Investigating the delivery of antimicrobial proteins and aminoglycoside antibiotics to the airways

Biopharmaceuticals are finding wide applications in the management of diverse disease conditions. Pulmonary delivery of proteins may constitute an effective and efficient non-invasive alternative to parenteral delivery, which is currently the main route of administration of biopharmaceutical drugs. A particular area, in which pulmonary delivery of peptides and proteins may find ready application, is in the local delivery of antimicrobial peptides and proteins to the airway, a measure that could potentially bring about improvements to currently available antipseudomonal therapies. This thesis has therefore sought to develop inhalable antimicrobial proteins in combination with antibiotics that have particularly good antimicrobial activity against Pseudomonas aeruginosa infections in the respiratory tract of people with cystic fibrosis (CF). Through process optimisation, a suitable spray drying method was developed and used for the preparation of active, inhalable dry powder formulations of the antimicrobial protein, lactoferrin, and aminoglycosides (tobramycin and gentamicin). The physicochemical properties, aerosolisation performance and the antibacterial properties of the various spray-dried formulations were assessed. In addition, a relevant in vitro cellular model was employed to investigate the potential cytotoxic and pro-inflammatory effects of the various formulations on four bronchial human epithelial cells together with their effectiveness at reducing bacterial colonies when administered on to biofilm co-cultured on the epithelial cells. It was found that following spray drying the particles obtained were mostly spherical, amorphous and possessed suitable aerosolisation characteristics. The various spray-dried antimicrobial proteins (lactoferrin or apo lactoferrin) and co-spray dried combinations of the proteins and aminoglycosides were found to exhibit bactericidal activity against planktonic and biofilms of P. aeruginosa. In general, the spray drying process was found not to significantly affect the antimicrobial activities of the protein. Treatment of the different bronchial epithelial cell lines with the antimicrobial formulations showed that the various formulations were non-toxic and that the co-spray dried combinations significantly reduced established P. aeruginosa biofilms on the four bronchial epithelial cells. Overall, the results from this thesis demonstrates that spray drying could potentially be employed to prepare inhalable antimicrobial agents comprised of proteins and antibiotics. These new combinations of proteins and aminoglycosides has promising applications in the management of P. aeruginosa in the airway of cystic fibrosis patients.

Factors determining disease duration in Alzheimer's disease:a postmortem study of 103 cases using the Kaplan-Meier estimator and Cox regression

Factors associated with duration of dementia in a consecutive series of 103 Alzheimer's disease (AD) cases were studied using the Kaplan-Meier estimator and Cox regression analysis (proportional hazard model). Mean disease duration was 7.1 years (range: 6 weeks-30 years, standard deviation = 5.18); 25% of cases died within four years, 50% within 6.9 years, and 75% within 10 years. Familial AD cases (FAD) had a longer duration than sporadic cases (SAD), especially cases linked to presenilin (PSEN) genes. No significant differences in duration were associated with age, sex, or apolipoprotein E (Apo E) genotype. Duration was reduced in cases with arterial hypertension. Cox regression analysis suggested longer duration was associated with an earlier disease onset and increased senile plaque (SP) and neurofibrillary tangle (NFT) pathology in the orbital gyrus (OrG), CA1 sector of the hippocampus, and nucleus basalis of Meynert (NBM). The data suggest shorter disease duration in SAD and in cases with hypertensive comorbidity. In addition, degree of neuropathology did not influence survival, but spread of SP/NFT pathology into the frontal lobe, hippocampus, and basal forebrain was associated with longer disease duration. © 2014 R. A. Armstrong.

What risk factors are associated with Alzheimer's disease?

A large number of risk factors have been associated with Alzheimer’s disease (AD). This article discusses the validity of the major risk factors that have been identified including age, genetics, exposure to aluminium, head injury, malnutrition and diet, mitochondrial dysfunction, vascular disease, immune system dysfunction, and infection. Rare forms of early-onset familial AD (FAD) are strongly linked to the presence of specific gene mutations, viz. mutations in amyloid precursor protein (APP) and presenilin (PSEN1/2) genes. By contrast, late-onset sporadic AD (SAD) is a multifactorial disorder in which age-related changes, genetic risk factors, such as allelic variation in apolipoprotein E (Apo E) gene, vascular disease, head injury and risk factors associated with diet, the immune system, mitochondrial function, and infection may all be involved. Life-style changes that may reduce the effect of these risk factors and therefore, the risk of AD are discussed.

Risk factors for Alzheimer's disease

A large number of possible risk factors have been associated with Alzheimer'sdisease (AD).This chapter discusses the validity of the major risk factors that have been identifiedincluding age, genetics, exposure to aluminum, head injury, malnutrition and diet,mitochondrial dysfunction, vascular disease, immune system dysfunction, and infectionand proposes a hypothesis to explain how these various risk factors may cause ADpathology.Rare forms of early-onset familial AD (FAD) are strongly linked to the presence ofspecific gene mutations, viz. mutations in amyloid precursor protein (APP) andpresenilin (PSEN1/2) genes. By contrast, late-onset sporadic AD (SAD) is amultifactorial disorder in which age-related changes, genetic risk factors, such as allelicvariation in apolipoprotein E (Apo E) gene, vascular disease, head injury and risk factorsassociated with diet, immune system, mitochondrial function, and infection may all beinvolved.These risk factors interact to increase the rate of normal aging (=allostatic load')which over a lifetime results in degeneration of neurons and blood vessels and as aconsequence, the formation of abnormally aggregated =reactive' proteins such as ß-amyloid (Aß) and tau leading to the development of senile plaques (SP) andneurofibrillary tangles (NFT) respectively. Life-style changes that may reduce theallostatic load and therefore, the risk of dementia are discussed.

Laminar distribution of β-amyloid (Aβ) peptide deposits in the frontal lobe in familial and sporadic Alzheimer's disease

To determine whether genetic factors influence frontal lobe degeneration in Alzheimer's disease (AD), the laminar distributions of diffuse, primitive, and classic β-amyloid (Aβ) peptide deposits were compared in early-onset familial AD (EO-FAD) linked to mutations of the amyloid precursor protein (APP) or presenilin 1 (PSEN1) gene, late-onset familial AD (LO-FAD), and sporadic AD (SAD). The influence of apolipoprotein E (Apo E) genotype on laminar distribution was also studied. In the majority of FAD and SAD cases, maximum density of the diffuse and primitive Aβ deposits occurred in the upper cortical layers, whereas the distribution of the classic Aβ deposits was more variable, either occurring in the lower layers, or a double-peaked (bimodal) distribution was present, density peaks occurring in upper and lower layers. The cortical layer at which maximum density of Aβ deposits occurred and maximum density were similar in EO-FAD, LO-FAD and SAD. In addition, there were no significant differences in distributions in cases expressing Apo E ε4 alleles compared with cases expressing the ε2 or ε3 alleles. These results suggest that gene expression had relatively little effect on the laminar distribution of Aβ deposits in the frontal lobe of the AD cases studied. Hence, the pattern of frontal lobe degeneration in AD is similar regardless of whether it is associated with APP and PSEN1, mutation, allelic variation in Apo E, or with SAD.

Fas-FasL expression and interactions in mouse tumor cell lines: Implications for tumor immune escape

The Fas system, comprising the Fas receptor (Fas/Apo-1/CD95) and its ligand, Fas ligand (FasL), is a central mediator of programmed cell death in various physiological and pathological processes. FasL exists as transmembrane and soluble forms and induces apoptosis on crosslinking with Fas receptor. Recent evidence indicated that tumor cells exploit this system for their immunologic escape that includes the loss of Fas and the gain of FasL expression. In the present study, nine mouse tumor cell lines of diverse origin were examined immunocytochemically for the expression of Fas and FasL. Nine of nine cell lines expressed FasL, and five of nine cell lines expressed Fas. FasL expression in these tumor cell lines was demonstrated to be functional by its induction of apoptosis in Fas-sensitive target cells in coculture experiments. These results suggest that FasL may be a prevalent mediator of immune privilege in mouse malignancies, and support the recently proposed "counterattack model" for local elimination of tumor-reactive immune cells by tumor cell-derived FasL.^ Culture supernatant of four cell lines expressing FasL showed cytotoxic effect on Fas-sensitive target cells, indicating the possibility of secreted FasL in the medium. The Fas-expressing cell lines were sensitized to anti-Fas antibody cytotoxicity following treatment with IL-2 and IFN-$\gamma$, suggesting cytokine stimulation as an effective target for future immunotherapeutic strategies. ^

A influência do conforto luminoso na satisfação dos profissionais que atuam no ginásio do Centro de Reabilitação Infantil, Natal-RN

This study addresses the environmental quality in therapeutic spaces for children's rehabilitation. The assumption that space is active and interfere in interpersonal relationships, highlights the importance of natural light to the hospital architecture, to foster the creation of environments that encourage and assist in the recovery of patients in the rehabilitation process. Therefore, interferes with health humanization through positive actions in the physiological and emotional effects of natural light, as facilitators of the health recovery process. In Brazil hospital openings systems projects are built exclusively to follow requirements of the local construction code which do not consider the landscape, but only ventilation and heat stroke; and the luminance levels are treated just as recommendations for artificial lighting. The National Policy for Healthcare Humanization presents the environmental comfort as a priority. However, it does not guidelines for achieving it. In this context this research aims to evaluate the lighting comfort in infant therapeutic areas from the professional satisfaction, in order to identify human preferences on the variables: technical and constructive aspects, relationship with the exterior, internal visual interface and quality elements. With this purpose it was adopted as research strategy the Post-Occupancy Evaluation (Technical Functional) through a multi method approach, which included a case study in the rehabilitation gym of Children Rehabilitation Center, at Natal, Rio Grande do Norte, and a reference study at SARAH Rehabilitation Center, Fortaleza Unit at Ceará, both in Brazil northeast. The results indicate that the definition of openings systems should consider external and internal factors to the building, as the natural landscape, the immediate surroundings and activities to be performed. The POE found out the preference of the professional visual privacy in detriment to other analyzed aspects. Thus, it is expected that this study can contribute to the discussion of luminous quality and generate inputs for future projects or renovations in the Children's Rehabilitation Centers, which should not be projected as hospitals

Data for the co-expression and purification of human recombinant CaMKK2 in complex with calmodulin in Escherichia coli

Calcium/calmodulin-dependent kinase kinase 2 (CaMKK2) has been implicated in a range of conditions and pathologies from prostate to hepatic cancer. Here, we describe the expression in Escherichia coli and the purification protocol for the following constructs: full-length CaMKK2 in complex with CaM, CaMKK2 'apo', CaMKK2 (165-501) in complex with CaM, and the CaMKK2 F267G mutant. The protocols described have been optimized for maximum yield and purity with minimal purification steps required and the proteins subsequently used to develop a fluorescence-based assay for drug binding to the kinase, "Using the fluorescent properties of STO-609 as a tool to assist structure-function analyses of recombinant CaMKK2"

Satuja

Imprimatur: I. U. Wallenius.

Muuttuva lapsen maailma. Minttu-kuvakirjat viidellä vuosikymmenellä 1978–2015.

Tutkimuksen tavoitteena on selvittää, miten kuvaa lapsuudesta rakennetaan verbaalisilla ja visuaalisilla elementeillä kuvakirjan tarjoamien mahdollisuuksien ja rajoitusten puitteissa. Tutkimuskohteinani toimivat Maikki Harjanteen Minttu-kuvakirjoista löytämäni kirjaparit. Samasta aiheesta kirjoitettuja kirjapareja ovat joulusta kertovat kirjat mintun joulu (1979) ja Mintun joulukirja (1989), lemmikin liittymistä Mintun perheeseen käsittelevät kirjat minttu saa koiran (1982) ja Minttu saa oman koiran (2007), mummon nimikkoteokset minttu mummolassa (1978) ja Minttu ja mummo (2010) sekä syntymäpäivistä kertovat kirjat mintun kiva päivä (1979) ja Mintun syntymäpäivät (2014). Tutkin siis, miten kuvaa lemmikeistä, koirista, mummoista, jouluista ja syntymäpäivistä rakennetaan näissä kuvakirjapareissa. Tutkin tutkimuskohteitani ikonoteksteinä eli teoksina, jotka muodostuvat sanan ja kuvan vuorovaikutuksessa. Kuvan ja sanan suhteessa tapahtuneen muutoksen määrittelyn apuna käytän Ulla Rhedinin kolmea kuvakirjakategoriaa sekä Maria Nikolajevan ja Carole Scottin kertovuuden aspektin varaan perustuvaa kuvakirjajaottelua. Representaation ymmärrän Mikko Lehtosen ja Stuart Hallin tapaan kaksisuuntaiseksi ilmiöksi, toisinnoksi, jolla on kuitenkin maailmaa muokkaava voima. Mintut ovat muuttuneet 70-luvun ensimmäisistä kuvakirjoista 2010-luvulle paljon niin visuaalisesti kuin verbaalisesti, mikä on vaikuttanut kirjojen tuottamaan lemmikki-, mummo-, joulu- ja syntymäpäiväkuvaan. Alkutuotannossa painottui kerronnan visuaalinen puoli ja lopputuotannossa tekstin osuus kerronnassa on kasvanut suureksi. Alkutuotannon Mintut pyrkivät mimeettiseen ja realistiseen kuvaukseen kun taas lopputuotanto saa karnevalismin ja nonsensen piirteitä. Samalla Minttujen arvot ovat säilyneet jokseenkin samoina: Vaikka Minttu-kirjoissa representoidaankin melko idyllisesti boheemia lapsiperheen arkea, löytyy niistä säröjä, joilla Mintut kyseenalaistavat valtakulttuuria. Mintut puhuvat vihreiden arvojen puolesta kulutuskulttuuria vastaan ja tekevät selväksi, ettei lapsi tarvitse leluja ja kalliita harrastuksia, vaan oman tärkeän aikuisen aikaa. Minttu-kuvakirjat tuottavat tietynlaista kuvaa lapsuudesta eri vuosikymmenillä. Kuvakirjat luovat omaa todellisuuttaan representoidessaan asioita, eikä niitä voida pitää vallan käytön suhteen neutraalina mediana. Niillä on agenda muuttaa maailmaa.

Ikuisesta runsaudensarvesta uppoaviin paratiisisaariin. Muuttuvan maailman kuvauksia suomalaisissa kaukomatkakertomuksissa

Nykypäivän matkakirjallisuuden suurimpia haasteita on se, miten yhdistää globalisoituvan maailman kuvaaminen lajityyppiin, jonka kuvastoa ovat perinteisesti hallinneet tallaamattomat polut, neitseelliset paratiisirannat sekä kulttuurien eksoottinen toiseus. Tämä tutkielma tarkastelee sitä, millaista kuvaa muuttuvasta maailmasta suomalaiset matkakirjailijat ovat luoneet. Tutkielma sijoittuu ekokriittisen sekä postkolonialistisen kirjallisuudentutkimuksen välimaastoon, jossa painottuu vahva kytkös konkreettiseen maailmaan. Kohdeteoksina on neljä kaukomatkakertomusta eri ajoilta: Tapio Hiisivaaran Myrkkynuolia, kahvia, banaaneja (1945), Tuomas Vennon Katoavaa Grönlantia (1960), Annikki Sankarin Afrikkalainen uni (1984) ja Pirkko Lindbergin SOS Tuvalu (2004). Kohdeteosten avulla käydään läpi eurooppalaisen maisemakuvauksen vanhoja traditioita, joita teokset sekä uusintavat että uudistavat. Matkailijan omat halut ja pelot ilmenevät usein matkakuvauksissa esimerkiksi paratiisien ja helvettien sekä ikuisen rauhan ja ajattomuuden teemoina, ja samankaltaisesti määrittyy myös matkailijan suhde muihin olentoihin. Matkalla tapahtuvat kohtaamiset rikkovat omien ennakkokäsitysten varaan rakennettuja olettamuksia, sillä äärimmäistä kuvaa voi pitää yllä vain, jos asioita tarkastelee etäältä. Kohdeteoksia vertailemalla piirtyy kuva maapallon muuttumisesta luonnonvaroiltaan yltäkylläisestä runsaudensarvesta saasteiden tahrimaksi maailmankyläksi. Pelko länsimaalaisen kulttuurin globaalista leviämisestä ilmenee kohdeteoksissa vahvana, mikä näkyy esimerkiksi primitiivisyyden ihannointina. Tulevaisuuden matkakuvauksissa ilmastonmuutos saattaa toisaalta voimistaa tätä vastakohta-asetelmaa, mutta toisaalta myös luoda yhtenäisempää, koko ihmiskunnan laajuista perspektiiviä. Yksi tärkeimmistä nykypäivän matkakirjallisuuden lajityyppiä uudistavista kerronnan tavoista on moniperspektiivisyys, joka nostaa esille myös kertojan kokemusmaailman ulkopuolisia näkökulmia. Sen valossa koskemattomien idyllienkään ei tarvitse olla matkakirjallisuudelta kiellettyjä aiheita, sillä ne voidaan nähdä vain yhtenä perspektiivin palasena sekä kaunokirjallisen ilmaisun tehokeinoina. Tutkielma osoittaa, miten konkreettisten paikkojen fiktiivisillä esityksillä on voima vahvistaa paikan tunnetta ja siten tuottaa läheistä suhdetta kaukaisiinkin paikkoihin.

Avaliação Pós-Ocupação da iluminação natural das salas dos setores de aulas teóricas da Universidade Federal do Rio Grande do Norte

The objective of this study is to assess the natural light of the classrooms sectors, at Universidade Federal do Rio Grande do Norte. It was applied the Post-Occupation Evaluation technique, by using questionnaires and brightness levels measurements inside the classrooms. In order to check the users satisfaction degree, it was initially done a general approach on the related aspects to natural light: their characteristics, availability, sources, opening systems and evaluating tools. It was also determined the necessary brightness levels for the activities development in the classroom and the Post-Occupation Evaluation technique used in the search analyses. Then, it was made the UFRN Campus` characterization; the models` definition which formed the data collection basis; the detailing of the procedures used in the research, the processing description and the data analysis. Subsequently, the results that clarify the issues raised were shown through quantitative and / or qualitative data analyses. This research notes a high level of satisfaction by the users, despite some problems such as the reflections occurrence on the board, the lack of uniformity and, occasionally, the brightness low levels

Percepção dos usuários nos espaços públicos: avaliação pós-ocupação em três praças de Natal-RN

The research investigates the acting and the importance for the users of the public squares located predominantly in residential areas. It presents the results of the posoccupation evaluations accomplished in three squares, whose physical, environmental characteristics, equipments and fumitures are different in its qualities and amounts, taking in consideration aspects related to the users' physical and psychological comfort and of the inhabitants of the I spill. The collection of data involved physical risings and of files, observations behavious, application of questionnaires and interviews, analyzed qualitative as quantitatively so much for a larger precision and validity of the investigation. The results were obtained through the relationship among the users' perception the environmental attributes and the different levels of apropriation/use of the studied places. They indicate that the aspects composicionais of physical order of the space affects the type and the intensity of use of the squares intimately, contributing positive or negatively for its valor. It is evidenced like this, that the low freqüentation of the public squares of Natal, is due mainly to referring aspects to the physical quality and the amount of the fumitures and urban equipments. It is ended that the investments and the physical planning of these public spaces should be based in the real knowledge of the aspirations of the population objective, in way to allow its largest use and valor

Système d'aide à la planification et à la conception d'une stratégie d'évaluation formative instrumentée

Le logiciel SysML (Système Mastery Learning) est le prototype d'un logiciel pour enseignants qui désirent implanter le Mastery Learning dans leurs cours. Appartenant à la famille des APO (Applications Pédagogiques de l'Ordinateur), ce logiciel de GPAQ (Génie Pédagogique Assisté par Ordinateur) vient répondre à des besoins d'encadrement et d'assistance formulés par les enseignants qui utilisent le Mastery Learning. Ce projet de recherche et de développement visait à produire un logiciel pour assister et encadrer les enseignants dans le processus de planification et de conception d'une stratégie instrumentée d'évaluation formative. Le but de SysML est de réduire la perte de temps lors de la conception des instruments et à rendre ces instruments de meilleure qualité. Le logiciel SysML a été développé suite à une analyse du problème et des besoins des enseignants et est basé sur des modèles pédagogiques éprouvés. Le logiciel est programmé dans l'environnement HyperCard sur ordinateur Macintosh . SysML a été expérimenté par une conseillère pédagogique et trois enseignants du réseau collégial. Les commentaires des expérimentateurs, recueillis lors des entrevues, sont positifs et nous indiquent qu'un tel logiciel serait bienvenu dans le monde de l'éducation. Par ailleurs, SysML ne serait pas limité aux utilisateurs du Mastery Learning mais serait ouvert aux enseignants qui veulent organiser, planifier un cours et une stratégie d'évaluation. L'essentiel du projet de recherche a été la production du logiciel. Tout le travail de recherche est dans le logiciel, ce document n'est que le rapport accompagnateur. On peut avoir une démonstration du fonctionnement du logiciel en s'adressant à l'auteur.

Narrative transportation in global brand content

People understand life and events around them through narratives. Narratives are a new way for marketers to convey messages to consumers about their brands and products. Brand narratives are an effective way to reach out to people due to their influential nature. Narratives have a power to change beliefs and attitudes, making them relevant and interesting for any marketer. The power of narratives has to do with narrative transportation, which narratives can trigger in people. A transported person is more likely to perceive brand or product in a more positive light. The creators of the narrative are able to influence the content of the narrative through message factors that work as antecedents for narrative transportation. This study explored narrative transportation qualities of well established advertisements. The study uses qualitative content analysis to analyze and identify narrative transportation antecedents among Cannes Lion grand prix winners in the film category between years 2005 to 2015 (15 in total). The narrative transportation antecedents are identifiable characters, imaginable plot and verisimilitude, which were used in the analysis of the data. The study analyzes the winners to make judgment on whether they can trigger narrative transportation or not. It was found that the Cannes Lion grand prix winner advertisement mostly had identifiable narratives in them. In most of the advertisement (ten out of fifteen) at least two out of three antecedents were found, thus most of them are able to trigger narrative transportation. The study also found that most narratives in the advertisement were able to be linked to the main brand narrative of the advertiser. In four of the advertisements the link to the brand narrative was not able to be established. The study concludes by discussing certain factors and aspects of the advertisements that were identified to further enhance the narrative transportation qualities of the advertisement.