Delta opioid-receptor function in the dorsal striatum plays a role in high levels of ethanol consumption in rats

Binge-like patterns of excessive drinking during young adulthood increase the propensity for alcohol use disorders (AUDs) later in adult life; however, the mechanisms that drive this are not completely understood. Previous studies showed that the δ-opioid peptide receptor (DOP-R) is dynamically regulated by exposure to ethanol and that the DOP-R plays a role in ethanol-mediated behaviors. The aim of this study was to determine the role of the DOP-R in high ethanol consumption from young adulthood through to late adulthood by measuring DOP-R-mediated [(35)S]GTPγS binding in brain membranes and DOP-R-mediated analgesia using a rat model of high ethanol consumption in Long Evans rats. We show that DOP-R activity in the dorsal striatum and DOP-R-mediated analgesia changes during development, being highest during early adulthood and reduced in late adulthood. Intermittent access to ethanol but not continuous ethanol or water from young adulthood leads to an increase in DOP-R activity in the dorsal striatum and DOP-R-mediated analgesia into late adulthood. Multiple microinfusions of naltrindole into the dorsal striatum or multiple systemic administration of naltrindole reduces ethanol consumption, and following termination of treatment, DOP-R activity in the dorsal striatum is attenuated. These findings suggest that DOP-R activity in the dorsal striatum plays a role in high levels of ethanol consumption and suggest that targeting the DOP-R is an alternative strategy for the treatment of AUDs.

The α5 neuronal nicotinic acetylcholine receptor subunits plays an important role in the sedative effects of ethanol but does not modulate consumption in mice

Alcohol use disorders (AUDs) are a major public health problem, and the few treatment options available to those seeking treatment offer only modest success rates. There remains a need to identify novel targets for the treatment of AUDs. The neuronal nicotinic acetylcholine receptors (nAChRs) represent a potential therapeutic target in the brain, as recent human genetic studies have implicated gene variants in the α5 nAChR subunit as high risk factors for developing alcohol dependence. Here, we evaluate the role of 5* nAChR for ethanol-mediated behaviors using α5+/+ and α5-/- mice. We characterized the effect of hypnotic doses of ethanol and investigated drinking behavior using an adapted Drinking-in-the Dark (DID) paradigm that has been shown to induce high ethanol consumption in mice. We found the α5 subunit to be critical in mediating the sedative effects of ethanol. The α5-/- mice showed slower recovery from ethanol-induced sleep, as measured by loss of righting reflex. Additionally the α5-/- mice showed enhanced impairment to ethanol-induced ataxia. We found the initial sensitivity to ethanol and ethanol metabolism to be similar in both α5+/+ and α5-/- mice. Hence the enhanced sedation is likely due to a difference in the acute tolerance of ethanol in mice deficient of the α5 subunit. However the α5 subunit did not play a role in ethanol consumption for ethanol concentrations ranging from 5% to 30% in the DID paradigm. Additionally, varenicline (Chantix®) was effective in reducing ethanol intake in α5-/- mice. Together, our data suggest that the α5 nAChR subunit is important for the sedative hypnotic doses of ethanol but does not play a role in ethanol consumption. Varenicline can be a treatment option even when there is loss of function of the α5 nAChR subunit.

Exo1 plays a major role in DNA end resection in humans and influences double-strand break repair and damage signaling decisions

The resection of DNA double-strand breaks (DSBs) to generate ssDNA tails is a pivotal event in the cellular response to these breaks. In the two-step model of resection, primarily elucidated in yeast, initial resection by Mre11-CtIP is followed by extensive resection by two distinct pathways involving Exo1 or BLM/WRN-Dna2. However, resection pathways and their exact contributions in humans in vivo are not as clearly worked out as in yeast. Here, we examined the contribution of Exo1 to DNA end resection in humans in vivo in response to ionizing radiation (IR) and its relationship with other resection pathways (Mre11-CtIP or BLM/WRN). We find that Exo1 plays a predominant role in resection in human cells along with an alternate pathway dependent on WRN. While Mre11 and CtIP stimulate resection in human cells, they are not absolutely required for this process and Exo1 can function in resection even in the absence of Mre11-CtIP. Interestingly, the recruitment of Exo1 to DNA breaks appears to be inhibited by the NHEJ protein Ku80, and the higher level of resection that occurs upon siRNA-mediated depletion of Ku80 is dependent on Exo1. In addition, Exo1 may be regulated by 53BP1 and Brca1, and the restoration of resection in BRCA1-deficient cells upon depletion of 53BP1 is dependent on Exo1. Finally, we find that Exo1-mediated resection facilitates a transition from ATM- to ATR-mediated cell cycle checkpoint signaling. Our results identify Exo1 as a key mediator of DNA end resection and DSB repair and damage signaling decisions in human cells.

The human topoisomerase I damage response plays a role in apoptosis

Previous studies have shown that human topoisomerase I cleavage complexes form as a response to various DNA damages in vivo, the so called human topoisomerase I “damage response”. It was suggested that this damage response may play a role in DNA repair as well as in apoptosis, but only very few investigations have been done and the significance of the damage response still remains unclear. Here we demonstrate that human topoisomerase I cleavage complexes induced by high doses of UV irradiation are highly stable for up to 48 h. Furthermore, we show that human topoisomerase I cleavage complexes correlate with apoptosis. However, at low UV doses the cleavage complex level was very low and the complexes were repaired. Surprisingly, we found that high levels of stable cleavage complexes were not only found in UV-irradiated cells but also in untreated cells that underwent apoptosis. A possible role of human topoisomerase I in apoptosis is discussed.

Calcitonin receptor plays a physiological role to protect against hypercalcemia in mice

It is well established that calcitonin is a potent inhibitor of bone resorption; however, a physiological role for calcitonin acting through its cognate receptor, the calcitonin receptor (CTR), has not been identified. Data from previous genetically modified animal models have recognized a possible role for calcitonin and the CTR in controlling bone formation; however, interpretation of these data are complicated, in part because of their mixed genetic background. Therefore, to elucidate the physiological role of the CTR in calcium and bone metabolism, we generated a viable global CTR knockout (KO) mouse model using the Cre/loxP system, in which the CTR is globally deleted by >94% but <100%. Global CTRKOs displayed normal serum ultrafiltrable calcium levels and a mild increase in bone formation in males, showing that the CTR plays a modest physiological role in the regulation of bone and calcium homeostasis in the basal state in mice. Furthermore, the peak in serum total calcium after calcitriol [1,25(OH)2D3]-induced hypercalcemia was substantially greater in global CTRKOs compared with controls. These data provide strong evidence for a biological role of the CTR in regulating calcium homeostasis in states of calcium stress.

The kiwifruit lycopene beta-cyclase plays a significant role in carotenoid accumulation in fruit

The composition of carotenoids, along with anthocyanins and chlorophyll, accounts for the distinctive range of colour found in the Actinidia (kiwifruit) species. Lutein and beta-carotene are the most abundant carotenoids found during fruit development, with beta-carotene concentration increasing rapidly during fruit maturation and ripening. In addition, the accumulation of beta-carotene and lutein is influenced by the temperature at which harvested fruit are stored. Expression analysis of carotenoid biosynthetic genes among different genotypes and fruit developmental stages identified Actinidia lycopene beta-cyclase (LCY-β) as the gene whose expression pattern appeared to be associated with both total carotenoid and beta-carotene accumulation. Phytoene desaturase (PDS) expression was the least variable among the different genotypes, while zeta carotene desaturase (ZDS), beta-carotene hydroxylase (CRH-β), and epsilon carotene hydroxylase (CRH-ε) showed some variation in gene expression. The LCY-β gene was functionally tested in bacteria and shown to convert lycopene and delta-carotene to beta-carotene and alpha-carotene respectively. This indicates that the accumulation of beta-carotene, the major carotenoid in these kiwifruit species, appears to be controlled by the level of expression of LCY-β gene.

Senataxin plays an essential role with DNA damage response proteins in meiotic recombination and gene silencing

Senataxin, mutated in the human genetic disorder ataxia with oculomotor apraxia type 2 (AOA2), plays an important role in maintaining genome integrity by coordination of transcription, DNA replication, and the DNA damage response. We demonstrate that senataxin is essential for spermatogenesis and that it functions at two stages in meiosis during crossing-over in homologous recombination and in meiotic sex chromosome inactivation (MSCI). Disruption of the Setx gene caused persistence of DNA double-strand breaks, a defect in disassembly of Rad51 filaments, accumulation of DNA:RNA hybrids (R-loops), and ultimately a failure of crossing-over. Senataxin localised to the XY body in a Brca1-dependent manner, and in its absence there was incomplete localisation of DNA damage response proteins to the XY chromosomes and ATR was retained on the axial elements of these chromosomes, failing to diffuse out into chromatin. Furthermore persistence of RNA polymerase II activity, altered ubH2A distribution, and abnormal XY-linked gene expression in Setx⁻/⁻ revealed an essential role for senataxin in MSCI. These data support key roles for senataxin in coordinating meiotic crossing-over with transcription and in gene silencing to protect the integrity of the genome.

A rare view: Barbara Hatley’s new book documents a career-long study of, and passion for performance in Central Java

Javanese Performances on an Indonesian Stage: Contesting Culture, Embracing Change, is Barbara Hatley’s first book about the performing arts in Indonesia, a topic that piqued her interest while undergoing a masters program at Yale University in the late 1960s. In this sense, it is a landmark study, for Hatley has since become very well known in Indonesianist circles, especially among those with an interest in matters of culture, popular and elite. Until recently, her writings on Indonesian performing arts have only been available in the form of journal articles and book chapters...

Destinations: 3 Large Cast Plays for Young People

This book is a collection of three large-cast plays written in response to a very specific problem. My work as a teacher of drama often required me to locate a script that would somehow miraculously work for a cast of unknown number and gender, and most likely uneven skills and enthusiasm, who I hadn’t even met yet. It’s a familiar dilemma for teachers and students of drama in education contexts, at whatever level you’re teaching. I’d first addressed this creative problem with scripts such as Gate 38 (2010). I had tried using scripts that already existed, but found they required such extensive editing to suit the parameters of cast and performance duration that I may as well have been writing them myself. Even in the setting of a closed studio, in altering these plays I felt I was bending the vision of the playwright, and certainly their narrative structure, out of shape. Everyone who’s attempted to stage a performance with a large cast of students in an educational setting knows it takes time to truly connect with a play, its social contexts, themes and characters. It also takes a lot of time to get on top of the practicalities of learning, rehearsing, directing and running a performance with young people. Often the curtain goes up on something unfinished and unstable. I was looking for ways to reduce the complexity of staging a script, while maintaining the potential of this process as a site of rich, enjoyable learning. Two of the plays (Duty Free and Please Be Seated) are comprised of multiple monologues, combined with music-driven ensemble sequences. The monologues enable individuals to develop and polish their own performances, work in small groups, and cut down on the laborious detail of directing naturalistic scenes based in character interaction. The third (Australian Drama) involves a lot of duologues, meaning that its rehearsal process can happily employ that mainstay of the drama classroom: small group work. There’s plenty of room to move in terms of gender-blind casting as well. Please be Seated is mainly young women. The scripts also contain ensemble-based interludes which are non-verbal, music driven, with a choreographic element. They have also springboarded further explorations in form. The ethical and aesthetic complexities of verbatim works; the interaction between music and theatre; and meta-concerns related to the performing of performance: ‘how can the act of acting ‘acted’. The narratives of all three of these plays are deliberately open, enabling the flexible casting and on-the hop editing that large-group, time-poor processes sometimes necessitate. Duty Free is about the overseas ‘adventures’ of young people. Please Be Seated is based in verbatim text about young people falling in and out of love. Australian Drama is about young people in a drama classroom trying to connect with each other and put their own shine on dull fragments of the theatrical canon. The plays were published as a collection in hardcopy and digital editions by Playlab Press in 2015. Please be Seated is a co-write with a large group. These co-author’s names are listed in the publication, and below in ‘additional information’.

Èto i to v povesti Staruha Daniila Harmsa

This dissertation focuses on the short story Starukha (The Old Woman), one of the last works of the Russian writer Daniil Kharms (1905-1942). The story, written in 1939, is analysed using the Kharmsian concepts èto and to (this and that) as a heuristic interpretative model. The first chapter gives a detailed analysis of this model, as well as a survey of the critical work done to date on Kharms and Starukha. In the second chapter the model is applied to study the different states of consciousness of the male protagonist. This is significant, because he is the "I" of the work, from whose point of view everything is being told. The third chapter takes a closer look at the reality of the world that exists independently of the consciousness of the protagonist. Physical objects can be said to bear - besides their everyday meaning - a hidden symbolic meaning. Similarly, the characters can be considered as representatives of everyday reality and otherworldliness. The fourth chapter deals with the narrative devices of Starukha. The problematics of the relation between fact and fiction plays an essential role in the story. Kharms's use of Ich-Erzählung and different tenses, which contributes to achieving a complicated elaboration of this kind of problematics, is examined in detail. The fifth chapter provides an intertextual reading of Starukha, based on its allusions to the Bible and the Christian tradition. As a result, the whole story can be seen as a kind of meditation on the Passion of Christ. The final chapter examines how the important Kharmsian concepts of the grotesque and the absurd manifest themselves in Starukha. The old woman represents in a grotesque way two opposite systems: the religious and the totalitarian. The absurdity of Starukha can be claimed to be illusory. Therefore, it is better to speak about paradoxicality. Starukha itself is a kind of paradox, in the sense that it tries to say something of the ultimate truth of reality, which inevitably remains ineffable.

Passion vallassa : Hermostunut aika Minna Canthin teoksissa

Under the power of passion. The age of nervousness in Minna Canth s works This research contemplates the psychology of Minna Canth s characters through the historical image of man in late 19th century Europe. The central operative term of the study is passion , understood as a twofold philosophical concept that includes both desire and suffering. The method of this study is historical and contextual. The study interprets the passions and the psychology of Canth s characters as they were understood in their own time. The indicator of the relevant contexts is the realist and naturalist genre of Canth s works. New research on the genre of the time is also the basis of a new kind of psychological approach to Canth s works. The most important context of passion in Canth s works is the positivistic and pathological image of man at the end of the 19th century. Then, passion was widely discussed, and was perceived as a physiological phenomenon that influenced humans neurologically and caused different kinds of physiological symptoms and nervous disorders. But at the same time, passion was understood as a manifestation of human instincts and drives. The naturalistic literature of the day aimed at creating deterministic studies of human morality and psychology following Émile Zola s application of experimental science methods in his writing. The pathological image of man is most explicitly manifested in Canth s formerly unknown short story Lääkäri (Doctor, 1891), in which a doctor who is interested in psychology visits a jail to meet a peculiar criminal, a girl who feels no remorse for her multiple crimes. In other works of Canth the medically motivated viewpoint is more hidden in the deterministic narrative and depiction of the characters. The present study approaches the passion in Minna Canth s works through five thematic chapters, in witch characters are interpreted suffering from blind love, ennui, crippling romantic idealism, melancholy, guilt and nostalgia, and their stories can be prescribed as medical histories which depict the born of the passion and its development towards ruin. All protagonists are also manifestations of their own time. Canth criticises the modern life and its demands as well as social defects through the tragic stories of individuals. The study demonstrates that Canth did not, like previous research has suggested, wait until the 1890s before writing works of a psychological nature but had already written according to the psychological paradigm of her time in Työmiehen vaimo (1885). The social and psychological interests intertwine in Canth s works and are not exclusionary as has formerly been interpreted. Canth is also critical of the medical power implicit in the naturalist experimental method and this shows itself especially in her depiction of working class women.

Quest for new materials: Inorganic chemistry plays a crucial role

There is an endless quest for new materials to meet the demands of advancing technology. Thus, we need new magnetic and metallic/semiconducting materials for spintronics, new low-loss dielectrics for telecommunication, new multi-ferroic materials that combine both ferroelectricity and ferromagnetism for memory devices, new piezoelectrics that do not contain lead, new lithium containing solids for application as cathode/anode/electrolyte in lithium batteries, hydrogen storage materials for mobile/transport applications and catalyst materials that can convert, for example, methane to higher hydrocarbons, and the list is endless! Fortunately for us, chemistry - inorganic chemistry in particular - plays a crucial role in this quest. Most of the functional materials mentioned above are inorganic non-molecular solids, while much of the conventional inorganic chemistry deals with isolated molecules or molecular solids. Even so, the basic concepts that we learn in inorganic chemistry, for example, acidity/basicity, oxidation/reduction (potentials), crystal field theory, low spin-high spin/inner sphere-outer sphere complexes, role of d-electrons in transition metal chemistry, electron-transfer reactions, coordination geometries around metal atoms, Jahn-Teller distortion, metal-metal bonds, cation-anion (metal-nonmetal) redox competition in the stabilization of oxidation states - all find crucial application in the design and synthesis of inorganic solids possessing technologically important properties. An attempt has been made here to illustrate the role of inorganic chemistry in this endeavour, drawing examples from the literature its well as from the research work of my group.

Sostdc1 Plays an Essential Role in Mammalian Tooth Patterning : Insight into the Rodent Dental Evolution

This thesis work focuses on the role of TGF-beta family antagonists during the development of mouse dentition. Tooth develops through an interaction between the dental epithelium and underlying neural crest derived mesenchyme. The reciprocal signaling between these tissues is mediated by soluble signaling molecules and the balance between activatory and inhibitory signals appears to be essential for the pattern formation. We showed the importance of Sostdc1 in the regulation of tooth shape and number. The absence of Sostdc1 altered the molar cusp patterning and led to supernumerary tooth formation both in the molar and incisor region. We showed that initially, Sostdc1 expression is in the mesenchyme, suggesting that dental mesenchyme may limit supernumerary tooth induction. We tested this in wild-type incisors by minimizing the amount of mesenchymal tissue surrounding the incisor tooth germs prior to culture in vitro. The cultured teeth phenocopied the extra incisor phenotype of the Sostdc1-deficient mice. Furthermore, we showed that minimizing the amount of dental mesenchyme in cultured Sostdc1-deficient incisors caused the formation of additional de novo incisors that resembled the successional incisor development resulting from activated Wnt signaling. Sostdc1 seemed to be able to inhibit both mesenchymal BMP4 and epithelial canonical Wnt signaling, which thus allows Sostdc1 to restrict the enamel knot size and regulate the tooth shape and number. Our work emphasizes the dual role for the tooth mesenchyme as a suppressor as well as an activator during tooth development. We found that the placode, forming the thick mouse incisor, is prone to disintegration during initiation of tooth development. The balance between two mesenchymal TGF-beta family signals, BMP4 and Activin is essential in this regulation. The inhibition of BMP4 or increase in Activin signaling led to the splitting of the large incisor placode into two smaller placodes resulting in thin incisors. These two signals appeared to have different effects on tooth epithelium and the analysis of the double null mutant mice lacking Sostdc1 and Follistatin indicated that these TGF-beta inhibitors regulate the mutual balance of BMP and Activin in vivo. In addition, this work provides an alternative explanation for the issue of incisor identity published in Science by Tucker et al. in 1998 and proposes that the molar like morphology that can be obtained by inhibiting BMP signaling is due to partial splitting of the incisor placodes and not due to change in tooth identity from the incisor to the molar. This thesis work presents possible molecular mechanisms that may have modified the mouse dental pattern during evolution leading to the typical rodent dentition of modern mouse. The rodent dentition is specialized for gnawing and consists of two large continuously growing incisors and toothless diastema region separating the molars and incisors. The ancestors of rodents had higher number of more slender incisors together with canines and premolars. Additionally, murine rodents, which include the mouse, have lost their ability for tooth replacement. This work has revealed that the inhibitory molecules appear to play a role in the tooth number suppression by delineating the spatial and temporal action of the inductive signals. The results suggest that Sostdc1 plays an essential role in several stages of tooth development through the regulation of both the BMP and Wnt pathway. The work shows a dormant sequential tooth forming potential present in wild type mouse incisor region and gives a new perspective on tooth suppression by dental mesenchyme. It reveals as well a novel mechanism to create a large mouse incisor through the regulation of mesenchymal balance between inductive and inhibitory signals.

Substrate specificity plays an important role in uncoupling the catalytic and scaffolding activities of rat testis DNA topoisomerase IIalpha

Topoisomerase II (topo II) is a dyadic enzyme found in all eukaryotic cells. Topo II is involved in a number of cellular processes related to DNA metabolism, including DNA replication, recombination and the maintenance of genomic stability. We discovered a correlation between the development of postnatal testis and increased binding of topo IIalpha to the chromatin fraction. We used this observation to characterize DNA-binding specificity and catalytic properties of purified testis topo IIalpha. The results indicate that topo IIalpha binds a substrate containing the preferred site with greater affinity and, consequently, catalyzes the conversion of form I to form IV DNA more efficiently in contrast to substrates lacking such a site. Interestingly, topo IIalpha displayed high-affinity and cooperativity in binding to the scaffold associated region. In contrast to the preferred site, however, high-affinity binding of topo IIalpha to the scaffold-associated region failed to result in enhanced catalytic activity. Intriguingly, competition assays involving scaffold-associated region revealed an additional DNA-binding site within the dyadic topo IIalpha. These results implicate a dual role for topo IIalpha in vivo consistent with the notion that its sequestration to the chromatin might play a role in chromosome condensation and decondensation during spermatogenesis.

The prankster’s passion: Using theatrical terms to unpack players motivations in popular social performance

Amongst social players, the prank, as a social performance form, holds a lot of potential to impact on personal, relational and social status within a group or between one group and another group. More than simply showing off, a prank in the strictest definition of the term, is a social performance in which one player, a prankster, deploys mischief, trickery or deceit, to cause a moment of anxiety, fear or anger about a happening for another spectator-become-collaborating-player, a prankee – to enhance social bonds, entertain, or comment on a social, cultural or political phenomenon. During a prank, the prankster’s ability to be creative, clever or culturally astute, and the prankee’s ability to be duped, be a good sport, play along, or even play/pay the prankster back, both become fodder for other spectators and society to scrutinize. In Australia, pranking traditions are popular with many social groups, from the community-building pranks of footballers, bucks parties and ‘drop bear’ tales told to tourists, to the more controversial pranks of radio shock jocks, activists and artists. In this paper, I consider whether theatrical terms – theoretical terms from the stage such as actor, acting, objective, arc, performance, audience and emotion, such as those offered by Joseph Roach – are useful in understanding the passion some social players show for pranksterism. Are theatrical terms such as Roach’s as useful as analysts of social self-performance such as Erving Goffman suggest they are? Do they assist in understanding the personal actions, reactions and emotions of prankster and prankee? Do they assist in understanding the power relations between prankster and prankee? Do they assist in understanding the relation between the prank – be it an everyday prank amongst families, friends and coworkers, an entertainment program prank of the sort seen on Prank Patrol, Punked or Scare Tactics, or an activist pranks perpetrated by a guerrilla artist, ‘jammers’ or ‘hackers’ intent on turning dominant social systems back on themselves – the social players, and the public sphere in which the prank takes place? I reflect on how reading pranks as performances, by players, for highly participatory audiences, helps understand why they are so prevalent, and so recurrent across times, cultures and contexts, and also so controversial when not performed well enough – or when performed too well – prompting outrage from the prankster, prankee or society as passionate as any debate about a performance by players in a theatre.