The Role of TRPM4 Channel in Pseudomonas aeruginosa Infection and the Regulation of the Inflammatory Response in Airway Epithelial Cells

Bacterial pneumonia is among the leading causes of death worldwide. The emerging resistant strains require the development of new treatment strategies. Ion channels play a role in many signaling pathways of the cell. TRPM4 belongs to the family of TRP-channels (transient receptor potential ion channels), which are membrane bound ion channels expressed in a wide variety of cells. The activity of TRPM4 is induced by rising Ca2+ concentrations in the cytosol. The channel is permissive for Na+ and K+ ions, which lead to a depolarization of the cellular membrane. However, activated by Ca2+, TRPM4 is not permissive for Ca2+. The goal of this work is to investigate the influence of the activity of TRPM4 on the pulmonary innate immune response in the course of bacterial infection. After detecting the expression of TRPM4 in the human bronchial epithelial cell line NCI-H292, the cytotoxicity of the pharmacologic TRPM4 inhibitor 9-Phenanthrol (9-Ph.) and bacterial stimulation with heat inactivated Pseudomonas aeruginosa (Ps.a.) was investigated. The stimulation with heat inactivated Ps.a. or 9-Ph. in the IC50-range had no cytotoxic effects on the cells for at least 18 hr. Most interestingly, the release of IL-6, TNF-α, CXCL2, and S100A8 was inhibited after the pre-treatment with 9-Ph. and stimulation with Ps.a.. This effect was dependent on the concentration of 9-Ph. and was not observed at concentrations of 1/4 - 1/8 of the IC50 value of 9 Ph. Furthermore, the inhibition was not detected on transcriptional level, except for S100A8, whose transcription was also inhibited by the pre-incubation with 9-Ph. These findings were confirmed by TRPM4-specific siRNA, which also resulted in significantly less concentrations of IL-6 after the stimulation with Ps.a.. This indicates that the function of TRPM4 was necessary for the export of inflammatory mediators. This was proved by FACS-Analysis and ELISA. The bronchial-epithelial cell line NCI-H292 and human primary airway epithelial cells were also stimulated with TLR1/2, TLR4, and TLR5-ligands. These experiments showed that the treatment with distinct TLR-ligands can also be influenced by the inhibition of TRPM4 and that the effects observed so far are not due to the inhibition of a certain TLR-signaling pathway. The antimicrobial activity of airway-epithelial cells is mostly mediated by the expression of antimicrobial peptides. The expression of the antimicrobial peptide hBD-2 (human beta-defensin-2) was highly induced in differentiated human primary airway epithelial cells after the stimulation with live Ps.a.. This correlated with increased concentrations of hBD-2 in the cell culture supernatant and increased antimicrobial activity of the cells. In contrast, the inhibition of TRPM4 significantly downregulated the expression of hBD2, the concentration in cell culture supernatant, and the antimicrobial activity of the cells. This indicates a direct connection of the activity of TRPM4 to the innate immune response of the lung. To investigate the influence of TRPM4 on the release of inflammatory cytokines in different cell types and species, monocyte derived macrophages from the bone marrow of mice and the human monocyte cell line U937 were used. Again, the inhibition of TRPM4 resulted in a significantly decreased release of IL-6, TNF-α, CXCL1, and CXCL2. These results were also confirmed with macrophages isolated from TRPM4-deficient mice. To further explore these findings, the lungs of TRPM4-deficient mice were stimulated with heat inactivated Ps.a.. TRPM4-deficient mice showed less inflammation than the corresponding wildtype control animals. These findings are based upon less influx of neutrophilic granulocytes into the lung and lower concentrations of IL-6, TNF-a, CXCL1, and CXCL2 in the broncho-alveolar lavage (BALF) of the TRPM4-deficient animals. Furthermore, the expression of mBD-4, the murine homologue of the human beta-defensin-2, was significantly reduced in the TRPM4-deficient mice. The stimulation of differentiated primary murine tracheal epithelial cells with heat inactivated Ps.a. induced a higher release of KC and MIP-2 if TRPM4 was inhibited. Furthermore, the stimulation of TRPM4-deficient mice with LPS resulted in an increased inflammatory response from the TRPM4-knockout mice compared to the wildtype controls. These differences may result from the different stimulations or the reactivity of the different cell types and possible variations in the immune response of human and mouse cells. These results show that the function of TRPM4 is important for the regulation of inflammation and the innate immune response of the lung

A city wihtin a city : from urbanization stress to the relief realm

LAUREA MAGISTRALELa città di Berlino è sempre stata una fonte di ispirazione per architetti e urbanisti di tutto il mondo, con la sua ricca storia, la variegata scena culturale e il tessuto urbano unico. Una delle teorie più interessanti e rilevanti sulla forma urbana di Berlino è l'ipotesi "Città nella città; Berlino come arcipelago verde" proposta da Rem Koolhaas e Oswald Mathias Ungers. Questa ipotesi suggerisce che Berlino sia una città composta da una serie di isole isolate o "arcipelaghi", ciascuna con il proprio carattere, la propria storia e il proprio paesaggio. Sulla base di ciò, la tesi propone la progettazione di un progetto di "città nella città" situato al confine tra Berlino e la campagna del Brandeburgo. Questo progetto mira ad affrontare alcune delle sfide urbanistiche più urgenti che la città deve affrontare oggi, tra cui la crisi abitativa, la necessità di trovare un equilibrio tra individualismo e collettivismo e la perdita di identità all'interno del tessuto urbano. In linea con l'ipotesi "Città nella città; Berlino come arcipelago verde", il progetto cerca di creare una serie di isole interconnesse o "arcipelaghi" all'interno del complesso edilizio più ampio. Ogni isola ha il suo carattere, la sua funzione e la sua identità distinti ed è collegata alle altre attraverso una rete di percorsi e spazi pubblici. Il progetto consiste in tre grandi “arcipelaghi” di complessi edilizi orizzontali con funzioni miste, comprendenti strutture residenziali, commerciali, educative e culturali. Ogni isola è organizzata attorno a un cortile centrale, che funge da spazio di raccolta e oasi verde all'interno dell'edificio. Il cortile è circondato da fitte aree verdi, che esaltano gli aspetti sociali e spirituali del progetto e forniscono un collegamento con il paesaggio circostante. In breve, la "Città nella città" cerca di offrire un nuovo modello di sviluppo urbano che concilia le esigenze spesso contrastanti di individualismo e collettivismo, sostenibilità e crescita, contemporaneità e modernità, densità e spazio aperto, identità locale e connettività globale . Basandosi sulla ricca eredità della storia e della cultura urbana di Berlino, ma guardando anche alle sfide e alle opportunità del 21° secolo, questo progetto rappresenta un approccio audace e visionario alla progettazione e pianificazione urbana.The city of Berlin has always been a source of inspiration for architects and urban planners around the world, with its rich history, diverse cultural scene, and unique urban fabric. One of the most interesting and relevant theories about Berlin's urban form is the "City within a city; Berlin as green archipelago" hypothesis proposed by Rem Koolhaas and Oswald Mathias Ungers. This hypothesis suggests that Berlin is a city made up of a series of isolated islands or "archipelagos", each with its own distinct character, history, and landscape. Based on that, the thesis proposes the design of a "city within a city" project located on the border of Berlin and the Brandenburg countryside. This project aims to address some of the most pressing urbanization challenges facing the city today, including the housing crisis, the need to find a balance between individualism and collectivism, and the loss of identity within the urban fabric. In line with the "City within a city; Berlin as green archipelago" hypothesis, the project seeks to create a series of interconnected islands or "archipelagos" within the larger building complex. Each island has its own distinct character, function, and identity, and is connected to the others through a network of pathways and public spaces. The project consists of three large horizontal building complex “archipelagos” with mixed-use functions, including residential, commercial, educational, and cultural facilities. Each island is organized around a central courtyard, which serves as a gathering space and green oasis within the building. The courtyard is surrounded by dense green areas, which enhance the social and spiritual aspects of the project and provide a connection to the surrounding landscape. In short, the “City within a City” seeks to offer a new model for urban development that reconciles the often-competing necessities of individualism and collectivism, sustainability and growth, contemporary and modernity, density and open space, and local identity and global connectivity. By building on the rich legacy of Berlin's urban history and culture, while also looking forward to the challenges and opportunities of the 21st century, this project represents a bold and visionary approach to urban design and planning

The compatibility of Yusuf Al Qaradawi\u27s wasateyya school of thought with the second generation of the Muslim Brotherhood

This paper aims to study the influence of the Wasateyya (centrist) school of thought on the Muslim Brotherhood\u27s second generation of leaders. The study shows that the Wasateyya school and its advocates within the Muslim Brotherhood were unsuccessful in bringing about a meaningful change in the direction of moderateness within the movement. The research assesses the influence of the Wasateyya school of thought by evaluating the compatibility of Qaradawi\u27s ideology with that of four members of the middle generation of the Brotherhood; Abd El Moniem Abul Fotouh, Essam el Eryan, Khairat al Shater, and Mohamed Morsi. The thesis is composed of seven large sections; sections one to three introduce the research question, methodology, and limitations. Section four is a literature review, section five addresses the relationship between Qaradawi and the second generation of the MB and defines ideological and political Wasateyya, section six is about the case study of the four leaders, and finally section seven concludes the main findings of the study

Low Complexity Selective Adaptive Multicarrier DS-CDMA Receiver

In this paper, selective adaptive (SA) receiver for Multicarrier Direct Sequence Code Division Multiple Access (MC DS-CDMA) system is presented. This receiver has high performance and at the same time reduces the multiple access interference (MAI) of the MC DS-CDMA) system with low computational complexity. The performance of SA receiver is measured in terms of the bit error rate (BER). An upper bound expression of the BER for the SA receiver under Rayleigh fading channel condition is derived and validated by computer simulations. Moreover, the implementation complexities of the SA receiver is compared with the Adaptive Parallel Interference Cancellation (APIC) receiver.

Recursive Detection of M-Ary Signals over Fast Varying Mobile Communication Channel

          Mobile radio is characterized by a fast time varying channel. Conventional detectors which designed optimal for non-fading channel exhibit a limited performance in fast time varying channel. In this paper a recursive detector for M-ary signals over fast time varying mobile communication channel is introduced. The proposed detector continuously estimates the channel directly within the metric calculation of the log-likelihood function in a recursive manner. The estimation of the channel is performed by the covariance form of the recursive least square approach. The performance of the detector is evaluated in terms of the misdetection probability. The effects of timing and phase offsets on the performance of detector are examined by simulation. Simulation results show that the proposed detector can accommodate the fast time varying channel with adequate performance

The Effect of NiNO3 on Optical Properties for (PVA- PEG) Composites Polymers

The present paper is aimed to modification of the optical properties of poly-vinyl alcohol and poly-ethylene glycol with different concentrations of ( NiNO3). The absorption and transmission spectra have been recorded in the wavelength range     (200-800)nm . The absorption coefficient and energy gap of the indirect allowed and forbidden transition have been determined, Also, extinction coefficient, index coefficient, real and imaginary part of dielectric constant have been calculated. Keywords: optical properties, polymer composites, optical constants, polyethylene glycol