Proinflammatory Microenvironment During Kingella kingae Infection Modulates Osteoclastogenesis

Kingella kingae is an emerging pathogen that causes septic arthritis, osteomyelitis, and bacteremia in children from 6 to 48 months of age. The presence of bacteria within or near the bone is associated with an inflammatory process that results in osteolysis, but the underlying pathogenic mechanisms involved are largely unknown. To determine the link between K. kingae and bone loss, we have assessed whether infection per se or through the genesis of a pro-inflammatory microenvironment can promote osteoclastogenesis. For that purpose, we examined both the direct effect of K. kingae and the immune-mediated mechanism involved in K. kingae-infected macrophage-induced osteoclastogenesis. Our results indicate that osteoclastogenesis is stimulated by K. kingae infection directly and indirectly by fueling a potent pro-inflammatory response that drives macrophages to undergo functional osteoclasts via TNF-α and IL-1β induction. Such osteoclastogenic capability of K. kingae is counteracted by their outer membrane vesicles (OMV) in a concentration-dependent manner. In conclusion, this model allowed elucidating the interplay between the K. kingae and their OMV to modulate osteoclastogenesis from exposed macrophages, thus contributing to the modulation in joint and bone damage.

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Brucella abortus Proliferates in Decidualized and Non-Decidualized Human Endometrial Cells Inducing a Proinflammatory Response

Pathogens ◽

10.3390/pathogens9050369 ◽

2020 ◽

Vol 9 (5) ◽

pp. 369

Author(s):

Lucía Zavattieri ◽

Mariana C. Ferrero ◽

Iván M. Alonso Paiva ◽

Agustina D. Sotelo ◽

Andrea M. Canellada ◽

...

Keyword(s):

Brucella Abortus ◽

Membrane Vesicles ◽

Outer Membrane Vesicles ◽

Dependent Manner ◽

Proinflammatory Response ◽

Endometrial Stromal Cells ◽

Endometrial Cells ◽

Tnf Α ◽

Proinflammatory Responses ◽

Intracellular Proliferation

Brucella spp. have been associated with abortion in humans and animals. Although the mechanisms involved are not well established, it is known that placental Brucella infection is accompanied by inflammatory phenomena. The ability of Brucella abortus to infect and survive in human endometrial stromal cells (T-HESC cell line) and the cytokine response elicited were evaluated. B. abortus was able to infect and proliferate in both non-decidualized and decidualized T-HESC cells. Intracellular proliferation depended on the expression of a functional virB operon in the pathogen. B. abortus internalization was inhibited by cytochalasin D and to a lower extent by colchicine, but was not affected by monodansylcadaverine. The infection did not induce cytotoxicity and did not alter the decidualization status of cells. B. abortus infection elicited the secretion of IL-8 and MCP-1 in either decidualized or non-decidualized T-HESC, a response also induced by heat-killed B. abortus and outer membrane vesicles derived from this bacterium. The stimulation of T-HESC with conditioned media from Brucella-infected macrophages induced the production of IL-6, MCP-1 and IL-8 in a dose-dependent manner, and this effect was shown to depend on IL-1β and TNF-α. The proinflammatory responses of T-HESC to B. abortus and to factors produced by infected macrophages may contribute to the gestational complications of brucellosis.

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Bioactivity-Guided Extract Optimization of Osmanthus fragrans var. aurantiacus Leaves and Anti-Inflammatory Activities of Phillyrin

Plants ◽

10.3390/plants10081545 ◽

2021 ◽

Vol 10 (8) ◽

pp. 1545

Author(s):

Hwa-Young Song ◽

Da-Eun Jeong ◽

Mina Lee

Keyword(s):

Biological Activities ◽

Radical Scavenging ◽

No Production ◽

Dependent Manner ◽

Optimal Method ◽

Concentration Dependent Manner ◽

Osmanthus Fragrans ◽

Tnf Α ◽

Anti Inflammatory ◽

Extracellular Regulated Kinase

The aim of this study was to identify the optimal extraction conditions for leaves of Osmanthus fragrans var. aurantiacus. Inhibitory effects of various extracts on NO production were compared. Antioxidant evaluations for total phenol and flavonoid contents were carried out using various extracts of O. fragrans var. aurantiacus leaves obtained under optimal extraction conditions that showed the greatest effect on NO production. The optimal method for extracting O. fragrans var. aurantiacus leaves resulted in an extract named OP OFLE. OP OFLE showed DPPH and ABTS radical scavenging activities in a concentration-dependent manner. Phillyrin (PH) was isolated as a major compound from OP OFLE by HPLC/DAD analysis. OP OFLE and PH reduced inducible nitric oxide (iNOS) and cyclooxygenase (COX)-2 protein expression and downregulated proinflammatory cytokines such as interleukin (IL)-1β, IL-6, IL-8, and tumor necrosis factor (TNF)-α in LPS-stimulated RAW 264.7 and HT-29 cells. To determine the signal pathway involved in the inhibition of NO production, a Western blot analysis was performed. Results showed that OP OFLE decreased phosphorylation of extracellular regulated kinase (pERK) 1/2 and the expression of nuclear factor-kappa B (NF-κB). Our results suggest that extracts of O. fragrans var. aurantiacus leaves and its major components have biological activities such as antioxidative and anti-inflammatory properties.

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Effects of irbesartan on phenotypic alterations in monocytes and the inflammatory status of hypertensive patients with left ventricular hypertrophy

BMC Cardiovascular Disorders ◽

10.1186/s12872-021-02004-7 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Jingsi Zhang ◽

Lina Yang ◽

Yanchun Ding

Keyword(s):

Ventricular Hypertrophy ◽

Target Organ Damage ◽

Organ Damage ◽

Left Ventricular ◽

Target Organ ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

Hypertensive Patients ◽

Inflammatory Status ◽

Tnf Α

Abstract Background Circulating monocytes and tissue macrophages play complex roles in the pathogenesis of hypertension and the resulting target organ damage. In this study, we observed alterations in the monocyte phenotype and inflammatory state of hypertensive patients with left ventricular hypertrophy (LVH) and studied the effects of irbesartan in these patients. This study might reveal a novel mechanism by which irbesartan alleviates LVH, and it could provide new targets for the prevention and treatment of hypertensive target organ damage. Methods CD163 and CD206 expression on monocytes and IL-10 and TNF-α levels in the serum of hypertensive patients with or without LVH and of healthy volunteers were detected. Furthermore, we treated monocytes from the LVH group with different concentrations of irbesartan, and then, CD163, CD206, IL-10 and TNF-α expression was detected. Results We found, for the first time, that the expression of CD163, CD206 and IL-10 in the LVH group was lower than that in the non-LVH group and healthy control group, but the TNF-α level in the LVH group was significantly higher. Irbesartan upregulated the expression of CD163 and CD206 in hypertensive patients with LVH in a concentration-dependent manner. Irbesartan also increased the expression of IL-10 and inhibited the expression of TNF-α in monocyte culture supernatants in a concentration-dependent manner. Conclusions Our data suggest that inflammation was activated in hypertensive patients with LVH and that the monocyte phenotype was mainly proinflammatory. The expression of proinflammatory factors increased while the expression of anti-inflammatory factors decreased. Irbesartan could alter the monocyte phenotype and inflammatory status in hypertensive patients with LVH. This previously unknown mechanism may explain how irbesartan alleviates LVH. Trail registration The study protocols were approved by the Ethical Committee of the Second Affiliated Hospital of Dalian Medical University. Each patient signed the informed consent form.

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Modulatory Effects of Osthole on Lipopolysaccharides-Induced Inflammation in Caco-2 Cell Monolayer and Co-Cultures with THP-1 and THP-1-Derived Macrophages

Nutrients ◽

10.3390/nu13010123 ◽

2020 ◽

Vol 13 (1) ◽

pp. 123

Author(s):

Natalia K. Kordulewska ◽

Justyna Topa ◽

Małgorzata Tańska ◽

Anna Cieślińska ◽

Ewa Fiedorowicz ◽

...

Keyword(s):

Gene Expression ◽

Inflammatory Reaction ◽

Wide Spectrum ◽

Cell Monolayer ◽

In Vivo Studies ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

Tnf Α

Lipopolysaccharydes (LPS) are responsible for the intestinal inflammatory reaction, as they may disrupt tight junctions and induce cytokines (CKs) secretion. Osthole has a wide spectrum of pharmacological effects, thus its anti-inflammatory potential in the LPS-treated Caco-2 cell line as well as in Caco-2/THP-1 and Caco-2/macrophages co-cultures was investigated. In brief, Caco-2 cells and co-cultures were incubated with LPS to induce an inflammatory reaction, after which osthole (150–450 ng/mL) was applied to reduce this effect. After 24 h, the level of secreted CKs and changes in gene expression were examined. LPS significantly increased the levels of IL-1β, -6, -8, and TNF-α, while osthole reduced this effect in a concentration-dependent manner, with the most significant decrease when a 450 ng/mL dose was applied (p < 0.0001). A similar trend was observed in changes in gene expression, with the significant osthole efficiency at a concentration of 450 ng/μL for IL1R1 and COX-2 (p < 0.01) and 300 ng/μL for NF-κB (p < 0.001). Osthole increased Caco-2 monolayer permeability, thus if it would ever be considered as a potential drug for minimizing intestinal inflammatory symptoms, its safety should be confirmed in extended in vitro and in vivo studies.

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Cell surface interactions in conjugation: Tetrahymena ciliary membrane vesicles

Molecular and Cellular Biology ◽

10.1128/mcb.4.4.681-687.1984 ◽

1984 ◽

Vol 4 (4) ◽

pp. 681-687

Author(s):

B Love ◽

M B Rotheim

Keyword(s):

Cell Surface ◽

Mating Type ◽

Mammalian Cells ◽

Membrane Vesicles ◽

Surface Interactions ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

Ciliary Membrane ◽

Adhesion Sites ◽

Cell Surface Interactions

Tetrahymena ciliary membrane vesicles are shown to interact with preconjugant cells in a mating type-specific way. When cells are treated with vesicles of a different mating type before mixing for conjugation, cell pairing is enhanced, and the normal prepairing period is partially eliminated. This enhancement is mating type specific since it is not observed after pretreatment of cells with vesicles of their own mating type. In contrast, when vesicles are added at the time of mixing of two starved cultures, cell pairing is delayed in a concentration-dependent manner. By varying the conditions, we demonstrated enhancement or inhibition, or both. These results are interpreted in terms of two independent interactions of cells with vesicles. We suggest that first, vesicles substitute for another cell in cell-cell prepairing interaction and second, vesicles compete for adhesion sites produced during the prepairing period. Finally, the data presented are summarized within a speculative framework that calls attention to potential analogies with hormone-receptor signaling in mammalian cells.

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Interactions of Dichlorodiphenyltrichloroethane (DDT) and Dichlorodiphenyldichloroethylene (DDE) With Skeletal Muscle Ryanodine Receptor Type 1

Toxicological Sciences ◽

10.1093/toxsci/kfz120 ◽

2019 ◽

Vol 170 (2) ◽

pp. 509-524

Author(s):

Kim M Truong ◽

Gennady Cherednichenko ◽

Isaac N Pessah

Keyword(s):

Skeletal Muscle ◽

Ryanodine Receptor ◽

Membrane Vesicles ◽

Receptor Type ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

Voltage Gated Sodium Channels ◽

Muscle Health ◽

Living Organisms

Abstract Dichlorodiphenyltrichloroethane (DDT) and its metabolite dichlorodiphenyldichloroethylene (DDE) are ubiquitous in the environment and detected in tissues of living organisms. Although DDT owes its insecticidal activity to impeding closure of voltage-gated sodium channels, it mediates toxicity in mammals by acting as an endocrine disruptor (ED). Numerous studies demonstrate DDT/DDE to be EDs, but studies examining muscle-specific effects mediated by nonhormonal receptors in mammals are lacking. Therefore, we investigated whether o,p′-DDT, p,p′-DDT, o,p′-DDE, and p,p′-DDE (DDx, collectively) alter the function of ryanodine receptor type 1 (RyR1), a protein critical for skeletal muscle excitation-contraction coupling and muscle health. DDx (0.01–10 µM) elicited concentration-dependent increases in [3H]ryanodine ([3H]Ry) binding to RyR1 with o,p′-DDE showing highest potency and efficacy. DDx also showed sex differences in [3H]Ry-binding efficacy toward RyR1, where [3H]Ry-binding in female muscle preparations was greater than male counterparts. Measurements of Ca2+ transport across sarcoplasmic reticulum (SR) membrane vesicles further confirmed DDx can selectively engage with RyR1 to cause Ca2+ efflux from SR stores. DDx also disrupts RyR1-signaling in HEK293T cells stably expressing RyR1 (HEK-RyR1). Pretreatment with DDx (0.1–10 µM) for 100 s, 12 h, or 24 h significantly sensitized Ca2+-efflux triggered by RyR agonist caffeine in a concentration-dependent manner. o,p′-DDE (24 h; 1 µM) significantly increased Ca2+-transient amplitude from electrically stimulated mouse myotubes compared with control and displayed abnormal fatigability. In conclusion, our study demonstrates DDx can directly interact and modulate RyR1 conformation, thereby altering SR Ca2+-dynamics and sensitize RyR1-expressing cells to RyR1 activators, which may ultimately contribute to long-term impairments in muscle health.

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Naturally Occurring IgG Antibodies Provide Innate Protection against Vibrio cholerae Bacteremia by Recognition of the Outer Membrane Protein U

Journal of Innate Immunity ◽

10.1159/000443646 ◽

2016 ◽

Vol 8 (3) ◽

pp. 269-283 ◽

Cited By ~ 17

Author(s):

Kyaw Min Aung ◽

Annika E. Sjöström ◽

Ulrich von Pawel-Rammingen ◽

Kristian Riesbeck ◽

Bernt Eric Uhlin ◽

...

Keyword(s):

Membrane Protein ◽

Outer Membrane ◽

Vibrio Cholerae ◽

Outer Membrane Protein ◽

Membrane Vesicles ◽

Outer Membrane Vesicles ◽

Immune Defense ◽

Dependent Manner ◽

Naturally Occurring ◽

Serum Killing

Cholera epidemics are caused by Vibrio cholerae serogroups O1 and O139, whereas strains collectively known as non-O1/non-O139 V. cholerae are found in cases of extraintestinal infections and bacteremia. The mechanisms and factors influencing the occurrence of bacteremia and survival of V. cholerae in normal human serum have remained unclear. We found that naturally occurring IgG recognizing V. cholerae outer membrane protein U (OmpU) mediates a serum-killing effect in a complement C1q-dependent manner. Moreover, outer membrane vesicles (OMVs) containing OmpU caused enhanced survival of highly serum-sensitive classical V. cholerae in a dose-dependent manner. OMVs from wild-type and ompU mutant V. cholerae thereby provided a novel means to verify by extracellular transcomplementation the involvement of OmpU. Our data conclusively indicate that loss, or reduced expression, of OmpU imparts resistance to V. cholerae towards serum killing. We propose that the difference in OmpU protein levels is a plausible reason for differences in serum resistance and the ability to cause bacteremia observed among V. cholerae biotypes. Our findings provide a new perspective on how naturally occurring antibodies, perhaps induced by members of the microbiome, may play a role in the recognition of pathogens and the provocation of innate immune defense against bacteremia.

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Inhibition of Na+/Ca2+ exchange in renal BLMV by IP3 depends on site of action and direction of Ca2+ flux

AJP Renal Physiology ◽

10.1152/ajprenal.1994.266.5.f785 ◽

1994 ◽

Vol 266 (5) ◽

pp. F785-F790 ◽

Cited By ~ 1

Author(s):

C. L. Fraser ◽

C. Cummings ◽

G. Cassafer

Keyword(s):

Plasma Membrane ◽

Basolateral Membrane ◽

Membrane Vesicles ◽

Dependent Manner ◽

The Novel ◽

Concentration Dependent Manner ◽

Site Of Action ◽

Brain Synaptosomes ◽

Membrane Bound ◽

Inside Out

It has previously been shown in synaptosomes that inositol 1,4,5-trisphosphate (1,4,5-IP3) inhibits Ca2+ transport by the plasma membrane-bound Na+/Ca2+ exchanger. The present study was therefore designed to determine if the effect of 1,4,5-IP3 was dependent on its site of action at the plasma membrane or on the direction of Ca2+ flux. To investigate this possibility, studies were performed in basolateral membrane vesicles (BLMV) isolated from rat renal cortex. As with synaptosomes, Ca2+ transport was inhibited by 1,4,5-IP3 in a concentration-dependent manner. At a concentration of 10(-6) M, 1,4,5-IP3 significantly (P < 0.005) inhibited Ca2+ transport by 36%. When Ca2+ transport was carried out in inside-out vesicles, 10(-6) M 1,4,5-IP3 significantly (P < 0.002) increased the degree of inhibition by an additional 75% (63 vs. 36%). However, 1,4,5-IP3 had no significant effect on Ca2+ transport in inside-out vesicles when Ca2+ flux was reversed (i.e., Ca2+ efflux). These data in renal BLMV confirm the novel action of 1,4,5-IP3 on the Na+/Ca2+ exchanger previously described in brain synaptosomes. These results also suggest that the action of 1,4,5-IP3 depends on both its site of action at the plasma membrane and on the direction of Ca2+ flux.

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Increased Anti-Inflammatory Effects on LPS-Induced Microglia Cells by Spirulina maxima Extract from Ultrasonic Process

Applied Sciences ◽

10.3390/app9102144 ◽

2019 ◽

Vol 9 (10) ◽

pp. 2144 ◽

Cited By ~ 5

Author(s):

Woon Yong Choi ◽

Jae-Hun Sim ◽

Jung-Youl Lee ◽

Do Hyung Kang ◽

Hyeon Yong Lee

Keyword(s):

Mrna Expression ◽

Dependent Manner ◽

Spirulina Maxima ◽

Concentration Dependent Manner ◽

Quantitative Correlation ◽

Tnf Α ◽

Ultrasonic Process ◽

Anti Inflammatory ◽

Low Cytotoxicity ◽

High Concentrations

The Spirulina maxima exact from a non-thermal ultrasonic process (UE) contains 17.5 mg/g of total chlorophyll, compared to 6.24 mg/g of chlorophyll derived from the conventional 70% ethanol extraction at 80 °C for 12 h (EE). The UE also showed relatively low cytotoxicity against murine microglial cells (BV-2) and inhibited the production of the inflammatory mediators, NO and PGE2. The UE also effectively suppresses both mRNA expression and the production of pro-inflammatory cytokines, such as TNF-α, IL-6 and IL-1β, in a concentration-dependent manner. Notably, TNF-α gene and protein production were most strongly down-regulated, while IL-6 was the least affected by all ranges of treatment concentrations. This work first demonstrated a quantitative correlation between mRNA expression and the production of cytokines, showing that suppression of TNF-α gene expression was most significantly correlated with its secretion. These results clearly proved that the anti-inflammatory effects of Spirulina extract from a nonthermal ultrasonic process, which yielded high concentrations of intact forms of chlorophylls, were increased two-fold compared to those of conventional extracts processed at high temperature.

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Modulatory Effect of Chelidonium majus Extract and Its Alkaloids on LPS-Stimulated Cytokine Secretion in Human Neutrophils

Molecules ◽

10.3390/molecules25040842 ◽

2020 ◽

Vol 25 (4) ◽

pp. 842 ◽

Cited By ~ 1

Author(s):

Sylwia Zielińska ◽

Monika Ewa Czerwińska ◽

Magdalena Dziągwa-Becker ◽

Andrzej Dryś ◽

Mariusz Kucharski ◽

...

Keyword(s):

Biological Activities ◽

Cytokine Secretion ◽

Human Neutrophils ◽

Root Extract ◽

Dependent Manner ◽

Chelidonium Majus ◽

Concentration Dependent Manner ◽

Cytotoxic Effects ◽

Tnf Α ◽

High Cytotoxicity

Due to certain differences in terms of molecular structure, isoquinoline alkaloids from Chelidonium majus engage in various biological activities. Apart from their well-documented antimicrobial potential, some phenanthridine and protoberberine derivatives as well as C. majus extract present with anti-inflammatory and cytotoxic effects. In this study, the LC–MS/MS method was used to determine alkaloids, phenolic acids, carboxylic acids, and hydroxybenzoic acids. We investigated five individually tested alkaloids (coptisine, berberine, chelidonine, chelerythrine, and sanguinarine) as well as C. majus root extract for their effect on the secretion of IL-1β, IL-8, and TNF-α in human polymorphonuclear leukocytes (neutrophils). Berberine, chelidonine, and chelerythrine significantly decreased the secretion of TNF-α in a concentration-dependent manner. Sanguinarine was found to be the most potent inhibitor of IL-1β secretion. However, the overproduction of IL-8 and TNF-α and a high cytotoxicity for these compounds were observed. Coptisine was highly cytotoxic and slightly decreased the secretion of the studied cytokines. The extract (1.25–12.5 μg/mL) increased cytokine secretion in a concentration-dependent manner, but an increase in cytotoxicity was also noted. The alkaloids were active at very low concentrations (0.625–2.5 μM), but their potential cytotoxic effects, except for chelidonine and chelerythrine, should not be ignored.

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