The importance of localised delivery of chemotherapeutic drugs for cancer treatment and specifically solid tumours has been widely reported. In this study, the anticancer drug N4-myristoyl gemcitabine (a lipophilic form of gemcitabine) was formulated as organogel to achieve a localised depot delivery. Thus, the first goal of this study was to evaluate the suitability of the oragnogel for intartumoural injection and this attained by investigating the thermostability and elasticity of the organogel. Further to this, the second goal was to slow the release of N4-myristoyl gemcitabine from the organogel. Accomplishment of these two goals will guarantee a better efficacy of cancer treatment by obtaining direct contact of the organogel containing the N4-myristoyl gemcitabine with the cancerous cells. The studies herein selected the 12-hydroxystearic acid (12-HSA) as the gelator and using 2 types of solvents the liquid part of the organogel. The first type of solvent was a series of oils which were soybean oil (SO), medium chain triglyceride (MCT), glyceryl tributyrate (TGB) and glyceryl triacetate (GTA) whilst, the second type of solvent was propylene glycol (PG). Initially thermal stability was screened using table top rheology and DSC from 0.5% to 5% w/w 12-HSA in different oils. Also to test the mechanical strength of the organogels, amplitude sweep, frequency sweep, time dependant recovery and creep and recovery tests were executed to differentiate between the organogels. The best organogels were the 5% w/w 12-HSA in SO and MCT due to their highest thermal stability, denser scaffolds, thixotropic behaviour and were the least compliant. The same experiments were utilised to evaluate the selected range of 0.5% to 14% w/w 12-HSA in PG. 14% w/w 12-HSA in PG was selected again due to its higher thermal stability, thixotropic behaviour and was less compliant compared to other concentrations of 12-HSA in PG. Drug release from the selected organogels was then carried out. The cumulative percentage released from 0.5% and 0.3% w/w N4-myristoyl gemcitabine in 5% w/w 12-HSA/MCT organogels as a solid organogel was 18.95% and 26.62% after 30 days whilst for the organogel liquefied with N-methyl pyrrolidone (NMP), the cumulative percentage released was 35.02% and 34.37% within the same frame time. Further to this, a sample and separate release method was used to study the liquefied form of the 5% w/w 12-HSA/MCT. Also, this method revealed that the 5% w/w 12-HSA/MCT organogels gave a slow release of N4-myristoyl gemcitabine and 56.18% and 70.07% was released from the 0.5% and 0.3% w/w selected organogels respectively within 30 days. For the 14% w/w 12-HSA in PG organogel, the cumulative percentage released for 0.5% and 0.3% w/w N4-myristoyl gemcitabine in 14% w/w 12-HSA/PG organogels was 26% and 40% respectively after 30 days. To conclude, our selected organogels (5% w/w 12-HSA/MCT and 14% w/w 12-HSA/PG) met the goal of our work firstly, by showing the strength and the elasticity to be injected. Secondly, they were able to slow down the release of N4-myristoyl gemcitabine.

Made available in DSpace on 2012-09-05T18:23:45Z (GMT). No. of bitstreams: 2 license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) 167.pdf: 2685354 bytes, checksum: b69f7bb07b7fdf66098c0af6cb293825 (MD5) Previous issue date: 2003 / No presente trabalho descrevemos o processo de racionalização social, caracterizado pela expansão de uma forma de razão instrumental e sua produção, enfatizando a convergência de organizações profissionais e ideologias técnicas. Esta racionalidade é fundamentada em mitos modernos, símbolos e ritos de tecnologia, que produzem as distorções, aqui descritas. O processo de cientificismo cultural e consumismo originou construções teóricas de controle e dominação, como o behaviorismo, a cibernética social (capítulo II) e a Health Information for Consumers (capítulo IX). Nos ambientes hospitalares descrevemos deslocamentos de atos cotidianos para terrenos técnicos, assim como a idéia de saúde como um item manipulável por especialistas a ser consumido por pessoas comuns. As organizações hospitalares como comunidades lingüísticas (e inspiradas por esta racionalidade instrumental), são estruturadas em equipes de especialistas que contam com parcos recursos comunicativos (capítulo V to VIII). Os atos de fala comuns são substituídos por próteses de comunicação , concentradas em sistemas semânticos técnicos, extraídos por estas equipes de suas referências culturais específicas (capítulo V). As debilidades destes sistemas de comunicação estão inscritas em seus típicos mecanismos de argumentação e suas fontes implícitas de referência e validação. A formalização destes sistemas é descrita como modelos comunicativos (capítulo VII). Descrevemos a cacofonia da mídia causada pela super-exposição dos experts (em convergência com conteúdos linguísticos reprimidos, como a morte) que degenera em patologias sociais no campo comunicativo. Tais distorções refletem vieses econômicos, científicos e instrumentais, os quais validam a informação sobre saúde como um objeto a ser transferido sem qualquer tipo de pesquisa de recepção de suas mensagens. Neste sentido destacamos como as TCI (Tecnologias de Informação e Comunicação) têm coberto domínios de informação médica cada vez mais amplos, produzindo novos campos de especialização conhecidos como Telemedicina, Cyber-Medicina e o Consumo de Informação em Saúde (capítulo IX).Usamos a teoria habermasiana da ação comunicativa como base teórica para observação destes limitados sistemas de pensamento. Propomos uma ética do discurso baseada na necessidade das conexões intersubjetivas (a atenuar os pressupostos de univocidade das mensagens) que estreitaria os laços de confiança entre profissionais e seus pacientes.

Extrusion based three dimensional (3D) printing is defined as a process used to make a 3D object layer by layer directly from a computer aided device (CAD). The application of extrusion based 3D printing process to manufacture functional oral solid tablets with relatively complex geometries is demonstrated in this thesis. In Chapter 3 the viability of using a basic desktop 3D printer (Fab@Home) to print functional guaifenesin bilayer tablets (GBTs) is demonstrated. Guaifenesin is an over the counter (OTC) water soluble medicine used as expectorant for reduction of chest congestion caused by common cold and infections in respiratory system. The bilayer tablets were printed using the standard pharmaceutical excipients; hydroxypropyl methyl cellulose (HPMC) 2208, 2910, sodium starch glycolate (SSG), microcrystalline cellulose (MCC) and polyacrylic acid (PAA) in order mimic the commercial model formulation (Mucinex®) guaifenesin extended-release bilayer tablets. The 3D printed guaifenesin bilayer tablets (GBTs) were evaluated for mechanical properties as a comparison to the commercial GBTs and were found to be within acceptable range as defined by the international standards stated in the USP. Drug releases from the 3D printed GBTs were decreased as the amount of HPMC 2208 increased due to the increased wettability, swelling properties and gel barrier formation of the HPMC. The 3D printed GBTs also showed, as required, two release profiles: immediate release (IR) from the top layer containing disintegrants; SSG and MCC and sustained release (SR) profile from the lower layer containing HPMC 2208. The kinetic drug release data from the 3D printed and commercial GBTs were best modelled using the Korsmeyer–Peppas model with n values between 0.27 and 0.44. This suggests Fickian diffusion drug release through a hydrated HPMC gel layer. Other physical characterisations: X-Ray Powder Diffraction (XRPD), Attenuated Total Reflectance Fourier Transform Infrared Spectroscopy (ATR-FTIR), and Differential Scanning Calorimetry (DSC) showed that there was no detectable interaction between guaifenesin and the used excipients in both 3D printed and commercial GBTs. A more complex printer (RegenHu 3D bioprinter) was subsequently used to print complex multi-active tablets containing captopril, nifedipine, and glipizide as a model therapeutic combination. These drugs are frequently used to treat hypertension and diabetes mellitus. The 3D printed tablets were evaluated for drug release and showed that captopril was released by osmosis through permeable cellulose acetate (CA) film and both glipizide and nifedipine were released by diffusion through the hydrophilic HPMC 2208 matrix. According to XRPD and ATR-FTIR results, there was no detectable interaction between the actives and the used excipients. In the final experimental chapter, a combined treatment regimen: atenolol, ramipril, hydrochlorothiazide (anti-hypertensive medications), pravastatin (cholesterol lowering agent), and aspirin (anti-platelets) were printed into more complex geometry (polypill) using the RegenHu 3D bioprinter. This combined drug regimen is manufactured by Cadila Pharmaceuticals Limited as a capsule formulation under the trade name of Polycap™ and is currently the only polypill formulation commercially available and is used to treat and prevent cardiovascular diseases. The printed polypills were characterized for drug release using USP dissolution testing and showed the intended immediate and sustained release profiles based upon the active/excipient ratio used. Aspirin and hydrochlorothiazide were immediately released after the polypill contacted the dissolution medium, and atenolol, ramipril, and pravastatin were released over a period of 12 hrs. XRPD and ATR-FTIR showed that there was no detectable interaction between the actives and the used excipients. In this work, extrusion based 3D printing technique was used to print oral solid dosage forms with complex and well-defined geometries and function. The technology of 3D printing could offer the opportunity to print oral tablets with high and precise drug dosing and controlled drug release profiles tailored for sub-populations or individuals. If the manufacturing and regulatory issues associated with 3DP can be resolved such personalised medicine delivered by 3D printing could improve patient compliance and provide more effective treatment regimes.

Successful tissue repair requires orchestrating a range of biochemical and biophysical factors to direct cell differentiation towards tissue specific lineages. Biodegradable poly DL lactic acid-co-glycolic acid (PDLLGA) microspheres have been reported as a promising injectable cell delivery system with controllable growth factor release potential for different tissue engineering applications. Injectable PDLLGA microspheres have been shown to form highly porous scaffolds at body temperature with a mechanical strength comparable to bone tissues. However, as the elastic properties of the injury microenvironment were shown to have a pivotal role on directing stem cell lineage specification, this work has proposed photo-crosslinkable gelatine methacrylate (gel-MA) hydrogels as promising surface coatings with tunable elastic properties. Moreover, as PDLLGA microspheres have limited functional groups on their surface, this work has evaluated different surface modification and grafting approaches to enable proper grafting of thick gel-MA hydrogel layer to the surface. Surface adsorption, surface entrapment, and oxygen plasma treatment approaches have been proposed and evaluated to modify the surface of PDLLGA microspheres with high density of gel-MA molecules. Surface analytical techniques such as ToF SIMs and XPS have been used to evaluate and quantify the density of gel-MA molecules on the surface, while fluorescent and scanning electron microscopies have been used to visualise the fluorescent deposition of fit-C gel-MA to the surface. Later, grafting-to and encapsulation approaches have been investigated to graft a thick layer (10-20 μm thick) of gel-MA hydrogel to the surface of PDLLGA microspheres following modification with gel-MA. Fluorescein isothiocyanate labelled human serum albumin Fit-C HSA has been loaded into PDLLGA microspheres as a model protein to study its release behaviour from the proposed system. Release data have shown a comparable release profile between PDLLGA microspheres before and after coating with the hydrogel layer suggesting no adverse effect of the proposed coating approach on the release behaviour. Gel-MA hydrogels with tunable elastic properties have been prepared and analysed using texture analyser and atomic force microscopy (AFM). Hydrogels have been later imaged with focused ion beam scanning electron microscope (FIB-SEM) using a novel approach to capture the hydrated structure of the hydrogel. Data obtained from the texture analyser using the compression and indentation mode tests have shown that the elastic modulus values were significantly higher than the values obtained from tension mode tests. In comparison, the values obtained from the texture analyser with the tension mode test were comparable with the values obtained using the AFM nano-indentation tests. This has been explained with the poroelastic behaviour of the hydrated hydrogel structure where a micron size pores have been observed. To verify findings, human mesenchymal stem cells have been cultured on the surface of gel-MA hydrogels to study their phenotypic behaviour and stained with anti-osteogenic or anti-neurogenic immunofluorescent markers to define their fate accordingly. Images have shown that cells cultured on hydrogels with AFM analysed elastic values of (~26, ~9.3, and ~0.1 KPa) have committed to a phenotypic behaviour related to the elastic modulus values of bone, muscle, and neuronal tissues respectively. In comparison, the elastic modulus values obtained from gel-MA hydrogel microbeads with AFM have been notably higher and appeared to be dependent on the cross-linking temperatures. Finally, the proposed cell delivery system can be used to control the chemical and the mechanical properties of the stem cell microenvironment which may pave the way towards directing stem cell differentiation into tissue specific cell lineages for different tissue repair applications.

Background and purpose: Angelicae Sinensis Raidx (Chinese Angelica, Dang Gui, DG), the dry root of Angelica sinensis (Oliv.) Diels, is one of the most popular herbs used around the world. It has been named as the “female ginseng and served as an indispensable herb to treat many obstetrical and gynecological diseases. Traditionally, DG was recommended to pregnant women to ease delivery and to eliminate complications. It is believed that the body of DG (Dang Gui Shen, DGS) is superior in nourishing blood, while the tail of DG (Dang Gui Wei, DGW) is commonly used to remove blood stagnation. Clinically, DG is commonly combined with Paeoniae Radix Alba (White Peony Root, Bai Shao, BS) and Rehmanniae Radix (Unprocessed Rehmannia Root, Sheng Di Huang, SDH) to treat disorders during pregnancy as it may not only the strengthen therapeutic effects but also eliminate adverse effects caused by each single herb. However, it is contradictory that DG may increase the risk of miscarriages reported by previous studies: the use of DGS among pregnant women, while avoiding using DGW has always been recommended since ancient times to avoid miscarriage. To date, there is no clear evidence to identify the safety of DG in pregnant women and to support the theory that different pharmaceutical effects are attributed to chemical difference between DGS and DGW. Furthermore, little is known regarding the specific effects of DG on fetal bone while limited research has been done to explore herb-herb interactions between DG and other herbs. The aims of this project are (1) to identify the safety of DG in maternal and fetal health; (2) to compare the chemical composition of DGS and DGW and their cytotoxicity; (3) to analyze the integrated role of herb-pair (DG plus BS or SDH); (4) to investigate the mechanism of specific impact of herb-herb interaction emphasis on embryonic development. Based on the theory of traditional Chinese medicine, our project is believed to provide experimental evidence to rationalize clinical use of DG in pregnant women. Method: (1) For the herbal quality control, aqueous extracts of DG, DGS, DGW, BS and SDH were prepared respectively, and their reference marker compounds were quantitatively authenticated by HPLC. In addition, pesticide residues and heavy metals in DG extract were examined by GC-MS and ICP-MS. Moreover, comparison of composition of DGS and DGW extract in terms of main constituents was performed by GC-MS and LC-MS analysis. (2) In-vivo mouse study (Segment II study), pregnant mice were randomly assigned into different dosage groups: oral administration of either distilled water as negative control, or DG extract of 2, 8, 16, 32 g/kg/day, or BS extract of 2, 16, 32 g/kg/day, or SDH extract of 2, 16, 32 g/kg/day, or DG (32 g/kg/day) plus BS (32 g/kg/day), or DG (32 g/kg/day) plus SDH (32 g/kg/day), respectively from the gestation day (GD) 6 to 15; another group mice were treated with vitamin A (200,000 IU) on the GD7, 9 and 11 as positive control. The mice were sacrificed for assessing parameters on GD18. (3) In-vitro assay using embryonic stem cell (ESC) and fibroblast 3T3 cell was conducted to investigate the cytotoxicity of DG, Z-LIG, FA, DGS, DGW, BS and SDH by MTT test, according to European Centre for the Validation of Alternative Methods. (4) For mechanistic study of DG impacts and herb-herb interactions, the expression of a characteristic set of bone formation/resorption markers, and some site-specific bone regulatory factors in fetal tissues and amniotic fluids on the GD15 were measured by ELISA. Result: (1) In the study to evaluate the safety of DG extract, maternal body weight (BW), gravid uterine weight, corrected BW change, live fetus/litter, mean fetal body weight in the group of DG (32 g/kg/day) were significantly lower than those of the negative control (p < 0.05); while resorption site/litter, post-implantation loss (PIL)/litter, percentage of abnormal skeleton were significantly higher than those of the negative control (p < 0.05). Although there was no statistical difference between IC50 values of ESCs (IC50 ESC) and 3T3 cells (IC50 3T3) after treatment with DG, Z-LIG and FA samples respectively, the IC50 Z-LIG was significantly less than IC50 FA in both ESCs and 3T3 cells (p < 0.05). It was indicated that DG extract (32 g/kg/day) might result in adverse impacts to maternal function and fetal development in mice. Z-LIG in DG extracts might be less safe compared to FA in in-vitro cultured cells and its potential impacts should be further examined its potential impacts in in-vivo studies. (2) In the study to compare the composition of main constituents from DGS and DGW water extract, HPLC quantitative analysis indicated that the ratio of FA and Z-LIG between extract from DGS and DGW is 1:1.83 and 1:1.35, respectively. Sathulenol (1), 3-butylphthalide (2), Z-butylidenephthalide (3), benzeneacetic acid (4), Z-LIG (5) and E-LIG (6) were identified by GC-MS analysis. The peak area of compound 5 in DGW extract was close to 5 times of that in DGS extract. The amounts of compound 2 and 3 in DGW extract were respectively over 20 times and 2 times higher than that in DGS extract, respectively. Except for compound 3, 5, 6, additional three compounds: coniferyl ferulate (7), FA (8), senkyunolide A (9) were identified by LC-MS analysis. The amount of compound 3, 5, 6, 7, 8, and 9 in DGW extract was higher than that in DGS extract. The peak area of compound 3 and 5 in DGW extract was over 2 times of that in the DGS extract. In MTT assay, the effect of DGS and DGW water extract on inhibition of cell viability of cultured ESCs and 3T3 cells was in a dose-dependent manner, respectively. The difference between IC50 ESC and IC50 3T3 after DGS extract treatment was statistically significance (p < 0.05), however no statistical significance was identified in DGW (p > 0.05). Both IC50 ESC and IC50 3T3 values of DGW were much lower than those of DGS (p < 0.05). (3) In the study to evaluate the role of DG plus BS or SDH, expectedly DG extract (32 g/kg/day) resulted in significant abnormalities in maternal and fetal parameters when compared with the negative control. Whereas BS or SDH extracts at a dosage of 2, 16, or 32 g/kg/day did not result in any adverse effect for both maternal health and embryonic development. There was no statistically significant difference between the IC50 ESC and IC50 3T3 value in the cytotoxicity assays of BS or SDH extracts (p > 0.05). It was indicated that the use of BS or SDH extract should be safer than DG extract in pregnant mice. More importantly, the treatment with DG plus BS or DG plus SDH extract could significantly correct abnormalities caused by DG extract alone as seen in the corrected BW change, mean fetal body weight, live fetus/litter (%), resorption site/litter (%), PIL/litter (%), skeletal variation (%), etc. (p < 0.05) in pregnant mice. (4) In the study to analyze the mechanism of herb-herb interactions, the mean values of PICP, ALP-Bone, osteocalcin, BMPs and GDF-5 in fetal tissues were significantly lower in mice treated with DG extract (32g/kg/day) alone when compared with the negative control (p < 0.05); while there was no significant difference among the mice treated respectively with BS, SDH, DG plus BS and DG plus SDH extracts with the same dosage. The outcome was similar to those of the negative control (p > 0.05). In addition, there were no significant differences in the mean value of ICTP in both fetal tissues and amniotic fluids among all mice groups (p > 0.05). Conclusion: (1) High dosage and long-term use of DG water extract may result in adverse effects on embryonic development including fetal bone malformations, hence its use is considered as not safe in pregnant women. As DG extract in this study was not contaminated by pesticide residues and heavy metals, the embryonic toxicity of DG extract can be considered as due to the intrinsic constituents of the herb. (2) As seen in cytotoxicity assay, that water extract of DGW had the lower IC50 value, hence it is believed that the higher phthalides level (3-butylphthalide, Z-butylidenephthalide, senkyunolide A Z-LIG and E-LIG) contributes to a more toxicity on both ESC and 3T3 cells. (3) Herb-pair extract of DG plus BS or SDH could significantly correct abnormalities caused by DG extract alone in pregnant mice. Therefore, herb BS or SDH not only has beneficial effects when used for treating pregnant disorders safety, but also has attenuated effects for DG when used together as herb-pair extract. (4) At the molecular biomarker level, DG extract might significantly affect bone formation rather than bone resorption. However, it could be ameliorated when applied combination with either BS or SDH. These results should be valuable for further analysis on the integrated effects of herb-herb interactions and complex mechanism of formula therapies in Chinese herbal medicine.

No description available.

The National Health Service (NHS) focuses on quality of care as a priority. With the NHS planning to go paperless by 2018, more hospitals in England are making the transition from paper to electronic prescribing (ePrescribing) systems. The aim of this programme of work was to understand and explore the influence different in-patient prescribing systems can have on key NHS healthcare professionals (doctors, nurses and pharmacists) working practices in England and quality healthcare. The programme of work, a three phase sequential design, used both qualitative and quantitative approaches. The first phase involved structured telephone interviews with chief pharmacists. Chief pharmacist interviews (n=65) focused upon the type of in-patient prescribing systems in use within each Trust and gained a management perspective of the different prescribing systems. Phases two and three were carried out at three acute NHS hospitals in England, at various stages of developing and implementing their prescribing systems. Phase two data were collected through multidisciplinary team (MDT) focus group discussions. The MDT discussions explored a number of areas associated with the prescribing systems in use: these included clinical workflow, communication, collaboration, patient safety and the use of a clinical indication on the prescription chart. Phase three data were collected using documentation analysis of the prescribing system and medical records, taken from patients cared for by the MDTs involved in phase two. Information extracted included any documentation made of a newly initiated medication, as well as the design of the prescribing system. The clarity and accuracy of documentation in the prescribing system and medical notes were compared to the GMC standards Good Practice in Prescribing Guidelines. Triangulation of data indicated how a change in prescribing system can impact upon individuals working practices by changing the design and clarity of the prescription chart, enforcing of regulations, accessibility and reliability, communication between key HCPs and the patient. These influences can be considered latent conditions in the systems that need addressing to prevent quality of patient care being compromised. The use of Socio-technical systems (STS) theory considered the interaction between humans and technology when using the prescribing systems. Understanding the issues where social and technical aspects interact in the prescribing system, emphasised where healthcare quality is impacted and therefore facilitated recommendations to improve working practices. The findings will help healthcare organisations to consider the impact a change in prescribing system can have on working practices and the latent failures that need consideration within the prescribing systems. The Electronic Prescribing and Medicines Administration (EPMA) system design must take into account the visual and physical needs of the user and consider how they can be improved to facilitate clinical workflow.

S. pneumoniae is one of the most significant human pathogens, causing high morbidity and mortality rates globally. Although there are vaccine available such as PPV 23, PCV7, PCV10, and PCV13, they are ineffective in some situations due to the differing epidemiology of various serotypes depending on the site of infection and the geographical location. Furthermore, they are expensive to produce and distribute. Universal research is presently concentrated on establishing other pneumococcalvaccine approaches such as using pneumococcal surface protein A (PspA) which relate to pathogenesis and are common to all serotypes. In this study polymeric nanoparticles (NPs) encapsulating PspA4Pro were incorporated into microcarriers using L-leucine and spray dried to produce nanocomposite micro#particles (NCMPs) dry powder for inhalation. Parameters for the preparation of protein-loaded polyester poly (Glycerol Adipate-co-ω-Pentadecalactone), (PGA-co-PDL) NCMPs were optimised using Taguchi design and BSA as a model protein, by the double emulsion solvent evaporation method followed by spray drying. Particle size was mainly affected by the polymer mass and small particle size ≤ 500nm was achieved. The most important factor for obtaining a high BSA loading was BSA concentration. The spray drying process was optimised to produce NCMPs with a porous corrugated surface, 50% yield, MMAD of 1.71±0.10 μm and FPF% of 78.57±0.1%. Adsorption of chitosan hydrochloride (CHL) onto PGA-co-PDL NPs can be used assuccessful strategies to produce cationic NPs. Cationic NPs were prepared with similarparticle size to anionic NPs ≤ 500nm. The In vitro aerosolisation performance ofcationic NPs/NCMPs showed FPF% of 46.79±11.21% and MMAD of 1.49±0.29 μm. Further cell viability studies on A549 cell line showed a good profile with a cell viability of 79±4.7% for anionic NPs/NCMPs and 78.85±9.96% for cationic xviii NPs/NCMPs at 2.5 mg/ml concentration after 24 h exposure. The previous results introduced a successful method for preparing anionic and cationic NPs/NCMPs for delivering PspA4Pro as dry powder via inhalation. The particle size of PspAPro4 loaded anionic NPs and cationic NPs were 310±25.3 nm and 409.7±49.5 nm, respectively, to be effectively taken up by dendritic cells (DCs). The PspA4Pro loading in anionic NPs was 65.73±5.6 μg/mg and in cationic NPs was 9.84±1.4 μg/mg. The PspA4Pro released from anionic and cationic NPs/NCMPs preserved its primary and secondary structure as evaluated by SDS-PAGE and circular dichroism. In vitro release studies showed that the anionic NPs/NCMPs formulations achieved a cumulative release of 21.01±1.5% while the cationic NPs/NCMPs formulation released 83.13 ±0.84% after 48 h. DCs uptake studies provide evidence of particles uptake by DCs upon incubation for 1 h as visualized by confocal microscopy. These results indicate the use of optimised methods for developing polymeric based NCMPs for vaccine delivery via inhalation against pneumococcal diseases.

Orientador: João Manuel Grisi Candeias / Resumo: Não disponível. / Abstract: From about 200 virus that cause respiratory infections, only 8 are responsible for severe illness in children, immunodeficient adults and elderly, inc1uding Adenovirus, Influenza A and B, Parainfluenza 1, 2 and 3, Human RespÍratory Syncytial Vírus (HRSV) and Human Metapneumovirus (hMPV). Three of these virus are responsible for a significant morbidity in elderly: Influenza, HRSV and hMPV. The nosocomial infection caused by these vÍruses can be fatal in hospitalized children and patients with other pathologies. With the advance of the of molecular biology techniques, the diagnosis and the characterization of these vírus became more effective. Beside vírus detection by PCR, isolation of vírus in specific cellular cultures to increase the amount of pathogens have been used in severallaboratories. Studies on prevalence of respÍratory vírus in elderly are rare. The objective of this research was to evaluate the occurrence of Influenza, HRSV and of the hMPV and the risk factors involved in the diseases caused by these vÍruses. Nasopharyngeal swabs were collected and used for inoculation in cells culture and dÍrect analysis by RT-PCR. The results from RT-PCR of Influenza Vírus and RSV were negative. We also tested the samples with GeneScan, that is a technique based on fluorescent primers specific to the studied vírus. Results for Flu virus and HRSV were the same ofRT-PCR, but 1 MPV sample was positive. 55,32% ofthe 47 studied elderly were vaccinated against Influenza; 14,9% had tabagism habits and 70,2% were women. These risk factors might had influenced in the absence of positive samples for the Influenza vírus andHRSV. / Mestre

Orientador: Osvaldo Malafaia / Inclui apęndice / Inclui bibliografia