Chemotactic Response Research Articles

Chemotaxis-driven stationary and oscillatory patterns in a diffusive HIV-1 model with CTL immune response and general sensitivity.

In this paper, a reaction-diffusion-chemotaxis HIV-1 model with a cytotoxic T lymphocyte (CTL) immune response and general sensitivity is investigated. We first prove the global classical solvability and L∞-boundedness for the considered model in a bounded domain with arbitrary spatial dimensions, which extends the previous existing results. Then, we apply the global existence result to the case with a linear proliferation immune response and an incidence rate. We study the spatiotemporal dynamics about the three types of spatially homogeneous steady states: infection-free steady state S0, CTL-inactivated infection steady state S1, and CTL-activated infection steady state S∗. Our analyses indicate that S0 is globally asymptotically stable if the basic reproduction number R0 is less than 1; if R0 is between 1 and a threshold, then S1 is globally asymptotically stable. However, if R0 is larger than the threshold, then the chemoattraction and chemorepulsion can destabilize S∗, and thus, a spatiotemporal pattern forms as the chemotactic sensitivity crosses certain critical values. We obtain two kinds of important patterns, which are induced by chemotaxis: stationary Turing pattern and irregular oscillatory pattern. We also find that different chemotactic response functions can affect system's dynamics. Based on some empirical parameter values, numerical simulations are given to illustrate the effectiveness of the theoretical predications.

Certain Tomato Root Exudates Induced by Pseudomonas stutzeri NRCB010 Enhance Its Rhizosphere Colonization Capability

Plant growth-promoting rhizobacteria (PGPR) can colonize plant root surfaces or form biofilms to promote plant growth and enhance plant resistance to harsh external environments. However, plant-PGPR interactions, especially chemical signaling molecules, are poorly understood. This study aimed to gain an in-depth understanding of the rhizosphere interaction mechanisms between PGPR and tomato plants. This study found that inoculation with a certain concentration of Pseudomonas stutzeri significantly promoted tomato growth and induced significant changes in tomato root exudates. Furthermore, the root exudates significantly induced NRCB010 growth, swarming motility, and biofilm formation. In addition, the composition of the root exudates was analyzed, and four metabolites (methyl hexadecanoate, methyl stearate, 2,4-di-tert-butylphenol, and n-hexadecanoic acid) significantly related to the chemotaxis and biofilm formation of NRCB010 were screened. Further assessment showed that these metabolites positively affected the growth, swarming motility, chemotaxis, or biofilm formation of strain NRCB010. Among these, n-hexadecanoic acid induced the most remarkable growth, chemotactic response, biofilm formation, and rhizosphere colonization. This study will help develop effective PGPR-based bioformulations to improve PGPR colonization and crop yields.

Integrated interactome and transcriptome analysis reveals key host factors critical for SARS-CoV-2 infection

The coronavirus disease 2019 (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has seriously threatened global public health and caused huge economic losses. Omics studies of SARS-CoV-2 can help understand the interaction between the virus and host, thereby providing a new perspective in guiding the intervention and treatment of the SARS-CoV-2 infection. Since large amount of SARS-CoV-2 omics data have been accumulated in public databases, this study aimed to identify key host factors involved in SARS-CoV-2 infection through systematic integration of transcriptome and interactome data. By manually curating published studies, we obtained a comprehensive SARS-CoV-2-human protein-protein interactions (PPIs) network, comprising 3591 human proteins interacting with 31 SARS-CoV-2 viral proteins. Using the RobustRankAggregation method, we identified 123 multiple cell line common genes (CLCGs), of which 115 up-regulated CLCGs showed host enhanced innate immunity and chemotactic response signatures. Combined with network analysis, co-expression and functional enrichment analysis, we discovered four key host factors involved in SARS-CoV-2 infection: IFITM1, SERPINE1, DDX60, and TNFAIP2. Furthermore, SERPINE1 was found to facilitate SARS-CoV-2 replication, and can alleviate the endoplasmic reticulum (ER) stress induced by ORF8 protein through interaction with ORF8. Our findings highlight the importance of systematic integration analysis in understanding SARS-CoV-2-human interactions and provide valuable insights for future research on potential therapeutic targets against SARS-CoV-2 infection.

Severe CD8+ T Lymphopenia in WHIM Syndrome Caused by Selective Sequestration in Primary Immune Organs.

Warts, hypogammaglobulinemia, infections, and myelokathexis (WHIM) syndrome is an ultra-rare combined primary immunodeficiency disease caused by heterozygous gain-of-function mutations in the chemokine receptor CXCR4. WHIM patients typically present with recurrent acute infections associated with myelokathexis (severe neutropenia due to bone marrow retention of mature neutrophils). Severe lymphopenia is also common, but the only associated chronic opportunistic pathogen is human papillomavirus and mechanisms are not clearly defined. In this study, we show that WHIM mutations cause more severe CD8 than CD4 lymphopenia in WHIM patients and WHIM model mice. Mechanistic studies in mice revealed selective and WHIM allele dose-dependent accumulation of mature CD8 single-positive cells in thymus in a cell-intrinsic manner due to prolonged intrathymic residence, associated with increased CD8 single-positive thymocyte chemotactic responses invitro toward the CXCR4 ligand CXCL12. In addition, mature WHIM CD8+ T cells preferentially home to and are retained in the bone marrow in mice in a cell-intrinsic manner. Administration of the specific CXCR4 antagonist AMD3100 (plerixafor) in mice rapidly and transiently corrected T cell lymphopenia and the CD4/CD8 ratio. After lymphocytic choriomeningitis virus infection, we found no difference in memory CD8+ T cell differentiation or viral load between wild-type and WHIM model mice. Thus, lymphopenia in WHIM syndrome may involve severe CXCR4-dependent CD8+ T cell deficiency resulting in part from sequestration in the primary lymphoid organs, thymus, and bone marrow.

Rhizobacterial Colonization and Management of Bacterial Speck Pathogen in Tomato by Pseudomonas spp.

Plants and soil microorganisms interact at every stage of growth. Pseudomonas spp. are highly regarded for their ability to increase crop production and protection from diseases. The aim of this study is to understand the mechanisms of the rhizobacterial colonization of tomato roots via chemotaxis assay and the activation of tomato resistance against the pathogenic bacterium, Pseudomonas syringae pv. tomato DC3000 (Pst). The capillary assay was used to evaluate the chemotaxis response of PGPRs (plant growth-promoting rhizobacteria). The activities of defense enzymes and the expressions of PR (pathogenesis-related) genes were measured using real-time qPCR. Chemotactic responses to malic and citric acids (the most important root exudates found in different plant species) at low concentrations varied substantially among the rhizobacterial isolates (63 species). Beneficial isolates including Pseudomonas resinovorans A5, P. vranovensis A30, P. resinovorans A28, P. umsongensis O26, P. stutzeri N42, and P. putida T15 reacted well to different concentrations of root exudates. P. putida T15 demonstrated the most potent anti-Pst activity. At three and six days after inoculation, the greatest levels of polyphenol oxidase and peroxidase activity were reported in the A5 and T15 groups. In tomato, transcript levels of four PR (pathogenesis-related) genes were elevated by rhizobacterial treatments. PGPR isolates alone or in combination with BABA (β-amino butyric acid) up-regulated the transcriptions of PR1, PR2, LOX, and PAL genes. Treatments with N42 and T15 resulted in the greatest improvements in tomato growth and yield traits. In conclusion, the results explain the mechanisms of rhizobacterial colonization for the improved management of Pst. Rhizobacterial isolates play a role in tomato’s resistance to Pst via salicylic acid and jasmonic acid pathways.

Palbociclib blocks neutrophilic phosphatidylinositol 3-kinase activity to alleviate psoriasiform dermatitis.

Neutrophilic inflammation is a critical pathogenic factor in psoriasis. The therapeutic applicability of palbociclib, a cyclin-dependent kinase 4 and 6 (CDK4/6) inhibitor clinically used to treat cancer, in the treatment of neutrophil-associated psoriasis remains undefined. In this study, we evaluated the therapeutic potential and pharmacological effect of palbociclib on neutrophil-associated psoriasiform dermatitis. The anti-inflammatory effects of palbociclib were determined in activated human neutrophils. The therapeutic feasibility of palbociclib in psoriasis was demonstrated in a mouse model of imiquimod-induced psoriasiform dermatitis. The in vitro enzymatic assays and in silico analyses were used to identify the underlying pharmacological mechanisms. This study found that palbociclib inhibited neutrophilic inflammation, including superoxide anion generation, reactive oxygen species (ROS) formation, elastase degranulation and chemotactic responses. The mechanistic studies identified that the anti-inflammatory effects of palbociclib involved the targeting of phosphatidylinositol 3-kinase (PI3K) but not CDK4/6 in human neutrophils. Palbociclib preferentially targeted the p110δ catalytic subunit of PI3K and thereby blocked signalling via the PI3K/protein kinase B (Akt) pathway. Furthermore, topical application of palbociclib significantly ameliorated imiquimod-induced psoriasiform dermatitis in mice, including psoriatic symptoms, neutrophil infiltration, Akt activation and cytokine up-regulation. This is the first study to demonstrate that palbociclib can potentially be used to treat neutrophil-associated psoriasiform dermatitis through the targeting of neutrophilic PI3K activity. Our findings prompt further research to explore the potential of palbociclib and PI3K in psoriasis and other inflammatory diseases.

Immune phenotype is differentially affected by changing the type of bovine respiratory disease vaccine administered at revaccination in beef heifers.

During preconditioning, modified-live vaccines are frequently administered to beef calves before weaning. In this study, we began to characterize the immune phenotype of calves that received a modified-live vaccination at 3-4 months of age and then either received the same modified-live or an inactivated vaccine upon arrival at the feedlot (weaning) and 28 days post-arrival (booster). Innate and adaptive immune measures were assessed before revaccination and 14 and 28 days post. Heifers that received three doses of the modified-live vaccine exhibited a relatively balanced immune response based on increases in mean cytokine concentrations (IL-17, IL-21) and total immunoglobulin-G (IgG) and subsets IgG1 and IgG2, which are related to both arms of the adaptive immune system. Conversely, heifers that received one dose of modified live and two doses of the inactivated vaccine had a more robust neutrophil chemotactic response and greater serum-neutralizing antibody titers, resulting in an enhanced innate immune and a skewed proinflammatory response. These results indicate that the revaccination protocol used after initial vaccination with a modified-live vaccine differentially influences the immune phenotype of beef calves, with three doses of modified live inducing potentially immune homeostasis and a combination of modified live and inactivated vaccines inducing a skewed immune phenotype. However, more research is needed to determine the protective efficacy of these vaccination protocols against disease.

Enhancement of neutrophil chemotaxis by trans-anethole-treated Staphylococcus aureus strains.

This study aimed to analyze the chemotactic response of differentiated HL-60 neutrophil-like (dHL-60) cells to trans-anethole (TA)-treated Staphylococcus aureus strains. Special attention was paid to evaluate the influence of TA on the chp gene expression level, as well as molecular docking and molecular dynamics (MD) simulation studies on interactions of TA with chemotaxis inhibitory protein of S. aureus (CHIPS). The following parameters were studied: susceptibility to TA using the agar diffusion method, the chp gene detection and its expression under TA influence, and clonal diversity of S. aureus strains using molecular techniques. Furthermore, a chemotactic response of dHL-60 cells to TA-treated S. aureus using Boyden chamber assay was detected and molecular modeling using both the docking methodology and unbiased MD simulations was conducted. It was found that TA showed antibacterial activity against all strains. Three genotypes and one unique pattern were distinguished among the strains. 50% of the isolates were chp-positive. It was observed that TA reduced/inhibited chp gene expression in most S. aureus strains. Enhanced chemotactic response of dHL-60 cells to TA-treated S. aureus strains was also noted. This correlation was similar for both chp-positive and chp-negative strains. Both molecular docking and MD simulations studies confirmed that TA is preferentially bound in the complement component 5a/CHIPS interface interaction region and can interfere with any processes exploiting this binding cavity. It has been proven that dHL-60 cells exhibited a higher chemotactic response to TA-treated S. aureus strains in comparison to non-treated bacteria, regardless of the achieved expression of the chp gene or its lack. Nevertheless, further analyses are required to understand this mechanism better.

Abstract 4193: N-NOSE technology based on C. elegans olfaction. Multi-cancer early detection with high sensitivity using urine. Pancreatic-cancer-specific early detection with high sensitivity using urine

Abstract N-NOSE (Nematode Nose) is a cancer screening test based on the chemotactic characteristics of the nematode C. elegans. C. elegans shows avoidance of the urine of healthy individuals while displaying a chemotactic attraction toward the urine of patients with 15 types of cancer (stomach, colon-rectum, lung, breast, pancreas, liver, prostate, uterus, esophagus, gallbladder, bile duct, kidney, urinary bladder, ovary, oropharynx) rendering N-NOSE a primary multi-cancer screening test. N-NOSE has a high sensitivity of 87.5% and 90.2% specificity, on average, from early stage 0-I. N-NOSE is non-invasive because it relies on a urine sample and is affordable (JPY 13,800, ~USD 100).N-NOSE by Hirotsu Bio Science has been available in Japan since late 2020 and is a commercial hit with over 250,000 screenings performed, rapidly increasing. In this presentation, we outline the N-NOSE technology and business in Japan. We review N-NOSE clinical research and critical findings compared to competitors that highlight its excellent performance as a non-invasive primary cancer screening for early detection. N-NOSE is a multi-cancer early detection test. Next, we introduce an evolution of N-NOSE for identifying the cancer type. Pancreatic cancer is the 12th most common cancer worldwide but remains one the deadliest cancer with a 5-year relative survival rate at only 11%, all stages combined. An affordable, non-invasive, and highly sensitive early-stage pancreatic cancer screening test is sorely needed. As an evolution of N-NOSE we genetically engineered C. elegans by knocking out in AWC olfactory neurons a specific olfactory receptor (GPCR), triggering a chemotactic response particular to the only pancreatic cancer urine specimen at an early stage. In this study, we outline, for the first time, the critical discovery of cr-4 GPCR in AWC, which is crucial for a specific chemotaxis response to the urine of patients with pancreatic cancer. Next, we briefly conclude by outlining the link between N-NOSE primary cancer screening followed by N-NOSE cancer type identification, effectively completing the whole cancer screening and identification process in a non-invasive way. Citation Format: Eric Di Luccio, Takaaki Hirotsu. N-NOSE technology based on C. elegans olfaction. Multi-cancer early detection with high sensitivity using urine. Pancreatic-cancer-specific early detection with high sensitivity using urine. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4193.

Epigallocatechin-3-Gallate Prevents the Acquisition of a Cancer Stem Cell Phenotype in Ovarian Cancer Tumorspheres through the Inhibition of Src/JAK/STAT3 Signaling

Three-dimensional tumorsphere cultures recapitulate the expression of several cancer stem cell (CSC) biomarkers and represent an effective in vitro platform to screen the anti-CSC properties of drugs. Whereas ovarian carcinoma is among the leading causes of death for women, ovarian CSC (OvCSC), a highly malignant subpopulation of ovarian cancer cells, is thought to be responsible for therapy resistance, metastasis, and tumor relapse. Epigallocatechin-3-gallate (EGCG), a diet-derived active polyphenol found in green tea leaves, can suppress ovarian cancer cell proliferation and induce apoptosis. However, its capacity to prevent the acquisition of cancer stemness traits in ovarian malignancies remains unclear. Here, we exploited the in vitro three-dimensional tumorsphere culture model to explore the capacity of EGCG to alter CSC biomarkers expression, signal transducing events and cell chemotaxis. Total RNA and protein lysates were isolated from human ES-2 ovarian cancer cell tumorspheres for gene assessment by RT-qPCR and protein expression by immunoblot. Real-time cell chemotaxis was assessed with xCELLigence. Compared with their parental adherent cells, tumorspheres expressed increased levels of the CSC markers NANOG, SOX2, PROM1, and Fibronectin. EGCG treatment reduced dose-dependently tumorspheres size and inhibited the transcriptional regulation of those genes. Src and JAK/STAT3 signaling pathways appeared to be relevant for CSC phenotype and chemotactic response. In conclusion, these data highlight and support the chemopreventive benefits of the diet-derived EGCG and its capacity to target intracellular transducing events that regulate the acquisition of an invasive CSC phenotype.

Determination of Hourly Distribution of Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) Using Sex Pheromone and Ultraviolet Light Traps in Protected Tomato Crops

Tuta absoluta (Meyrick), a leafminer that damages tomato leaves, terminal buds, flowers, and fruits, is a destructive tomato pest and is responsible for 80–100% of tomato yield losses globally. Different insect species have different courtship responses and phototropic flight rhythms. Improving the trapping effects of the sex pheromone and light traps is important for constructing an IPM system for T. absoluta. The present study explored the hourly distribution of T. absoluta adults caught by the sex pheromone (on the ground) and UV light (380 nm) traps in greenhouses over 24 h. The responses of males to sex pheromone (false female) lures were detected at dawn and early morning. The responses lasted for 3 h, from 05:30 (1 h before sunrise) to 08:30 (2 h after sunrise), and 95.8% of the males were caught during this period. The peak of the male responses to the sex pheromone was detected at 07:30 (from 06:30 to 07:30, 1 h after sunrise), and 80.8% of the males were caught during this period. The flight of male (proportion of 54.3%) and female (45.7%) adults toward the UV light traps occurred from 19:30 (time of sunset) to 06:30 (time of sunrise), lasted for 11 h, and exhibited a scotophase rhythm; 97.4% of the adults were caught during this period. The peak of adults flying toward the UV light traps occurred at 21:30 (from 20:30 to 21:30, 2 h after sunset). The rhythms of males’ responses to the sex pheromone and of the adults’ flight toward the UV lights can help to reveal the mechanisms of chemotactic and phototactic responses and may play a significant role in constructing an IPM system for this pest.

Colonization Mechanism of Endophytic Enterobacter cloacae TMX-6 on Rice Seedlings Mediated by Organic Acids Exudated from Roots

Small molecular organic acids (SMOAs) in root exudates are critical for plant-microbe interaction, especially under environmental stresses. However, the dominant organic acids driving the process and promoting the colonization are unclear. Here, using a target metabolomics, 20 main SMOAs of rice root exudates were identified and analyzed in control and 10 mg/L thiamethoxam-treated groups. The composition of these SMOAs differed significantly between the two treatments. Among which, malic acid, citric acid, succinic acid, and proline induced a chemotactic response, swimming ability, and biofilm formation of Enterobacter cloacae TMX-6 in a dose-dependent manner. The maximal chemotactic response of TMX-6 was induced by proline at 10 mg/L, and a strong chemotactic response was even observed at 0.01 mg/L. The recruitment assay confirmed that the addition of these four compounds promoted the colonization of TMX-6. The results provide insight for directional regulation of plant-microbe interactions for beneficial outcomes.

Chemoreceptors from the commensal gut Roseburia rectibacter bind to mucin and trigger chemotaxis.

Chemotaxis is crucial for bacterial adherence and colonization of the host gastrointestinal tract. Previous studies have demonstrated that chemotaxis affects the virulence of causative pathogens and the infection in the host. However, the chemotactic abilities of non-pathogenic and commensal gut bacteria have rarely been explored. We observed that Roseburia rectibacter NSJ-69 exhibited flagella-dependent motility and chemotaxis to a variety of molecules, including mucin and propionate. A genome-wide analysis revealed that NSJ-69 has 28 putative chemoreceptors, 15 of which have periplasmic ligand-binding domains (LBDs). These LBD-coding genes were chemically synthesized and expressed heterologously in Escherichia coli. Intensive screening of ligands revealed four chemoreceptors bound to mucin and two bound to propionate. When expressed in Comamonas testosteroni or E. coli, these chemoreceptors elicited chemotaxis toward mucin and propionate. Hybrid chemoreceptors were constructed, and results showed that the chemotactic responses to mucin and propionate were dependent on the LBDs of R. rectibacter chemoreceptors. Our study identified and characterized R. rectibacter chemoreceptors. These results will facilitate further investigations on the involvement of microbial chemotaxis in host colonization.

Resistance to Human Immunodeficiency Virus 1 Infection Conferred by a Compound CCR5Δ32 and CCR5 C20S Heterozygote.

We analyzed findings in a same-gender couple discordant in their human immunodeficiency virus (HIV) status. The HIV+ partner was homozygous for CCR5 while his receptive HIV- partner was a CCR5Δ32 heterozygote with a C20S missense mutation in his CCR5 allele. The cells from the HIV- partner showed significant resistance to R5 fusion/infection and had no chemotactic response to CCL4 (macrophage inflammatory protein 1β). We demonstrated abundant CCR5-specific RNA in the HIV- partner's cells but no detectable CCR5 protein. CCR5 promoter region cloned from each partner's DNA indicated no significant impact on RNA transcription. The compound effect of CCR5Δ32 and C20S mutation impaired CCR5 coreceptor function and conferred resistance to HIV-1.

2-Arachidonoylglycerol Modulates CXCL12-Mediated Chemotaxis in Mantle Cell Lymphoma and Chronic Lymphocytic Leukemia

To survive chemotherapy, lymphoma cells can relocate to protective niches where they receive support from the non-malignant cells. The biolipid 2-arachidonoylglycerol (2-AG), an agonist for the cannabinoid receptors CB1 and CB2, is released by stromal cells in the bone marrow. To investigate the role of 2-AG in lymphoma, we analyzed the chemotactic response of primary B-cell lymphoma cells enriched from peripheral blood of twenty-two chronic lymphocytic leukemia (CLL) and five mantle cell lymphoma (MCL) patients towards 2-AG alone and/or to the chemokine CXCL12. The expression of cannabinoid receptors was quantified using qPCR and the protein levels visualized by immunofluorescence and Western blot. Surface expression of CXCR4, the main cognate receptor to CXCL12, was analyzed by flow cytometry. Phosphorylation of key downstream signaling pathways activated by 2-AG and CXCL12 were measured by Western blot in three MCL cell lines and two primary CLL samples. We report that 2-AG induces chemotaxis in 80% of the primary samples, as well as 2/3 MCL cell lines. 2-AG induced in a dose-dependent manner, the migration of JeKo-1 cell line via CB1 and CB2. 2-AG affected the CXCL12-mediated chemotaxis without impacting the expression or internalization of CXCR4. We further show that 2-AG modulated p38 and p44/42 MAPK activation. Our results suggest that 2-AG has a previously unrecognized role in the mobilization of lymphoma cells by effecting the CXCL12-induced migration and the CXCR4 signaling pathways, however, with different effects in MCL compared to CLL.

Kinetic Asymmetry versus Dissipation in the Evolution of Chemical Systems as Exemplified by Single Enzyme Chemotaxis.

Single enzyme chemotaxis is a phenomenon by which a nonequilibrium spatial distribution of an enzyme is created and maintained by concentration gradients of the substrate and product of the catalyzed reaction. These gradients can arise either naturally through metabolism or experimentally, e.g., by flow of materials through microfluidic channels or by use of diffusion chambers with semipermeable membranes. Numerous hypotheses regarding the mechanism of this phenomenon have been proposed. Here, we discuss a mechanism based solely on diffusion and chemical reaction and show that kinetic asymmetry, a difference in the transition state energies for dissociation/association of substrate and product, and diffusion asymmetry, a difference in the diffusivities of the bound and free forms of the enzyme, are the determinates of the direction of chemotaxis and can result in either positive or negative chemotaxis, both of which have been demonstrated experimentally. Exploration of these fundamental symmetries that govern nonequilibrium behavior helps to distinguish between possible mechanisms for the evolution of a chemical system from initial to the steady state and whether the principle that determines the direction a system shifts when exposed to an external energy source is based on thermodynamics or on kinetics with the latter being supported by the results of the present paper. Our results show that, while dissipation ineluctably accompanies nonequilibrium phenomena, including chemotaxis, systems do not evolve to maximize or minimize dissipation but rather to attain greater kinetic stability and accumulate in regions where their effective diffusion coefficient is as small as possible. The chemotactic response to the chemical gradients formed by other enzymes participating in a catalytic cascade provides a mechanism for forming loose associations known as metabolons. Significantly, the direction of the effective force due to these gradients depends on the kinetic asymmetry of the enzyme and so can be nonreciprocal, where one enzyme is attracted to another enzyme, but the other enzyme is repelled by the one, in seeming contradiction to Newtons third law. This nonreciprocity is an important ingredient in the behavior of active matter.

Watermelon Root Exudates Enhance Root Colonization of Bacillus amyloliquefaciens TR2.

Bacillus amyloliquefaciens TR2, one of plant growth-promoting rhizobacteria (PGPR), is capable of colonizing plant roots in a large population size. However, the interaction of watermelon root exudates and colonization of the strain TR2 has not yet been clearly elucidated. In this investigation, we demonstrated that B. amyloliquefaciens TR2 promoted watermelon plants growth and exhibited biocontrol efficacy against watermelon Fusarium wilt under greenhouse conditions. Collected watermelon root exudates significantly induced chemotaxis, swarming motility, and biofilm formation of the strain TR2. We also tested the components of root exudates (organic acids: malic acid, citric acid, succinic acid, and fumaric acid; amino acids: methionine, glutamic acid, alanine, and aspartic acid; phenolic acid: benzoic acid) and the results showed that a majority of these compounds could promote chemotactic response, swarming motility, and biofilm formation in a different degree. Benzoic acid induced the strongest chemotactic response; however, the swarming motility and biofilm formation of the strain TR2 were maximumly enhanced by supplement of fumaric acid and glutamic acid, respectively. In addition, the root colonization examination indicated that the population of B. amyloliquefaciens TR2 colonized on watermelon root surfaces was dramatically increased by adding concentrated watermelon root exudates. In summary, our studies provide evidence suggesting that root exudates are important for colonization of B. amyloliquefaciens TR2 on plant roots and help us to understand the interaction between plants and beneficial bacteria.

Oscillatory ERK Signaling and Morphology Determine Heterogeneity of Breast Cancer Cell Chemotaxis via MEK-ERK and p38-MAPK Signaling Pathways.

Chemotaxis, regulated by oscillatory signals, drives critical processes in cancer metastasis. Crucial chemoattractant molecules in breast cancer, CXCL12 and EGF, drive the activation of ERK and Akt. Regulated by feedback and crosstalk mechanisms, oscillatory signals in ERK and Akt control resultant changes in cell morphology and chemotaxis. While commonly studied at the population scale, metastasis arises from small numbers of cells that successfully disseminate, underscoring the need to analyze processes that cancer cells use to connect oscillatory signaling to chemotaxis at single-cell resolution. Furthermore, little is known about how to successfully target fast-migrating cells to block metastasis. We investigated to what extent oscillatory networks in single cells associate with heterogeneous chemotactic responses and how targeted inhibitors block signaling processes in chemotaxis. We integrated live, single-cell imaging with time-dependent data processing to discover oscillatory signal processes defining heterogeneous chemotactic responses. We identified that short ERK and Akt waves, regulated by MEK-ERK and p38-MAPK signaling pathways, determine the heterogeneous random migration of cancer cells. By comparison, long ERK waves and the morphological changes regulated by MEK-ERK signaling, determine heterogeneous directed motion. This study indicates that treatments against chemotaxis in consider must interrupt oscillatory signaling.

The Two Chemotaxis Gene Clusters of Ensifer alkalisoli YIC4027T, a Symbiont of Sesbania cannabina, Play Different Roles in Chemotaxis and Competitive Nodulation

Ensifer alkalisoli YIC4027T is a dominant rhizobium that has been isolated from the root nodules of Sesbania cannabina. Motility and chemotaxis are critical to maintaining competitiveness in establishing the symbiotic relationship. E. alkalisoli carries two gene clusters, che1 and che2, containing chemotaxis-related gene homologues. To determine the respective role of each gene cluster, we constructed mutants and compared them with the wild type in a free-living state and in symbiosis with the host plant. A swimming analysis revealed that the che1 cluster was the major pathway controlling the chemotaxis and swimming bias, while the che2 cluster had a minor role in these behaviors. However, the Δche2 mutant was impaired in exopolysaccharide (EPS) production. During symbiosis, the Δche1 mutant was more severely impaired in its competitive root colonization and nodulation ability than the Δche2 mutant. Taken together, our data strongly suggested that both of the che clusters contribute to the competitive symbiotic association, the che1-like homologue being the main regulator of the chemotactic response and the che2 cluster regulating EPS production. These data illustrated a novel strategy of motile rhizobia bacteria to utilize the two pathways containing the homologous genes to enhance the efficiency of nodule formation by regulating distinct motility parameters or other cellular functions.

The Effects of Pregnancy on the Pulmonary Immune Response in a Mouse Model of LPS-Induced Acute Lung Injury.

This study evaluated the effect of pregnancy on the pulmonary innate immune response in a mouse model of acute lung injury (ALI) using nebulized lipopolysaccharide (LPS). Pregnant (day 14) C57BL/6NCRL mice and nonpregnant controls received nebulized LPS for 15 minutes. Twenty-four hours later, mice were euthanized for tissue harvest. Analysis included blood and bronchoalveolar lavage fluid (BALF) differential cell counts, whole-lung inflammatory cytokine transcription levels by reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR), and whole-lung vascular cell adhesion molecule 1 (VCAM-1), intercellular adhesion molecule 1 (ICAM-1), and BALF albumin by western blot. Mature bone marrow neutrophils from uninjured pregnant and nonpregnant mice were examined for chemotactic response using a Boyden chamber and for cytokine response to LPS by RT-qPCR. In LPS-induced ALI, pregnant mice had higher BALF total cell (p < 0.001) and neutrophil counts (p < 0.001) as well as higher peripheral blood neutrophils (p < 0.01) than nonpregnant mice, but a similar increase (as compared with unexposed mice) in airspace albumin levels. Whole-lung expression of interleukin 6, tumor necrosis factor-α (TNF-α), and keratinocyte chemoattractant (CXCL1) was also similar. In vitro, marrow-derived neutrophils from pregnant and nonpregnant mice had similar chemotaxis to CXCL1 and N-formylmethionine-leucyl-phenylalanine, but neutrophils from pregnant mice expressed lower levels of TNF (p < 0.001) and CXCL1 (p < 0.01) after LPS stimulation. In uninjured mice, VCAM-1 was higher in lungs from pregnant versus nonpregnant mice (p < 0.05). In this model, pregnancy is associated with an augmented lung neutrophil response to ALI without increased capillary leak or whole-lung cytokine levels relative to the nonpregnant state. This may stem from increased peripheral blood neutrophil response and intrinsically increased expression of pulmonary vascular endothelial adhesion molecules. Differences in lung innate cell homeostasis may affect the response to inflammatory stimuli and explain severe lung disease in respiratory infection during pregnancy. · Inhalation of LPS in midgestation versus virgin mice is associated with increased neutrophilia.. · This occurs without a comparative increase in cytokine expression.. · This may be explained by pregnancy-enhanced pre-exposure expression of VCAM-1 and ICAM-1..