Effect of atorvastatin and methotrexate on solid Ehrlich tumor.

Cellular analysis is a central concept for both biology and medicine Over the past two decades, acoustofluidic technologies, which marry acoustic waves with microfluidics, have significantly contributed to the development of innovative approaches for cellular analysis Acoustofluidic technologies enable precise manipulations of cells and the fluids that confine them, and these capabilities have been utilized in many cell analysis applications In this review article, we examine various applications where acoustofluidic methods have been implemented, including cell imaging, cell mechanotyping, circulating tumor cell phenotyping, sample preparation in clinics, and investigation of cell-cell interactions and cell-environment responses We also provide our perspectives on the technological advantages, limitations, and potential future directions for this innovative field of methods

Acoustofluidic methods in cell analysis.

Vasoactive intestinal peptide suppresses the NLRP3 inflammasome activation in lipopolysaccharide-induced acute lung injury mice and macrophages

Atorvastatin in combination with inhaled beclometasone modulates inflammatory sputum mediators in smokers with asthma

Mast cells (MCs), the ‘first responders’ in brain injury, are able to disrupt the blood–brain barrier (BBB), but the underlying mechanism is not well understood. Tryptase is the most abundant MC secretory product. Protease-activated receptor 2 (PAR-2) has been identified as a specific receptor for tryptase, which is abundantly expressed in brain microvascular endothelial cells. The BBB comprises brain microvascular endothelial cells that display specialised molecular properties essential for BBB function and integrity. Therefore, the purpose of the present study was to investigate the effects of tryptase on mouse brain microvascular endothelial cell line bEnd3 and its potential mechanisms of action. Induction of mouse brain microvascular endothelial cell activation by tryptase was examined. Then, mouse brain microvascular endothelial cells were pretreated with a PAR-2 antagonist and stimulated with tryptase. Cellular activation, proinflammatory cytokine production, expression of PAR-2, Toll-like receptors (TLRs) and mitogen-activated protein kinases (MAPK), nuclear factor kappa B (NF-kappa B) phosphorylation were assessed. Tryptase upregulated the production of VCAM-1, MMPs (MMP9 and MMP2), TLR4 and TNF-α and downregulated the expression of the tight junction proteins occludin and claudin-5 in mouse brain microvascular endothelial cell. Among the MAPK and NF-kappa B pathway, ERK and NF-kappa B were activated by tryptase. All of these effects could be eliminated by the PAR-2 inhibitor. Based on our findings, we conclude that tryptase can trigger brain microvascular endothelial cell activation and proinflammatory mediator release. These findings may further clarify the involvement and mechanism of tryptase in BBB disruption.

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Effect of tryptase on mouse brain microvascular endothelial cells via protease-activated receptor 2.

Background: tryptase, the most abundant protease of the human mast cell, has been implicated as a key mediator of allergic inflammation that acts through activation of PAR2. Objectives: to investigate the contribution of PAR2 in the pro-inflammatory actions mediated by tryptase in a mice model. Methods: we have injected recombinant human βII-tryptase into the peritoneum of PAR2-deficient and wild-type C57BL/6 mice. After 6, 12 and 24 hours mice were euthanized, peritoneal lavage performed and inflammatory changes investigated. Results: tryptase stimulated an increase in neutrophil numbers in the peritoneum, but responses did not differ between PAR2-deficient and wild-type mice. Heat-inactivation of tryptase or pre-incubation with a selective tryptase inhibitor reduced neutrophilia, but neutrophil accumulation was not elicited with a peptide agonist of PAR2 (SLIGRL-NH2 ). Zymography indicated that tryptase stimulated the release of matrix metalloproteinases (MMP) 2 and 9 in the peritoneum of both mouse strains. Studies involving immunomagnetic isolation of neutrophils suggested that neutrophils represent the major cellular source of tryptase-induced MMP2 and MMP9. At 24 h after tryptase injection there was increased microvascular leakage as indicated by high levels of albumin in peritoneal lavage fluid, and this appeared to be partially abolished by heat-inactivating tryptase or addition of a protease inhibitor. There was no corresponding increase in levels of histamine or total protein. The extent of tryptase-induced microvascular leakage or gelatinase release into the peritoneum did not differ between PAR2-deficient and wild-type mice. Conclusions: our findings indicate that tryptase is a potent stimulus for neutrophil accumulation, MMP release and microvascular leakage. Though these actions required an intact catalytic site, the primary mechanism of tryptase in vivo would appear to involve processes independent of PAR2. This article is protected by copyright. All rights reserved.

Neutrophilia, gelatinase release and microvascular leakage induced by human mast cell tryptase in a mouse model: lack of a role of protease activated receptor 2 (PAR2)

Study of Atorvastatin in experimental allergic airway inflammation in mice.

Background: VIP is a gastrointestinal peptide hormone which regulates cell proliferation and differentiation in many cell types. In vitro, hepatocytes have limited viability and proliferative capacity. Although several growth factors such as epidermal growth factor (EGF) have been studied, role of VIP is still unclear. We have investigated the effect of VIP on proliferation of human hepatocytes. METHODS: Human hepatocytes were isolated by two-steps collagenase protocol from liver specimens obtained from patients undergoing liver surgery. Cells were grown in Williams’ E media on collagen coated culture plate. Following 4 to 6 hours of cell plating, treatment with VIP or EGF was started and continued for 3 or 5 days. DNA replication was investigated by measuring Bromodeoxyuridine, BrdU incorporation using immunoflorescent staining. In parallel experiments; 1, 3 or 5 days total RNA was isolated and RT–PCR was performed. In the cell culture supernatant, urea and albumin concentrations were detected. RESULTS: VIP was able to increase total number of hepatocytes and number of proliferating cells in a dose dependent manner markedly at day 3 of treatment. Treatment with VIP was associated with an increase in mRNA expression of ki-67 and H3 genes in a dose dependent process. Although mRNA expression of albumin gene was increased significantly with EGF, no marked alteration was found with VIP treatment. With increasing time, addition of VIP was associated with a decrease in albumin and urea secretion from liver cells. CONCLUSIONS: VIP was able to induce proliferation of human hepatocytes but with little effects on hepatocytes differentiation.

/pdf/vasoactive-intestinal-peptide-induces-proliferation-of-human-4u16oi2u07.pdf

Vasoactive intestinal peptide induces proliferation of human hepatocytes.

Ultrasonic standing wave systems have previously been used for the generation of 3D constructs for a range of cell types. In the present study, we cultured cells from the human hepatoma Huh7 cell line in a Bulk Acoustic Wave field and studied their viability, their functions, and their response to the anti-cancer drug, 5 Fluorouracil (5FU). We found that cells grown in the acoustofluidic bioreactor (AFB) expressed no reduction in viability up to 6 h of exposure compared to those cultured in a conventional 2D system. In addition, constructs created in the AFB and subsequently cultured outside of it had improved functionality including higher albumin and urea production than 2D or pellet cultures. The viability of Huh7 cells grown in the ultrasound field to 5FU anti-cancer drug was comparable to that of cells cultured in the 2D system, showing rapid diffusion into the aggregate core. We have shown that AFB formed 3D cell constructs have improved functionality over the conventional 2D monolayer and could be a promising model for anti-cancer drug testing.

/pdf/generation-of-functional-hepatocyte-3d-discoids-in-an-37tdvm3i9z.pdf

Generation of functional hepatocyte 3D discoids in an acoustofluidic bioreactor.

O17 Mast cell tryptase and carboxypeptidase A3 (CPA3) as markers for predicting susceptibility to severe allergic drug reactions Abadalkareem Rana S1, Lau Laurie C K1, Attia Enas2, Abdelmotelb Ahmed1, Zhou Xiaoying3, Eren Efrem1, Goda Sayed4, Walls Andrew F1 1University of Southampton, Southampton, United Kingdom 2Queen Medical Hospital, Doha, Qatar 3Changzhou University, Changzhou, China 4Anti-Doping Laboratory, Doha, Qatar

Ahmed Abdelmotelb

Papers

Neutrophilia, gelatinase release and microvascular leakage induced by human mast cell tryptase in a mouse model: lack of a role of protease activated receptor 2 (PAR2)

Study of Atorvastatin in experimental allergic airway inflammation in mice.

Vasoactive intestinal peptide induces proliferation of human hepatocytes.

Generation of functional hepatocyte 3D discoids in an acoustofluidic bioreactor.

Mast cell Tryptase and Carboxypeptidase A3 (CPA3) as Markers for Predicting Susceptibility to Severe Allergic Drug Reactions