GPR87 is an overexpressed G-protein coupled receptor in squamous cell carcinoma of the lung

Lung cancer is the leading cause of cancer death worldwide. The overall 5-year survival after therapy is about 16% and there is a clear need for better treatment options, such as therapies targeting specific molecular structures. G-protein coupled receptors (GPCRs), as the largest family of cell surface receptors, represent an important group of potential targets for diagnostics and therapy. We therefore used laser capture microdissection and GPCR-focused Affymetrix microarrays to examine the expression of 929 GPCR transcripts in tissue samples of 10 patients with squamous cell carcinoma and 7 with adenocarcinoma in order to identify novel targets in non-small cell lung carcinoma (NSCLC). The relative gene expression levels were calculated in tumour samples compared to samples of the neighbouring alveolar tissue in every patient. Based on this unique study design, we identified 5 significantly overexpressed GPCRs in squamous cell carcinoma, in the following decreasing order of expression: GPR87 > CMKOR1 > FZD10 > LGR4 > P2RY11. All are non-olfactory and GRAFS (glutamate, rhodopsin, adhesion, frizzled/taste2, secretin family) classified. GPR87, LGR4 and CMKOR1 are orphan receptors. GPR87 stands out as a candidate for further target validation due to its marked overexpression and correlation on a mutation-based level to squamous cell carcinoma.

Automated design of self-stabilization

Self-stabilization is a property of a distributed system such that, regardless of the legitimacy of its current state, the system behavior shall eventually reach a legitimate state and shall remain legitimate thereafter. The elegance of self-stabilization stems from the fact that it distinguishes distributed systems by a strong fault tolerance property against arbitrary state perturbations. The difficulty of designing and reasoning about self-stabilization has been witnessed by many researchers; most of the existing techniques for the verification and design of self-stabilization are either brute-force, or adopt manual approaches non-amenable to automation. In this dissertation, we first investigate the possibility of automatically designing self-stabilization through global state space exploration. In particular, we develop a set of heuristics for automating the addition of recovery actions to distributed protocols on various network topologies. Our heuristics equally exploit the computational power of a single workstation and the available parallelism on computer clusters. We obtain existing and new stabilizing solutions for classical protocols like maximal matching, ring coloring, mutual exclusion, leader election and agreement. Second, we consider a foundation for local reasoning about self-stabilization; i.e., study the global behavior of the distributed system by exploring the state space of just one of its components. It turns out that local reasoning about deadlocks and livelocks is possible for an interesting class of protocols whose proof of stabilization is otherwise complex. In particular, we provide necessary and sufficient conditions – verifiable in the local state space of every process – for global deadlock- and livelock-freedom of protocols on ring topologies. Local reasoning potentially circumvents two fundamental problems that complicate the automated design and verification of distributed protocols: (1) state explosion and (2) partial state information. Moreover, local proofs of convergence are independent of the number of processes in the network, thereby enabling our assertions about deadlocks and livelocks to apply on rings of arbitrary sizes without worrying about state explosion.

ANKLE IMPEDANCE AND ANKLE ANGLES DURING STEP TURN AND STRAIGHT WALK: IMPLICATIONS FOR THE DESIGN OF A STEERABLE ANKLE-FOOT PROSTHETIC ROBOT

During locomotion, turning is a common and recurring event which is largely neglected in the current state-of-the-art ankle-foot prostheses, forcing amputees to use different steering mechanisms for turning, compared to non-amputees. A better understanding of the complexities surrounding lower limb prostheses will lead to increased health and well-being of amputees. The aim of this research is to develop a steerable ankle-foot prosthesis that mimics the human ankle mechanical properties. Experiments were developed to estimate the mechanical impedance of the ankle and the ankles angles during straight walk and step turn. Next, this information was used in the design of a prototype, powered steerable ankle-foot prosthesis with two controllable degrees of freedom. One of the possible approaches in design of the prosthetic robots is to use the human joints’ parameters, especially their impedance. A series of experiments were conducted to estimate the stochastic mechanical impedance of the human ankle when muscles were fully relaxed and co-contracting antagonistically. A rehabilitation robot for the ankle, Anklebot, was employed to provide torque perturbations to the ankle. The experiments were performed in two different configurations, one with relaxed muscles, and one with 10% of maximum voluntary contraction (MVC). Surface electromyography (sEMG) was used to monitor muscle activation levels and these sEMG signals were displayed to subjects who attempted to maintain them constant. Time histories of ankle torques and angles in the lateral/medial (LM) directions, inversion-eversion (IE), and dorsiflexionplantarflexion (DP) were recorded. Linear time-invariant transfer functions between the measured torques and angles were estimated providing an estimate of ankle mechanical impedance. High coherence was observed over a frequency range up to 30 Hz. The main effect of muscle activation was to increase the magnitude of ankle mechanical impedance in all degrees of freedom of the ankle. Another experiment compared the three-dimensional angles of the ankle during step turn and straight walking. These angles were measured to be used for developing the control strategy of the ankle-foot prosthesis. An infrared camera system was used to track the trajectories and angles of the foot and leg. The combined phases of heel strike and loading response, mid stance, and terminal stance and pre-swing were determined and used to measure the average angles at each combined phase. The Range of motion (ROM) in IE increased during turning while ML rotation decreased and DP changed the least. During the turning step, ankle displacement in DP started with similar angles to straight walk and progressively showed less plantarflexion. In IE, the ankle showed increased inversion leaning the body toward the inside of the turn. ML rotation initiated with an increased medial rotation during the step turn relative to the straight walk transitioning to increased lateral rotation at the toe off. A prototype ankle-foot prosthesis capable of controlling both DP and IE using a cable driven mechanism was developed and assessed as part of a feasibility study. The design is capable of reproducing the angles required for straight walk and step turn; generates 712N of lifting force in plantarflexion, and shows passive stiffness comparable to a nonload bearing ankle impedance. To evaluate the performance of the ankle-foot prosthesis, a circular treadmill was developed to mimic human gait during steering. Preliminary results show that the device can appropriately simulate human gait with loading and unloading the ankle joint during the gait in circular paths.

DISSECTION OF STRESS RESPONSE NETWORKS REGULATING MULTIPLE STRESSES IN RICE

Important food crops like rice are constantly exposed to various stresses that can have devastating effect on their survival and productivity. Being sessile, these highly evolved organisms have developed elaborate molecular machineries to sense a mixture of stress signals and elicit a precise response to minimize the damage. However, recent discoveries revealed that the interplay of these stress regulatory and signaling molecules is highly complex and remains largely unknown. In this work, we conducted large scale analysis of differential gene expression using advanced computational methods to dissect regulation of stress response which is at the heart of all molecular changes leading to the observed phenotypic susceptibility. One of the most important stress conditions in terms of loss of productivity is drought. We performed genomic and proteomic analysis of epigenetic and miRNA mechanisms in regulation of drought responsive genes in rice and found subsets of genes with striking properties. Overexpressed genesets included higher number of epigenetic marks, miRNA targets and transcription factors which regulate drought tolerance. On the other hand, underexpressed genesets were poor in above features but were rich in number of metabolic genes with multiple co-expression partners contributing majorly towards drought resistance. Identification and characterization of the patterns exhibited by differentially expressed genes hold key to uncover the synergistic and antagonistic components of the cross talk between stress response mechanisms. We performed meta-analysis on drought and bacterial stresses in rice and Arabidopsis, and identified hundreds of shared genes. We found high level of conservation of gene expression between these stresses. Weighted co-expression network analysis detected two tight clusters of genes made up of master transcription factors and signaling genes showing strikingly opposite expression status. To comprehensively identify the shared stress responsive genes between multiple abiotic and biotic stresses in rice, we performed meta-analyses of microarray studies from seven different abiotic and six biotic stresses separately and found more than thirteen hundred shared stress responsive genes. Various machine learning techniques utilizing these genes classified the stresses into two major classes' namely abiotic and biotic stresses and multiple classes of individual stresses with high accuracy and identified the top genes showing distinct patterns of expression. Functional enrichment and co-expression network analysis revealed the different roles of plant hormones, transcription factors in conserved and non-conserved genesets in regulation of stress response.

Treatment of recurrent rejection in heart transplantation: cytolytic therapy or bolus steroids?

OBJECTIVES: The treatment of recurrent rejection in heart transplant recipients has been a controversial issue for many years. The intent of this retrospective study was to perform a risk-benefit analysis between treatment strategies with bolus steroids only versus anti-thymocyte globulins (RATG; 1.5 mg/kg q 4 days). METHODS: Between 1986 and 1993, 69 of 425 patients (17 male, 52 female; mean age 44 +/- 11 years) who had more than one rejection/patient per month (rej/pt per mo) in the first 3 postoperative months were defined as recurrent rejectors. RESULTS: Repetitive methylprednisolone bolus therapy (70 mg/kg q 3 days) was given in 27 patients (group M; 1.4 +/- 0.2 rej/pt per mo) and RATG therapy for one of the rejection episodes of the 42 remaining patients (group A; 1.5 +/- 0.2 rej/pt per mo). The quality of triple drug immunosuppression in the two study groups was comparable. The rejection-free interval (RFI) following RATG treatment in group A was 21.6 +/- 10 days and 22 +/- 11 in group M. In group M, 3 of 27 patients (11%) had a rejection treatment-related infection (2 bacterial; 1 viral) versus 6 of the 42 patients of group A (14.2%; bacterial 1, viral 5). During postoperative months 3-24, 0.15 +/- 0.12 rej/pat per mo were observed in group M and 0.21 +/- 0.13 rej/pat per mo in group A (n.s.). In this 21-month period cytolytic therapy for rejection was initiated in 8 of the remaining 21 patients of group M (38%) and 15 of the remaining 37 patients of group A (40.5%). The absolute survival and the individual causes of death were not affected by the type of initial treatment of recurrent rejection. The actuarial freedom of graft atherosclerosis is comparable in the two groups with 78% in group A versus 79% in group M free of graft atherosclerosis at 3 years postoperatively. CONCLUSIONS: A comparison of cytolytic therapy versus repeated applications of bolus steroids for treatment of recurrent rejection reveals no significant difference in the long-term patient outcome with respect to the incidence of future rejection episodes and survival.

OCT4 expression in human non-small cell lung cancer: implications for therapeutic intervention

Here we investigate the expression of OCT4 human lung adenocarcinoma and bronchioloalveolar carcinoma (BAC) tumor biopsies and tumor-derived primary cell cultures. OCT4 has been detected in several human tumors suggesting a potentially critical role in tumorigenesis. We assessed the presence of OCT4 in clinical tumor samples of both adenocarcinoma and BAC at the cellular and transcriptional levels, respectively. Furthermore, we evaluated tumor-derived cell cultures for potential differences in OCT4 expression. Immunohistochemical analysis depicted OCT4 in 2 of 8 adenocarcinoma tumor samples and 3 of 5 BAC tumor samples, with no apparent difference in the degree of expression among the sections examined. These results were validated by transcript analysis. Flow cytometric assessment of 11 adenocarcinoma-derived cell cultures and 3 BAC-derived cell cultures revealed significantly higher OCT4 expression in adenocarcinoma tumors compared to their normal counterparts. This, however, was not observed in the BAC cultures. Comparative studies of OCT4 in adenocarcinoma and BAC tumor cell cultures demonstrated a dramatically higher expression in the former. The expression of OCT4 may represent a specific and effective target for therapeutic intervention in adenocarcinoma and BAC. In addition, the aberrant expression and distribution of OCT4 may indicate important parameters concerning the differences between adenocarcinoma and BAC.

Use of concatamers to study GABAA receptor architecture and function: application to delta-subunit-containing receptors and possible pitfalls

Many membrane proteins, including the GABA(A) [GABA (gamma-aminobutyric acid) type A] receptors, are oligomers often built from different subunits. As an example, the major adult isoform of the GABA(A) receptor is a pentamer built from three different subunits. Theoretically, co-expression of three subunits may result in many different receptor pentamers. Subunit concatenation allows us to pre-define the relative arrangement of the subunits. This method may thus be used to study receptor architecture, but also the nature of binding sites. Indeed, it made possible the discovery of a novel benzodiazepine site. We use here subunit concatenation to study delta-subunit-containing GABA(A) receptors. We provide evidence for the formation of different functional subunit arrangements in recombinant alpha(1)beta(3)delta and alpha(6)beta(3)delta receptors. As with all valuable techniques, subunit concatenation has also some pitfalls. Most of these can be avoided by carefully titrating and minimizing the length of the linker sequences joining the two linked subunits and avoiding inclusion of the signal sequence of all but the N-terminal subunit of a multi-subunit construct. Maybe the most common error found in the literature is that low expression can be overcome by simply overloading the expression system with genetic information. As some concatenated constructs result by themselves in a low level of expression, this erroneous assembly leading to receptor function may be promoted by overloading the expression system and leads to wrong conclusions.

Statistical analysis of multichannel scalp field data

High density spatial and temporal sampling of EEG data enhances the quality of results of electrophysiological experiments. Because EEG sources typically produce widespread electric fields (see Chapter 3) and operate at frequencies well below the sampling rate, increasing the number of electrodes and time samples will not necessarily increase the number of observed processes, but mainly increase the accuracy of the representation of these processes. This is namely the case when inverse solutions are computed. As a consequence, increasing the sampling in space and time increases the redundancy of the data (in space, because electrodes are correlated due to volume conduction, and time, because neighboring time points are correlated), while the degrees of freedom of the data change only little. This has to be taken into account when statistical inferences are to be made from the data. However, in many ERP studies, the intrinsic correlation structure of the data has been disregarded. Often, some electrodes or groups of electrodes are a priori selected as the analysis entity and considered as repeated (within subject) measures that are analyzed using standard univariate statistics. The increased spatial resolution obtained with more electrodes is thus poorly represented by the resulting statistics. In addition, the assumptions made (e.g. in terms of what constitutes a repeated measure) are not supported by what we know about the properties of EEG data. From the point of view of physics (see Chapter 3), the natural “atomic” analysis entity of EEG and ERP data is the scalp electric field

Democracy Concepts of the Fundamentalist Parties of Algeria and Tunisia – Claim and Reality

This article discusses democratic elements in early Islamic sources and in the programs of the Algerian FIS (Front Islamique du Salut) and ANNAHDA in Tunesia. According to historic writings, Islam includes the principles of democratic consensus, consultation, and freedom of opinion, and an understanding that the sources of Islamic jurisdiction are subject to interpretation, that the sharia can be changed, and that religious authorities’ power to issue instructions on worldly matters is limited. These are the type of expectations that fundamentalist parties arouse when they speak of an Islamic caliphate as a state system. Against this background, an examination of the political system proposed until 1992 by the Algerian FIS shows that this system would have resulted in a very restrictive form of Islam. An investigation of the political system of the Tunisian fundamentalist leader Rached al-Ghannouchi reveals that the system he proposes may be designated as an Islamic democracy, since it takes into account separation of powers and pluralism of political parties. The head of state would be subject to the law in the same manner as the people. However, it is no liberal democracy, as he categorically rejects secularism, intends to punish apostates, and is only willing to allow political parties that are based on the religion of Islam. His state would only be a state of those citizens who follow Islam, completely neglecting secularist groups. Social conflicts and unrest are thus predetermined.

Efforts and Models of Education for Parents: the Danish Approach

The report examines the relationship between day care institutions, schools and so called “parents unfamiliar to education” as well as the relationship between the institutions. With in Danish public and professional discourse concepts like parents unfamiliar to education are usually referring to environments, parents or families with either no or just very restricted experience of education except for the basic school (folkeskole). The “grand old man” of Danish educational research, Prof. Em. Erik Jørgen Hansen, defines the concept as follows: Parents who are distant from or not familiar with education, are parents without tradition of education and by that fact they are not able to contribute constructively in order to back up their own children during their education. Many teachers and pedagogues are not used to that term; they rather prefer concepts like “socially exposed” or “socially disadvantaged” parents or social classes or strata. The report does not only focus on parents who are not capable to support the school achievements of their children, since a low level of education is usually connected with social disadvantage. Such parents are often not capable of understanding and meeting the demands from side of the school when sending their children to school. They lack the competencies or the necessary competence of action. For the moment being much attention is done from side of the Ministries of Education and Social Affairs (recently renamed Ministry of Welfare) in order to create equal possibilities for all children. Many kinds of expertise (directions, counsels, researchers, etc.) have been more than eager to promote recommendations aiming at achieving the ambitious goal: 2015 95% of all young people should complement a full education (classes 10.-12.). Research results are pointing out the importance of increased participation of parents. In other word the agenda is set for ‘parents’ education’. It seems necessary to underline that Danish welfare policy has been changing rather radical. The classic model was an understanding of welfare as social assurance and/or as social distribution – based on social solidarity. The modern model looks like welfare as social service and/or social investment. This means that citizens are changing role – from user and/or citizen to consumer and/or investor. The Danish state is in correspondence with decisions taken by the government investing in a national future shaped by global competition. The new models of welfare – “service” and “investment” – imply severe changes in hitherto known concepts of family life, relationship between parents and children etc. As an example the investment model points at a new implementation of the relationship between social rights and the rights of freedom. The service model has demonstrated that weakness that the access to qualified services in the field of health or education is becoming more and more dependent of the private purchasing power. The weakness of the investment model is that it represents a sort of “The Winner takes it all” – since a political majority is enabled to make agendas in societal fields former protected by the tripartite power and the rights of freedom of the citizens. The outcome of the Danish development seems to be an establishment of a political governed public service industry which on one side are capable of competing on market conditions and on the other are able being governed by contracts. This represents a new form of close linking of politics, economy and professional work. Attempts of controlling education, pedagogy and thereby the population are not a recent invention. In European history we could easily point at several such experiments. The real news is the linking between political priorities and exercise of public activities by economic incentives. By defining visible goals for the public servants, by introducing measurement of achievements and effects, and by implementing a new wage policy depending on achievements and/or effects a new system of accountability is manufactured. The consequences are already perceptible. The government decides to do some special interventions concerning parents, children or youngsters, the public servants on municipality level are instructed to carry out their services by following a manual, and the parents are no longer protected by privacy. Protection of privacy and minority is no longer a valuable argumentation to prevent further interventions in people’s life (health, food, school, etc.). The citizens are becoming objects of investment, also implying that people are investing in their own health, education, and family. This means that investments in changes of life style and development of competences go hand in hand. The below mentioned programmes are conditioned by this shift.

Impact of SLM build parameters on the surface quality

Additive manufacturing by melting of metal powders is an innovative method to create one-offs and customized parts. Branches like dentistry, aerospace engineering and tool making were indicated and the manufacturing methods are established. Besides all the advantages, like freedom of design, manufacturing without a tool and the reduction of time-to-market, there are however some disadvantages, such as reproducibility or the surface quality. The surface quality strongly depends on the orientation of the component in the building chamber, the process parameters which are laser power and exposure time, but also on the so-called “hatch”-strategy, which includes the way the laser exposes the solid areas. This paper deals with the investigation and characterization of the surface quality of generated parts produced by SLM. Main process parameters including part orientation, part size and hatch strategies are investigated and monitored. The outcome is a recommendation of suitable hatch strategies depending on desired part properties. This includes metered values and takes into account process stability and reproducibility.

Who Is Afraid of School Choice?

This study uses survey data to investigate attitudes among Swiss voters to different models offering more freedom of choice in the educational system. There is a clear opposition to the use of taxpayer money to fund private schools, while free choice between public schools seems to appeal to a majority. The opinions appear to be based on a rational calculation of personal utility. For both types of choice, approval rates are lower for middle to high-income groups and individuals with a teaching qualification. Furthermore, residents of small to medium-sized towns are opposed to more school choice. On the support side, approval rates for private school choice are higher among parents of school-age children and residents in urban areas. The results also indicate differences between the country's language regions, attributable to intercultural differences in what people consider the role of the state.

miR-21 and its target gene CCL20 are both highly overexpressed in the microenvironment of colorectal tumors: significance of their regulation

Recently, we reported a functional interaction between miR-21 and its identified chemokine target CCL20 in colorectal cancer (CRC) cell lines. Here, we investigated whether such functional interactions are permitted at the cellular level which would require an inverse correlation of expression and also co-expression of miR-21 and CCL20 in the same cell. Expression profiling was performed using qPCR, and ELISA, in situ hybridization and immunohistochemistry were applied for the presentation of their cellular localization. We demonstrated that miR-21 as well as CCL20 were both significantly upregulated in CRC tissues; thus, showing no antidromic expression pattern. This provided an initial clue that miR-21 and CCL20 may not be expressed in the same cell. In addition, we located miR-21 expression at the cellular level predominantly in stromal cells such as tumor-associated fibroblasts and to a minor degree in immune cells such as macrophages and lymphocytes. Likewise, CCL20 expression was primarily detected in tumor-infiltrating immune cells. Thus, investigating the cellular localization of miR-21 and its target CCL20 revealed that both molecules are expressed predominantly in the microenvironment of CRC tumors.

Multidrug resistance protein 4 (MRP4) expression in prostate cancer is associated with androgen signaling and decreases with tumor progression

Multidrug resistance protein 4 (MRP4) is a transmembrane transport protein found in many cell types and is involved in substrate-specific transport of endogenous and exogenous substrates. Recently, it has shown to be expressed in prostate cancer cell lines and to be among the most commonly upregulated transcripts in prostate cancer, although a comprehensive expression analysis is lacking so far. We aimed to investigate its expression by immunohistochemistry in a larger cohort of neoplastic and nonneoplastic prostate tissues (n = 441) and to correlate its expression with clinicopathological parameters including PSA-free survival times and molecular correlates of androgen signaling (androgen receptor (AR), prostate-specific antigen (PSA), and forkhead box A (FoxA)). MRP4 is widely expressed in benign and neoplastic prostate epithelia, but its expression gradually decreases during tumor progression towards castrate-resistant disease. Concordantly, it correlated with conventional prognosticators of disease progression and-within the group of androgen-dependent tumors-with AR and FoxA expression. Moreover, lower levels of MRP4 expression were associated with shorter PSA relapse-free survival times in the androgen-dependent group. In benign tissues, we found zone-dependent differences of MRP4 expression, with the highest levels in the peripheral and central zones. Although MRP4 is known to be regulated in prostate cancer, this study is the first to demonstrate a gradual downregulation of MRP4 protein during malignant tumor progression and a prognostic value of this loss of expression.

Dynamics of a minimal model of interlocked positive and negative feedback loops of transcriptional regulation by cAMP-response element binding proteins.

cAMP-response element binding (CREB) proteins are involved in transcriptional regulation in a number of cellular processes (e.g., neural plasticity and circadian rhythms). The CREB family contains activators and repressors that may interact through positive and negative feedback loops. These loops can be generated by auto- and cross-regulation of expression of CREB proteins, via CRE elements in or near their genes. Experiments suggest that such feedback loops may operate in several systems (e.g., Aplysia and rat). To understand the functional implications of such feedback loops, which are interlocked via cross-regulation of transcription, a minimal model with a positive and negative loop was developed and investigated using bifurcation analysis. Bifurcation analysis revealed diverse nonlinear dynamics (e.g., bistability and oscillations). The stability of steady states or oscillations could be changed by time delays in the synthesis of the activator (CREB1) or the repressor (CREB2). Investigation of stochastic fluctuations due to small numbers of molecules of CREB1 and CREB2 revealed a bimodal distribution of CREB molecules in the bistability region. The robustness of the stable HIGH and LOW states of CREB expression to stochastic noise differs, and a critical number of molecules was required to sustain the HIGH state for days or longer. Increasing positive feedback or decreasing negative feedback also increased the lifetime of the HIGH state, and persistence of this state may correlate with long-term memory formation. A critical number of molecules was also required to sustain robust oscillations of CREB expression. If a steady state was near a deterministic Hopf bifurcation point, stochastic resonance could induce oscillations. This comparative analysis of deterministic and stochastic dynamics not only provides insights into the possible dynamics of CREB regulatory motifs, but also demonstrates a framework for understanding other regulatory processes with similar network architecture.