Cellular Defense Response Research Articles

Low Magnetic Field Exposure Alters Prostate Cancer Cell Properties.

Prostate cancer is the second most common neoplasia and fifth-leading cause of cancer death in men worldwide. Electromagnetic and magnetic fields have been classified as possible human carcinogens, but current understanding of molecular and cellular pathways involved is very limited. Effects due to extremely low magnetic/hypomagnetic fields (LMF) are furthermore poorly understood. Extracellular vesicles (EVs) are crucial mediators of cellular communication with multifaceted roles in cancer progression, including via transport and uptake of various protein and microRNA (miRNA) EV-cargoes. miRNAs regulate gene expression and are implicated in cancer-related processes such as proliferation, metastasis, and chemoresistance. This study investigated the effects of LMF exposure (20 nT) by magnetic shielding on the prostate cancer cell line PC3 compared to the prostate epithelial cell line PNT2 under short-term (4 h) conditions. We examined EV profiles following a 4 h LMF exposure alongside associated functional enrichment KEGG and GO pathways for the EV proteomes. The 4 h LMF exposure significantly reduced cellular EV release and modified PC3 EV cargoes to a more inflammatory and metastatic profile, with 16 Disease Pathways and 95 Human Phenotypes associated specifically with the LMF-treated PC3 EV proteomes. These included cancerous, metabolic, blood, skin, cardiac and skeletal Disease Pathways, as well as pain and developmental disorders. In the normal PNT2 cells, less EV protein cargo was observed following LMF exposure compared with cells not exposed to LMF, and fewer associated functional enrichment pathways were identified. This pointed to some differences in various cellular functions, ageing, defence responses, oxidative stress, and disease phenotypes, including respiratory, digestive, immune, and developmental pathways. Furthermore, we analysed alterations in matrix metalloproteinases (MMPs) and miRNAs linked to metastasis, as this is crucial in cancer aggressiveness. The 4 h LMF exposure caused a significant increase in MMP2 and MMP9, as well as in onco-miRs miR-155, miR-210, miR-21, but a significant reduction in tumour-suppressor miRs (miR-200c and miR-126) in the metastatic PC3 cells, compared with normal PNT2 cells. In addition, 4 h LMF exposure significantly induced cellular invasion of PC3 cells. Overall, our findings suggest that changes in magnetic field exposures modulate EV-mediated and miR-regulatory processes in PCa metastasis, providing a basis for exploring novel therapeutic strategies.

Dissecting the mechanism of synergistic interactions between Aspergillus fumigatus and the microalgae Synechocystis sp. PCC6803 under Cd(II) exposure: insights from untargeted metabolomics

Co-culturing fungi and microalgae may effectively remediate wastewater containing Cd and harvest microalgae. Nevertheless, a detailed study of the mechanisms underlying the synergistic interactions between fungi and microalgae under Cd(II) exposure is lacking. In this study, Cd(II) exposure resulted in a significant enhancement of antioxidants, such as glutathione (GSH), malondialdehyde (MDA), hydrogen peroxide (H2O2) and superoxide dismutase (SOD) compared to the control group, suggesting that the cellular antioxidant defense response was activated. Extracellular proteins and extracellular polysaccharides of the symbiotic system were increased by 60.61 % and ,24.29 %, respectively, after Cd(II) exposure for 72 h. The adsorption behavior of Cd(II) was investigated using three-dimensional fluorescence excitation-emission matrix (3D-EEM), fourier transform infrared spectroscopy (FTIR), and scanning electron microscope (SEM). Metabolomics results showed that the TCA cycle provided effective material and energy supply for the symbiotic system to resist the toxicity of Cd(II); Proline, histidine, and glutamine strengthened the synergistic adsorption capacity of the fungus and microalgae. Overall, the theoretical foundation for a deep comprehension of the beneficial interactions between fungi and microalgae under Cd(II) exposure and the role of the fungal-algal symbiotic system in the management of heavy metal pollution is provided by this combined physiological and metabolomic investigation.

Abstract 4507: Discovery of a smart molecular glue: A small-molecular compound selectively degrades Nrf2 in KEAP1-mutated tumor cells, by restoring the broken KEAP1 mutant/Nrf2 interaction

Abstract Overcoming therapeutic resistance without incurring prohibitive normal tissue toxicity is a great challenge in anticancer therapies. Transcription factor NRF2 is a master regulator of cellular protective response, which provides protective effects in normal tissues. KEAP1, the endogenous NRF2 inhibitor, binds NRF2 and redirect it towards proteasome-dependent degradation. KEAP1/NRF2 interaction is therefore critical for maintaining NRF2 at a basal level. Accumulated studies revealed cancer cells could hijack NRF2 pathway to confer drug resistance. A number of clinically-relevant KEAP1 mutations were shown to disrupt the KEAP1/NRF2 interaction, leading to elevated NRF2 level and confer drug resistance. Here, by structure-based drug design approach, we discovered a small-molecule NRF2 inhibitor, R16, which selectively binds KEAP1 mutants and restores their NRF2-inhibitory function in tumor cells. We performed in silico screening against the NCI-open database. Top-ranking candidates were initially evaluated by ARE-luc reporter assays, leading to discovery of R16. Effect of R16 in restoring interaction between KEAP1 mutants and NRF2 were evaluated by BRET2 assays. Among 14 KEAP1 mutations that result in disruption of KEAP1/NRF2 interaction, R16 is active against 11 of them. A variety of assays, including tryptophan quenching, cellular target engagement, BRET2 and GST-pull down assays, indicated that R16 selectively engage KEAP1 mutants and restore their interaction with NRF2, leading to NRF2 degradation. R16 at 0.5 uM substantially sensitizes KEAP1-mutated tumor cells to cisplatin and gefitinib, with no obvious effect in wild-type KEAP1 cells. Importantly, R16 showed significant in-vivo efficacy in sensitizing A549 xenograft bearing KEAP1 G333C mutation to cisplatin, while it has no effect on xenograft with wild-type KEAP1. KEAP1-mutated tumors are emerging as a sub-group with severe resistance to currently available therapies. Here, we have identified a KEAP1 mutant-selective NRF2 inhibitor, which potently sensitizes KEAP1-mutated cells/tumors to anticancer agents, with little effects on the WT-KEAP1. Our work demonstrated it is feasible to restore the NRF2-inhibitory function of KEAP1 mutants, by targeting the mutants with compounds to repair the disrupted KEAP1 mutant/NRF2 interactions. The KEAP1 mutant-selectivity is critical, as NRF2 is a master regulator of cellular defense response in WT-KEAP1 normal tissues. Citation Format: Xiaohong Tian, Tahar Aboulkassim, Qiang Liu, Mark Hancock, Jian Hui Wu, Gerald Batist. Discovery of a smart molecular glue: A small-molecular compound selectively degrades Nrf2 in KEAP1-mutated tumor cells, by restoring the broken KEAP1 mutant/Nrf2 interaction [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 4507.

Unveiling Shared Immune Responses in Porcine Alveolar Macrophages during ASFV and PRRSV Infection Using Single-Cell RNA-seq.

African swine fever virus (ASFV) and porcine reproductive and respiratory syndrome virus (PRRSV) infections lead to severe respiratory diseases in pigs, resulting in significant economic losses for the global swine industry. While numerous studies have focused on specific gene functions or pathway activities during infection, an investigation of shared immune responses in porcine alveolar macrophages (PAMs) after ASFV and PRRSV infections was lacking. In this study, we conducted a comparison using two single-cell transcriptomic datasets generated from PAMs under ASFV and PRRSV infection. Pattern recognition receptors (PRRs) RIG-I (DDX58), MDA5 (IFIH1), and LGP2 (DHX58) were identified as particularly recognizing ASFV and PRRSV, triggering cellular defense responses, including the upregulation of four cytokine families (CCL, CXCL, IL, and TNF) and the induction of pyroptosis. Through weighted gene co-expression network analysis and protein-protein interaction analysis, we identified thirteen gene and protein interactions shared by both scRNA-seq analyses, suggesting the ability to inhibit both ASFV and PRRSV viral replication. We discovered six proteins (PARP12, PARP14, HERC5, DDX60, RSAD2, and MNDA) in PAMs as inhibitors of ASFV and PRRSV replication. Collectively, our findings showed detailed characterizations of the immune responses in PAMs during ASFV and PRRSV infections, which may facilitate the treatments of these viral diseases.

Mosquito midgut stem cell cellular defense response limits Plasmodium parasite infection

A novel cellular response of midgut progenitors (stem cells and enteroblasts) to Plasmodium berghei infection was investigated in Anopheles stephensi. The presence of developing oocysts triggers proliferation of midgut progenitors that is modulated by the Jak/STAT pathway and is proportional to the number of oocysts on individual midguts. The percentage of parasites in direct contact with enteroblasts increases over time, as progenitors proliferate. Silencing components of key signaling pathways through RNA interference (RNAi) that enhance proliferation of progenitor cells significantly decreased oocyst numbers, while limiting proliferation of progenitors increased oocyst survival. Live imaging revealed that enteroblasts interact directly with oocysts and eliminate them. Midgut progenitors sense the presence of Plasmodium oocysts and mount a cellular defense response that involves extensive proliferation and tissue remodeling, followed by oocysts lysis and phagocytosis of parasite remnants by enteroblasts.

In-depth transcriptomic analysis of Anopheles gambiae hemocytes uncovers novel genes and the oenocytoid developmental lineage

BackgroundHemocytes are immune cells that patrol the mosquito hemocoel and mediate critical cellular defense responses against pathogens. However, despite their importance, a comprehensive transcriptome of these cells was lacking because they constitute a very small fraction of the total cells in the insect, limiting the study of hemocyte differentiation and immune function.ResultsIn this study, an in-depth hemocyte transcriptome was built by extensive bulk RNA sequencing and assembly of hemocyte RNAs from adult A. gambiae female mosquitoes, based on approximately 2.4 billion short Illumina and about 9.4 million long PacBio high-quality reads that mapped to the A. gambiae PEST genome (P4.14 version). A total of 34,939 transcripts were annotated including 4,020 transcripts from novel genes and 20,008 novel isoforms that result from extensive differential splicing of transcripts from previously annotated genes. Most hemocyte transcripts identified (89.8%) are protein-coding while 10.2% are non-coding RNAs. The number of transcripts identified in the novel hemocyte transcriptome is twice the number in the current annotation of the A. gambiae genome (P4.14 version). Furthermore, we were able to refine the analysis of a previously published single-cell transcriptome (scRNAseq) data set by using the novel hemocyte transcriptome as a reference to re-define the hemocyte clusters and determine the path of hemocyte differentiation. Unsupervised pseudo-temporal ordering using the Tools for Single Cell Analysis software uncovered a novel putative prohemocyte precursor cell type that gives rise to prohemocytes. Pseudo-temporal ordering with the Monocle 3 software, which analyses changes in gene expression during dynamic biological processes, determined that oenocytoids derive from prohemocytes, a cell population that also gives rise to the granulocyte lineage.ConclusionA high number of mRNA splice variants are expressed in hemocytes, and they may account for the plasticity required to mount efficient responses to many different pathogens. This study highlights the importance of a comprehensive set of reference transcripts to perform robust single-cell transcriptomic data analysis of cells present in low abundance. The detailed annotation of the hemocyte transcriptome will uncover new facets of hemocyte development and function in adult dipterans and is a valuable community resource for future studies on mosquito cellular immunity.

Signaling Pathway Governed by Lipid Derived Molecules as Secondary Messenger

Research shown that the cell death, particularly apoptosis, can extend beyond single cell boundaries. Gap junctions IP3 diffusion, and sphingolipids play significant roles in membrane biology and regulation of cell function. S1P plays crucial role in the cardiovascular and immune systems, serving as a mediator of signaling during cell migration, differentiation, proliferation, and apoptosis. Intestinal phospholipid metabolism, including 1B phospholipase A2 and autotaxin-mediated pathways, contributes to cardiometabolic diseases through multiple mechanisms. A potential strategy for treating cardiovascular and metabolic diseases is the therapeutic suppression of1B phospholipase A2 and autotaxin in the gastrointestinal tract. Cellular stress signalling, inflammation, resolution, and host defence responses are all significantly influenced by lysophospholipids such LPA and S1P. New therapies for cancer, vascular diseases, fibrotic disorders, and autoimmune diseases have been made possible by developments in lysophospholipid research.

Exogenous γ-aminobutyric acid improves the photosynthesis efficiency, soluble sugar contents, and mineral nutrients in pomegranate plants exposed to drought, salinity, and drought-salinity stresses

Backgroundγ-aminobutyric acid (GABA), as a regulator of many aspects of plant growth, has a pivotal role in improving plant stress resistance. However, few studies have focused on the use of GABA in increasing plants’ resistance to interactional stresses, such as drought-salinity. Therefore, the focus of this study was to examine the effect of foliar application of GABA (0, 10, 20, and 40 mM) on growth indices and physio-biochemical parameters in plants of two pomegranate cultivars, ‘Rabab’ and ‘Atabaki’ exposed to drought, salinity, and drought-salinity.ResultsUnder stress conditions, the photosynthetic capacity of two pomegranate cultivars, including transpiration rate, net photosynthetic rate, intercellular carbon dioxide concentration, stomatal conductance of water vapour, and mesophyll conductance, was significantly reduced. This resulted in a decrease in root morphological traits such as fresh and dry weight, diameter, and volume, as well as the fresh and dry weight of the aerial part of the plants. However, the application of GABA reversed the negative effects caused by stress treatments on growth parameters and maintained the photosynthetic capacity. GABA application has induced the accumulation of compatible osmolytes, including total soluble carbohydrate, starch, glucose, fructose, and sucrose, in charge of providing energy for cellular defense response against abiotic stresses. Analysis of mineral nutrients has shown that GABA application increases the absorption of potassium, potassium/sodium, magnesium, phosphorus, manganese, zinc, and iron. As concentration increased up to 40 mM, GABA prevented the uptake of toxic ions, sodium and chloride.ConclusionsThese findings highlight the potential of GABA as a biostimulant strategy to enhance plant stress tolerance.

Alteration in the callogenesis, tropane alkaloid formation, and gene expression in Hyoscyamus niger under clinorotation.

This study aimed to investigate the effects of clinorotation induced by 2-D clinostat on the growth, tropane alkaloid production, gene expression, antioxidant capacity, and cellular defense responses in the callus tissue of Hyoscyamus niger. Callus induction was conducted by putting hypocotyl explants in the MS culture medium supplemented with 1 mgL-1 2,4-D and 1 mgL-1 BAP growth regulators. The sub-cultured calli were placed on a clinostat for 0, 3, 7, and 10 days (2.24 × 10-5 g on the edge of the callus ring). Clinorotation significantly increased callus fresh weight, dry weight, protein, carbohydrate, and proline contents compared to the control, and their maximum contents were obtained after 7 and 10 days. H2O2 level enhanced under clinorotation with a 76.3% rise after 10 days compared to control and positively affected the atropine (77.1%) and scopolamine (69.2%) productions. Hyoscyamine 6-beta hydroxylase and putrescine N-methyltransferase gene expression involved in the tropane alkaloid biosynthesis were upregulated markedly with 14.2 and 17.1-folds increase after 10 days of clinorotation, respectively. The expressions of jasmonic acid, mitogen-activated protein kinase, and ethylene-responsive element-binding transcription factor were upregulated, and the activity of peroxidase and catalase showed a 72.7 and 80% rise after 10 days. These findings suggest that microgravity can enhance callogenesis by stimulating the ROS level, which can impact the antioxidant enzymes, tropane alkaloid formation, and gene expression.

Label-free Raman microspectroscopic imaging with chemometrics for cellular investigation of apple ring rot and nondestructive early recognition using near-infrared reflection spectroscopy with machine learning

Apple ring rot caused by Botryosphaeria dothidea can cause fruit decay during the growth and storage stages of apple fruit. Understanding the infection process and cellular defense response at the cellular micro-level holds immense importance in the field of prevention and control. Consequently, there is a pressing need to develop suitable chemical imaging analysis methods. Here we proposed a label-free, high-throughput imaging method for cellular investigation of apple fruit ring rot infected by Botryosphaeria dothidea, based on confocal Raman microspectroscopic imaging technology combined with multivariate curve resolution-alternating least squares algorithm (MCR-ALS). We conducted Raman measurements on every apple fruit and obtain an image cube. This cube was then unfolded into an augmented matrix in a column-wise manner. We proceeded with simultaneous MCR-ALS analysis, resolving the single-substance spectrum and concentration profile from the mixed signals. Lastly, the accurate and pure molecular imaging of low methoxyl pectin, high methoxyl pectin, cellulose, lignin, and phenols were realized by refolding the resolved concentration data to construct the composition image. Thereafter, we realized the study of the spatial-temporal changes distribution of the above substances in the cuticle and cell wall of green and red apples at different stages of infection. The imaging method proposed in this paper is expected to provide a chemical imaging strategy for studying pathogen infection process and fruit defense response at the cellular level. In addition, by utilizing a fiber-optic probe near-infrared reflection spectrometer in conjunction with machine learning, we developed a rapid and non-destructive classification method. This method allows for the timely identification of apples exhibiting early infection by Botryosphaeria dothidea. Notably, both principal component analysis-quadratic discriminant analysis and support vector machine achieved a classification accuracy of 100%.

P-372 RNA sequencing studies evaluating oocyte quality in women with endometriosis: a meta-analysis

Abstract Study question Can a network-based integrative analysis (NBIA) meta-analysis approach be used to identify clinically relevant cumulus-oocyte complex biological pathway and gene targets in women with endometriosis? Summary answer Bi-level meta-analysis (BLMA) of multiple RNAseq studies shows enrichment of both inflammatory and mitochondrial pathway-related differential gene expression within oocyte, granulosa and cumulus cells. What is known already Endometriosis-related infertility is multifactorial, and studies suggest that impaired oocyte quality plays a significant role. The close relationship between granulosa, cumulus cells and developing oocytes makes them ideal candidates for studying non-invasive biomarkers for oocyte quality. As high-throughput sequencing becomes more accessible, the volume of RNAseq molecular data in public databases such as Gene Expression Omnibus (GEO) has increased. Combined analysis of multiple datasets theoretically increases study power due to larger number of samples. Comparison of differentially expressed genes (DEGs) and biological pathways across multiple independent studies therefore requires a ‘meta-analysis’ framework that can effectively integrate data from different sources. Study design, size, duration We identified relevant RNAseq datasets through NCBI public functional genomics data repository Gene Expression Omnibus (GEO). Datasets were split into “test” and “control” groups referring to models of oocyte incompetence and competence. Accession numbers GSEXXX or PRJNXXX represent unique identifiers for RNAseq data files contained within a given NCBI GEO series or BioProject record, respectively. Five human RNAseq datasets (PRJNA514416, GSE168214, PRJNA216966, PRJNA727838 and GSE81579) were identified and DEG lists generated for each individual dataset. Participants/materials, setting, methods Normalisation and differential expression between RNAseq datasets was carried out in R using the Bioconductor DESeq2 package. DEGs were identified as those whose P-value was adjusted using a false discovery rate less than 0.5, adjusted using Benjamini-Hochberg procedure. Log2 fold-change ratio (logFC) of each DEG was used to identify significantly up and downregulated genes. The NBIA pipeline (Nguyen et al, 2020) examines over-representation of significantly impacted genes in a pathway as per KEGG pathway analysis. Main results and the role of chance Upregulated DEGs were identified in mitochondrial membrane ATP synthase pathways, (ATP5G1, ATP5G2, ATP5J2 and ATP5H). KEGG Pathway terms “Parkinson’s”, “Huntington’s”, and “Alzheimer disease” were also enriched, due to many genes partaking in oxidative phosphorylation in the context of these pathologies. Functional analysis also revealed significantly down-regulated genes related to inflammatory pathways (CXCL1, 2, 3, 5,6 and 8), which have a role in innate immunity. Although surprising, in our analysis, downregulated functions based on KEGG functional pathway analysis, observed that innate immune response, cellular defense response, chemokine-mediated signaling were significantly downregulated. BLMA over-representation analysis (ORA) and impact analysis (IA) analysis showed that the Amoebiasis pathway was enriched expressed across all datasets. Genes involved in this pathway include interleukin (IL), CXCL1, CXCL2, CXCL3, SERPINB, TGFB1 and TGFB2, and its biological process is related to natural killer (NK) cell mediated cytotoxicity. Overall, the enrichment of immune and inflammation-related pathways revealed by BLMA meta-analysis underscores the relevance of inflammation to follicular function. Indeed, ovulation is inextricably linked with inflammation. Other significantly enriched pathways highlighted by this meta-analysis are related to infection (path:hsa05134 Legionellosis; path:hsa05140 Leishmaniasis) immune response (path:hsa04145 Phagosome; path:hsa04610 Complement and coagulation cascades) and inflammation (path:hsa05323 Rheumatoid arthritis). Limitations, reasons for caution Methods used availed of publicly available GEO datasets and bioinformatics software (Bioconductor, R and DAVID). Although useful tools, these approaches are open to inter-user variation. Cell types compared across studies were oocytes, granulosa and cumulus cells. Controlled ovarian stimulation may also be open to influence based on stimulation protocol used. Wider implications of the findings Enrichment of inflammatory and mitochondrial pathways in our GO analysis is compelling. Given the role of mitochondria in oocyte developmental competence, this reinforces the role of impaired oocyte competence in women with endometriosis. Our study identifies several genes that require further elucidation as potential predictors of oocyte quality in endometriosis. Trial registration number N/A

Bioactivity-guided fractionation of Helicteres angustifolia L. extract and its molecular evidence for tumor suppression.

Helicteres angustifolia L. (Helicteres angustifolia) has been commonly used in folk medicine to treat cancer; however, its mechanisms of action remain obscure. In our earlier work, we reported that aqueous extract of H. angustifolia root (AQHAR) possesses attractive anticancer properties. In the present study, we isolated five ethanol fractions from AQHAR and investigated their therapeutic efficacy in human non-small cell lung cancer (NSCLC) cells. The results showed that among the five fractions, the 40% ethanol fraction (EF40) containing multiple bioactive compounds exhibited the best selective killing effect on NSCLC cells with no obvious toxicity to normal human fibroblasts. Mechanistically, EF40 reduced the expression of nuclear factor-E2-related factor 2 (Nrf2), which is constitutively expressed at high levels in many types of cancers. As a result, Nrf2-dependent cellular defense responses are suppressed, leading to the intracellular accumulation of reactive oxygen species (ROS). Extensive biochemical analyses revealed that EF40 caused cell cycle arrest and apoptosis through activation of the ROS-mediated DNA damage response. Furthermore, treatment with EF40 compromised NSCLC cell migration, as evidenced by the downregulation of matrix metalloproteinases (MMPs) and heterogeneous nuclear ribonucleoprotein K (hnRNP-K). In vivo studies using A549 xenografts in nude mice also revealed significant suppression of tumor growth and lung metastasis in the treated group. We propose that EF40 may serve as a potential natural anti-NSCLC drug that warrants further mechanistic and clinical attention.

Bacillus thuringiensis exopolysaccharides induced systemic resistance against Sclerotinia sclerotiorum in Brassica campestris L.

Bacillus thuringiensis is known as an entomopathogenic bacterium active against different agricultural pests. Recently, some novel functions have been explored, including plant growth-promoting activities. In our previous work, B. thuringiensis strains were found to effectively suppress Sclerotinia sclerotiorum by inducing systemic resistance (ISR) in Brassica campestris (L.). However, the mechanism by which exogenous signals from B. thuringiensis are perceived by plants is still not clear. Here, based on challenge-inoculation assays, we found that exopolysaccharides (EPSs) of B. thuringiensis strains could limit the spread of S. sclerotiorum by ISR in B. campestris. We purified and characterized the Bt 4F5 EPS, and the molecular weight was 66.2 kDa. The main monosaccharide units of Bt 4F5 EPS were determined to be mannose, GluA, glucose and galactose. Cellular defense response markers such as defense-related enzymes were induced after the 4F5 EPSs pretreatment in the leaves. Moreover, quantitative real-time PCR results indicated that the defense-related genes HEL and PDF1.2 were upregulated, suggesting an activation of the JA/ET signaling pathways by Bt 4F5 EPS. In addition, complete genome sequencing of the Bt 4F5 strain revealed twenty-one candidate genes involved in exopolysaccharide synthesis in chromosome. In summary, the EPSs of Bt strains play an important role in micro-associated molecular pattern (MAMP) perception during the process of Bt-triggered systemic resistance.

Seed Bio-priming of wheat with a novel bacterial strain to modulate drought stress in Daegu, South Korea.

Wheat is one of the major cereal crop grown food worldwide and, therefore, plays has a key role in alleviating the global hunger crisis. The effects of drought stress can reduces crop yields by up to 50% globally. The use of drought-tolerant bacteria for biopriming can improve crop yields by countering the negative effects of drought stress on crop plants. Seed biopriming can reinforce the cellular defense responses to stresses via the stress memory mechanism, that its activates the antioxidant system and induces phytohormone production. In the present study, bacterial strains were isolated from rhizospheric soil taken from around the Artemisia plant at Pohang Beach, located near Daegu, in the South Korea Republic of Korea. Seventy-three isolates were screened for their growth-promoting attributes and biochemical characteristics. Among them, the bacterial strain SH-8 was selected preferred based on its plant growth-promoting bacterial traits, which are as follows: abscisic acid (ABA) concentration = 1.08 ± 0.05 ng/mL, phosphate-solubilizing index = 4.14 ± 0.30, and sucrose production = 0.61 ± 0.13 mg/mL. The novel strain SH-8 demonstrated high tolerance oxidative stress. The antioxidant analysis also showed that SH-8 contained significantly higher levels of catalase (CAT), superoxide dismutase (SOD), and ascorbic peroxidase (APX). The present study also quantified and determined the effects of biopriming wheat (Triticum aestivum) seeds with the novel strain SH-8. SH-8 was highly effective in enhancing the drought tolerance of bioprimed seeds; their drought tolerance and germination potential (GP) were increased by up to 20% and 60%, respectively, compared with those in the control group. The lowest level of impact caused by drought stress and the highest germination potential, seed vigor index (SVI), and germination energy (GE) (90%, 2160, and 80%, respectively), were recorded for seeds bioprimed with with SH-8. These results show that SH-8 enhances drought stress tolerance by up to 20%. Our study suggests that the novel rhizospheric bacterium SH-8 (gene accession number OM535901) is a valuable biostimulant that improves drought stress tolerance in wheat plants and has the potential to be used as a biofertilizer under drought conditions.

Proteomic profiling of eIF3a conditional knockout mice.

Eukaryotic translation initiation factor 3 subunit A (eIF3a) is the largest subunit of the eukaryotic translation initiation factor 3 (eIF3). eIF3a plays an integral role in protein biosynthesis, hence impacting the onset, development, and treatment of tumors. The proteins regulated by eIF3a are still being explored in vivo. In this study, a Cre-loxP system was used to generate eIF3a conditional knockout mice. Tandem mass tag (TMT) labeling with LC-MS/MS analysis was used to identify differentially expressed proteins (DEPs) in fat, lungs, skin, and spleen tissue of the eIF3a knockout mice and controls. Bioinformatics analysis was then used to explore the functions and molecular signaling pathways of these protein landscapes. It was observed that eIF3a is essential for life sustenance. Abnormal tissue pathology was found in the lungs, fat, skin, spleen, and thymus. In total, 588, 210, 324, and 944 DEPs were quantified in the lungs, fat, skin, and spleen, respectively, of the eIF3a knockout mice as compared to the control. The quantified differentially expressed proteins were tissue-specific, except for eight proteins shared by the four tissues. A broad range of functions for eIF3a, including cellular signaling pathway, immune response, metabolism, defense response, phagocytes, and DNA replication, has been revealed using bioinformatics analysis. Herein, several pathways related to oxidative stress in the Kyoto Encyclopedia of Genes and Genomes (KEGG) database, including nitrogen metabolism, peroxisome, cytochrome P450 drug metabolism, pyruvate metabolism, PPAR signaling pathway, phospholipase D signaling pathway, B-cell receptor signaling pathway, ferroptosis, and focal adhesion, have been identified. Collectively, this study shows that eIF3a is an essential gene for sustaining life, and its downstream proteins are involved in diverse novel functions beyond mRNA translational regulation.

Transcriptomic analysis reveals shared gene signatures and molecular mechanisms between obesity and periodontitis.

Both obesity (OB) and periodontitis (PD) are chronic non-communicable diseases, and numerous epidemiological studies have demonstrated the association between these two diseases. However, the molecular mechanisms that could explain the association between OB and PD are largely unclear. This study aims to investigate the common gene signatures and biological pathways in OB and PD through bioinformatics analysis of publicly available transcriptome datasets. The RNA expression profile datasets of OB (GSE104815) and PD (GSE106090) were used as training data, and GSE152991 and GSE16134 as validation data. After screening for differentially expressed genes (DEGs) shared by OB and PD, gene enrichment analysis, protein-protein interaction (PPI) network construction, GeneMANIA analysis, immune infiltration analysis and gene set enrichment analysis (GSEA) were performed. In addition, receiver operating characteristic (ROC) curves were used to assess the predictive accuracy of the hub gene. Finally, we constructed the hub gene-associated TF-miRNA-mRNA regulatory network. We identified a total of 147 DEGs shared by OB and PD (38 down-regulated and 109 up-regulated). Functional analysis showed that these genes were mainly enriched in immune-related pathways such as B cell receptor signalling, leukocyte migration and cellular defence responses. 14 hub genes (FGR, MNDA, NCF2, FYB1, EVI2B, LY86, IGSF6, CTSS, CXCR4, LCK, FCN1, CXCL2, P2RY13, MMP7) showed high sensitivity and specificity in the ROC curve analysis. The results of immune infiltration analysis showed that immune cells such as macrophages, activated CD4 T cells and immune B cells were present at high infiltration levels in both OB and PD samples.The results of GeneMANIA analysis and GSEA analysis suggested that five key genes (FGR, LCK, FYB1, LY86 and P2RY13) may be strongly associated with macrophages. Finally, we constructed a TF-miRNA-mRNA regulatory network consisting of 233 transcription factors (TFs), 8 miRNAs and 14 mRNAs based on the validated information obtained from the database. Five key genes (FGR, LCK, FYB1, LY86, P2RY13) may be important biomarkers of OB and PD. These genes may play an important role in the pathogenesis of OB and PD by affecting macrophage activity and participating in immune regulation and inflammatory responses.

Morphological and functional characterization of the oyster Crassostrea gasar circulating hemocytes: Cell types and phagocytosis activity

Hemocytes are the circulating cells of the hemolymph of oysters and are responsible for numerous physiological functions, including immune defense. The oyster Crassostrea gasar is a native species inhabiting mangrove habitat and is of great commercial interest, cultured throughout the Brazilian coast, mainly in the north and northeast. Despite its commercial importance, little is known about its immunological aspects and defense cells, the hemocytes. This work aimed to morphologically characterize hemocytes of the oyster C. gasar and to study one of the main cellular defense response, phagocytosis, using light microscopy and flow cytometry. The results showed the presence of six hemocyte populations in C. gasar hemolymph. These comprise of large and small granulocytes, large and small hyalinocytes, blast-like cells and a rare type classified as vesicular or serous hemocytes. Hyalinocytes were highly abundant and the most heterogeneous cell population, while small granulocytes, along with vesicular hemocytes were the less abundant population. Hemocytes of C. gasar oysters demonstrated capabilities to phagocytose three different types of particles tested: zymosan A, latex particles and Escherichia coli, indicating a broad defense capacity. The zymosan A were the most engulfed particles, followed by beads, mainly phagocytized by granulocytes, the most phagocytic cells, and finally E. coli, which were the least phagocytized. This study is the first characterization of C. gasar oyster hemocytes and will support future studies that aim to understand the participation of different hemocyte types in defense responses against pathogens and/or environmental changes.

Evidence for Extensive Duplication and Subfunctionalization of FCRL6 in Armadillo (Dasypus novemcinctus).

The control of infections by the vertebrate adaptive immune system requires careful modulation to optimize defense and minimize harm to the host. The Fc receptor-like (FCRL) genes encode immunoregulatory molecules homologous to the receptors for the Fc portion of immunoglobulin (FCR). To date, nine different genes (FCRL1-6, FCRLA, FCRLB and FCRLS) have been identified in mammalian organisms. FCRL6 is located at a separate chromosomal position from the FCRL1-5 locus, has conserved synteny in mammals and is situated between the SLAMF8 and DUSP23 genes. Here, we show that this three gene block underwent repeated duplication in Dasypus novemcinctus (nine-banded armadillo) resulting in six FCRL6 copies, of which five appear functional. Among 21 mammalian genomes analyzed, this expansion was unique to D. novemcinctus. Ig-like domains that derive from the five clustered FCRL6 functional gene copies show high structural conservation and sequence identity. However, the presence of multiple non-synonymous amino acid changes that would diversify individual receptor function has led to the hypothesis that FCRL6 endured subfunctionalization during evolution in D. novemcinctus. Interestingly, D. novemcinctus is noteworthy for its natural resistance to the Mycobacterium leprae pathogen that causes leprosy. Because FCRL6 is chiefly expressed by cytotoxic T and NK cells, which are important in cellular defense responses against M. leprae, we speculate that FCRL6 subfunctionalization could be relevant for the adaptation of D. novemcinctus to leprosy. These findings highlight the species-specific diversification of FCRL family members and the genetic complexity underlying evolving multigene families critical for modulating adaptive immune protection.

Host-pathogen interaction in arthropod vectors: Lessons from viral infections.

Haematophagous arthropods can harbor various pathogens including viruses, bacteria, protozoa, and nematodes. Insects possess an innate immune system comprising of both cellular and humoral components to fight against various infections. Haemocytes, the cellular components of haemolymph, are central to the insect immune system as their primary functions include phagocytosis, encapsulation, coagulation, detoxification, and storage and distribution of nutritive materials. Plasmatocytes and granulocytesare also involved in cellular defense responses. Blood-feeding arthropods, such as mosquitoes and ticks, can harbour a variety of viral pathogens that can cause infectious diseases in both human and animal hosts. Therefore, it is imperative to study the virus-vector-host relationships since arthropod vectors are important constituents of the ecosystem. Regardless of the complex immune response of these arthropod vectors, the viruses usually manage to survive and are transmitted to the eventual host. A multidisciplinary approach utilizing novel and strategic interventions is required to control ectoparasite infestations and block vector-borne transmission of viral pathogens to humans and animals. In this review, we discuss the arthropod immune response to viral infections with a primary focus on the innate immune responses of ticks and mosquitoes. We aim to summarize critically the vector immune system and their infection transmission strategies to mammalian hosts to foster debate that could help in developing new therapeutic strategies to protect human and animal hosts against arthropod-borne viral infections.

Hydralazine Revives Cellular and Ocular Lens Health-Span by Ameliorating the Aging and Oxidative-Dependent Loss of the Nrf2-Activated Cellular Stress Response.

A major hallmark of aging-associated diseases is the inability to evoke cellular defense responses. Transcriptional protein Nrf2 (nuclear factor erythroid-derived 2-related factor) plays a pivotal role in the oxidative stress response, cellular homeostasis, and health span. Nrf2's activation has been identified as a therapeutic target to restore antioxidant defense in aging. Here, we demonstrated that FDA-approved drug, hydralazine (Hyd), was a reactivator of the Nrf2/ARE (antioxidant response element) pathway in various ages and types of mouse (m) or human (h) lens epithelial cells (LECs) and mice lenses in-vitro/in-vivo. This led to Hyd-driven abatement of carbonyls, reduced reactive oxygen species (ROS), and reduced 4-HNE/MDA-adducts with cytoprotection, and extended lens healthspan by delaying/preventing lens opacity against aging/oxidative stress. We elucidated that Hyd activated the protective signaling by inducing Nrf2 to traverse from the cytoplasm to the nucleus and potentiated the ARE response by direct interaction of Nrf2 and ARE sequences of the promoter. Loss-of-function study and cotreatment of Hyd and antioxidant, N-acetyl cysteine (NAC) or Peroxiredoxin (Prdx)6, specified that Nrf2/ARE-driven increase in the promoter activity was Hyd-dependent. Our study provides proof-of concept evidence and, thereby, paves the way to repurposing Hyd as a therapeutic agent to delay/prevent aging and oxidative-related disorders.