Changes In Expression Of Related Genes Research Articles

Dynamic regulation of CeA gene expression during acute and protracted abstinence from chronic binge drinking of male and female C57BL/6J mice

Dynamic regulation of CeA gene expression during acute and protracted abstinence from chronic binge drinking of male and female C57BL/6J mice

Abstract 944: Pre-clinical activity of the Wnt pathway inhibitor RXC004 in combination with MAPK pathway inhibitors in GI cancer models

Abstract Background: Gastrointestinal (GI) cancers cause more cancer deaths than any other body system, with 3.4 million related deaths in 2018. Wnt pathway activation is common in GI cancers, with the pathogenic upstream Wnt pathway variant sub-population (RNF43 loss of function/RSPO gain of function) showing a combined prevalence of 4.7% in GI cancer patients; pre-clinically this sub-population shows exquisite sensitivity to RXC004, a small molecule Porcupine inhibitor. A striking co-occurrence of upstream Wnt pathway variants with MAPK pathway driver mutations (KRAS, BRAF, NRAS) was detected in 77% of Microsatellite stable (MSS) GI cancers, suggesting co-inhibiting these pathways could enhance clinical activity in these patients. Methods: In vitro proliferation assays were performed in pre-clinical models of upstream Wnt pathway mutated MSS GI cancer (SNU-1411, HPAF-II and AsPC-1) using Wnt (RXC004) or MAPK (Trametinib) pathway inhibitors at multiple concentrations as single agents or in combination. Control cells with downstream Wnt pathway mutations (WiDr and HCT116) were also tested. Synergistic combination effects were detected by BLISS scores. RXC004 (1.5mg/kg QD) and Trametinib (0.3mg/kg QD) were evaluated in an RSPO-fusion xenograft model (SNU-1411) as single agents or in combination. Efficacy was measured by tumour volume and weight; PD markers were assessed in end of study tumour samples. Relative AXIN2 and DUSP6 gene expression changes were determined upon inhibitor treatment by quantitative Polymerase Chain Reaction (qPCR). Results: RXC004 in combination with Trametinib demonstrated strong anti-proliferative synergy in SNU-1411 cells and moderate synergy/additivity in HPAF-II and AsPC-1 cells; no synergy was observed in control cells. In vivo, RXC004 and Trametinib monotherapy led to significant moderate tumour growth control (p<0.05) at the doses tested, whereas their combination led to significantly enhanced efficacy (p<0.05). Gene expression analysis demonstrated a reciprocal Wnt/MAPK signaling mechanism; RXC004 strongly suppressed Wnt signaling (AXIN2) but increased MAPK signaling (DUSP6), whilst Trametinib strongly suppressed MAPK signaling (DUSP6) but increased Wnt signaling (AXIN2). Combination of RXC004 and Trametinib sustained inhibition of both Wnt and MAPK signalling. Similar effects were observed with inhibitors of other MAPK pathway components. Conclusion: Wnt and MAPK pathways are frequently co-activated in GI cancers, particularly in the upstream Wnt pathway mutated sub-population. Pre-clinically we show that Wnt and MAPK pathways act as potential reciprocal resistance mechanisms following single agent inhibition of either pathway; co-inhibition of these pathways leads to synergistic effects in vitro and enhanced efficacy in vivo. This provides a clear rationale to assess this combination approach in the clinic. Citation Format: Simon A. Woodcock, Dorottya Keppel, Catherine Eagle, James Kelly, Eimear Flanagan, Emma Bishop, Inder Bhamra, Clifford D. Jones, Richard Armer, Jane Robertson, Caroline Phillips. Pre-clinical activity of the Wnt pathway inhibitor RXC004 in combination with MAPK pathway inhibitors in GI cancer models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 944.

Offspring's own serotonin transporter genotype, independently from the maternal one, increases anxiety- and depression-like behavior and alters neuroplasticity markers in rats

IntroductionDevelopmental changes due to early life variations in the serotonin system affect stress-related behavior and neuroplasticity in adulthood. These outcomes can be caused both by offspring's own and maternal serotonergic genotype. We aimed to dissociate the contribution of the own genotype from the influences of mother genotype. MethodsSixty-six male homozygous (5-HTT−/−) and heterozygous (5-HTT+/−) serotonin transporter knockout and wild-type rats from constant 5-HTT genotype mothers crossed with varying 5-HTT genotype fathers were subjected to tests assessing anxiety- and depression-like behaviors. Additionally, we measured plasma corticosterone levels and mRNA levels of BDNF, GABA system and HPA-axis components in the prelimbic and infralimbic cortex. Finally, we assessed the effect of paternal 5-HTT genotype on these measurements in 5-HTT+/− offspring receiving their knockout allele from their mother or father. Results5-HTT−/− offspring exhibited increased anxiety- and depression-like behavior in the elevated plus maze and sucrose preference test. Furthermore, Bdnf isoform VI expression was reduced in the prelimbic cortex. Bdnf isoform IV and GABA related gene expression was also altered but did not survive false discovery rate (FDR) correction. Finally, 5-HTT+/− offspring from 5-HTT−/− fathers displayed higher levels of anxiety- and depression-like behavior and changes in GABA, BDNF and HPA-axis related gene expression not surviving FDR correction. LimitationsOnly male offspring was tested. ConclusionsOffspring's own 5-HTT genotype influences stress-related behaviors and Bdnf isoform VI expression, independently of maternal 5-HTT genotype. Paternal 5-HTT genotype separately influenced these outcomes. These findings advance our understanding of the 5-HTT genotype dependent susceptibility to stress-related disorders.

Toxicity of Three Optical Brighteners: Potential Pharmacological Targets and Effects on Caenorhabditis elegans.

Optical brighteners (OBs) have become an integral part of our daily lives and culture, with a growing number of applications in various fields. Most industrially produced OBs are derived from stilbene, which has been found in environmental matrices. The main objectives for this work are as follows: first, to identify protein targets for DAST, FB-28, and FB-71, and second, to assess their effects in some behaviors physiologic of Caenorhabditis elegans. To achieve the first objective, each OB was tested against a total of 844 human proteins through molecular docking using AutoDock Vina, and affinities were employed as the main criteria to identify potential target proteins for the OB. Molecular dynamics simulations took and validated the best 25 docking results from two protein databases. The highest affinity was obtained for the Hsp70-1/DAST, CD40 ligand/FB-71, and CD40 ligand/FB-28 complexes. The possible toxic effects that OBs could cause were evaluated using the nematode C. elegans. The lethality, body length, locomotion, and reproduction were investigated in larval stage L1 or L4 of the wild-type strain N2. In addition, transgenic green fluorescent protein (GFP) strains were employed to estimate changes in relative gene expression. The effects on the inhibition of growth, locomotion, and reproduction of C. elegans nematodes exposed to DAST, FB-71, and FB-28 OBs were more noticeable with respect to lethality. Moreover, an interesting aspect in OB was increased the expression of gpx-4 and sod-4 genes associated with oxidative stress indicating a toxic response related to the generation of reactive oxygen species (ROS). In all cases, a clear concentration-response relationship was observed. It is of special attention that the use of OBs is increasing, and their different sources, such as detergents, textiles, plastics, and paper products, must also be investigated to characterize the primary emissions of OBs to the environment and to develop an adequate regulatory framework.

Distinctive Patterns of 5-Methylcytosine and 5-Hydroxymethylcytosine in Schizophrenia.

Schizophrenia is a highly heritable neuropsychiatric disorder characterized by cognitive and social dysfunction. Genetic, epigenetic, and environmental factors are together implicated in the pathogenesis and development of schizophrenia. DNA methylation, 5-methycytosine (5mC) and 5-hydroxylcytosine (5hmC) have been recognized as key epigenetic elements in neurodevelopment, ageing, and neurodegenerative diseases. Recently, distinctive 5mC and 5hmC pattern and expression changes of related genes have been discovered in schizophrenia. Antipsychotic drugs that affect 5mC status can alleviate symptoms in patients with schizophrenia, suggesting a critical role for DNA methylation in the pathogenesis of schizophrenia. Further exploring the signatures of 5mC and 5hmC in schizophrenia and developing precision-targeted epigenetic drugs based on this will provide new insights into the diagnosis and treatment of schizophrenia.

Association between gut health and gut microbiota in a polluted environment

Animals host complex bacterial communities in their gastrointestinal tracts, with which they share a mutualistic interaction. The numerous effects these interactions grant to the host include regulation of the immune system, defense against pathogen invasion, digestion of otherwise undigestible foodstuffs, and impacts on host behaviour. Exposure to stressors, such as environmental pollution, parasites, and/or predators, can alter the composition of the gut microbiome, potentially affecting host-microbiome interactions that can be manifest in the host as, for example, metabolic dysfunction or inflammation. However, whether a change in gut microbiota in wild animals associates with a change in host condition is seldom examined. Thus, we quantified whether wild bank voles inhabiting a polluted environment, areas where there are environmental radionuclides, exhibited a change in gut microbiota (using 16S amplicon sequencing) and concomitant change in host health using a combined approach of transcriptomics, histological staining analyses of colon tissue, and quantification of short-chain fatty acids in faeces and blood. Concomitant with a change in gut microbiota in animals inhabiting contaminated areas, we found evidence of poor gut health in the host, such as hypotrophy of goblet cells and likely weakened mucus layer and related changes in Clca1 and Agr2 gene expression, but no visible inflammation in colon tissue. Through this case study we show that inhabiting a polluted environment can have wide reaching effects on the gut health of affected animals, and that gut health and other host health parameters should be examined together with gut microbiota in ecotoxicological studies.

Cell-type brain-region specific changes in prefrontal cortex of a mouse model of alcohol dependence

The prefrontal cortex is a crucial regulator of alcohol drinking, and dependence, and other behavioral phenotypes associated with AUD. Comprehensive identification of cell-type specific transcriptomic changes in alcohol dependence will improve our understanding of mechanisms underlying the excessive alcohol use associated with alcohol dependence and will refine targets for therapeutic development. We performed single nucleus RNA sequencing (snRNA-seq) and Visium spatial gene expression profiling on the medial prefrontal cortex (mPFC) obtained from C57BL/6 J mice exposed to the two-bottle choice-chronic intermittent ethanol (CIE) vapor exposure (2BC-CIE, defined as dependent group) paradigm which models phenotypes of alcohol dependence including escalation of alcohol drinking. Gene co-expression network analysis and differential expression analysis identified highly dysregulated co-expression networks in multiple cell types. Dysregulated modules and their hub genes suggest novel understudied targets for studying molecular mechanisms contributing to the alcohol dependence state. A subtype of inhibitory neurons was the most alcohol-sensitive cell type and contained a downregulated gene co-expression module; the hub gene for this module is Cpa6, a gene previously identified by GWAS to be associated with excessive alcohol consumption. We identified an astrocytic Gpc5 module significantly upregulated in the alcohol-dependent group. To our knowledge, there are no studies linking Cpa6 and Gpc5 to the alcohol-dependent phenotype. We also identified neuroinflammation related gene expression changes in multiple cell types, specifically enriched in microglia, further implicating neuroinflammation in the escalation of alcohol drinking. Here, we present a comprehensive atlas of cell-type specific alcohol dependence mediated gene expression changes in the mPFC and identify novel cell type-specific targets implicated in alcohol dependence.

IMD signaling in the gut and the brain modulates Amyloid-beta-induced deficits in Drosophila

AimsEvidence indicates accumulating Aβ peptides in brain activates immune responses in neuronal and peripheral system, which may collaboratively influence pathogenesis of Alzheimer's disease (AD). We aim to investigate whether regulating intestinal innate immune signaling ameliorates Aβ-induced impairments in Drosophila melanogaster. Main methodsQuantitative polymerase chain reaction (qPCR) was used to observe expression changes of innate immune responses related genes in brain and in gut under the circumstance of Aβ overexpressing in nerve system. Aversive olfactory conditioning and survival assay were used to investigate effects of modulating Attacin-A (AttA) and Dpitercin-A (DptA). Fluorometric assays of respiratory burst activity was introduced to explore whether reducing oxidative stress enables overexpressing intestinal AttA and DptA to reverse Aβ-induced deficits. Key findingsIn vivo genetic analysis revealed that accumulating Aβ42 in neurons modulates innate immune signaling of the IMD pathway both in the brain and in the gut. Increased expression levels of the intestinal AttA and DptA improved learning performance and extended the lifespan of Aβ42 flies. The administration of apramycin led to alleviations of Aβ-induced behavioral changes, indicating that gram-negative bacteria are associated with the development of Aβ-induced pathologies. Further analysis showed that the neural expression of Aβ42 increased oxidative stress in the gut, which disrupted intestinal integrity and decreased learning performance. In addition, increased levels of AMPs targeting gram-negative bacteria and antioxidants reduced oxidative stress in the gut and reversed Aβ-induced behavioral damage. SignificanceThese findings suggest that innate immune responses in the gut play a pivotal role in modulating Aβ-induced pathologies.

Differences of cadmium uptake and accumulation in roots of two maize varieties (Zea mays L.).

Different maize varieties respond differentially to cadmium (Cd) stress. As the first organ in contact with the soil, the response of the root is particularly important. However, the physiological mechanisms that determine the response are not well defined. Here, we compared the differences in Cd-induced related gene expression, ionic homeostasis, and ultrastructural changes in roots of Cd-tolerant maize variety (XR57) and Cd-sensitive maize variety (LY296), and assessed their effects on Cd uptake and accumulation. Our findings indicate that XR57 absorbed a significantly lower Cd than LY296 did, and that the expression levels of genes related to Cd uptake (ZmNRAMP5 and ZmZIP4) and efflux (ZmABCG4) in the root were consistent with the Cd absorption at the physiological levels. Compared with LY296, the lower Cd concentration in the roots of XR57 caused less interference with the ion balance. Transmission electron microscope images revealed that the roots from XR57 exposed to Cd had developed thicker cell walls than LY296. In addition, the large increase ZmABCC1 and ZmABCC2 expression levels in XR57 mediated the appearance of numerous electron-dense granules in the vacuoles present in the roots. As a result, the high Cd tolerance of XR57 is the result of a multi-level response that involves increased resistance to Cd uptake, a stronger capacity for vacuolar regionalization, and the formation of thicker cell walls. These findings may provide a theoretical basis for maize cultivation in Cd-contaminated areas.

Polysaccharides from Platycodon grandiflorus attenuates high-fat diet induced obesity in mice through targeting gut microbiota

The root of Platycodon grandiflorus (PG), abundant in soluble polysaccharides, has a long history in traditional Asian diets and herbal medicine due to its anti-inflammatory activity and anti-obesity effects. Our previous study was the first to establish a link between the beneficial effects of PG and changes in the gut microbiota, and suggested potential roles that the polysaccharide components play. However, more evidence was needed to understand the anti-obesity functions of polysaccharides from PG (PS) and their relationship with the regulation of the gut microbiota. In this study, we first performed an experiment to explore the anti-obesity activities of PS: Male C57BL/6 mice (six-weeks-old) were fed either a standard control diet (CON), or a high-fat diet (HFD) to induce obesity, or a HFD supplemented with PS (HFPS) for 8 weeks. Body weight and food intake were monitored throughout. Lipid metabolism were determined and related gene expression changes in adipose tissues were analyzed by RNA-seq. Amplicon sequencing of the bacterial 16 S rRNA gene was used to explore gut microbiota structure in fecal samples. Then, we performed the second experiment to explore whether the anti-obesity activities of PS were dependent on the regulation of the gut microbiota: Male C57BL/6 mice (six-weeks-old), treated with an antibiotic cocktail to reduce the gut microbial load, were fed either a HFD (A-HFD) or a HFPS (A-HFPS) diet for 8 weeks. Finally, we used in vitro fermentation experiments to verify the effects of PS on the growth and metabolic activities of the gut microbes. We found that PS significantly reduced HFD-induced weight gain and excessive fat accumulation, changed the expression of key genes involved in lipid metabolism, and attenuated HFD-induced changes in the gut microbiota. However, PS did not affect fat accumulation or lipid metabolism in the gut microbiota depleted mice. Overall, our results show that PS has significant effects on the gut microbiota in the mouse model, and the anti-obesity effects of PS are mediated via changes in the gut microbiota composition and metabolic activity.

Effects of Salinity Stress on Growth and Physiological Parameters and Related Gene Expression in Different Ecotypes of Sesuvium portulacastrum on Hainan Island.

We conducted a study to examine the growth and physiological changes in 12 different ecotypes of Sesuvium portulacastrum collected from Hainan Island in China. These ecotypes were subjected to different concentrations (0, 200, 400, and 600 mmol/L) of sodium chloride (NaCl) salt stress for 14 days. We also analyzed the expression of metabolic genes related to stress response. Under low salt stress, indicators such as plant height in region K (0 mmol/L: 45% and highest at 200 mmol/L: 80%), internode length (0 mmol/L: 0.38, 200 mmol/L: 0.87, 400 mmol/L: 0.25, and 600 mmol/L: 1.35), as well as leaf area, relative water content, fresh weight, and dry weight exhibited an overall increasing trend with the increase in salt concentration. However, as the salt concentration increased, these indicators showed a decreasing trend. Proline and malondialdehyde contents increased with higher salt concentrations. When the NaCl concentration was 400 mmol/L, MDA content in the leaves was highest in the regions E (196.23%), F (94.28%), J (170.10%), and K (136.08%) as compared to the control group, respectively. Most materials demonstrated a significant decrease in chlorophyll a, chlorophyll b, and total chlorophyll content compared to the control group. Furthermore, the ratio of chlorophyll a to chlorophyll b (Rab) varied among different materials. Using principal component analysis, we identified three ecotypes (L from Xinglong Village, Danzhou City; B from Shuigoupo Village, Lingshui County; and J from Haidongfang Park, Dongfang City) that represented high, medium, and low salt tolerance levels, respectively, based on the above growth and physiological indexes. To further investigate the expression changes of related genes at the transcriptional level, we employed qRT-PCR. The results showed that the relative expression of SpP5CS1, SpLOX1, and SpLOX1 genes increased with higher salt concentrations, which corresponded to the accumulation of proline and malondialdehyde content, respectively. However, the relative expression of SpCHL1a and SpCHL1b did not exhibit a consistent pattern. This study contributes to our understanding of the salt tolerance mechanism in the true halophyte S. portulacastrum, providing a solid theoretical foundation for further research in this field.

Changes in volatile profile and related gene expression during senescence of tobacco leaves.

Volatile organic compounds are critical for food flavor and play important roles in plant-plant interactions and plants' communications with the environment. Tobacco is well-studied for secondary metabolism and most of the typical flavor substances in tobacco leaves are generated at the mature stage of leaf development. However, the changes in volatiles during leaf senescence are rarely studied. The volatile composition of tobacco leaves at different stages of senescence was characterized for the first time. Comparative volatile profiling of tobacco leaves at different stages was performed using solid-phase microextraction coupled with gas chromatography/mass spectrometry. In total, 45 volatile compounds were identified and quantified, including terpenoids, green leaf volatiles (GLVs), phenylpropanoids, Maillard reaction products, esters, and alkanes. Most of the volatile compounds showed differential accumulation during leaf senescence. Some terpenoids, including neophytadiene, β-springene, and 6-methyl-5-hepten-2-one, increased significantly with the progress of leaf senescence. Hexanal and phenylacetaldehyde also showed increased accumulation in leaves during senescence. The results from gene expression profiling indicated that genes involved in metabolism of terpenoids, phenylpropanoids, and GLVs were differentially expressed during leaf yellowing. Dynamic changes in volatile compounds during tobacco leaf senescence are observed and the integration of gene-metabolites datasets offers important readouts for the genetic control of volatile production during the process of leaf senescence. © 2023 Society of Chemical Industry.

The alteration of proteins and metabolites in leaf apoplast and the related gene expression associated with the adaptation of Ammopiptanthus mongolicus to winter freezing stress

Ammopiptanthus mongolicus, an evergreen broad-leaved plant, can tolerate severe freezing stress (temperatures as low as −20 °C in winter). The apoplast is the space outside the plasma membrane that plays an important role in plant responses to environmental stress. Here, we investigated, using a multi-omics approach, the dynamic alterations in the levels of proteins and metabolites in the apoplast and related gene expression changes involved in the adaptation of A. mongolicus to winter freezing stress. Of the 962 proteins identified in the apoplast, the abundance of several PR proteins, including PR3 and PR5, increased significantly in winter, which may contribute to winter freezing-stress tolerance by functioning as antifreeze proteins. The increased abundance of the cell-wall polysaccharides and cell wall-modifying proteins, including PMEI, XTH32, and EXLA1, may enhance the mechanical properties of the cell wall in A. mongolicus. Accumulation of flavonoids and free amino acids in the apoplast may be beneficial for ROS scavenging and the maintenance of osmotic homeostasis. Integrated analyses revealed gene expression changes associated with alterations in the levels of apoplast proteins and metabolites. Our study improved the current understanding of the roles of apoplast proteins and metabolites in plant adaptation to winter freezing stress.

Combination of furosemide, gold, and dopamine as a potential therapy for breast cancer.

Breast cancer is one of the leading causes of death in women worldwide. Initially, it develops in the epithelium of the ducts or lobules of the breast glandular tissues with limited growth and the potential to metastasize. It is a highly heterogeneous malignancy; however, the common molecular mechanisms could help identify new targeted drugs for treating its subtypes. This study uses computational drug repositioning approaches to explore fresh drug candidates for breast cancer treatment. We also implemented reversal gene expression and gene expression-based signatures to explore novel drug candidates computationally. The drug activity profiles and related gene expression changes were acquired from the DrugBank, PubChem, and LINCS databases, and then in silico drug screening, molecular dynamics (MD) simulation, replica exchange MD simulations, and simulated annealing molecular dynamics (SAMD) simulations were conducted to discover and verify the valid drug candidates. We have found that compounds like furosemide, gold, and dopamine showed significant outcomes. Furthermore, the expression of genes related to breast cancer was observed to be reversed by these shortlisted drugs. Therefore, we postulate that combining furosemide, gold, and dopamine would be a potential combination therapy measurement for breast cancer patients.

Characteristic changes in cell wall metabolism and related gene expression in different parts of Tarocco blood orange pulp during cold storage

Characteristic changes in cell wall metabolism and related gene expression in different parts of Tarocco blood orange pulp during cold storage

Expression profile of genes regulating cellular response to cytokine stimulus in Sjögren’s syndrome

Abstract The existence of abnormal cytokine profile in Sjögren’s syndrome (SS) may contribute to find potential SS biomarkers and accompanying comorbidities. We aimed to investigate and compare gene expressions in the labial salivary glands of SS and of healthy subjects (HS) by means of microarray analysis. The study group comprised 8 SS patients and 8 HS in microarray analysis. The relative gene expression changes were validated with real time quantitative polymerase chain reaction in 25 SS and 20 HS. Among the differently expressed genes belonging to the “cellular response to cytokine stimulus”, “cytokine-mediated signaling pathway” and “innate immune response” ontology groups with a fold change >1.5 and with a p value < 0.05, the adiponectin, C1Q and collagen domain containing (ADIPOQ), S100 calcium binding protein A9 (S100A9), leptin receptor (LEPR), and fatty acid binding protein 4 (FABP4) gene expressions in SS group were higher than in HS. Increased expression of the assessed genes are determined by the presence of SS and the accompanying inflammation and may be a valuable diagnostic biomarker of SS. Their selected systemic effects may suggest their role as potential prognostic biomarkers of the risk of accelerated atherosclerosis and cardiovascular complications in SS.

Antagonism of tomato spotted wilt virus against tomato yellow leaf curl virus in Nicotiana benthamiana detected by transcriptome analysis.

Tomato spot wilt virus (TSWV) and tomato yellow leaf curl virus (TYLCV) are highly harmful viruses in agricultural production, which can cause serious economic losses to crops and even devastating consequences for vegetable yield in some countries and regions. Although the two viruses belong to different families and have different transmission vectors, they share most hosts. This study aimed to examine the transcriptomic expression of single and mixed inoculations of TSWV and TYLCV, leading to antagonism using high-throughput RNA sequencing. We confirmed the single and mixed infections of these viruses in Nicotiana benthamiana (N. benthamiana) by artificial inoculation. And the expression changes of related genes and their biological functions and pathways during the mixed infection of TSWV and TYLCV were analyzed by comparative transcriptome. Basically, similar symptoms were observed in the plants singly infected with TSWV and co-infected with TYLCV; the symptoms of TYLCV in the co-infected plants were not obvious compared with single TYLCV infections. When inoculated with TYLCV, the accumulation of the virus significantly reduced in single and mixed infections with TSWV; the TSWV accumulated slightly less in co-infection with TYLCV, whereas this reduction was much smaller than that of TYLCV. The results suggested that TSWV had an antagonistic effect on the accumulation of TYLCV in N. benthamiana. It mainly focused on the changes in unique differentially expressed genes (DEGs) caused by the co-infection of TSWV and TYLCV. The eight pathways enriched by upregulated DEGs mainly included amino acid biosynthesis, citrate cycle (or tricarboxylic acid cycle, TCA cycle), and so on. However, only pentose phosphate pathway (PPP) and peptidoglycan biosynthesis could be downregulated in the Kyoto Encyclopedia of Genes and Genomes pathway in which peptidoglycan biosynthesis was involved in upregulated and downregulated pathways. The antagonistic effect of TSWV on TYLCV in N.benthamiana and the change trends and specific pathways of DEGs in this process were found. Our study provided new insights into the host regulation and competition between viruses in response to TSWV and TYLCV mixed infection.

Influence of Sex and Strain on Hepatic and Adipose Tissue Trace Element Concentrations and Gene Expression in C57BL/6J and DBA/2J High Fat Diet Models.

The objective of this study was to determine the influence of sex and strain on the dysregulation of trace element concentration and associative gene expression due to diet induced obesity in adipose tissue and the liver. Male and female C57BL/6J (B6J) and DBA/2J (D2J) were randomly assigned to a normal-fat diet (NFD) containing 10% kcal fat/g or a mineral-matched high-fat diet (HFD) containing 60% kcal fat/g for 16 weeks. Liver and adipose tissue were assessed for copper, iron, manganese, and zinc concentrations and related changes in gene expression. Notable findings include three-way interactions of diet, sex, and strain amongst adipose tissue iron concentrations (p = 0.005), adipose hepcidin expression (p = 0.007), and hepatic iron regulatory protein (IRP) expression (p = 0.012). Cd11c to Cd163 ratio was increased in adipose tissue due to HFD amongst all biological groups except B6J females, for which tissue iron concentrations were reduced due to HFD (p = 0.002). Liver divalent metal transporter 1 (DMT-1) expression was increased due to HFD amongst B6J males (p < 0.005) and females (p < 0.004), which coincides with the reduction in hepatic iron concentrations found in these biological groups (p < 0.001). Sex, strain, and diet affected trace element concentration, the expression of genes that regulate trace element homeostasis, and the expression of macrophages that contribute to tissue iron-handling in adipose tissue. These findings suggest that sex and strain may be key factors that influence the adaptive capacity of iron mismanagement in adipose tissue and its subsequent consequences, such as insulin resistance.

Transcriptome analyses show changes in heat-stress related gene expression in tomato cultivar ‘Moneymaker’ under high temperature

Transcriptome analyses show changes in heat-stress related gene expression in tomato cultivar ‘Moneymaker’ under high temperature

P443 Altered expression of fungal CotH, human glucose-regulated protein 78 (GRP78), and predicted miRNAs in macrophages and model diabetic mice infected with Rhizopus oryzae

Poster session 3, September 23, 2022, 12:30 PM - 1:30 PMObjectives Rhizopus oryzae is one of the most common causes of mucormycosis. Among the virulence factors of the Mucorales, CotH protein has recently been identified, which causes the invasion of R. oryzae into endothelial cells. In this study, we aimed to examine the reaction between GRP78 at the level of human cells and different groups of mice and CotH3 at the surface of the R. oryzae hyphae. We evaluated the relative expression of GRP78 and CotH3 genes and changes in the expression of some miRNAs that target the human GRP78 gene.MethodsIn this study, the relative changes in gene expression were studied. In three groups (1) Macrophages derived from human monocytes: monocytes from the blood of healthy donors were isolated using Ficoll and in RPMI 1640 medium containing FCS 10% and with penicillin-streptomycin after 2 weeks were differentiated into macrophages. Two groups were investigated, including control and infected with R. oryzae hyphae, for 6 and 16 hours after infection. (2) Hematogenous dissemination mucormycosis model: Seven groups of male BALb/c mice were examined in control, infected, and treated groups with Liposomal amphotericin B. (3) Human mucormycosis: in this study, two samples of patients with rhinocerebral mucormycosis with diabetes mellitus treated and untreated with liposomal amphotericin B were examined. Total RNA extraction and cDNA synthesis were performed from the studied samples. The relative expression changes of the target genes and miRNAs were evaluated using real-time PCR carried out using Sybrgreen-based detection methods.ResultsMonocyte-derived macrophages had a steady pattern in relative changes in gene expression. An increase in expression of two genes, GRP78 and CotH3, was observed in the samples, and all miRNAs targeted by the GRP78 gene included hsa - miR-16-5p; hsa -miR-335-5p and hsa -miR-93-3p showed a decreasing pattern. In the mice mucormycosis model, relative gene expression changes were observed, and mmu-miR-181b-5p showed increased expression deviation in all groups. The clinical sample of diabetic patients with untreated rhinocerebral mucormycosis also had a consistent pattern of GRP78 and CotH3 increased gene expression. The hsa-miR-16-5p and hsa-miR-335-5p have increased expression, while hsa-miR-93-3p decreased expression.ConclusionAfter validation, these micro RNAs can be used as valuable markers in mucormycosis detection and treatment processes.