Analysis of the Expression Pattern of Wnt Receptor Frizzled 3

Wnt signaling plays a vital role in many developmental processes. Wnt signaling has been implicated in neural crest induction and cell differentiation among other functions. In mice Wnts comprise a family of nineteen glycoproteins that bind to Frizzled (Fzd) receptors and LRP5/6 co-receptors. This activates beta-catenin, which translocates into the nucleus and acts as a transcription factor, resulting in differential gene expression. Specifically, Fzd 3 enhances Wnt 1 signaling. Wnt 1 and Fzd 3 are involved in neural crest induction and in neural crest-derived melanocyte development. We analyzed the expression pattern ofFzd 3 and the LRP 5/6 by in situ hybridization inmouse embryos. Our data suggests a role for these genes in neural crest induction and in melanocyte differentiation in the murine system. Results show Fzd 3 expression in the anterior part of the neural tube and in the hindbrain, while LRP 5 is expressed in the anterior part of the neural tube, in the hindbrain, and in the eye. We conclude that Fzd 3 and LRP 5 are expressed in the neural crest. In addition, Fzd 3 might act as the receptor while LRP 5 might act as the co-receptor for Wntl signaling in the murine system.

Analysis and comparison of the moral development of students required to graduate with an ethics course

Accounting students become practitioners facing ethical decision-making challenges that can be subject to various interpretations; hence, the profession is concerned with the appropriateness of their decisions. Moral development of these students has implications for a profession under legal challenges, negative publicity, and government scrutiny. Accounting students moral development has been studied by examining their responses to moral questions in Rest's Defining Issues Test (DIT), their professional attitudes on Hall's Professionalism Scale Dimensions, and their ethical orientation-based professional commitment and ethical sensitivity. This study extended research in accounting ethics and moral development by examining students in a college where an ethics course is a requirement for graduation. Knowledge of differences in the moral development of accounting students may alert practitioners and educators to potential problems resulting from a lack of ethical understanding as measured by moral development levels. If student moral development levels differ by major, and accounting majors have lower levels than other students, the conclusion may be that this difference is a causative factor for the alleged acts of malfeasance in the profession that may result in malpractice suits. The current study compared 205 accounting, business, and nonbusiness students from a private university. In addition to academic major and completion of an ethics course, the other independent variable was academic level. Gender and age were tested as control variables and Rest's DIT score was the dependent variable. The primary analysis was a 2x3x3 ANOVA with post hoc tests for results with significant p-value of less than 0.05. The results of this study reveal that students who take an ethics course appear to have a higher level of moral development (p=0.013), as measured by the (DIT), than students at the same academic level who have not taken an ethics course. In addition, a statistically significant difference (p=0.034) exists between freshmen who took an ethics class and juniors who did not take an ethics class. For every analysis except one, the lower class year with an ethics class had a higher level of moral development than the higher class year without an ethics class. These results appear to show that ethics education in particular has a greater effect on the level of moral development than education in general. Findings based on the gender specific analyses appear to show that males and females respond differently to the effects of taking an ethics class. The male students do not appear to increase their moral development level after taking an ethics course (p=0.693) but male levels of moral development differ significantly (p=0.003) by major. Female levels of moral development appear to increase after taking an ethics course (p=0.002). However, they do not differ according to major (p=0.0 97). These findings indicate that accounting students should be required to have a class in ethics as part of their college curriculum. Students with an ethics class have a significantly higher level of moral development. The challenges facing the profession at the current time indicate that public confidence in the reports of client corporations has eroded and one way to restore this confidence could be to require ethics training of future accountants.

Observation of manganese coating and encrustation in different environments of the North-Western Atlantic ocean

The major topographic features, or provinces, beyond the continental slope off the Atlantic coast of the United States are (1) Sohm Plain, (2) Hatteras Plain, (3) Nares Plain, (4) Blake Basin, (5) Blake Plateau-Bahama Banks, and (6) Bermuda Rise. The whole of the described area is commonly referred to as the North American Basin. This basin is bounded on the north by Newfoundland Ridge and on the south by Puerto Rico Trench. Topographic features of note within the basin are the divide and the area of depressions between Sohm and Hatteras Plains, the sharply crested Blake Ridge, and the Puerto Rico Ridge. Recently accumulated data on deep-sea oores has given good evidence that the silt and sand covering the abyssal plains are displaced continental sediments in a virtually quartz-free oceanic environment. These sediments were deposited on a primary volcanic bottom. The primary or volcanic bottom is characterized by abyssal hills and seamounts, and the sediment bottom is characterized by abyssal plains, which extend seaward from the continental margins. On the Blake Plateau, bottom photographs and dredge hauls in the axis of the stream show that locally sediment has been removed and the bottom is paved with crusts and nodules of manganese. Photographs and dredged samples from the outer part of the New England Seamount, Chain and Caryn Peak also indicate extensive encrustations of manganese oxide which acts as a binding agent in areas of ooze or other organic debris and thus helps to stabilize the bottom.

Function of the PDLIM2 protein in oncogenic Wnt signalling pathways

Aberrant regulation of the Wnt signalling pathway is a recurrent theme in cancer biology. Hyper activation due to oncogenic mutations and paracrine activity has been found in both colon cancer and breast cancer, and continues to evolve as a central mechanism in oncogenesis. PDLIM2, a cytoskeletal PDZ protein, is an IGF-1 regulated gene that is highly expressed in cancer cell lines derived from metastatic tumours. Suppression of PDLIM2 inhibits polarized cell migration, reverses the Epithelial to Mesenchymal transition (EMT) phenotype, suppresses the transcription of β-catenin target genes, and regulates gene expression of key transcription factors in EMT. This thesis investigates the mechanism by which PDLIM2 contributes to the maintenance of Wnt signalling in cancer cells. Here we show that PDLIM2 is a critical regulator of the Wnt pathway by regulating β-catenin at the adherens juctions, as also its transcriptional activity by the interaction of PDLIM2 with TCF4 at the nucleus. Evaluation of PDLIM2 in macrophages and co-culture studies with cancer cells and fibroblasts showed the influence exerted on PDLIM2 by paracrine cues. Thus, PDLIM2 integrates cytoskeleton signalling with gene expression by modulating the Wnt signalling pathway and reconciling microenvironmental cues with signals in epithelial cells. Negative correlation of mRNA and protein levels in the triple negative breast cancer cell BT549 suggests that PDLIM2 is part of a more complex mechanism that involves transcription and posttranslational modifications. GST pulldown studies and subsequent mass spectrometry analysis showed that PDLIM2 interacts with 300 proteins, with a high biological function in protein biosynthesis and Ubiquitin/proteasome pathways, including 13 E3 ligases. Overall, these data suggest that PDLIM2 has two distinct functions depending of its location. Located at the cytoplasm mediates cytoskeletal re-arrangements, whereas at the nucleus PDLIM2 acts as a signal transduction adaptor protein mediating transcription and ubiquitination of key transcription factors in cancer development.

Electrophysiological study of the effects of arginine vasopressin in the rat juxtacapsular nucleus of the bed nucleus of the stria terminalis

Arginine vasopressin (AVP), a nine amino acid neuropeptide (CYFQNCPRG- NH2) fulfills a dual function: (i) in the periphery, AVP acts as a peptide hormone and (ii) in the CNS, AVP is a neuromodulatory peptide. AVP produces its effects through 3 AVP receptors (AVPRs). AVPR1a and AVPR1b are expressed in the CNS and periphery, whilst AVPR2 is not found centrally but instead solely expressed in the kidneys. Recent evidence revealed a high density of AVP-binding sites in the juxtacapsular nucleus of the bed nucleus of the stria terminalis (jxBNST). While in other regions of the brain, AVP acts at AVPRs to regulate an array of biological processes, including male-typical social behaviours, social memory, stress adaptation, fear, anxiety, and fluid homeostasis, its role in the jxBNST remains elusive. Furthermore, the neurophysiological properties of AVP in the jxBNST are unknown so this study aimed to examine how AVP modulates synaptic transmission in the rat jxBNST. The BNST being one of the most notable sexually dimorphic brain regions and AVPR expression being influenced by gonadal steroids, we investigated the putative influence of sex on the modulatory effects of AVP in the jxBNST. Finally, due to AVP being released at a substantially higher concentration following periods of water deprivation, we examined changes in AVPs modulatory role following water deprivation. Male and female Long Evans rats were euthanized and brain slice whole-cell voltage-clamp electrophysiology was done in the jxBNST to measure the effects of AVP on synaptic transmission of GABA synapses. Exogenous application of AVP produced three responses; either postsynaptic long-term potentiation (LTP) of GABAA-inhibitory postsynaptic currents (IPSC), postsynaptic long-term depression (LTD) of GABAA-IPSC, or no change in GABAA-IPSC amplitudes. Interestingly, the proportion of neurons responding in each of these ways did not differ between sexes and within females was not estrous cycle-dependent. Finally, although not statistically significant, 24-hour water deprivation abolished GABAA-LTD, an effect that was not a consequence of social isolation. Taken together, our data show that AVP modulates GABAA synaptic transmission in the jxBNST in fluid homeostasis- but not sex-dependent manner.

Floristic And Ethnobotanical Studies Of Medicinal Plants Of The City Of El -Jadida (MOROCCO)

Abstract: BRIGUICHE. H, ZIDANE. L. Floristic And Ethnobotanical Studies Of Medicinal Plants Of The City Of El -Jadida (MOROCCO). In the framework of the ethnobotanical studies on medicinal plants undertaken by the Laboratory of Biodiversity and Natural Resources of the Faculty of Sciences of Kenitra (Morocco), we are interested in the area of El Jadida which presents a rather important floristic richness thanks to changes in its ecological conditions By using 204 questionnaire, the ethnobotanical surveys were conducted in the field during the years 2012-2013. The location of the different sampling sites was determined by the stratified sampling method. The analysis of the results obtained from the questionnaires and forms using statistical processing allowed us to identify 70 plant species distributed in 69 genera and 37 families. These results also show that most of these species are mainly used in the care of the digestive system and respiratory system. The seed is the most used part in local traditional medicines and the decoction is the most frequent mode with a rate of 31%. The species Origanum compactum is the most used by the population of the city of El Jadida 47 quotes.  

Acts of Solidarity: Crossing and Reiterating Israeli-Palestinian Frontiers

In academic and public discourses on the Zionist-Palestinian conflict prevails still a ‘methodological nationalism’ based on a separatist imagination that overshadows the existence and role of Israeli-Palestinian forms of communality and solidarity. This article analyzes micropolitical practices that cross existing frontiers both within Israel and between occupied Palestinian territories and Israel. Through recent conceptualizations of ‘acts’, I read these ethnographic episodes in their intentional and performative dimension. What is the role of these ‘acts’? What are their effects on both the participants and the wider public? Through two interconnected cases, different functions of acts are explored. The first case relates to encounters between Israelis and Palestinian in the embattled city of Hebron in the occupied Palestinian territories; the second investigates moments of a Gandhi-inspired peace march at the ‘internal’ frontier of the Israeli Negev desert. The ethnographic perspective reveals what lies behind and beneath the acts, going beyond the obvious structures of power of the conflict. Acts function primarily as a valve of catharsis for the participants themselves, both overcoming and reproducing hegemonic discursive elements of the conflict. Paradoxically, acts of solidarity are often crucial in creating public knowledge about the conflict in more sectarian terms. 

Laser-Driven Ion Accelerators: State of the Art and Applications

Laser-plasma based accelerators of protons and heavier ions are a source of potential interest for several applications, including in the biomedical area. While the potential future use in cancer hadrontherapy acts as a strong aspirational motivation for this research field, radiobiology employing laser-driven ion bursts is alreadyan active field of research. Here we give a summary of the state of the art in laser driven ion acceleration, of the main challenges currently faced by the research inthis field and of some of the current and future strategies for overcoming them.

Prognostic role of the LCS6 KRAS variant in locally advanced rectal cancer: results of the EXPERT-C trial.

Background: Lethal-7 (let-7) is a tumour suppressor miRNA which acts by down-regulating several oncogenes including KRAS. A single-nucleotide polymorphism (rs61764370, T > G base substitution) in the let-7 complementary site 6 (LCS-6) of KRAS mRNA has been shown to predict prognosis in early-stage colorectal cancer (CRC) and benefit from anti-epidermal growth factor receptor monoclonal antibodies in metastatic CRC. Patients and methods: We analysed rs61764370 in EXPERT-C, a randomised phase II trial of neoadjuvant CAPOX followed by chemoradiotherapy, surgery and adjuvant CAPOX plus or minus cetuximab in locally advanced rectal cancer. DNA was isolated from formalin-fixed paraffin-embedded tumour tissue and genotyped using a PCR-based commercially available assay. Kaplan–Meier method and Cox regression analysis were used to calculate survival estimates and compare treatment arms. Results: A total of 155/164 (94.5%) patients were successfully analysed, of whom 123 (79.4%) and 32 (20.6%) had the LCS-6 TT and LCS-6 TG genotype, respectively. Carriers of the G allele were found to have a statistically significantly higher rate of complete response (CR) after neoadjuvant therapy (28.1% versus 10.6%; P = 0.020) and a trend for better 5-year progression-free survival (PFS) [77.4% versus 64.5%: hazard ratio (HR) 0.56; P = 0.152] and overall survival (OS) rates (80.3% versus 71.9%: HR 0.59; P = 0.234). Both CR and survival outcomes were independent of the use of cetuximab. The negative prognostic effect associated with KRAS mutation appeared to be stronger in patients with the LCS-6 TT genotype (HR PFS 1.70, P = 0.078; HR OS 1.79, P = 0.082) compared with those with the LCS-6 TG genotype (HR PFS 1.33, P = 0.713; HR OS 1.01, P = 0.995). Conclusion: This analysis suggests that rs61764370 may be a biomarker of response to neoadjuvant treatment and an indicator of favourable outcome in locally advanced rectal cancer possibly by mitigating the poor prognosis of KRAS mutation. In this setting, however, this polymorphism does not appear to predict cetuximab benefit.

Molecular Mechanism of the Response to Hydrogen Sulfide in C. elegans

Thesis (Ph.D.)--University of Washington, 2016-08

Spiroacetals in the Colonization Behaviour of the Coffee Berry Borer: A 'Push-Pull' System

Coffee berries are known to release several volatile organic compounds, among which is the spiroacetal, conophthorin, an attractant for the coffee berry borer Hypothenemus hampei. Elucidating the effects of other spiroacetals released by coffee berries is critical to understanding their chemo-ecological roles in the host discrimination and colonization process of the coffee berry borer, and also for their potential use in the management of this pest. Here, we show that the coffee berry spiroacetals frontalin and 1,6-dioxaspiro [4.5] decane (referred thereafter as brocain), are also used as semiochemicals by the coffee berry borer for host colonization. Bioassays and chemical analyses showed that crowding coffee berry borers from 2 to 6 females per berry, reduced borer fecundity, which appeared to correlate with a decrease in the emission rates of conophthorin and frontalin over time. In contrast, the level of brocain did not vary significantly between borer-uninfested and infested berries. Brocain was attractive at lower doses, but repellent at higher doses while frontalin alone or in a blend was critical for avoidance. Field assays with a commercial attractant comprising a mixture of ethanol and methanol (1:1), combined with frontalin, confirmed the repellent effect of this compound by disrupting capture rates of H. hampei females by 77% in a coffee plantation. Overall, our results suggest that the levels of frontalin and conophthorin released by coffee berries determine the host colonization behaviour of H. hampei, possibly through a 'push-pull' system, whereby frontalin acts as the 'push' (repellent) and conophthorin acting as the 'pull' (attractant). Furthermore, our results reveal the potential use of frontalin as a repellent for management of this coffee pest.

Investigation of the role of long non-coding RNAs in oncogene induced cellular senescence

Cellular senescence is a stable arrest of cell proliferation induced by several factors such as activated oncogenes, oxidative stress and shortening of telomeres. Senescence acts as a tumour suppression mechanism to halt the progression of cancer. However, senescence may also impact negatively upon tissue regeneration, thus contributing to the effects of ageing. The eukaryotic genome is controlled by various modes of transcriptional and translational regulation. Focus has therefore centred on the role of long non- coding RNAs (lncRNAs) in regulating the genome. Accordingly, understanding how lncRNAs function to regulate the senescent genome is integral to improving our knowledge and understanding of tumour suppression and ageing. Within this study, I set out to investigate the expression of lncRNAs’ expression within models of senescence. Through a custom expression array, I have shown that expression of multiple different lncRNAs is up-regulated and down regulated in IMR90 replicative senescent fibroblasts and oncogene-induced senescent melanocytes. LncRNA expression was determined to be specific to stable senescence-associated cell arrest and predominantly within the nucleus of senescent cells. In order to examine the function of lncRNA expression in senescence, I selected lncRNA transcript ENST0000430998 (lncRNA_98) to focus my investigations upon. LncRNA_98 was robustly upregulated within multiple models of senescence and efficiently depleted using anti-sense oligonucleotide technology. Characterisation and unbiased RNA-sequencing of lncRNA_98 deficient senescent cells highlighted a list of genes that are regulated by lncRNA_98 expression in senescent cells and may regulate aspects of the senescence program. Specifically, the formation of SAHF was impeded upon depletion of lncRNA_98 expression and levels of total pRB protein expression severely decreased. Validation and recapitulation of consequences of pRB depletion was confirmed through lncRNA_98 knock-out cells generated using CRISPR technology. Surprisingly, inhibition of ATM kinase functions permitted the restoration of pRB protein levels within lncRNA_98 deficient cells. I propose that lncRNA_98 antagonizes the ability of ATM kinase to downregulate pRB expression at a post-transcriptional level, thereby potentiating senescence. Furthermore, lncRNA expression was detected within fibroblasts of old individuals and visualised within senescent melanocytes in human benign nevi, a barrier to melanoma progression. Conversely, mining of 337 TCGA primary melanoma data sets highlighted that the lncRNA_98 gene and its expression was lost from a significant proportion of melanoma samples, consistent with lncRNA_98 having a tumour suppressor functions. The data presented in this study illustrates that lncRNA_98 expression has a regulatory role over pRB expression in senescence and may regulate aspects of tumourigenesis and ageing.

The metabolic and epigenetic effects of the isocitrate dehydrogenase-1 mutation in glioma

Cancer cells have been noted to have an altered metabolic phenotype for over ninety years. In the presence of oxygen, differentiated cells predominately utilise the tricarboxylic acid (TCA) cycle and oxidative phosphorylation to efficiently produce energy and the metabolites necessary for protein and lipid synthesis. However, in hypoxia, this process is altered and cells switch to a higher rate of glycolysis and lactate production to maintain their energy and metabolic needs. In cancer cells, glycolysis is maintained at a high rate, even in the presence of oxygen; a term described as “aerobic glycolysis”. Tumour cells are rapidly dividing and have a much greater need for anabolism compared to normal differentiated cells. Rapid glucose metabolism enables faster ATP production as well as a greater redistribution of carbons to nucleotide, protein, and fatty acid synthesis, thus maximising cell growth. Recently, other metabolic changes, driven by mutations in genes related to the TCA cycle, indicate an alternative role for metabolism in cancer, the “oncometabolite”. This is where a particular metabolite builds up within the cell and contributes to the tumorigenic process. One of these genes is isocitrate dehydrogenase (IDH) IDH is an enzyme that forms part of the tricarboxylic acid (TCA) cycle and converts isocitrate to α-ketoglutarate (α-KG). It exists in three isoforms; IDH1, IDH2 and IDH3 with the former present in the cytoplasm and the latter two in the mitochondria. Point mutations have been identified in the IDH1 and IDH2 genes in glioma which result in a gain of function by converting α-KG to 2-hydroxyglutarate (2HG), an oncometabolite. 2HG acts as a competitive inhibitor of the α-KG dependent dioxygenases, a superfamily of enzymes that are involved in numerous cellular processes such as DNA and histone demethylation. It was hypothesised that the IDH1 mutation would result in other metabolic changes in the cell other than 2HG production, and could potentially identify pathways which could be targeted for therapeutic treatment. In addition, 2HG can act as a potential competitive inhibitor of α-KG dependent dioxygenases, so it was hypothesised that there would be an effect on histone methylation. This may alter gene expression and provide a mechanism for tumourogenesis and potentially identify further therapeutic targets. Metabolic analysis of clinical tumour samples identified changes associated with the IDH1 mutation, which included a reduction in α-KG and an increase in GABA, in addition to the increase in 2HG. This was replicated in several cell models, where 13C labelled metabolomics was also used to identify a possible increase in metabolic flux from glutamate to GABA, as well as from α-KG to 2HG. This may provide a mechanism whereby the cell can bypass the IDH1 mutation as GABA can be metabolised to succinate in the mitochondria by GABA transaminase via the GABA shunt. JMJ histone demethylases are a subset of the α-KG dependent dioxygenases, and are involved in removing methyl groups from histone tails. Changes in histone methylation are associated with changes in gene expression depending on the site and extent of chemical modification. To identify whether the increase in 2HG and fall in α-KG was associated with inhibition of histone demethylases a histone methylation screen was used. The IDH1 mutation was associated with an increase in methylation of H3K4, which is associated with gene activation. ChiP and RNA sequencing identified an increase in H3K4me3 at the transcription start site of the GABRB3 subunit, resulting in an increase in gene expression. The GABRB3 subunit forms part of the GABA-A receptor, a chloride channel, which on activation can reduce cell proliferation. The IDH1 mutation was associated with an increase in GABA and GABRB3 subunit of the GABA-A receptor. This raises the possibility of GABA transaminase as a potential therapeutic target. Inhibition of this enzyme could reduce GABA metabolism, potentially reducing any beneficial effect of the GABA shunt in IDH1 mutant tumours, and increasing activation of the GABA-A receptor by increasing the concentration of GABA in the brain. This in turn may reduce cell proliferation, and could be achieved by using Vigabatrin, a GABA transaminase inhibitor licensed for use in epilepsy.

Every nation under heaven: the importance of the biblical concept of nations for contemporary mission practice

Is there a concept of nationhood in the Bible that can provide us with a framework for cross-cultural Christian mission? This thesis argues that current evangelical missiology has accepted too willingly the categories of the secular Enlightenment understanding of ethnicity and nationhood, and that it needs to rethink its understanding of nations from a biblical standpoint. While the pressures of globalisation are seen by some as rapidly eclipsing the nation-state, this thesis will argue that we need to move beyond the narrower secular categories of citizenship, political power and the boundaries of the state to recover a more biblical understanding of nationhood. By reference to Genesis 10-11, Acts 2:1-11 and those passages in the Book of Revelation that discuss the destiny of the nations, it will show that the biblical understanding of nations includes deeper ideas of shared history, culture and language as the essential components of nationhood. It will explain how nations are part of the created order, and explore the impact of the Babel narrative on our understanding of nations in relation to God. It will demonstrate that Pentecost did not reverse the curse of Babel, but served rather to honour the dignity and value of nations and their languages. It will also argue that nations have a destiny in the New Creation according to the Book of Revelation. This biblical concept of nationhood has significant implications in several areas: the development of a public theology; a Christian response to nationalism; the question of how urban mission fits within mission to the nations; and the importance of indigenous languages in cross-cultural mission, especially in the multicultural cities of Europe.

The insect repellent N,N-diethyl-m-toluamide (DEET) induces angiogenesis via allosteric modulation of the M3 muscarinic receptor in endothelial cells

International audience