Mode of action of FMRFamide-like peptide on drosophila body wall muscle conractions

Neuropeptides are the largest group of signalling chemicals that can convey the information from the brain to the cells of all tissues. DPKQDFMRFamide, a member of one of the largest families of neuropeptides, FMRFamide-like peptides, has modulatory effects on nerve-evoked contractions of Drosophila body wall muscles (Hewes et aI.,1998) which are at least in part mediated by the ability of the peptide to enhance neurotransmitter release from the presynaptic terminal (Hewes et aI., 1998, Dunn & Mercier., 2005). However, DPKQDFMRFamide is also able to act directly on Drosophila body wall muscles by inducing contractions which require the influx of extracellular Ca 2+ (Clark et aI., 2008). The present study was aimed at identifying which proteins, including the membrane-bound receptor and second messenger molecules, are involved in mechanisms mediating this myotropic effect of the peptide. DPKQDFMRFamide induced contractions were reduced by 70% and 90%, respectively, in larvae in which FMRFamide G-protein coupled receptor gene (CG2114) was silenced either ubiquitously or specifically in muscle tissue, when compared to the response of the control larvae in which the expression of the same gene was not manipulated. Using an enzyme immunoassay (EIA) method, it was determined that at concentrations of 1 ~M- 0.01 ~M, the peptide failed to increase cAMP and cGMP levels in Drosophila body wall muscles. In addition, the physiological effect of DPKQDFMRFamide at a threshold dose was not potentiated by 3-lsobutyl-1-methylxanthine, a phosphodiesterase inhibitor, nor was the response to 1 ~M peptide blocked or reduced by inhibitors of cAMP-dependent or cGMP-dependent protein kinases. The response to DPKQDFMRFamide was not affected in the mutants of the phosholipase C-~ (PLC~) gene (norpA larvae) or IP3 receptor mutants, which suggested that the PLC-IP3 pathway is not involved in mediat ing the peptide's effects. Alatransgenic flies lacking activity of calcium/calmodul in-dependent protein kinase (CamKII showed an increase in muscle tonus following the application of 1 JlM DPKQDFMRFamide similar to the control larvae. Heat shock treatment potentiated the response to DPKQDFMRFamide in both ala1 and control flies by approximately 150 and 100 % from a non heat-shocked larvae, respectively. Furthermore, a CaMKII inhibitor, KN-93, did not affect the ability of peptide to increase muscle tonus. Thus, al though DPKQDFMRFamide acts through a G-protein coupled FMRFamide receptor, it does not appear to act via cAMP, cGMP, IP3, PLC or CaMKl1. The mechanism through which the FMRFamide receptor acts remains to be determined.

Mode of action of a Drosophila FMRFamide in inducing muscle contraction

Drosophila melanogaster is a model system for examining the mechanisms of action of neuropeptides. DPKQDFMRFamide was previously shown to induce contractions in Drosophila body wall muscle fibres in a Ca(2+)-dependent manner. The present study examined the possible involvement of a G-protein-coupled receptor and second messengers in mediating this myotropic effect after removal of the central nervous system. DPKQDFMRFamide-induced contractions were reduced by 70% and 90%, respectively, in larvae with reduced expression of the Drosophila Fmrf receptor (FR) either ubiquitously or specifically in muscle tissue, compared with the response in control larvae in which expression was not manipulated. No such effect occurred in larvae with reduced expression of this gene only in neurons. The myogenic effects of DPKQDFMRFamide do not appear to be mediated through either of the two Drosophila myosuppressin receptors (DmsR-1 and DmsR-2). DPKQDFMRFamide-induced contractions were not reduced in Ala1 transgenic flies lacking activity of calcium/calmodulin-dependent protein kinase (CamKII), and were not affected by the CaMKII inhibitor KN-93. Peptide-induced contractions in the mutants of the phospholipase C-β (PLCβ) gene (norpA larvae) and in IP3 receptor mutants were similar to contractions elicited in control larvae. The peptide failed to increase cAMP and cGMP levels in Drosophila body wall muscles. Peptide-induced contractions were not potentiated by 3-isobutyl-1-methylxanthine, a phosphodiesterase inhibitor, and were not antagonized by inhibitors of cAMP-dependent or cGMP-dependent protein kinases. Additionally, exogenous application of arachidonic acid failed to induce myogenic contractions. Thus, DPKQDFMRFamide induces contractions via a G-protein coupled FMRFamide receptor in muscle cells but does not appear to act via cAMP, cGMP, IP3, PLC, CaMKII or arachidonic acid.

A study of thermal decompositions of an allylic hydroperoxide /|nby Thomas A. McCarrick. -- 260 St. Catharines [Ont.] : Dept. of Chemistry, Brock University,

The thermal decomposition of 2,3-di~ethy l - J-hydr operox y- 1 - butene , p r epared f rol") singl e t oxygen, has been studied i n three solvents over the tempe r a ture r ange from 1500e to l o00e and t!1e i 111 t ial ~oncentrfttl nn r Ange from O. 01 M to 0.2 M. Analys i s of the kine tic data ind ica te s i nduced homolysis as the n ost probRble mode of d e composition, g iving rise to a 3/2 f S order dependence upon hy d.roperoxide concent :r8.tl on . Experimental activation e nergies for the decomposition were f ound to be between 29.5 kcsl./raole and 30.0 k cal./mole .• \,iith log A factors between 11 . 3 and 12.3. Product studies were conducted in R variety of solvents a s well as in the pr esence of a variety of free r adical initiators . Investigation of the kinetic ch a in length indicated a chain length of about fifty. A degenerat i ve chain branching mechanism 1s proposed which predicts the multi t ude of products which Rre observed e xperimentally as well as giving activation energies and log A factors si~il a r to those found experimentally .

Out of the darkness and into the light : a study of inauthentic and authentic speech

Introduction Man can be described as the being who shows himself in speech, and from birth to death is continually speaking. Communication is so close to us, so woven into our very being, that we have little understanding of the way it is constituted; for it is as hard to obtain distance from communication as it is to obtain distance from ourselves. All communication is not alike. There are two basic modesl of communication, the inauthentic and the authentic, between which there occurs a constant tension. It is in the inauthentic mode, points out Heidegger, that we find ourselves "proximately and for the most part"; 1. Being and Time, pg. 68 Dasein decides as to the way it will comport itself in taking up its task of having being as an issue for it. " •.• it~, in its very being 'choose' itself and win itself; it can also lose itself and never win itself or only "seem" to do so. But only in so far as it is essentially something which can be authentic--that is, something of its own--can it have lost itself and not yet won itself." 2. therefore Heidegger also terms it "everydayness".2 Caught up in the world of everydayness, our speaking covers over and conceals3 our rootedness in being, leaving us in the darkness of untruth. The image of darkness may be inferred from Heidegger's use of the image of "clearing,,4 to depict being as 2. ibid. pg. 69 "Dasein's average everydayness, however, is not to be taken as a mere 'aspect'. Here too, and even in the mode of inauthenticity, the structure of existentiality lies ~ priori and here too Dasein's being is an issue for it in a definite way; and Dasein comports itself towards it the mode of average everydayness, even if this is only the mode of fleeing in the face of it and forgetfulness thereof." 3. ibid. pg. 59 "covering over" and "concealing" are 1;yays Dasein tries to flee its task of having being as an issue for itself. " ••• This being can be covered up so extensively that it becomes forgotten and no question arises about it or its meaning ••• n How everyday speaking accomplishes this will be taken up in detail in the second chapter which explores Dasein's everyday speech. 4. ibid, pg. 171 lI ••• we have in mind nothing other than the Existential - ontological structure of this entity (Dasein), that it is in such a way as to be its 'there'. To say that it is -' illuminated' [tlerleuchtet"] means that as Being-in-theworld it is cleared [gelichtetJ in itself7 not through any other entity, but in such a way that it is itself the clearing. Only for an entity which is eXistentially cleared in this way does what is present-at-hand become accessible in the light or hidden in the dark •••• " 3 dis-coveredness and truth. Our first task will be to explore the nature of communication in general and then to explore each of the modes manifested in turn. The structure of the inauthentic mode of communication can be explored by asking the following questions: What is this speaking about? Who is it that is speaking and who is spoken to? Does this speaking show man in his speech? The authentic mode is distinguished by the rarity with which we encounter it; as the inauthentic conceals, so the authentic reveals our rootedness in being. Yet this rarity makes it difficult to delineate its elusive structure clearly. Its constituent elements can be brought into focus by asking the same questions of this mode that we previously asked of the inauthentic mode. Our initial response to the disclosure of the authentic mode is to attempt to abandon the inauthentic mode and leave the darkness behind dwelling only in the "lighted place". All through the ages, some men pushing this to extreme, have, upon uncovering their relatedness to being, experienced a deep longing to dwell in such a "place" of pure truth and oft times denigrated or attempted to exclude the everyday world. Such 4. flight is twice mistaken: first it atbempts to fix truth as unchanging and static and secondly, it opposes this to untruth which it seeks to abolish. This is both the wrong view of truth and the wrong view of untruth as Heidegger points out in The Origin of The-Work of Art: The Way-to-be of truth, i.e., of discoveredness, is under the sway of refusal. But this refusal is no lack or privation, as if truth could be simply discoveredness rid of all covers. If it could be that, it would no longer be itself . ••• Truth in its way-to-be is untruth.5 Pure light is not the nature of Being nor is pure unconcealedness possible for man. Failure to remember this is the failure to realize that communication destroys itself in such flight because it no longer maintains the contingency of its task, i.e., the dis-closedness of being. We are reminded of the strong attraction this flight from darkness held for Plato. Light, truth and Being are all beyond the darkness and have nothing to do with it. In Book VII of the R~public, Socrates' explanation of the Allegory of the Cave to Glaucon points to a decided preference men have for the "lighted place". 5. The Origin Of The Work Of Art, pg. 42 5. Come then, I said, and join me in this further thought, and do not be surprised that those who attained to this height are not willing to occupy themselves with the affairs of men, but their souls ever feel the upward urge and yearning for that sojourn above. For this, I take it, is likely if in this point too the likeliness of our image holds. 6 Despite the attraction to pure truth, human communication is more complex than putting down one mode of communication and picking up another. Due to the fact that we are always on the way, the title of my thesis will have to be amended: OUT OF THE DARKNESS AND INTO THE LIGHT--AGAIN AND AGAIN. It must be this way because this is what it means to be human. This is the point made by Mephisto to Faust in pointing out that man, standing between God and the devil, needs both darkness and light: Er findet sich in einem ewigen Gl~t Uns hat er in die Finsternis gebracht, Und euch taugt einzig Tag und Nacht. 7 6. Republic z (517 c & d) It should be noted however, that while the philosopherking must be compelled to return to the cave for purely political reasons, once he has taken adequate view of the "brightest region of being" he has the full truth and his return to darkness adds nothing to the truth. 7. Faust, pg. 188 6. This thesis proposes to examine the grounds that give rise to communication, uncovering the structure of its inauthentic and authentic modes and paying close attention to tpeir interrelationship and to their relationship to language as "the house of Being": language that both covers and opens up man's rootedness in Being, transforming him as he moves along his way, taking up his "ownmost task" of becoming who he is. roots. He is the being who shows himself inn that reflects his forgetfulness or remembrance of his rootedness in being. Man comes into an already existent world and is addressedl through things in the world which are c

Bullying in the Same-Sex Friendships of Young Adolescent Girls: A Qualitative Study

This qualitative, narrative study explored the bullying experiences of young adolescent girls within their same-sex dyadic and group friendships. The participants were 5 female students, ages 11 and 12 years old, from 1 private, religious school in southern Ontario. Each girl participated in an audiotaped, 30-minute, personal interview based on an unstructured interview protocol. Interview transcripts were analyzed for bullying behaviors using Marini and Dane's (2008) subtypes of bullying, including the form, function, and involvement in bullying. Interview transcripts were also analyzed for common and emerging themes using aspects of L. M., Brown and Gilligan's (1992) "Listener's Guide." The findings of this study suggested that within their same-sex friendships girls assume the roles of all participants in bullying, including bullies, victims, bystanders, and bully-victims. The findings also suggested that bullying behaviors within young adolescent girls' same-sex friendships are mainly indirect in their mode of attack and that they are both proactive and reactive. The bully behaviors identified in this study were used to inform the major themes or salient features within the dynamics of girls' same-sex friendships also identified. These themes included acceptance, intimacy, negotiation, inclusion/exclusion, moral character judgements, and power. The findings of this study will be used to inform current theory, personal and professional practice, as well as future research.