Oxylipins , Soil , Plant Roots , Soil Microbiology , Rhizosphere , Biofilms
4.
Microbiol Resour Announc ; 12(12): e0063323, 2023 Dec 14.
Article in English | MEDLINE | ID: mdl-37982654

ABSTRACT

We report the complete genome sequence of Methylomonas sp. UP202 isolated from an urban waterway sediment in Singapore. The genome contains genes involved in methane, methanol, formaldehyde, and formate oxidation. It also contains genes utilizing various nitrogen sources such as nitrogen, nitrate, nitrite, urea, and ammonium.

5.
mSystems ; 8(4): e0038723, 2023 08 31.
Article in English | MEDLINE | ID: mdl-37409823

ABSTRACT

There are gaps in our understandings on how did the evolutionary relationships among members of the phytobiomes shape their ability to produce tremendously complex specialized metabolites under the influence of plant host. To determine these relationships, we investigated the phylogenetic conservation of biosynthetic gene clusters (BGCs) on a global collection of 4,519 high-quality and nonredundant (out of 12,181) bacterial isolates and metagenome-assembled genomes from 47 different plant hosts and soil, by adopting three independent phylogenomic approaches (D-test, Pagel's λ, and consenTRAIT). We report that the BGCs are phylogenetically conserved to varying strengths and depths in their different classes. We show that the ability to produce specialized metabolites qualifies as a complex trait, and the depth of conservation is equivalent to ecologically relevant complex microbial traits. Interestingly, terpene and aryl polyene BGCs had the strongest phylogenetic conservation in the phytobiomes, but not in the soil microbiomes. Furthermore, we showed that terpenes are largely uncharacterized in phytobiomes and pinpointed specific clades that harbor potentially novel terpenes. Taken together, this study sheds light on the evolution of specialized metabolites' biosynthesis potential in phytobiomes under the influence of plant hosts and presents strategies to rationally guide the discovery of potentially novel classes of metabolites. IMPORTANCE This study expands our understandings of the biosynthetic potential of phytobiomes by using such worldwide and extensive collection of microbiomes from plants and soil. Apart from providing such vital resource for the plant microbiome researchers, this study provides fundamental insights into the evolution of biosynthetic gene clusters (BGCs) in phytobiomes under the influence of plant host. Specifically, we report that the strength of phylogenetic conservation in microbiomes varies for different classes of BGCs and is influenced as a result of plant host association. Furthermore, our results indicate that biosynthetic potential of specialized metabolites is deeply conserved equivalent to other complex and ecologically relevant microbial traits. Finally, for the most conserved class of specialized metabolites (terpenes), we identified clades harboring potentially novel class of molecules. Future studies could focus on plant-microbe coevolution and interactions through specialized metabolites building upon these findings.


Subject(s)
Metagenome , Terpenes , Phylogeny , Secondary Metabolism/genetics , Metagenome/genetics , Multigene Family/genetics
6.
Sci Data ; 10(1): 373, 2023 06 08.
Article in English | MEDLINE | ID: mdl-37291271

ABSTRACT

Tropical peatlands in South-East Asia are some of the most carbon-dense ecosystems in the world. Extensive repurposing of such peatlands for forestry and agriculture has resulted in substantial microbially-driven carbon emissions. However, we lack an understanding of the microorganisms and their metabolic pathways involved in carbon turnover. Here, we address this gap by reconstructing 764 sub-species-level genomes from peat microbiomes sampled from an oil palm plantation located on a peatland in Indonesia. The 764 genomes cluster into 333 microbial species (245 bacterial and 88 archaeal), of which, 47 are near-complete (completeness ≥90%, redundancy ≤5%, number of unique tRNAs ≥18) and 170 are substantially complete (completeness ≥70%, redundancy ≤10%). The capacity to respire amino acids, fatty acids, and polysaccharides was widespread in both bacterial and archaeal genomes. In contrast, the ability to sequester carbon was detected only in a few bacterial genomes. We expect our collection of reference genomes to help fill some of the existing knowledge gaps about microbial diversity and carbon metabolism in tropical peatlands.


Subject(s)
Carbon , Ecosystem , Soil , Agriculture , Carbon/analysis , Indonesia , Soil/chemistry , Microbiota
7.
J Fungi (Basel) ; 9(4)2023 Mar 29.
Article in English | MEDLINE | ID: mdl-37108875

ABSTRACT

Soil-borne beneficial microbes establish symbioses with plant hosts and play key roles during growth and development therein. In this study, two fungal strains, FLP7 and B9, were isolated from the rhizosphere microbiome associated with Choy Sum (Brassica rapa var. parachinensis) and barley (Hordeum vulgare), respectively. Sequence analyses of the internal transcribed spacer and 18S ribosomal RNA genes combined with colony and conidial morphology identified FLP7 and B9 to be Penicillium citrinum strains/isolates. Plant-fungus interaction assays revealed that isolate B9 showed significant growth promotion effects in Choy Sum plants cultivated in normal soil, as well as under phosphate-limiting conditions. In comparison to the mock control, B9-inoculated plants showed a 34% increase in growth in aerial parts and an 85% upsurge in the fresh weight of roots when cultivated in sterilized soil. The dry biomass of such fungus-inoculated Choy Sum increased by 39% and 74% for the shoots and roots, respectively. Root colonization assays showed that P. citrinum associates directly with the root surface but does not enter or invade the root cortex of the inoculated Choy Sum plants. Preliminary results also indicated that P. citrinum can promote growth in Choy Sum via volatile metabolites too. Interestingly, we detected relatively higher amounts of gibberellins and cytokinins in axenic P. citrinum culture filtrates through liquid chromatography-mass spectrometry analyses. This could plausibly explain the overall growth induction in P. citrinum-inoculated Choy Sum plants. Furthermore, the phenotypic growth defects associated with the Arabidopsis ga1 mutant could be chemically complemented by the exogenous application of P. citrinum culture filtrate, which also showed accumulation of fungus-derived active gibberellins. Our study underscores the importance of transkingdom beneficial effects of such mycobiome-assisted nutrient assimilation and beneficial fungus-derived phytohormone-like metabolites in the induction of robust growth in urban farmed crops.

8.
Sci Total Environ ; 851(Pt 1): 158141, 2022 Dec 10.
Article in English | MEDLINE | ID: mdl-35995171

ABSTRACT

Food security is critical and has become a global concern with many of our basic food crops growing in areas with high drought risk. To improve soil water holding capacity, hydrogels are a promising solution. However, the current ones are mostly derived from petroleum products and are environmental unsustainable. In this study, the main objective is to determine if bio-based hydrogel can help in the growth of leafy vegetables while minimizing water use under field conditions. To achieve this, we developed an okara-derived hydrogel (Ok-PAA; OP) from by-products of bean curd and soybean milk production. We incorporated OP into soil and assessed the growth performance of leafy vegetables. We observed that vegetables grown with 0.2% (w/v) OP in soil with a watering frequency of 7 times per week resulted in >60 % and 35 % yield increase for the common Asian leafy vegetables, choy sum (CS) and pak choi (PC), respectively, as compared to without hydrogel supplementation. Both vegetables produced larger leaf areas (20-40 % increment) in the presence of the hydrogel as compared to those without. In addition, with OP amendment, the irrigation water use efficiency improved >60 % and 30 % for CS and PC, respectively. It is estimated that with the use of the hydrogel, a reduction in watering frequency from 21 times to 7 times per week could be achieved, and based on a per hectare estimation, this would result in 196,000 L of water saving per crop cycle. Statistical analysis and modelling further confirmed vegetables grown with 0.2 % (w/v) OP and with a watering frequency of 7 times per week showed the best growth performance and water use efficiency. Such a waste-to-resource approach offers a plant-based soil supplement for crop growers, contributes to waste valorization, and enhances the growth of plants especially under water-limited conditions.


Subject(s)
Fabaceae , Petroleum , Hydrogels , Soil , Glycine max , Vegetables , Water
9.
Metabolites ; 12(5)2022 Apr 29.
Article in English | MEDLINE | ID: mdl-35629910

ABSTRACT

Rapid proliferation of cyanobacteria in both benthic and suspended (planktonic) habitats is a major threat to environmental safety, as they produce nuisance compounds such as cytotoxins and off-flavors, which degrade the safety and quality of water supplies. Temperature and light irradiance are two of the key factors in regulating the occurrence of algal blooms and production of major off-flavors. However, the role of these factors in regulating the growth and metabolism is poorly explored for both benthic and planktonic cyanobacteria. To fill this gap, we studied the effects of light and temperature on the growth and metabolic profiling of both benthic (Hapalosiphon sp. MRB220) and planktonic (Planktothricoides sp. SR001) environmental species collected from a freshwater reservoir in Singapore. Moreover, this study is the first report on the metabolic profiling of cyanobacteria belonging to two different habitats in response to altered environmental conditions. The highest growth rate of both species was observed at the highest light intensity (100 µmol photons/m²/s) and at a temperature of 33 °C. Systematic metabolite profiling analysis suggested that temperature had a more profound effect on metabolome of the Hapalosiphon, whereas light had a greater effect in the case of Planktothricoides. Interestingly, Planktothricoides sp. SR001 showed a specialized adaptation mechanism via biosynthesis of arginine, and metabolism of cysteine and methionine to survive and withstand higher temperatures of 38 °C and higher. Hence, the mode of strategies for coping with different light and temperature conditions was correlated with the growth and alteration in metabolic activities for physiological and ecological adaptations in both species. In addition, we putatively identified a number of unique metabolites with a broad range of antimicrobial activities in both species in response to both light and temperature. These metabolites could play a role in the dominant behavior of these species in suppressing competition during bloom formation. Overall, this study elucidated novel insights into the effects of environmental factors on the growth, metabolism, and adaptation strategies of cyanobacteria from two different habitats, and could be useful in controlling their harmful effects on human health and environmental concerns.

10.
Indian J Microbiol ; 61(3): 283-290, 2021 Sep.
Article in English | MEDLINE | ID: mdl-34294994

ABSTRACT

Environmental bioremediation relies heavily on the realized potential of efficient bioremediation agents or microbial strains of interest. Identifying suitable microbial agents for plant biomass waste valorization requires (i) high-quality genome assemblies to predict the full metabolic and functional potential, (ii) accurate mapping of lignocellulose metabolizing enzymes. However, fragmented nature of the sequenced genomes often limits the prediction ability due to breaks occurring in coding sequences. To address these challenges and as part of our ongoing agri-culturomics efforts, we have performed a hybrid genome assembly using Illumina and Nanopore reads with modified assembly protocol, for a novel Streptomyces strain isolated from the rhizosphere niche of green leafy vegetables grown in a commercial urban farm. High-quality genome was assembled with the size of 8.6 Mb in just two contigs with N50 of 8,542,030 and coverage of 383X. This facilitated identification and complete arrangement of approximately 248 CAZymes and 38 biosynthetic gene clusters in the genome. Multiple gene clusters consisting of cellulases and hemicellulases associated with substrate recognition domain were identified in the genome. Genes for lignin, chitin, and even some aromatic compounds degradation were found in the Streptomyces sp. genome which makes it a promising candidate for lignocellulosic waste valorization. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12088-021-00935-5.

11.
Front Robot AI ; 8: 572243, 2021.
Article in English | MEDLINE | ID: mdl-34124169

ABSTRACT

Using a team of robots for estimating scalar environmental fields is an emerging approach. The aim of such an approach is to reduce the mission time for collecting informative data as compared to a single robot. However, increasing the number of robots requires coordination and efficient use of the mission time to provide a good approximation of the scalar field. We suggest an online multi-robot framework m-AdaPP to handle this coordination. We test our framework for estimating a scalar environmental field with no prior information and benchmark the performance via field experiments against conventional approaches such as lawn mower patterns. We demonstrated that our framework is capable of handling a team of robots for estimating a scalar field and outperforms conventional approaches used for approximating water quality parameters. The suggested framework can be used for estimating other scalar functions such as air temperature or vegetative index using land or aerial robots as well. Finally, we show an example use case of our adaptive algorithm in a scientific study for understanding micro-level interactions.

12.
Plant Cell Physiol ; 62(2): 248-261, 2021 May 11.
Article in English | MEDLINE | ID: mdl-33475132

ABSTRACT

The Casparian strip (CS) constitutes a physical diffusion barrier to water and nutrients in plant roots, which is formed by the polar deposition of lignin polymer in the endodermis tissue. The precise pattern of lignin deposition is determined by the scaffolding activity of membrane-bound Casparian Strip domain proteins (CASPs), but little is known of the mechanism(s) directing this process. Here, we demonstrate that Endodermis-specific Receptor-like Kinase 1 (ERK1) and, to a lesser extent, ROP Binding Kinase1 (RBK1) are also involved in regulating CS formation, with the former playing an essential role in lignin deposition as well as in the localization of CASP1. We show that ERK1 is localized to the cytoplasm and nucleus of the endodermis and that together with the circadian clock regulator, Time for Coffee (TIC), forms part of a novel signaling pathway necessary for correct CS organization and suberization of the endodermis, with their single or combined loss of function resulting in altered root microbiome composition. In addition, we found that other mutants displaying defects in suberin deposition at the CS also display altered root exudates and microbiome composition. Thus, our work reveals a complex network of signaling factors operating within the root endodermis that establish both the CS diffusion barrier and influence the microbial composition of the rhizosphere.


Subject(s)
Arabidopsis/metabolism , Microbiota , Plant Roots/metabolism , Rhizosphere , Signal Transduction , Arabidopsis Proteins/metabolism , Nuclear Proteins/metabolism , Plant Roots/microbiology , Signal Transduction/physiology
13.
Front Microbiol ; 11: 539921, 2020.
Article in English | MEDLINE | ID: mdl-33178143

ABSTRACT

Microorganisms are critically important for the function of surface water ecosystems but are frequently subjected to anthropogenic disturbances at either acute (pulse) or long-term (press) scales. Response and recovery of microbial community composition and function following pulse disturbance is well-studied in controlled, laboratory scale experiments but is less well-understood in natural environments undergoing continual press disturbance. The objectives of this study were to determine the drivers of sediment microbial compositional and functional changes in freshwaters receiving continual press disturbance from agricultural land runoff and to evaluate the ability of the native microbial community to resist disturbance related changes as a proxy for freshwater ecosystem health. Freshwater sediments were collected seasonally over 1 year in Kewaunee County, Wisconsin, a region impacted by concentrated dairy cattle farming, manure fertilization, and associated agricultural runoff which together serve as a press disturbance. Using 16S rRNA gene amplicon sequencing, we found that sediments in locations strongly impacted by intensive agriculture contain significantly higher abundances (p < 0.01) of the genera Thiobacillus, Methylotenera, Crenotrhix, Nitrospira, and Rhodoferax compared to reference sediments, and functions including nitrate reduction, nitrite reduction, and nitrogen respiration are significantly higher (p < 0.05) at locations in close proximity to large farms. Nine species-level potential human pathogens were identified in riverine sediments including Acinetobacer lwoffi and Arcobacter skirrowii, two pathogens associated with the cattle microbiome. Microbial community composition at locations in close proximity to intensive agriculture was not resistant nor resilient to agricultural runoff disturbance within 5 months post-disturbance but did reach a new, stable microbial composition. From this data, we conclude that sediment microbial community composition is sensitive and shifts in response to chemical and microbial pollution from intensive agriculture, has a low capacity to resist infiltration by non-native, harmful bacteria and, overall, the natural buffering capacity of freshwater ecosystems is unable to fully resist the impacts from agricultural press disturbance.

14.
Sci Data ; 7(1): 278, 2020 08 25.
Article in English | MEDLINE | ID: mdl-32843634

ABSTRACT

The genome sequences of many microbial species from the phytobiomes of several leafy Asian greens remain unknown. Here, we address this gap by reconstructing 910 prokaryotic draft genomes from 24 leaf, 65 root, 12 soil, and 6 compost metagenomes from the seedling and adult developmental stages of three leafy Asian greens - Brassica rapa var. parachinensis, Brassica oleracea var. alboglabra and Amaranthus spp. - grown in a commercial, soil-based urban farm. Of these, 128 are near-complete (>90% completeness, <5% redundancy), 540 are substantially complete (≥70% completeness, <10%, redundancy), while the rest have a completeness ≥50% and redundancy <10%. The draft genomes together span 292 bacterial and 3 archaeal species, a subset of which are from underrepresented genus-level lineages in public databases. We expect our dataset to facilitate a wide range of comparative studies that seek to understand the different functional aspects of vegetable crop phytobiomes and for devising new strategies for microbial cultivation in the future.


Subject(s)
Amaranthus/microbiology , Brassica/microbiology , Metagenome , Plant Leaves/microbiology , Crops, Agricultural/microbiology , Farms
15.
J Agric Food Chem ; 68(46): 13241-13246, 2020 Nov 18.
Article in English | MEDLINE | ID: mdl-32364750

ABSTRACT

Okara (Ok) or soybean residue is produced as a byproduct from the soybean milk and soybean curd industries world wide, most of which is disposed or burned as waste. It is important to explore the possibilities to convert okara to useful materials, because okara is a naturally renewable bioresource. Here, we report the chemical modification of okara by grafting poly(acrylic acid) (PAA) onto the backbones of okara in water medium and the characterization of the Ok-PAA graft copolymers. It was found that the received okara mainly contained insoluble contents in water. The insoluble okara component Ok(Ins) was suspended in water and activated with ammonium persulfate as an initiator, followed by grafting PAA through a free radical polymerization. After the graft polymerization, the product (Ok-PAA) was separated into precipitate and supernatant, which were dried to give Ok-PAA(pre) and Ok-PAA(sup), respectively. It was found that PAA was grafted on Ok backbones and co-precipitated with the insoluble Ok. In addition, Ok-PAA(sup) was found to be translucent as a result of the grafting of PAA. Further, the successful grafting of PAA onto okara backbones was proven by Fourier transform infrared, thermogravimetric analysis, and microscopic measurements. Ok-PAA(sup) dispersed in water formed nanoparticles with an average diameter of 420 nm, while Ok-PAA(pre) was clustered coarse particles in water. The rheological data including the storage modulus, loss modulus, and viscosity indicated that the Ok-PAA product was a viscoelastic gel-like material with potential for agricultural and environmental applications.


Subject(s)
Acrylic Resins/chemistry , Free Radicals/chemistry , Plant Proteins/chemistry , Polysaccharides/chemistry , Waste Products/analysis , Biomass , Nanoparticles/chemistry , Particle Size , Polymerization , Rheology , Soy Foods , Viscosity
16.
Metabolites ; 10(5)2020 May 15.
Article in English | MEDLINE | ID: mdl-32429044

ABSTRACT

Plant-based diets (PBDs) are associated with environmental benefits, human health promotion and animal welfare. There is a worldwide shift towards PBDs, evident from the increased global demand for fresh plant-based products (PBPs). Such shifts in dietary preferences accompanied by evolving food palates, create opportunities to leverage technological advancements and strict quality controls in developing PBPs that can drive consumer acceptance. Flavor, color and texture are important sensory attributes of a food product and, have the largest influence on consumer appeal and acceptance. Among these, flavor is considered the most dominating quality attribute that significantly affects overall eating experience. Current state-of-art technologies rely on physicochemical estimations and sensory-based tests to assess flavor-related attributes in fresh PBPs. However, these methodologies often do not provide any indication about the metabolic features associated with unique flavor profiles and, consequently, can be used in a limited way to define the quality attributes of PBPs. To this end, a systematic understanding of metabolites that contribute to the flavor profiles of PBPs is warranted to complement the existing methodologies. This review will discuss the use of metabolomics for evaluating flavor-associated metabolites in fresh PBPs at post-harvest stage, alongside its applications for quality assessment and grading. We will summarize the current research in this area, discuss technical challenges and considerations pertaining to sampling and analytical techniques, as well as s provide future perspectives and directions for government organizations, industries and other stakeholders associated with the quality assessment of fresh PBPs.

17.
Food Res Int ; 125: 108565, 2019 11.
Article in English | MEDLINE | ID: mdl-31554083

ABSTRACT

Inflammation has been revealed to play a central role in the onset and progression of many illnesses. Nuclear magnetic resonance (NMR) based metabolomics method was adopted to evaluate the effects of Phoenix dactylifera seeds, in particular the Algerian date variety of Deglet on the metabolome of the LPS-IFN-γ-induced RAW 264.7 cells. Variations in the extracellular and intracellular profiles emphasized the differences in the presence of tyrosine, phenylalanine, alanine, proline, asparagine, isocitrate, inosine and lysine. Principal component analysis (PCA) revealed noticeable clustering patterns between the treated and induced RAW cells based on the metabolic profile of the extracellular metabolites. However, the effects of treatment on the intracellular metabolites appears to be less distinct as suggested by the PCA and heatmap analyses. A clear group segregation was observed for the intracellular metabolites from the treated and induced cells based on the orthogonal partial least squares-discriminant analysis (OPLS-DA) score plot. Likewise, 11 of the metabolites in the treated cells were significantly different from those in the induced groups, including amino acids and succinate. The enrichment analysis demonstrated that treatment with Deglet seed extracts interfered with the energy and of amino acids metabolism. Overall, the obtained data reinforced the possible application of Deglet seeds as a functional food with anti-inflammatory properties.


Subject(s)
Metabolomics/methods , Phoeniceae , Plant Extracts/pharmacology , Proton Magnetic Resonance Spectroscopy/methods , Seeds , Animals , Lipopolysaccharides , Mice , Principal Component Analysis , RAW 264.7 Cells
19.
Anal Biochem ; 576: 20-32, 2019 07 01.
Article in English | MEDLINE | ID: mdl-30970239

ABSTRACT

The variation in the extracellular metabolites of RAW 264.7 cells obtained from different passage numbers (passage 9, 12 and 14) was examined. The impact of different harvesting protocols (trypsinization and scraping) on recovery of intracellular metabolites was then assessed. The similarity and variation in the cell metabolome was investigated using 1H NMR metabolic profiling modeled using multivariate data analysis. The characterization and quantification of metabolites was performed to determine the passage-related and harvesting-dependent effects on impacted metabolic networks. The trypsinized RAW cells from lower passages gave higher intensities of most identified metabolites, including asparagine, serine and tryptophan. Principal component analysis revealed variation between cells from different passages and harvesting methods, as indicated by the formation of clusters in score plot. Analysis of S-plots revealed metabolites that acted as biomarkers in discriminating cells from different passages including acetate, serine, lactate and choline. Meanwhile lactate, glutamine and pyruvate served as biomarkers for differentiating trypsinized and scraped cells. In passage-dependent effects, glycolysis and TCA cycle were influential, whereas glycerophospholipid metabolism was affected by the harvesting method. Overall, it is proposed that typsinized RAW cells from lower passage numbers are more appropriate when conducting experiments related to NMR metabolomics.


Subject(s)
Magnetic Resonance Spectroscopy/methods , Metabolomics/methods , Animals , Biomarkers/metabolism , Mice , RAW 264.7 Cells
20.
Metabolites ; 9(4)2019 Apr 18.
Article in English | MEDLINE | ID: mdl-31003499

ABSTRACT

The use of multiple omics techniques (i.e., genomics, transcriptomics, proteomics, and metabolomics) is becoming increasingly popular in all facets of life science. Omics techniques provide a more holistic molecular perspective of studied biological systems compared to traditional approaches. However, due to their inherent data differences, integrating multiple omics platforms remains an ongoing challenge for many researchers. As metabolites represent the downstream products of multiple interactions between genes, transcripts, and proteins, metabolomics, the tools and approaches routinely used in this field could assist with the integration of these complex multi-omics data sets. The question is, how? Here we provide some answers (in terms of methods, software tools and databases) along with a variety of recommendations and a list of continuing challenges as identified during a peer session on multi-omics integration that was held at the recent 'Australian and New Zealand Metabolomics Conference' (ANZMET 2018) in Auckland, New Zealand (Sept. 2018). We envisage that this document will serve as a guide to metabolomics researchers and other members of the community wishing to perform multi-omics studies. We also believe that these ideas may allow the full promise of integrated multi-omics research and, ultimately, of systems biology to be realized.

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