Metabolic Processes Are Differentially Regulated During Wild-Type and Attenuated Dengue Virus Infection in Aedes aegypti

Successful completion of the dengue virus (DENV) life cycle in its mosquito vectors is important for efficient human–mosquito–human cycle of transmission, but the virus–mosquito interactions that underpin this critical event are poorly defined. To understand the virus–host interactions that determine viral infection by Aedes aegypti, the principal DENV vector, the authors compared transcriptomic changes in the head/thorax of the mosquito after intrathoracic infection with the wild-type DENV2 16681 strain and its attenuated derivative, PDK53. Using high-throughput RNA-sequencing, the authors identified 1,629 differentially expressed genes (DEGs) during 16681 infection, compared with only 22 DEGs identified during PDK53 infection, indicating that 16681 infection triggers a more robust host transcriptomic response compared with PDK53 infection. The authors further found that 16681 infection, but not PDK53 infection, altered metabolism in these heads/thoraces. Altogether, our findings reveal differential regulation of metabolic processes during wild-type and attenuated DENV infection, and suggest the need for future work to study the role of metabolic processes in determining DENV infection and replication in its mosquito vectors.

Transcriptome Profiling Based on Larvae at Different Time Points After Hatching Provides a Core Set of Gene Resource for Understanding the Metabolic Mechanisms of the Brood-Care Behavior in Octopus ocellatus

The metabolic processes of organisms are very complex. Each process is crucial and affects the growth, development, and reproduction of organisms. Metabolism-related mechanisms in Octopus ocellatus behaviors have not been widely studied. Brood-care is a common behavior in most organisms, which can improve the survival rate and constitution of larvae. Octopus ocellatus carried out this behavior, but it was rarely noticed by researchers before. In our study, 3,486 differentially expressed genes (DEGs) were identified based on transcriptome analysis of O. ocellatus. We identify metabolism-related DEGs using GO and KEGG enrichment analyses. Then, we construct protein–protein interaction networks to search the functional relationships between metabolism-related DEGs. Finally, we identified 10 hub genes related to multiple gene functions or involved in multiple signal pathways and verified them using quantitative real-time polymerase chain reaction (qRT-PCR). Protein–protein interaction networks were first used to study the effects of brood-care behavior on metabolism in the process of growing of O. ocellatus larvae, and the results provide us valuable genetic resources for understanding the metabolic processes of invertebrate larvae. The data lay a foundation for further study the brood-care behavior and metabolic mechanisms of invertebrates.

Intermittent compressive force induces cell cycling and reduces apoptosis in embryoid bodies of mouse induced pluripotent stem cells

In vitro manipulation of induced pluripotent stem cells (iPSCs) by environmental factors is of great interest for three-dimensional (3D) tissue/organ induction. The effects of mechanical force depend on many factors, including force and cell type. However, information on such effects in iPSCs is lacking. The aim of this study was to identify a molecular mechanism in iPSCs responding to intermittent compressive force (ICF) by analyzing the global gene expression profile. Embryoid bodies of mouse iPSCs, attached on a tissue culture plate in 3D form, were subjected to ICF in serum-free culture medium for 24 h. Gene ontology analyses for RNA sequencing data demonstrated that genes differentially regulated by ICF were mainly associated with metabolic processes, membrane and protein binding. Topology-based analysis demonstrated that ICF induced genes in cell cycle categories and downregulated genes associated with metabolic processes. The Kyoto Encyclopedia of Genes and Genomes database revealed differentially regulated genes related to the p53 signaling pathway and cell cycle. qPCR analysis demonstrated significant upregulation of Ccnd1, Cdk6 and Ccng1. Flow cytometry showed that ICF induced cell cycle and proliferation, while reducing the number of apoptotic cells. ICF also upregulated transforming growth factor β1 (Tgfb1) at both mRNA and protein levels, and pretreatment with a TGF-β inhibitor (SB431542) prior to ICF abolished ICF-induced Ccnd1 and Cdk6 expression. Taken together, these findings show that TGF-β signaling in iPSCs enhances proliferation and decreases apoptosis in response to ICF, that could give rise to an efficient protocol to manipulate iPSCs for organoid fabrication.

Changes in the concentration of methane in the ecosystem in vitro against the background of Asteraceae family plants biomass

The assessment of Asteráceae Family Plants (rhizomes and roots of elecampane and wormwood) influence on the process of methane formation in the rumen ecosystem and metabolic processes was carried out. Studies (in vitro) were carried out using ANKOM Daisy II incubator (modifications D200 and D200I) according to a specialized method. Rumen contents were obtained from beef bulls with chronic rumen fistula. Gas analysis of air and volatile fatty acids samples was performed by gas chromatography. The results of the study showed that different dosages of phytobiotic preparations did not significantly affect the characteristics of fermentation in vitro. Phytobiotic preparations of elecampane and wormwood reduce the production of methane in the ruminal fluid, which may be associated with various active components or dosages of their administration.

Investigation of changes in rat’s blood metabolomic profile, caused by lead exposure

Introduction. The prevalence of lead in the environment, due to human production and economic activities, and the xenobiotic nature of the element substantiate the relevance of studying the changes caused by the action of this metal. Materials and methods. A non-target metabolomic screening of the blood of rats exposed to intraperitoneal administration of lead acetate by HPLC-mass spectrometry was carried out. The expression of the selected masses was compared with those for the control group of animals. The masses that significantly changed the intensity compared to the control were subjected to fragmentation to obtain characteristic fragments. The annotation of metabolites was performed by searching in MS/MS databases and by comparison with in silico fragmentation spectra. The involvement of annotated metabolites in metabolic processes was established by literature analyzing. Results. Non-target metabolomic screening revealed 37 m/z values for the exposed group, significantly changing the intensity compared to the control. Annotation using fragmentation spectra and in silico fragmentation allows establishing the structure of eight metabolites, including an epoxy derivative of linolic acid, 15-hydroxyeicosatetraenoic acid, four oxo- and hydroxyacylcarnitine derivatives of long-chain fatty acids, one acylcarnitine derivatives of medium-chain fatty acids and one lysophosphoserine. Conclusion. Analyzing the literature, the known functions of the identified metabolites were established and attributed to the known metabolic processes. So, oxo- and hydroxyacylcarnitines are derivatives for intermediate products of β-oxidation fatty acids - it is increased concentration compared to the control indicates a violation of this process under the influence of oxidative stress caused by lead. Epoxy and 15-hydroxy derivatives of fatty acids (increased content relative to the control group) act as regulatory metabolites (vasodynamic activity), on the one hand, and markers of lead-induced hypoxia on the other hand. The increase of the concentration for the lysophosphatidylserine derivative indicates the intensification of apoptotic processes in the organism of the exposed group in contrast to the control.

The phenomenon of reversible myometrial dysfunction and delayed rehabilitation of uterine contractive ability

The author introduces the conception o f potentially reversible myometrial dysfunction with unaffected main physiological function o f myometrium (viability o f myometrium). This dysfunction is connected with the disturbances o f uterine haemodynamics. The phenomenon o f reversible myometrial dysfunction reflects the process o f prolonged decreased contractile ability o f the uterus.The therapy o f reversible myometrial dysfunction phenomenon should be directed to blood flow restoration under the conditions o f uterine hypoperfusion.The special treatment is not required fo r myometrium with reserved main physiological functions (tonus, excitability) because restoration o f myometrial contractile ability improves spontaneously in case o f blood flow restoration.With the aim o f prophylaxis o f myometrial dysfunction and delayed rehabilitation o f the uterine contractile function administration o f Ca antagonists, beta-adrenomymetics, antioxidants and preparations, which improve myometrial metabolic processes, is recommended before the expected delivery.

The EBNA2-EBF1 complex promotes oncogenic MYC expression levels and metabolic processes required for cell cycle progression of Epstein-Barr virus-infected B cells

Epstein-Barr virus (EBV) is a human tumor virus, which preferentially infects resting human B cells. Upon infection in vitro, EBV activates and immortalizes these cells. The viral latent protein EBV nuclear antigen (EBNA) 2 is essential for B cell activation and immortalization; it targets and binds the cellular and ubiquitously expressed DNA binding protein CBF1, thereby transactivating a plethora of viral and cellular genes. In addition, EBNA2 uses its N-terminal dimerization (END) domain to bind early B cell factor (EBF) 1, a pioneer transcription factor specifying the B cell lineage. We found that EBNA2 exploits EBF1 to support key metabolic processes and to foster cell cycle progression of infected B cells in their first cell cycles upon activation. An α1-helix within the END domain was found to promote EBF1 binding. EBV mutants lacking the α1-helix in EBNA2 can infect and activate B cells efficiently, but the activated cells fail to complete the early S phase of their initial cell cycle. Expression of MYC, target genes of MYC and E2F as well as multiple metabolic processes linked to cell cycle progression are impaired in EBV∆α1 infected B cells. Our findings indicate that EBF1 controls B cell activation via EBNA2 and, thus, has a critical role in regulating the cell cycle of EBV infected B cells. This is a function of EBF1 going beyond its well-known contribution to B cell lineage specification.

Whole Genome Methylation Analysis Reveals Role of DNA Methylation in Cow’s Ileal and Ileal Lymph Node Responses to Mycobacterium avium subsp. paratuberculosis Infection

Johne’s Disease (JD), caused by Mycobacterium avium subsp paratuberculosis (MAP), is an incurable disease of ruminants and other animal species and is characterized by an imbalance of gut immunity. The role of MAP infection on the epigenetic modeling of gut immunity during the progression of JD is still unknown. This study investigated the DNA methylation patterns in ileal (IL) and ileal lymph node (ILLN) tissues from cows diagnosed with persistent subclinical MAP infection over a one to 4 years period. DNA samples from IL and ILLN tissues from cows negative (MAPneg) (n = 3) or positive for MAP infection (MAPinf) (n = 4) were subjected to whole genome bisulfite sequencing. A total of 11,263 and 62,459 differentially methylated cytosines (DMCs), and 1259 and 8086 differentially methylated regions (DMRs) (FDR<0.1) were found between MAPinf and MAPneg IL and ILLN tissues, respectively. The DMRs were found on 394 genes (denoted DMR genes) in the IL and on 1305 genes in the ILLN. DMR genes with hypermethylated promoters/5′UTR [3 (IL) and 88 (ILLN)] or hypomethylated promoters/5′UTR [10 (IL) and 25 (ILLN)] and having multiple functions including response to stimulus/immune response (BLK, BTC, CCL21, AVPR1A, CHRNG, GABRA4, TDGF1), cellular processes (H2AC20, TEX101, GLA, NCKAP5L, RBM27, SLC18A1, H2AC20BARHL2, NLGN3, SUV39H1, GABRA4, PPA1, UBE2D2) and metabolic processes (GSTO2, H2AC20, SUV39H1, PPA1, UBE2D2) are potential DNA methylation candidate genes of MAP infection. The ILLN DMR genes were enriched for more biological process (BP) gene ontology (GO) terms (n = 374), most of which were related to cellular processes (27.6%), biological regulation (16.6%), metabolic processes (15.4%) and response to stimulus/immune response (8.2%) compared to 75 BP GO terms (related to cellular processes, metabolic processes and transport, and system development) enriched for IL DMR genes. ILLN DMR genes were enriched for more pathways (n = 47) including 13 disease pathways compared with 36 enriched pathways, including 7 disease/immune pathways for IL DMR genes. In conclusion, the results show tissue specific responses to MAP infection with more epigenetic changes (DMCs and DMRs) in the ILLN than in the IL tissue, suggesting that the ILLN and immune processes were more responsive to regulation by methylation of DNA relative to IL tissue. Our data is the first to demonstrate a potential role for DNA methylation in the pathogenesis of MAP infection in dairy cattle.