Molecular characterization of Arabidopsis thaliana PUF proteins - binding specificity and target candidates

PUF proteins regulate both stability and translation through sequence-specific binding to the 3` UTR of target mRNA transcripts. Binding is mediated by a conserved PUF domain, which contains eight repeats of approximately 36 amino acids each. Found in all eukaryotes, they have been related to several developmental processes. Analysis of the 25 Arabidopsis Pumilio (APUM) proteins presenting PUF repeats reveals that 12 (APUM-1 to APUM-12) have a PUF domain with 50-75% similarity to the Drosophila PUF domain. Through three-hybrid assays, we show that APUM-1 to APUM-6 can bind specifically to the Nanos response element sequence recognized by Drosophila Pumilio. Using an Arabidopsis RNA library in a three-hybrid screening, we were able to identify an APUM-binding consensus sequence. Computational analysis allowed us to identify the APUM-binding element within the 3` UTR in many Arabidopsis transcripts, even in important mRNAs related to shoot stem cell maintenance. We demonstrate that APUM-1 to APUM-6 are able to bind specifically to APUM-binding elements in the 3` UTR of WUSCHEL, CLAVATA-1, PINHEAD/ZWILLE and FASCIATA-2 transcripts. The results obtained in the present study indicate that the APUM proteins may act as regulators in Arabidopsis through an evolutionarily conserved mechanism, which may open up a new approach for investigating mRNA regulation in plants.

Argininosuccinate Synthetase Is a Functional Target for a Snake Venom Anti-hypertensive Peptide ROLE IN ARGININE AND NITRIC OXIDE PRODUCTION

Bj-BPP-10c is a bioactive proline-rich decapeptide, part of the C-type natriuretic peptide precursor, expressed in the brain and in the venom gland of Bothrops jararaca. We recently showed that Bj-BPP-10c displays a strong, sustained anti-hypertensive effect in spontaneous hypertensive rats (SHR), without causing any effect in normotensive rats, by a pharmacological effect independent of angiotensin-converting enzyme inhibition. Therefore, we hypothesized that another mechanism should be involved in the peptide activity. Here we used affinity chromatography to search for kidney cytosolic proteins with affinity for Bj-BPP-10c and demonstrate that argininosuccinate synthetase (AsS) is the major protein binding to the peptide. More importantly, this interaction activates the catalytic activity of AsS in a dose-dependent manner. AsS is recognized as an important player of the citrulline-NO cycle that represents a potential limiting step in NO synthesis. Accordingly, the functional interaction of Bj-BPP-10c and AsS was evidenced by the following effects promoted by the peptide: (i) increase of NO metabolite production in human umbilical vein endothelial cell culture and of arginine in human embryonic kidney cells and (ii) increase of arginine plasma concentration in SHR. Moreover, alpha-methyl-DL-aspartic acid, a specific AsS inhibitor, significantly reduced the anti-hypertensive activity of Bj-BPP-10c in SHR. Taken together, these results suggest that AsS plays a role in the anti-hypertensive action of Bj-BPP-10c. Therefore, we propose the activation of AsS as a new mechanism for the anti-hypertensive effect of Bj-BPP-10c in SHR and AsS as a novel target for the therapy of hypertension-related diseases.

Under flow impedimetric measurements using magnetic particles for label-free detection affinity target

The necessity to adapt sensors based on electrochemical techniques for high throughput analysis control increases the interest to develop new analytical systems able to perform measurements under buffer now. In this report we explored the possibility of employing a new system to make impedimetric measurements to detect the interaction between proteins and small molecules. The well-known biotin-streptavidin interaction was adopted to evaluate the proposed assembly. This system allows us to perform experiments under flow. Magnetic beads functionalized with streptavidin were used and first characterized using AFM and FTIR. Non-faradic impedance spectroscopy allowed the detection of the biotin-streptavidin interaction. Using our new system and under a flow of PBS buffer, 5 10-5 M of biotin was detected with a stable signal. (c) 2007 Elsevier B.V. All rights reserved.

Oroidin Inhibits the Activity of the Multidrug Resistance Target Pdr5p from Yeast Plasma Membranes

Oroidin was isolated from the marine sponge Agelas sventres and inhibited the activity and function of Pdr5p, an enzyme responsible for the multidrug resistance phenotype in Saccharomyces cerevisiae. This compound may help in the development of new drugs that reverse this dangerous phenotype of pathogenic yeast and fungi.

Integration of Ligand- and Target-Based Virtual Screening for the Discovery of Cruzain Inhibitors

A myriad of methods are available for virtual screening of small organic compound databases. In this study we have successfully applied a quantitative model of consensus measurements, using a combination of 3D similarity searches (ROCS and EON), Hologram Quantitative Structure Activity Relationships (HQSAR) and docking (FRED, FlexX, Glide and AutoDock Vina), to retrieve cruzain inhibitors from collected databases. All methods were assessed individually and then combined in a Ligand-Based Virtual Screening (LBVS) and Target-Based Virtual Screening (TBVS) consensus scoring, using Receiving Operating Characteristic (ROC) curves to evaluate their performance. Three consensus strategies were used: scaled-rank-by-number, rank-by-rank and rank-by-vote, with the most thriving the scaled-rank-by-number strategy, considering that the stiff ROC curve appeared to be satisfactory in every way to indicate a higher enrichment power at early retrieval of active compounds from the database. The ligand-based method provided access to a robust and predictive HQSAR model that was developed to show superior discrimination between active and inactive compounds, which was also better than ROCS and EON procedures. Overall, the integration of fast computational techniques based on ligand and target structures resulted in a more efficient retrieval of cruzain inhibitors with desired pharmacological profiles that may be useful to advance the discovery of new trypanocidal agents.

CDK9 a potential target for drug development

The family of Cyclin-Dependent Kinases (CDKs) can be subdivided into two major functional groups based on their roles in cell cycle and/or transcriptional control. CDK9 is the catalytic subunit of positive transcription elongation factor b (P-TEFb). CDK9 is the kinase of the TAK complex (Tat-associated kinase complex), and binds to Tat protein of HIV, suggesting a possible role for CDK9 in AIDS progression. CDK9 complexed with its regulatory partner cyclin T1, serves as a cellular mediator of the transactivation function of the HIV Tat protein. P-TEFb is responsible for the phosphorylation of the carboxyl-terminal domain of RNA Pol II, resulting in stimulation of transcription. Furthermore, the complexes containing CDK9 induce the differentiation in distinct tissue. The CDK9/cyclin T1 complex is expressed at higher level in more differentiated primary neuroectodermal and neuroblastoma tumors, showing a correlation between the kinase expression and tumor differentiation grade. This may have clinical and therapeutical implications for these tumor types. Among the CDK inhibitors two have shown to be effective against CDK9: Roscovitine and Flavopiridol. These two inhibitors prevented the replication of human immunodeficiency virus (HIV) type 1 by blocking Tat transactivation of the HIV type 1 promoter. These compounds inhibit CDKs by binding to the catalytic domain in place of ATP, preventing transfer of a phosphate group to the substrate. More sensitive therapeutic agents of CDK9 can be designed, and structural studies can add information in the understanding of this kinase. The major features related to CDK9 inhibition will be reviewed in this article.

Understanding Foreign Masters : the examples of Rabindranath Tagore and Fyodor Dostoyevskiy

The Indian author Rabindranath Tagore was received like royalty during his visits to the West after winning the Nobel Prize in 1913. Dreams of foreign cultures offered a retreat from a complicated age. In a time when the West appeared to be living under threat of disintegration and when industrialism seemed like a cul-de-sac, he appeared to offer the promise of a return to a lost paradise, a spiritual abode that is superior to the restless Western culture. However, Tagore’s popularity faded rapidly, most notably in England, the main target of his criticism. Soon after Tagore had won the Nobel Prize, the English became indignant at Tagore’s anti-colonial attitude.Tagore visited Sweden in 1921 and 1926 and was given a warm reception. His visits to Sweden can be seen as an episode in a longer chain of events. It brought to life old conceptions of India as the abode of spirituality on earth. Nevertheless, interest in him was a relatively short-lived phenomenon in Sweden. Only a few of his admirers in Sweden appreciated the complexity of Tagore’s achievements. His “anathema of mammonism”, as a Swedish newspaper called it, was not properly received. After a steady stream of translations his popularity flagged towards the end of the 1920s and then almost disappeared entirely. Tagores visits in Sweden gave an indication that India was on the way to liberate itself from its colonial legacy, which consequently contributed to the waning of his popularity in the West. In the long run, his criticism of the drawbacks in the western world became too obvious to maintain permanent interest. The Russian author Fyodor Dostoyevskiy’s Crime and Punishment (1866) has enticed numerous interpretations such as the purely biographical approach. In the nervous main character of the novel, the young student Raskolnikov, one easily recognizes Dostoyevskiy himself. The novel can also be seen as a masterpiece of realistic fiction. It gives a broad picture of Saint Petersburg, a metropolis in decay. Crime and Punishment can also be seen as one of the first examples of a modern psychological novel, since it is focused on the inner drama of its main character, the young student Raskolnikov. His actions seem to be governed by mere coincidences, dreams and the spur of the moment. it seems fruitful to study the novel from a psychoanalytical approach. In his book Raskolnikov: the way of the divided towards unity in Crime and Punishment (1982), a Swedish scholar, Owe Wikström, has followed this line of interpretation all the way to Freud’s disciple C G Jung. In addition to this, the novel functions as an exciting crime story. To a large extent it is Viktor Sjklovskij and other Russian formalists from the 1920s and onwards who have taught the western audience to understand the specific nature of the crime story. The novel could be seen as a story about religious conversion. Like Lasarus in the Bible (whose story attracts a lot of attention in the novel) Raskolnikov is awakened from the dead, and together with Sonja he starts a completely new life. The theme of conversion has a special meaning for Dostoyevskiy. For him the conversion meant an acknowledgement of the specific nature of Russia itself. Crime and punishment mirrors the conflict between traditional Russian values and western influences that has been obvious in Russia throughout the history of the country. The novel reflects a dialogue that still continues in Russian society. The Russian literary historian Mikhail Bakhtin, who is probably the most famous interpreter of the works of Dostoyevskiy, has become famous precisely by emphasizing the importance of dialogues in novels like Crime and Punishment. According to Bakhtin, this novel is characterized by its multitude of voices. Various ideas are confronted with each other, and each one of them is personified by one of the characters in the novel. The author has resigned from his position as the superior monitor of the text, and he leaves it to the reader to decide what interpretation is the correct one..The aim of the present study is thus to analyze the complex reactions in the west to Tagore’s visits in Sweden and to Fyodor Dostoyevskiys novel Crime and Punishment.. This leads to more general conclusions on communication between cultures.

Movies, Mind and Meaning : Studying Audience and Favourite Films

Construction of identity and meaning is becoming increasingly important in both media studies and religion scholarship. (Lövheim, 2004) Meaning construction outside traditional religion has become more interesting for religious studies and what individuals in the audience do with all messages circulated through media in everyday life has attended increasing interest within media studies (Stout and Buddenbaum, 2001). Motion pictures, soap operas and advertising are all examples of media contents which generate ideas among its audience which to a various degree are used as resources within the construction of identity (Jansson, 2001). The investigation of what modern humankind’s world views look like and what components they are composed of, in this context seems to be an important topic of investigation (Holm and Björkqvist, 1996). The ways in which the development of media has effected the daily lives of individuals is interest as is the nature of the self and the ways in which the process of self-formation is affected by the profusion of mediated materials (Thompson, 1995). Film and religion are my interest within this larger frame. The topic is not exactly new but the combination of film and religion has during the last ten years resulted in a rapidly growing number of books by scholars interested in this field (Lyden, 2003). One growing focus is on the role that films can and do play within the emerging and developing valuesystem of people in the West today (Marsh, 2004). The British theologian Clive Marsh’s point of departure is very similar to my own. Viewers bring to a film life-experience, immediate concerns and worldviews and the exploration of this interplay between movies and the interpreting process of meaning making is the very focus in this paper. Theoretically, the semeiological model of Alf Linderman is combined with cultural cognitive approaches used by a number of Scandinavian media scholars developing perspectives in audience theory (Linderman, 1996, Höijer and Werner, 1998). 13 individuals, their favourite movie and what it means to them in their life My aim is to examine how individuals comprehend film and what the meaning process look like. In this paper I present the outcome of 13 interviews with young people about their favourite film. I suggest how it is possible to interpret how they interrelate film comprehension with their personal beliefs and their culturally constructed worldview from a sociocognitive point of view. Examples of films chosen range from Disneys Lion King (1994), sciencefiction and fantasy successes like The Matrix (1999) and Lord of the Rings (2001) or the next best movie ever according to www.IMdb.com The Shawshank Redemption (1994) as well as the Swedish blockbuster Så som i himmelen (2004), aka “As in Heaven”.

Movies and meaning : studying audience, fiction film and existential matters

In what ways and under what circumstances can a movie be a resource for individuals and their thoughts about existential matters? This central research question has been investigated using a both quantitative and qualitative approach. First, a questionnaire was distributed amongst 179 Swedish students to provide a preliminary overview of film habits. The questionnaire was also used as a tool for selecting respondents to individual interviews. Second, focus group and individual interviews were conducted, with viewers choosing their favourite movie of all time. In the study socio-cognitive theory and a schema-based theoretical tool is adopted to analyze how different viewers make use of movies as cultural products in an interplay between culture and cognition in three contexts; a socio-historic process, a socio-cultural interaction with the world and inner psychological processes. The viewers in the study seem to be inspired by movies as a mediated cultural resource, promoting the development of a personal moral framework with references to values deeply fostered by a humanistic tradition. It is argued that these findings support theories discussing individualised meaning making, developing ‘self-expression values’ and ‘altruistic individualism’ in contemporary western society.

Förpackningsdesign i genusperspektiv : En granskning av duschcremesförpackningar

Färg och form kan bidra till att skapa uppmärksamhet, förmedla budskap och skapa känslor som i sin tur kan öka sannolikheten av att kunden genomför köpet. Förpackningens färg och form bör synkronisera för att budskapen inte ska motsäga varandra, och det är mycket viktigt att designen lämpar sig till målgruppen. Färg kan ses som ett språk som kommunicerar med konsumenten, och att förstå detta språk kan vara ett starkt redskap inom marknadsföring och design. Det är vanligt att förpackningsdesign riktas mot något eller bägge könen och att ge produkten en karaktär av maskulint eller feminint kan vara ett effektivt tillvägagångssätt att särskilja produkten från resterande sortiment. I samband med färg och form är det nödvändigt att förstå konsumentens behov samt användning av produkten, och det är viktigt att skapa en god design riktad mot rätt målgrupp. Det är viktigt att förstå vad konsumenten attraheras till och vad som lockar till att genomföra ett köp. Syftet med denna studie var att ta reda på varför förpackningsdesign för duschcreme specificerade för kvinnor respektive män skiljer sig, och vad som anses vara kvinnligt och manligt när det kommer till färg och form, i samband med duschcremesförpackningar. För att ta reda på detta genomfördes en visuell innehållsanalys samt en enkätundersökning. Resultatet som genererades av dessa metoder sammanfattades i tabeller och diagram, som tydligt visar de färger som anses vara maskulina samt feminina. Mörka färger, som svart och blått, anses vara maskulina medan ljusa färger och rosa toner anses vara feminina. Mjuka grafiska element och former kopplas till femininitet medan motsatsen kopplas till maskulinitet. Slutsatsen av detta är att anledningen till att förpackningarna skiljer sig är för att lättare kommunicera med målgruppen, och i detta fall via färg och form.

English as the target language : A literature study on teachers’ and L2 learners’ language use in the upper elementary classroom

Even though English is a subject where Swedish pupils do well compared to pupils in other countries, research indicates that pupils are not always motivated to learn in the English classroom. Therefore, the aim of this study is to find research relating to the use of the target language in classrooms for pupils at the upper elementary level, particularly language learners in Sweden. The focus of this thesis is to find out what benefits and challenges accompany the use of the target language during English lessons, as well as what pupils’ opinions are on the consistent use of the target language in the classroom. This literature review of five research articles shows that it is beneficial for pupils’ language development to have lessons where mainly the target language is used. It is for example beneficial for pupils’ ability to speak, their pronunciation, vocabulary and ability to use language strategies. The results show that there are challenges as well, especially for the teachers, since use of the target language presumes that the teacher has good language skills and is capable of scaffolding each pupil at their individual level and in their zone of proximal development. Furthermore, there are challenges like differences in pupils’ skill level, creating tasks that both motivate and stimulate, and creating a safe learning environment. Even though the results in this thesis are limited, it is still obvious that it is an important area, where more research is necessary in order to assist teachers in how to teach English as effectively as possible.