Genetic reporter assays to study the regulation of interleukin-7 receptor alpha gene expression

Interleukin-7 (IL-7) signaling through IL-7 receptor controls survival and proliliferation of B- and T- lymphocytes at early stages of development. IL-7 is also a critical survival factor for naive T-lymphocytes and memory T-cells circulating in the peripheral organs. Throughout T-cell development, IL-7Ralpha expression on distinct T-cell populations is strictly regulated. However, the mechanisms that regulate differential expression of the IL-7Ralpha chain on T- lymphocytes are poorly defined. Here, we used bioinformatic tools to determine possible cis-acting regulatory elements at the downstream of mouse IL-7Ralpha gene and we tried to analyze the transcriptions! activities of these possible cis-regulatory elements in a mouse T lymphoma cell line by means of genetic reporter systems We took advantage of the luciferase assay system for functional analysis of noncoding sequences at the downstream of mouse IL-7Ralpha gene. We modified luciferase reporter vector by inserting weak promoter into the ups team of luciferase cDNA and cloned the mouse noncoding sequences into this modified vector. We electroporated mouse T lymphoma cells with luciferase reporter vectors to evaluate the effects of these noncoding regions on the transcription of luciferase cDNA. Although the luciferase activities in T lymphoma cell lysates were low due to low sensitivity of the detection system used, we observed that efficient transfections with luciferase reporter vectors resulted in high luciferase activities in a different cell line. We concluded that different reporter systems or different cell lines were required to determine possible transcriptional activities of the cloned mouse noncoding sequences

The tales of two sacred buildings in urban pattern of a transportation hub : the case of Üsküdar, İstanbul

Üsküdar is one of the oldest settlements in Istanbul dating back to the early Ottoman period and the primary transportation hub of the modern city. The area is at the intersection point of bulk sea transport and underground system besides heavy motor traffic and pedestrian movement. Being by-the-sea has supported the district's function in terms of transportation and recreation facilities. There is a great square, which is defined by significant historical monuments and underground stop entrances, allows considerable pedestrian flow through the area. The aim of this study is revealing the changes in the urban form of Üsküdar throughout time and the effects of two constant sacred buildings as a significant role on users' daily lives today. For this purpose, firstly the morphological changes in the urban pattern of Üsküdar are demonstrated on the building and plot differences in the urban tissue and space syntax as a quantitative method is used to define the configuration of the urban structure in relation with the two monumental historical mosques. Secondly, the architectural structures of the two mosques examined as different systems from courtyard and surrounding of the buildings. According to this perspective, visibility graph analysis and related data sets on user behaviour are used. Moreover, the pedestrian movement pattern data in the area is collected in three different time periods during the weekday, weekend and also pray time on Friday. To conclude, this study combines different data sets, observations and analytical methods to explain the morphological character of the historic area and the influence of the Friday mosques on users' everyday life independent from their functional usage

A Distinct Gene Module for Dysfunction Uncoupled from Activation in Tumor-Infiltrating T Cells

Reversing the dysfunctional T cell state that arises in cancer and chronic viral infections is the focus of therapeutic interventions; however, current therapies are effective in only some patients and some tumor types. To gain a deeper molecular understanding of the dysfunctional T cell state, we analyzed population and single-cell RNA profiles of CD8+tumor-infiltrating lymphocytes (TILs) and used genetic perturbations to identify a distinct gene module for T cell dysfunction that can be uncoupled from T cell activation. This distinct dysfunction module is downstream of intracellular metallothioneins that regulate zinc metabolism and can be identified at single-cell resolution. We further identify Gata-3, a zinc-finger transcription factor in the dysfunctional module, as a regulator of dysfunction, and we use CRISPR-Cas9 genome editing to show that it drives a dysfunctional phenotype in CD8+TILs. Our results open novel avenues for targeting dysfunctional T cell states while leaving activation programs intact

Induction and transcriptional regulation of the co-inhibitory gene module in T cells

Expression of co-inhibitory receptors, such as CTLA-4 and PD-1, on effector T cells is a key mechanism for ensuring immune homeostasis. Dysregulated co-inhibitory receptor expression on CD4+ T cells promotes autoimmunity while sustained overexpression on CD8+ T cells promotes T cell dysfunction or exhaustion, leading to impaired ability to clear chronic viral infections and cancer1,2. Here, we used RNA and protein expression profiling at single-cell resolution to identify a module of co-inhibitory receptors that includes not only several known co-inhibitory receptors (PD-1, Tim-3, Lag-3, and TIGIT), but also a number of novel surface receptors. We functionally validated two novel co-inhibitory receptors, Activated protein C receptor (Procr) and Podoplanin (Pdpn). The module of co-inhibitory receptors is co-expressed in both CD4+ and CD8+ T cells and is part of a larger co-inhibitory gene program that is shared by non-responsive T cells in multiple physiological contexts and is driven by the immunoregulatory cytokine IL-27. Computational analysis identified the transcription factors Prdm1 and c-Maf as cooperative regulators of the co-inhibitory module, which we validated experimentally. This molecular circuit underlies the co-expression of co-inhibitory receptors in T cells and identifies novel regulators of T cell function with the potential to regulate autoimmunity and tumor immunity

Prospects, Problems of Marketing Research and Data Mining in Turkey

The objective of this paper is to review and assess the methodological issues and problems in marketing research, data and knowledge mining in Turkey. As a summary, academic marketing research publications in Turkey have significant problems. The most vital problem seems to be related with modeling. Most of the publications had major weaknesses in modeling. There were also, serious problems regarding measurement and scaling, sampling and analyses. Analyses myopia seems to be the most important problem for young academia in Turkey. Another very important finding is the lack of publications on data and knowledge mining in the academic world

Recent Trends in Marketing Research in Turkey

The aim of this research is to gain insights in the methodological issues and problems of marketing research in Turkey. This research provides information and recommendations for the research methodology of marketing papers published in Turkish National Marketing Congresses' Proceedings between 2007 and 2009. The evaluation of marketing papers is done with content analyses method, in terms of methodological process such as research type, research modeling and hypothesis development, sampling, data collection and data analysis

The content analysis of online newspapers in Turkey and consumer perceptions

The newspapers are one of the main information systems within the community for transaction, sharing news and informing people. The printed word, especially newspapers, used to be a dominant medium for mass communications for a long time. The media and its ability to present news constantly changes, keeping up with technology. Technological developments create new mediums that make information transaction easier. Through the use of hyperlinks, text, still and moving images, audio and other multimedia presentation, news can be utilized to provide more in-depth coverage than in traditional print editions. The online media is one of the emerging sectors which is rapidly growing in numbers of audience and usage opportunities. In this regard, this new media is also attracted many research attention from many aspects such as content, interactivity, comparison of printed media, evaluation of consumers

Measuring brand image using a cognitive approach: Representing brands as a network in the Turkish airline industry

This study extends the previous brand image measurement processes by using a mixed (qualitative and quantitative) methodology, drawing upon a cognitive approach to represent consumers' minds as a network. Therefore, three consecutive studies have been designed from a sample of 1000 passengers to construct four distinct concept maps and a common one pertaining to four Turkish airline companies. The results show the importance of increasing flight comfort, training cabin crew, providing free appetizers, improving service quality, and ensuring timely departures in creating and sustaining a positive brand image. Academicians could use this methodology as an alternative technique to the traditional ones and practitioners could use it to evaluate brands' competitive advantages to create powerful brand images in consumers' minds