scholarly journals A Novel Standardized Cannabis sativa L. Extract and Its Constituent Cannabidiol Inhibit Human Polymorphonuclear Leukocyte Functions

2019 ◽  
Vol 20 (8) ◽  
pp. 1833 ◽  
Author(s):  
Alex Mabou Tagne ◽  
Franca Marino ◽  
Massimiliano Legnaro ◽  
Alessandra Luini ◽  
Barbara Pacchetti ◽  
...  
Keyword(s):  
Cell Migration ◽  
Oxidative Metabolism ◽  
Molecular Mechanisms ◽  
Cannabis Sativa ◽  
Biological Effects ◽  
Enzyme Linked Immunosorbent Assay ◽  
Boyden Chamber ◽  
Ros Production ◽  
Therapeutic Benefits ◽  
Tnf Α

Cannabis and cannabinoids offer significant therapeutic benefits for a wide scope of pathological conditions. Among them, the clinical issues rooted in inflammation stand out, nonetheless, the underlying mechanisms are not yet plainly understood. Circumstantial evidence points to polymorphonuclear leukocytes (PMN) as targets for the anti-inflammatory effects of cannabis. Therefore, we conducted this study to assess the effects of CM5, a novel Cannabis sativa L. extract standardized in 5% cannabidiol (CBD), on human PMN functions, including cell migration, oxidative metabolism and production of tumour necrosis factor (TNF)-α. We then sought to investigate whether such effects could be ascribed to its content in CBD. Cell migration was assessed by the Boyden chamber assay, oxidative metabolism by means of spectrofluorimetric measurement of reactive oxygen species (ROS) production, and TNF-α was measured by real time polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA). Results show that both CM5 and CBD inhibit PMN migration, ROS and TNF-α production, indicating that CBD may be the main item responsible for the effects of CM5. CM5 is however more potent than CBD on cell migration and TNF-α production, and less effective on ROS production, suggesting that beyond CBD, other components of the cannabis plant may contribute to the biological effects of the extract. As a whole, such results support the use of cannabis standardized extract and CBD to stem inflammation; however, they also warrant in-depth investigation of the underlying cellular and molecular mechanisms to better exploit their therapeutic potential.

scholarly journals TheBDKRB2 +9/-9 Polymorphisms Influence Pro-Inflammatory Cytokine Levels in Knee Osteoarthritis by Altering TLR-2 Expression: Clinical and in Vitro Studies

2016 ◽  
Vol 38 (3) ◽  
pp. 1245-1256 ◽  
Author(s):  
Shuo Chen ◽  
Lei Zhang ◽  
Ruonan Xu ◽  
Yunfan Ti ◽  
Yunlong Zhao ◽  
...  
Keyword(s):  
Knee Osteoarthritis ◽  
Inflammatory Cytokine ◽  
Molecular Mechanisms ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mrna Levels ◽  
Knee Oa ◽  
Tnf Α ◽  
Cytokine Levels ◽  
Underlying Mechanisms

Background/Aims: The bradykinin B2 receptor (BDKRB2) +9/-9 gene polymorphisms have been shown to be associated with the susceptibility and severity of osteoarthritis (OA); however, the underlying mechanisms are unclear. In this study, we investigated the correlation between the BDKRB2 +9/-9 polymorphisms and pro-inflammatory cytokine levels in OA and the molecular mechanisms involved. Methods: A total of 156 patients with primary knee OA and 121 healthy controls were enrolled. The BDKRB2 +9/-9 polymorphisms were genotyped. The tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, and IL-8 levels were determined using Enzyme-linked immunosorbent assay (ELISA). The toll-like receptor (TLR)-2 and TLR-4 mRNA levels were determined by quantitative real-time PCR. The basal and bradykinin-stimulated pro-inflammatory cytokine secretion in human OA synoviocytes and the involvement of TLR-2 and mitogen-activated protein kinases (MAPKs) were investigated. Results: The presence of -9 bp genotype is associated with higher TNF-α, IL-6, and IL-8 levels and higher TLR-2 expression in OA patients. The basal and bradykinin-induced TLR-2 expressions in human OA synoviocytes were significantly reduced by specific inhibitors of p38, JNK1/2, and ERK1/2. Both the B2 receptor antagonist MEN16132 and TLR-2 silencing inhibited IL-6 and IL-8 secretion in human OA synoviocytes. Conclusion: The data suggested that the BDKRB2 +9/-9 polymorphisms influence pro-inflammatory cytokine levels in knee osteoarthritis by altering TLR-2 expression.

scholarly journals Vitamin D Signaling in Inflammation and Cancer: Molecular Mechanisms and Therapeutic Implications

Molecules ◽  
2020 ◽  
Vol 25 (14) ◽  
pp. 3219 ◽  
Author(s):  
Ahmed El-Sharkawy ◽  
Ahmed Malki
Keyword(s):  
Vitamin D ◽  
Vitamin D Receptor ◽  
Anticancer Agents ◽  
Molecular Mechanisms ◽  
Biological Effects ◽  
Active Form ◽  
The Body ◽  
Skeletal Health ◽  
Therapeutic Benefits ◽  
Vitamin D Signaling

Vitamin D and its active metabolites are important nutrients for human skeletal health. UV irradiation of skin converts 7-dehydrocholesterol into vitamin D3, which metabolized in the liver and kidneys into its active form, 1α,25-dihydroxyvitamin D3. Apart from its classical role in calcium and phosphate regulation, scientists have shown that the vitamin D receptor is expressed in almost all tissues of the body, hence it has numerous biological effects. These includes fetal and adult homeostatic functions in development and differentiation of metabolic, epidermal, endocrine, neurological and immunological systems of the body. Moreover, the expression of vitamin D receptor in the majority of immune cells and the ability of these cells to actively metabolize 25(OH)D3 into its active form 1,25(OH)2D3 reinforces the important role of vitamin D signaling in maintaining a healthy immune system. In addition, several studies have showed that vitamin D has important regulatory roles of mechanisms controlling proliferation, differentiation and growth. The administration of vitamin D analogues or the active metabolite of vitamin D activates apoptotic pathways, has antiproliferative effects and inhibits angiogenesis. This review aims to provide an up-to-date overview on the effects of vitamin D and its receptor (VDR) in regulating inflammation, different cell death modalities and cancer. It also aims to investigate the possible therapeutic benefits of vitamin D and its analogues as anticancer agents.

scholarly journals Protective effects of 4, 5-O-dicaffeoylquinic acid against mouse sepsis via down-regulation of TNF-α, IL-6 and IL-8

2020 ◽  
Vol 19 (4) ◽  
pp. 715-720
Author(s):  
XiaoBo Wang ◽  
JianHua Wu ◽  
BuKao Ni
Keyword(s):  
Survival Rate ◽  
Mrna Expression ◽  
Molecular Mechanisms ◽  
Cecal Ligation ◽  
Enzyme Linked Immunosorbent Assay ◽  
Protective Effects ◽  
Dicaffeoylquinic Acid ◽  
Xanthium Sibiricum ◽  
Tnf Α

Purpose: To investigate the protective effects of 4, 5-O-dicaffeoylquinic acid (DCQA) isolated from Xanthium sibiricum Patr. against mouse sepsis caused by cecal ligation/puncture (CLP) in vivo, as well as the molecular mechanisms of action involved.Methods: DCQA (7.5, 15, and 30 mg/kg/day) were administered to the mice with sepsis and the survival rate was obtained. Tumor necrosis factor (TNF)-α and interleukin (IL)-6 were examined by enzyme-linked immunosorbent assay (ELISA). Subsequently, the lipopolysaccharide (LPS) neutralizing ability of DCQA (2, 4, and 8 μg/mL) was measured using limulus amebocyte lysate (LAL) test in vitro. Furthermore, the effect of DCQA (10, 20, and 40 μg/mL) on mRNA expression of TNF-α, IL-6, and IL-8 in LPS (100 ng/mL)-treated RAW 264.7 cells was assessed using quantitative real time-polymerase chain reaction (RT-qPCR) assay.Results: DCQA significantly improved the survival rate of mice with sepsis caused by CLP (35, 50, and 65 %, respectively vs. 15 % for control, p < 0.05). LPS levels fell on co-incubation with DCQA in vitro. Moreover, ELISA and RT-qPCR results revealed that DCQA treatment lowered tendency in the mRNA expression of TNF-α, IL-6, and IL-8 (p < 0.01).Conclusion: DCQA exhibits protective effects against sepsis in mice mediated by downregulating TNF-α, IL-6, and IL-8. Further studies, in animals and humans are requied to determine the safety and efficacy of DCQA in both animal and clinical management of sepsis. Keywords: Xanthium sibiricum, 4,5-O-Dicaffeoylquinic acid, Sepsis, Cecal ligation and puncture, Lipopolysaccharide, TNF-α, IL-6, IL-8

Role of midkine in cadmium-induced liver, heart and kidney damage

2010 ◽  
Vol 30 (5) ◽  
pp. 391-397 ◽  
Author(s):  
Nuray Yazihan ◽  
Mehtap Kacar Kocak ◽  
Ethem Akcil ◽  
Onur Erdem ◽  
Ahmet Sayal
Keyword(s):  
Messenger Rna ◽  
Caspase 3 ◽  
Biological Effects ◽  
Toxic Substance ◽  
Underlying Mechanism ◽  
Enzyme Linked Immunosorbent Assay ◽  
Activity Levels ◽  
Cd Toxicity ◽  
Tnf Α

Accumulation of the widespread environmental toxin cadmium (Cd) in tissues results in toxicity. Cd, which can induce a broad spectrum of biological effects, is a toxic substance and is associated with inflammation and apoptosis. Midkine (MK) has fibrinolytic, antiapoptotic, transforming, angiogenetic and chemotactic activities. After Cd toxicity, we found increased MK expression in liver cells in an in vitro cell culture model. The aim of this study was to determine the possibility of relationship between tissue MK expression levels, tumor necrosis factor α(TNF-α) levels and apoptosis in a chronic Cd toxicity model in rats. Male Wistar rats were exposed to Cd at the dose of 15 parts per million (ppm) for 8 weeks. MK levels were measured in kidney, heart and liver tissue by enzyme-linked-immunosorbent assay (ELISA). MK messenger RNA (mRNA) expression was evaluated by RT-PCR. Tissue apoptosis level was evaluated with tissue caspase-3 activity levels. Accumulation of Cd in liver is higher than the kidney and heart. Cd-treated rats had significantly higher tissue TNF-α and caspase-3 levels when compared with the control rats (p < 0.001). MK mRNA and protein levels were also significantly upregulated in the Cd-treated group (p < 0.05, p < 0.001, respectively). When compared with apoptosis in tissues, it was more prominent in the liver than kidney and heart. MK level is found increased 3, 1.7 and 1.3× folds in liver, kidney and heart, respectively. Our results showed that chronic Cd administration induces inflammation and apoptosis in rat liver, kidney and heart. MK involved in damage mechanisms of Cd-induced tissues. Further studies will show the underlying mechanism of increased MK expression in Cd toxicity.

scholarly journals Inhibitory Effect of Valencene on the Development of Atopic Dermatitis-Like Skin Lesions in NC/Nga Mice

2016 ◽  
Vol 2016 ◽  
pp. 1-11 ◽  
Author(s):  
In Jun Yang ◽  
Dong-Ung Lee ◽  
Heung Mook Shin
Keyword(s):  
Atopic Dermatitis ◽  
Proinflammatory Cytokines ◽  
Molecular Mechanisms ◽  
Biological Effects ◽  
Therapeutic Option ◽  
Skin Barrier ◽  
Skin Lesions ◽  
Cyperus Rotundus ◽  
Raw 264.7 Cells ◽  
Enzyme Linked Immunosorbent Assay

Valencene (VAL) isolated fromCyperus rotunduspossesses various biological effects such as antiallergic and antimelanogenesis activity. We investigated the effect of VAL on atopic dermatitis (AD) skin lesions and their molecular mechanisms. We topically applied VAL to 1-chloro-2,4-dinitrobenzene (DNCB) sensitized NC/Nga mice. Modified scoring atopic dermatitis index, scratching behavior, and histological/immunohistochemical staining were used to monitor disease severity. RT-PCR, western blotting, and enzyme-linked immunosorbent assay were used to determine the level of IgE, proinflammatory cytokines/chemokines production, and skin barrier proteins expression. Topical application of VAL significantly reduced AD-like symptoms and recovered decreased expression of filaggrin in DNCB-sensitized NC/Nga mice. The levels of serum IgE, IL-1β, IL-6, and IL-13 in skin/splenic tissue were reduced.In vitrostudies using TNF-αand IFN-γtreated HaCaT cells revealed that VAL inhibited the exaggerated expression of Th2 chemokines including TARC/CCL17, MDC/CCL22, and proinflammatory chemokines such as CXCL8, GM-CSF, and I-CAM through blockade of the NF-κB pathway. In addition, expression of the skin barrier protein, involucrin, was also increased by VAL treatment. VAL inhibited the production and expression of proinflammatory cytokines IL-1βand IL-6 in LPS-stimulated RAW 264.7 cells. These results suggest that VAL may serve as a potential therapeutic option for AD.

scholarly journals Kirenol inhibits TNF-α-induced proliferation and migration of HaCaT cells by regulating NF-κB pathway

2021 ◽  
Vol 13 (4) ◽  
pp. 44-53
Author(s):  
Jin Li ◽  
Fang Ren ◽  
Wenliang Yan ◽  
Hong Sang
Keyword(s):  
Cell Migration ◽  
Cell Viability ◽  
Signaling Pathway ◽  
Inflammatory Cytokines ◽  
Cell Counting ◽  
Enzyme Linked Immunosorbent Assay ◽  
Dependent Manner ◽  
Hacat Cells ◽  
Tnf Α

Psoriasis is a common chronic, inflammatory skin disease possessing properties of inflammatory cell infiltration and excessive proliferation of keratinocytes, the occurrence and development of which remain fully elucidated. Therefore, the study was designed to determine the effects of kirenol (50, 100 and 200 μg/mL) on Cultured Human Keratinocytes (cells) (HaCaT) in vitro and reveal its molecular mechanism. The in vitro psoriasis model was established utilizing tumor necrosis factor-α (TNF-α)-stimulated HaCaT cells. Kirenol, a diterpenoid compound, was applied at different concentrations (50, 100 and 200 μg/mL) to HaCaT cells for 24 h. The Cell Counting Kit-8 (CCK-8) and thymidine monobromodeoxyuridine (BrdU) assays were used to assess cell viability and proliferation, followed by assessment of cell migration by Transwell assay. Subsequently, inflammatory cytokines were measured by enzyme-linked immunosorbent assay (ELISA), and Western blot assay was used to evaluate expres-sions of p65, p-p65, IκBα and p-IκBα. Activities of superoxide dismutase (SOD), catalase (CAT), glutathione (GSH) and malondialdehyde (MDA) contents were measured spectrophotometrically. The results demonstrated that TNF-α induced a significant increase in cell viability and inflammatory cytokines, including expressions of Interleukin (IL)-6, IL-8, IL-22 and IL-1β in HaCaT cells, which was dose-dependently inhibited by kirenol. Similarly, TNF-α-induced cell migration was also suppressed by kirenol treatment. Furthermore, TNF-α stimuli induced the upregulation of phosphorylation levels of p65 and IκBα as well as p-p65–p65 and p-IκBα–IκBα ratios, whereas kirenol significantly suppressed the activation of cellular nuclear factor-kappa B (NF-κB) signaling pathway. In addition, kirenol significantly decreased the level of MDA but increased the levels of SOD, CAT and GSH in a dose-dependent manner. These results proposed that kirenol could inhibit the proliferation, migration, expression of inflammatory factors, and oxidative stress in HaCaT cells via suppressing NF-κB signaling pathway.

Platycodin D Protects Human Fibroblast Cells from Premature Senescence Induced by H2O2 through Improving Mitochondrial Biogenesis

Pharmacology ◽  
2020 ◽  
Vol 105 (9-10) ◽  
pp. 598-608 ◽  
Author(s):  
Chenyang Shi ◽  
Qin Li ◽  
Xinyue Zhang
Keyword(s):  
Protein Expression ◽  
Mitochondrial Biogenesis ◽  
Molecular Mechanisms ◽  
Enzyme Linked Immunosorbent Assay ◽  
Peroxisome Proliferator ◽  
Premature Senescence ◽  
Peroxisome Proliferator Activated Receptor ◽  
Platycodin D ◽  
Therapeutic Benefits

<b><i>Background:</i></b> Although Platycodin D (PLD) is the main active saponin of <i>Platycodon grandiflorum</i> (PG) and responsible for multiple therapeutic benefits, including antioxidant and antiaging, only few direct demonstrations have been reported on the role of PLD in antiaging process. The present investigation was carried out to elucidate the protection of PLD against aging in vitro and associated molecular mechanisms on H<sub>2</sub>O<sub>2</sub>-induced premature senescence model in human ­fetal lung diploid fibroblasts 2BS cells. <b><i>Methods:</i></b> The cellular morphology, cell cycle, and senescence-associated β-galactosidase activity assays were used for senescence-like phenotypes determination in the oxidant challenged model. The oxygen-free radicals reactive oxygen species (ROS), 4-hydroxynonenal (4-HNE), and malondialdehyde (MDA) determinations were estimated by enzyme-linked immunosorbent assay assay. The potential of the mitochondria mass and the mitochondrial membrane were used to observe the alteration of mitochondria. Western blot analysis was performed to determine the protein expression. <b><i>Results:</i></b> The results showed that PLD significantly reversed senescence-like phenotypes in the oxidant challenged model, as well as related molecules expression such as p53, p21, and p16. Moreover, PLD treatment significantly decreased the levels of ROS, 4-HNE, and MDA in H<sub>2</sub>O<sub>2</sub>-treated 2BS cells. The mechanisms responsible for the antioxidant and antiaging effects of PLD were investigated, we found that mitochondria under PLD conditions show increase membrane potential ratio and stimulate the proliferation of mitochondria mass. In addition, the protein expression of peroxisome proliferator activated receptor gamma coactivator 1α and its downstream targets, that is, nuclear respiratory factor and mitochondrial transcription factor A were also increased in mitochondrial biogenesis. <b><i>Conclusion:</i></b> These results indicated that PLD prevented H<sub>2</sub>O<sub>2</sub>-induced premature senescence in vitro by improving mitochondrial biogenesis to attenuate age-dependent endogenous oxidative damage. <b><i>Key Message:</i></b> The study revealed the antioxidant and antiaging potential of PLD against H<sub>2</sub>O<sub>2</sub>-induced premature senescence.

Is MK2 (mitogen-activated protein kinase-activated protein kinase 2) the key for understanding post-transcriptional regulation of gene expression?

2002 ◽  
Vol 30 (6) ◽  
pp. 959-963 ◽  
Author(s):  
A. Kotlyarov ◽  
M. Gaestel
Keyword(s):  
Transcriptional Regulation ◽  
Cell Migration ◽  
Protein Kinase ◽  
Molecular Mechanisms ◽  
Cytokine Expression ◽  
Mitogen Activated Protein Kinase ◽  
Tnf Α ◽  
Mitogen Activated Protein ◽  
Post Transcriptional Regulation

The phenotype of mitogen-activated protein kinase-activated protein kinase-2 (MK2) knockout mice revealed the essential role of this enzyme in post-transcriptional regulation of lipopolysaccharide-induced expression of cytokines such as tumour necrosis factor (TNF)-α, interleukin-6 and interferon-γ, at the level of mRNA stability and translation. In the case of TNF-α, this regulation depends on the AU-rich element in TNF-α mRNA. In addition to cytokine expression, MK2 is also essential for cell migration in vitro. Although the role of MK2 in cytokine expression depends mainly on catalytic activity, its role in cell migration is also dependent on a proline-rich N-terminal motif. However, the molecular mechanisms involved and the relevant protein targets for MK2 are not completely defined. Here we discuss the possible mechanisms by which two potential target proteins of MK2, small heat-shock protein 25/27 (Hsp25/27) and tristetraprolin, could contribute to our understanding of the above regulation.

scholarly journals The prolyl isomerase Pin1 acts as a novel molecular switch for TNF-α–induced priming of the NADPH oxidase in human neutrophils

Blood ◽  
2010 ◽  
Vol 116 (26) ◽  
pp. 5795-5802 ◽  
Author(s):  
Tarek Boussetta ◽  
Marie-Anne Gougerot-Pocidalo ◽  
Gilles Hayem ◽  
Silvia Ciappelloni ◽  
Houssam Raad ◽  
...  
Keyword(s):  
Nadph Oxidase ◽  
Conformational Changes ◽  
Molecular Mechanisms ◽  
Inflammatory Diseases ◽  
Nicotinamide Adenine Dinucleotide Phosphate ◽  
Human Neutrophils ◽  
Ros Production ◽  
Prolyl Isomerase ◽  
Tnf Α ◽  
Factor Α

Abstract Neutrophils play a key role in host defense by releasing reactive oxygen species (ROS). However, excessive ROS production by neutrophil nicotinamide adenine dinucleotide phosphate (NADPH) oxidase can damage bystander tissues, thereby contributing to inflammatory diseases. Tumor necrosis factor-α (TNF-α), a major mediator of inflammation, does not activate NADPH oxidase but induces a state of hyperresponsiveness to subsequent stimuli, an action known as priming. The molecular mechanisms by which TNF-α primes the NADPH oxidase are unknown. Here we show that Pin1, a unique cis-trans prolyl isomerase, is a previously unrecognized regulator of TNF-α–induced NADPH oxidase hyperactivation. We first showed that Pin1 is expressed in neutrophil cytosol and that its activity is markedly enhanced by TNF-α. Inhibition of Pin1 activity with juglone or with a specific peptide inhibitor abrogated TNF-α–induced priming of neutrophil ROS production induced by N-formyl-methionyl-leucyl-phenylalanine peptide (fMLF). TNF-α enhanced fMLF-induced Pin1 and p47phox translocation to the membranes and juglone inhibited this process. Pin1 binds to p47phox via phosphorylated Ser345, thereby inducing conformational changes that facilitate p47phox phosphorylation on other sites by protein kinase C. These findings indicate that Pin1 is critical for TNF-α–induced priming of NADPH oxidase and for excessive ROS production. Pin1 inhibition could potentially represent a novel anti-inflammatory strategy.

Far Infrared Technology (FIT) Therapy Patches, Protects from Inflammation, Oxidative Stress and Promotes Cellular Vitality

2020 ◽  
Vol 26 (34) ◽  
pp. 4323-4329
Author(s):  
Donatella Pastore ◽  
Francesca Pacifici ◽  
Giampaolo Ciao ◽  
Valentina Bedin ◽  
Guido Pasquantonio ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Endothelial Cells ◽  
Adhesion Molecule ◽  
Molecular Mechanisms ◽  
Far Infrared ◽  
Hek 293 ◽  
Markers Of Inflammation ◽  
Therapeutic Benefits ◽  
Tnf Α ◽  
Infrared Technology

Background: It is known from the most recent literature that far-infrared (FIR) radiations promote a broad spectrum of therapeutic benefits for cells and tissues. Objective: To identify molecular mechanisms by which FIT patches, as a far infrared technology, protects against damage caused by inflammatory process and oxidative stress. Methods: Endothelial cells (HUVEC, Human Umbilical Vein Endothelial Cells) were used as in vitro experimental model. HUVEC were stimulated with a pro-inflammatory cytokine, TNF-α, or hydrogen peroxide (H2O2) to induce oxidative stress. As markers of inflammation were evaluated: VCAM1 (Vascular Cell Adhesion Molecule 1), ICAM1 (Intercellular Adhesion Molecule 1) and E-Selectin by Western Blot analysis. Oxidative stress was assessed by cytofluorimetric analysis. The experiments were performed on control cells (no patch) or in cells treated with the FIT infrared technology applied on the basis of the culture plate. Results: HUVEC stimulated with TNF-α and treated with FIT patches had significant reduction of the expression of VCAM1, ICAM1 and E-Selectin (p<0.05). HUVEC stimulated with H2O2 and treated with FIT patches were significantly protected from oxidative stress (p <0.01) when compared to control cells. We measured cell viability and proliferation in HUVEC and HEK-293 (Human embryonic kidney cells) cells by MTT assay. HEK-293 and HUVEC treated with FIT patches showed a significantly higher percentage of basal vitality compared to control cells (p<0.0001 for HEK-293, p<0.05 for HUVEC). Conclusion: FIT therapy patches - infrared technology, through these protective mechanisms, could be used in all pathologies where an increase in inflammation, oxidative stress and degenerative state are present.

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