scholarly journals Phenolic and Non-Polar Fractions of Elaeagnus rhamnoides (L.) A. Nelson Extracts as Virulence- Modulators - In Vitro Study on Bacteria, Fungi and Epithelial Cells

2018 ◽  
Author(s):  
Barbara Różalska ◽  
Beata Sadowska ◽  
Jerzy Żuchowski ◽  
Marzena Więckowska-Szakiel ◽  
Aleksandra Budzyńska ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Artificial Saliva ◽  
Fibroblast Cell ◽  
Array Detector ◽  
Persistence Properties ◽  
Virulence Potential ◽  
Charged Aerosol ◽  
The Impact

Butanol extracts from leaves, twigs and fruits of Elaeagnus rhamnoides (L.) A. Nelson (sea buckthorn, SBT) were fractionated into phenolic and non-polar lipid components. Their chemical composition was analyzed using the Thermo Ultimate 3000RS chromatographic system, equipped with a diode array detector, a corona-charged aerosol detector, and coupled with a (Q-TOF) mass spectrometer. Assuming that an effect on natural microbiotaand host epithelial cells needs to be assessed, regardless of the purpose of using SBT formulations in vivo, the MIC/MBC/MFC of the fractions and reference phytocompounds were screened involving 17 species of Gram-positive and Gram-negative bacteria and Candida species. The impact of the fractions (subMIC) on the important in vivo persistence properties of S. aureus and C. albicans strains was evaluated. Tests for adhesion and biofilm formation on an abiotic surface and the surfaces conditioned with fibrinogen, collagen, plasma or artificial saliva showed the inhibitory activity of the fractions. The effects on FITC-labeled staphylococci adhesion to fibroblasts (HFF-1) and epithelial cells (Caco-2), and on fungal morphogenesis, indicated that SBT extracts have high anti-virulence potential. Cytotoxicity tests (MTT-reduction) on the standard fibroblast cell line showed variable biological safety of the fractions depending on their composition and concentration.

Cathepsin B promotes collagen biosynthesis driving Bronchiolitis Obliterans Syndrome

2020 ◽  
pp. 2001416
Author(s):  
Carmela Morrone ◽  
Natalia F. Smirnova ◽  
Aicha Jeridi ◽  
Nikolaus Kneidinger ◽  
Christine Hollauer ◽  
...  
Keyword(s):  
Lung Tissue ◽  
Cathepsin B ◽  
Bronchiolitis Obliterans ◽  
Major Complication ◽  
Fibroblast Cell ◽  
Bronchiolitis Obliterans Syndrome ◽  
Collagen Deposition ◽  
The Impact

Bronchiolitis obliterans syndrome (BOS) is a major complication after lung transplantation (LTx). BOS is characterised by massive peribronchial fibrosis, leading to air trapping induced pulmonary dysfunction. Cathepsin B, a lysosomal cysteine-protease, was shown to enforce fibrotic pathways in several diseases. However, the relevance of Cathepsin B in BOS progression has not yet been addressed. The aim of the study was to elucidate the function of Cathepsin B in BOS pathogenesis.We determined Cathepsin B levels in BAL fluid and lung tissue from healthy donors (HD) and BOS LTx patients. Furthermore, Cathepsin B activity was assessed via a FRET-based assay and protein expression was determined using Western blotting, ELISA, and immunostaining. To investigate the impact of Cathepsin B in the pathophysiology of BOS, we used an in-vivo orthotopic left-LTx mouse model. Mechanistic studies were performed in-vitro using macrophage and fibroblast cell lines.We found a significant increase of Cathepsin B activity in BALF and lung tissue from BOS patients, as well as in our murine model of lymphocytic bronchiolitis (LB). Moreover, Cathepsin B activity was associated with an increased biosynthesis of collagen, and negatively affected lung function. Interestingly, we observed that Cathepsin B was mainly expressed in macrophages that infiltrated areas characterised by a massive accumulation of collagen deposition. Mechanistically, macrophage-derived Cathepsin B contributed to TGF-β1-dependent activation of fibroblasts, and its inhibition reversed the phenotype.Infiltrating macrophages release active Cathepsin B promoting fibroblast-activation and subsequent collagen deposition, driving BOS. Cathepsin B represents a promising therapeutic target to prevent the progression of BOS.

scholarly journals The adaptive potential of a survival artist: characterization of thein vitrointeractions ofToxoplasma gondiitachyzoites with di-cationic compounds in human fibroblast cell cultures

Parasitology ◽  
2011 ◽  
Vol 139 (2) ◽  
pp. 208-220 ◽  
Author(s):  
CHRISTIAN KROPF ◽  
KARIM DEBACHE ◽  
CHRISTOPH RAMPA ◽  
FABIENNE BARNA ◽  
MICHELLE SCHORER ◽  
...  
Keyword(s):  
Drug Exposure ◽  
Environmental Changes ◽  
Negative Impact ◽  
Fibroblast Cell ◽  
Drug Candidate ◽  
Selective Toxicity ◽  
Cationic Compounds ◽  
The Impact

SUMMARYThe impact of di-cationic pentamidine-analogues againstToxoplama gondii(Rh- and Me49-background) was investigated. The 72 h-growth assays showed that the arylimidamide DB750 inhibited the proliferation of tachyzoites ofT. gondii RhandT. gondii Me49with an IC50of 0·11 and 0·13μm, respectively. Pre-incubation of fibroblast monolayers with 1μmDB750 for 12 h and subsequent culture in the absence of the drug also resulted in a pronounced inhibiton of parasite proliferation. However, upon 5–6 days of drug exposure,T. gondiitachyzoites adapted to the compound and resumed proliferation up to a concentration of 1·2μm. Out of a set of 32 di-cationic compounds screened forin vitroactivity againstT. gondii,the arylimidamide DB745, exhibiting an IC50of 0·03μmand favourable selective toxicity was chosen for further studies. DB745 also inhibited the proliferation of DB750-adaptedT. gondii(IC50=0·07μm). In contrast to DB750, DB745 also had a profound negative impact on extracellular non-adaptedT. gondiitachyzoites, but not on DB750-adaptedT. gondii. Adaptation ofT. gondiito DB745 (up to a concentration of 0·46μm) was much more difficult to achieve and feasible only over a period of 110 days. In cultures infected with DB750-adaptedT. gondiiseemingly intact parasites could occasionally be detected by TEM. This illustrates the astonishing capacity ofT. gondiitachyzoites to adapt to environmental changes, at least underin vitroconditions, and suggests that DB745 could be an interesting drug candidate for further assessments in appropriatein vivomodels.

scholarly journals Impact of Mating Type, Serotype, and Ploidy on the Virulence of Cryptococcus neoformans

2008 ◽  
Vol 76 (7) ◽  
pp. 2923-2938 ◽  
Author(s):  
Xiaorong Lin ◽  
Kirsten Nielsen ◽  
Sweta Patel ◽  
Joseph Heitman
Keyword(s):  
Cryptococcus Neoformans ◽  
Mating Type ◽  
Pathogenic Fungi ◽  
Mating Types ◽  
Type Locus ◽  
Virulence Potential ◽  
The Impact ◽  
Ad Hybrids

ABSTRACT Hybridization with polyploidization is a significant biological force driving evolution. The effect of combining two distinct genomes in one organism on the virulence potential of pathogenic fungi is not clear. Cryptococcus neoformans, the most common cause of fungal infection of the central nervous system, has a bipolar mating system with a and α mating types and occurs as A (haploid), D (haploid), and AD hybrid (mostly diploid) serotypes. Diploid AD hybrids are derived either from a-α mating or from unisexual mating between haploid cells. The precise contributions of increased ploidy, the effect of hybridization between serotypes A and D, and the combination of mating types to the virulence potential of AD hybrids have remained elusive. By using in vitro and in vivo characterization of laboratory-constructed isogenic diploids and AD hybrids with all possible mating type combinations in defined genetic backgrounds, we found that higher ploidy has a minor negative effect on virulence in a murine inhalation model of cryptococcosis. The presence of both mating types a and α in AD hybrids did not affect the virulence potential, irrespective of the serotype origin. Interestingly, AD hybrids with only one mating type behaved differently, with the virulence of αADα strains similar to that of other hybrids, while aADa hybrids displayed significantly lower virulence due to negative epistatic interactions between the Aa and Da alleles of the mating type locus. This study provides insights into the impact of ploidy, mating type, and serotype on virulence and the impact of hybridization on the fitness and virulence of a eukaryotic microbial pathogen.

scholarly journals Immune response and pathogen invasion at the choroid plexus in the onset of cerebral toxoplasmosis

2022 ◽  
Vol 19 (1) ◽  
Author(s):  
Caio Andreeta Figueiredo ◽  
Johannes Steffen ◽  
Lorena Morton ◽  
Sushmitha Arumugam ◽  
Oliver Liesenfeld ◽  
...  
Keyword(s):  
Immune Response ◽  
Endothelial Cells ◽  
Epithelial Cells ◽  
Choroid Plexus ◽  
The Body ◽  
Potential Contribution ◽  
Pathogen Invasion ◽  
The Impact

Abstract Background Toxoplasma gondii (T. gondii) is a highly successful parasite being able to cross all biological barriers of the body, finally reaching the central nervous system (CNS). Previous studies have highlighted the critical involvement of the blood–brain barrier (BBB) during T. gondii invasion and development of subsequent neuroinflammation. Still, the potential contribution of the choroid plexus (CP), the main structure forming the blood–cerebrospinal fluid (CSF) barrier (BCSFB) have not been addressed. Methods To investigate T. gondii invasion at the onset of neuroinflammation, the CP and brain microvessels (BMV) were isolated and analyzed for parasite burden. Additionally, immuno-stained brain sections and three-dimensional whole mount preparations were evaluated for parasite localization and morphological alterations. Activation of choroidal and brain endothelial cells were characterized by flow cytometry. To evaluate the impact of early immune responses on CP and BMV, expression levels of inflammatory mediators, tight junctions (TJ) and matrix metalloproteinases (MMPs) were quantified. Additionally, FITC-dextran was applied to determine infection-related changes in BCSFB permeability. Finally, the response of primary CP epithelial cells to T. gondii parasites was tested in vitro. Results Here we revealed that endothelial cells in the CP are initially infected by T. gondii, and become activated prior to BBB endothelial cells indicated by MHCII upregulation. Additionally, CP elicited early local immune response with upregulation of IFN-γ, TNF, IL-6, host-defence factors as well as swift expression of CXCL9 chemokine, when compared to the BMV. Consequently, we uncovered distinct TJ disturbances of claudins, associated with upregulation of MMP-8 and MMP-13 expression in infected CP in vivo, which was confirmed by in vitro infection of primary CP epithelial cells. Notably, we detected early barrier damage and functional loss by increased BCSFB permeability to FITC-dextran in vivo, which was extended over the infection course. Conclusions Altogether, our data reveal a close interaction between T. gondii infection at the CP and the impairment of the BCSFB function indicating that infection-related neuroinflammation is initiated in the CP.

scholarly journals Evaluating the Impact of Human Amnion Epithelial Cells on Angiogenesis

2016 ◽  
Vol 2016 ◽  
pp. 1-13 ◽  
Author(s):  
Dandan Zhu ◽  
Ruth Muljadi ◽  
Siow Teng Chan ◽  
Patricia Vosdoganes ◽  
Camden Lo ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Lung Injury ◽  
Gene Transcription ◽  
Lung Fibrosis ◽  
Human Amnion ◽  
The Impact ◽  
Underlying Pathology ◽  
Amnion Epithelial Cells

The effects of human amnion epithelial cells (hAECs) on angiogenesis remain controversial. It is yet unknown if the presence of inflammation and/or gestational age of hAEC donors have an impact on angiogenesis. In this study, we examined the differences between term and preterm hAECs on angiogenesisin vitroandin vivo. Conditioned media from term hAECs induced the formation of longer huVEC tubules on Matrigel. Both term and preterm hAECs expressedVEGFA,PDGFB, ANGPT1,andFOXC1, which significantly increased after TNFαand IFNγstimulation. In the presence of TNFαand IFNγ, coculture with term hAECs reduced gene transcription ofTie-2andFoxc1in huVECs, while coculture with preterm hAECs increased gene transcription ofPDGFRαandPDGFRβand reduced gene transcription ofFOXC1in huVECs.In vivoassessment of angiogenesis using vWF immunostaining revealed that hAEC treatment decreased angiogenesis in a bleomycin model of lung fibrosis but increased angiogenesis in a neonatal model of hyperoxia-induced lung injury. In summary, our findings suggested that the impact of hAECs on angiogenesis may be influenced by the presence of inflammation and underlying pathology.

scholarly journals Immune Response and Pathogen Invasion at the Choroid Plexus in the Onset of Cerebral Toxoplasmosis

2021 ◽  
Author(s):  
Caio Andreeta Figueiredo ◽  
JOhannes Steffen ◽  
Lorena Morton ◽  
Sushmita Arumugam ◽  
OLiver Liesenfeld ◽  
...  
Keyword(s):  
Immune Response ◽  
Endothelial Cells ◽  
Epithelial Cells ◽  
Choroid Plexus ◽  
The Body ◽  
Potential Contribution ◽  
Pathogen Invasion ◽  
The Impact

Abstract Background: Toxoplasma gondii ( T. gondii ) is a highly successful parasite being able to cross all biological barriers of the body, finally reaching the central nervous system (CNS). Previous studies have highlighted the critical involvement of the blood-brain barrier (BBB) during T. gondii invasion and development of subsequent neuroinflammation. Still, the potential contribution of the choroid plexus (CP), a main structure forming the blood-cerebrospinal fluid (CSF)-barrier (BCSFB) have not been addressed. Methods: To investigate T. gondii invasion and the onset of neuroinflammation, the CP and brain microvessels (BMV) were isolated and analysed for parasite burden. Additionally, immuno-stained brain sections and three dimensional whole mount preparations were evaluated for parasite localization and morphological alterations. Activation of choroidal and brain endothelial cells were characterized by flow cytometry. To evaluate the impact of early immune responses on CP and BMV, expression levels of inflammatory mediators, tight junctions (TJ) and matrix metalloproteinases (MMPs) were quantified. Additionally, FITC-dextran was applied to determine infection-related changes in BCSFB permeability. Finally, the response of primary CP epithelial cells to T. gondii parasites was tested in vitro . Results: Here we revealed that endothelial cells in the CP are initially infected by T. gondii, and become activated prior to BBB endothelial cells indicated by MHCII upregulation. Additionally, CP elicited early local immune response with upregulation of IFN-γ, TNF, IL-6, host-defence factors as well as swift expression of CXCL9 chemokine, when compared to the BMV. Consequently, we uncovered distinct TJ disturbances of claudins, associated with upregulation of MMP-8 and MMP-13 expression in infected CP in vivo , which was confirmed by in vitro infection of primary CP epithelial cells. Notably, we detected early barrier damage and functional loss by increased BCSFB permeability to FITC-dextran in vivo , which was extended over the infection course. Conclusions: Altogether, our data reveal a close interaction between T. gondii infection at the CP and the impairment of the BCSFB function indicating that infection-related neuroinflammation is initiated in the CP.

The impact of Zataria multiflora Boiss extract on in vitro and in vivo Th1/Th2 cytokine (IFN-γ/IL4) balance

2013 ◽  
Vol 150 (3) ◽  
pp. 1024-1031 ◽  
Author(s):  
Mohammad Hossein Boskabady ◽  
Sakine Shahmohammadi Mehrjardi ◽  
Abadorrahim Rezaee ◽  
Houshang Rafatpanah ◽  
Sediqeh Jalali
Keyword(s):  
Zataria Multiflora ◽  
Th2 Cytokine ◽  
Ifn Γ ◽  
The Impact

scholarly journals The impact of FGF19/FGFR4 signaling inhibition in antitumor activity of multi-kinase inhibitors in hepatocellular carcinoma

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Hiroaki Kanzaki ◽  
Tetsuhiro Chiba ◽  
Junjie Ao ◽  
Keisuke Koroki ◽  
Kengo Kanayama ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Molecular Mechanisms ◽  
Kinase Inhibitors ◽  
Progression Free Survival ◽  
Erk Signaling ◽  
Enzyme Linked Immunosorbent Assay ◽  
Antitumor Effects ◽  
The Impact

AbstractFGF19/FGFR4 autocrine signaling is one of the main targets for multi-kinase inhibitors (MKIs). However, the molecular mechanisms underlying FGF19/FGFR4 signaling in the antitumor effects to MKIs in hepatocellular carcinoma (HCC) remain unclear. In this study, the impact of FGFR4/ERK signaling inhibition on HCC following MKI treatment was analyzed in vitro and in vivo assays. Serum FGF19 in HCC patients treated using MKIs, such as sorafenib (n = 173) and lenvatinib (n = 40), was measured by enzyme-linked immunosorbent assay. Lenvatinib strongly inhibited the phosphorylation of FRS2 and ERK, the downstream signaling molecules of FGFR4, compared with sorafenib and regorafenib. Additional use of a selective FGFR4 inhibitor with sorafenib further suppressed FGFR4/ERK signaling and synergistically inhibited HCC cell growth in culture and xenograft subcutaneous tumors. Although serum FGF19high (n = 68) patients treated using sorafenib exhibited a significantly shorter progression-free survival and overall survival than FGF19low (n = 105) patients, there were no significant differences between FGF19high (n = 21) and FGF19low (n = 19) patients treated using lenvatinib. In conclusion, robust inhibition of FGF19/FGFR4 is of importance for the exertion of antitumor effects of MKIs. Serum FGF19 levels may function as a predictive marker for drug response and survival in HCC patients treated using sorafenib.

Three-dimensional Growth of Renal Epithelial Cells in Vitro: A Tool in Toxicity Testing

1993 ◽  
Vol 21 (2) ◽  
pp. 191-195 ◽  
Author(s):  
Knut-Jan Andersen ◽  
Erik Ilsø Christensen ◽  
Hogne Vik
Keyword(s):  
Epithelial Cells ◽  
Three Dimensional ◽  
Toxicity Testing ◽  
Renal Tissue ◽  
Epithelial Cell Line ◽  
Multicellular Spheroids ◽  
Renal Epithelial Cell ◽  
Renal Epithelial Cells

The tissue culture of multicellular spheroids from the renal epithelial cell line LLC-PK1 (proximal tubule) is described. This represents a biological system of intermediate complexity between renal tissue in vivo and simple monolayer cultures. The multicellular structures, which show many similarities to kidney tubules in vivo, including a vectorial water transport, should prove useful for studying the potential nephrotoxicity of drugs and chemicals in vitro. In addition, the propagation of renal epithelial cells as multicellular spheroids in serum-free culture may provide information on the release of specific biological parameters, which may be suppressed or masked in serum-supplemented media.

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