Synthesis and functionalization of hydroxyapatite nanoparticles

Graças aos desenvolvimentos na área da síntese de nanomaterais e às potentes técnicas de caracterização à nanoescala conseguimos hoje visualizar uma nanopartícula (NP) como um dispositivo de elevado potencial terapêutico. A melhoria da sua efectividade terapêutica requer no entanto o aprofundamento e sistematização de conhecimentos, ainda muito incipientes, sobre toxicidade, selectividade, efeitos colaterais e sua dependência das próprias características físico-químicas da NP em análise. O presente trabalho, elegendo como alvo de estudo uma substância considerada biocompatível e não tóxica, a hidroxiapatite (Hap), pretende dar um contributo para esta área do conhecimento. Definiram-se como metas orientadoras deste trabalho (i) estudar a síntese de nanoparticulas de Hap (Hap NP), e a modificação das características físico-químicas e morfológicas das mesmas através da manipulação das condições de síntese; (ii) estudar a funcionalização das Hap NP com nanoestruturas de ouro e com ácido fólico, para lhes conferir capacidades acrescidas de imagiologia e terapêuticas, particularmente interessantes em aplicações como o tratamento do cancro (iii) estudar a resposta celular a materiais nanométricos, com propriedades físico-químicas diversificadas. No que se refere à síntese de Hap NP, comparam-se dois métodos de síntese química distintos, a precipitação química a temperatura fisiológica (WCS) e a síntese hidrotérmica (HS), em meios aditivados com ião citrato. A síntese WCS originou partículas de tamanho nanométrico, com uma morfologia de agulha, pouco cristalinas e elevada área superficial especifica. A síntese HS à temperatura de 180ºC permitiu obter partículas de dimensões também nanométricas mas com área específica inferior, com morfologia de bastonete prismático com secção recta hexagonal e elevada cristalinidade. Com o objectivo de aprofundar o papel de algumas variáveis experimentais na definição das características finais das partículas de hidroxiapatite, designadamente o papel do ião citrato (Cit), variou-se a razão molar [Cit/Ca] da solução reagente e o tempo de síntese. Demonstrou-se que o ião citrato e outras espécies químicas resultantes da sua decomposição nas condições térmicas (180ºC) de síntese tem um papel preponderante na velocidade de nucleação e de crescimento dessas mesmas partículas e por conseguinte nas características físico-químicas das mesmas. Elevadas razões [Cit/Ca] originam partículas de dimensão micrométrica cuja morfologia é discutida no contexto do crescimento com agregação. Com o objectivo de avaliar a citotoxicidade in vitro das nanopartículas sintetizadas procedeu-se à esterilização das mesmas. O método de esterilização escolhido foi a autoclavagem a 121º C. Avaliou-se o impacto do processo de esterilização nas características das partículas, verificando-se contrariamente às partículas WCS, que as partículas HS não sofrem alterações significativas de morfologia, o que se coaduna com as condições de síntese das mesmas, que são mais severas do que as de esterilização. As partículas WCS sofrem processos de dissolução e recristalização que se reflectem em alterações significativas de morfologia. Este estudo demonstrou que a etapa de esterilização de nanopartículas para aplicações biomédicas, por autoclavagem, pode alterar substancialmente as propriedades das mesmas, sendo pois criticamente importante caracterizar os materiais após esterilização. Os estudos citotoxicológicos para dois tipos de partículas esterilizadas (HSster e WCSster) revelaram que ambas apresentam baixa toxicidade e possuem potencial para a modelação do comportamento de células osteoblásticas. Tendo em vista a funcionalização da superfície das Hap NP para multifunções de diagnóstico e terapia exploraram-se condições experimentais que viabilizassem o acoplamento de nanopartículas de ouro à superfície das nanopartículas de Hidroxiapatite (Hap-AuNP). Tirando partido da presença de grupos carboxílicos adsorvidos na superfície das nanopartículas de Hap foi possível precipitar partículas nanométricas de ouro (1,5 a 2,5 nm) na superfície das mesmas adaptando o método descrito por Turkevich. No presente trabalho as nanopartículas de Hap funcionaram assim como um template redutor do ouro iónico de solução, propiciando localmente, na superfície das próprias nanopartículas de Hap, a sua redução a ouro metálico. A nucleação do ouro é assim contextualizada pelo papel redutor das espécies químicas adsorvidas, designadamente os grupos carboxílicos derivados de grupos citratos que presidiram à síntese das próprias nanopartículas de Hap. Estudou-se também a funcionalização das Hap NP com ácido fólico (FA), uma molécula biologicamente interessante por ser de fácil reconhecimento pelos receptores existentes em células cancerígenas. Os resultados confirmaram a ligação do ácido fólico à superfície das diferentes partículas produzidas HS e Hap-AuNPs. Graças às propriedades ópticas do ouro nanométrico (efeito plasmão) avaliadas por espectroscopia vis-UV e às potencialidades de hipertermia local por conversão fototérmica, as nanoestruturas Hap-AuNPs produzidas apresentam-se com elevado interesse enquanto nanodispositivos capazes de integrar funções de quimio e terapia térmica do cancro e imagiologia. O estudo da resposta celular aos diversos materiais sintetizados no presente trabalho foi alvo de análise na tentativa de se caracterizar a toxicidade dos mesmos bem como avaliar o seu desempenho em aplicações terapêuticas. Demonstrou-se que as Hap NP não afectam a proliferação das células para concentrações até 500 g/ml, observando-se um aumento na expressão genética da BMP-2 e da fosfatase alcalina. Verificou-se também que as Hap NP são susceptíveis de internalização por células osteoblásticas MG63, apresentando uma velocidade de dissolução intracelular relativamente reduzida. A resposta celular às Hap-AuNP confirmou a não citotoxicidade destas partículas e revelou que a presença do ouro na superfície das Hap NP aumenta a taxa proliferação celular, bem como a expressão de parâmetros osteogénicos. No seu conjunto os resultados sugerem que os vários tipos de partículas sintetizadas no presente estudo apresentam também comportamentos interessantes para aplicações em engenharia de tecido ósseo.

Propriedades de lamas de salinas de Aveiro, tendo em vista aplicações dermoterapêuticas e dermocosméticas

Desde tempos históricos que diferentes tipos de lama são utilizados para aplicações externas no corpo humano, para fins terapêuticos e cosméticos. As lamas cuja beneficiação e caracterização físico-química são apresentadas nesta dissertação são formadas no ambiente hipersalino que existe nas salinas da Troncalhada e de São Tiago da Fonte, localizadas no estuário do rio Vouga, em Aveiro, Portugal. As salinas são constituídas por uma sequência de tanques onde, no período de verão, da água do mar e por evaporação natural, se produz sal marinho que precipita no último tanque (o cristalizador) do qual é extraído. Na base dos tanques ocorrem lamas que correspondem a sedimentos constituídos por material argiloso de cor preto-cinzento e que incorporam argila, silte, areia, bioclastos, sal, matéria orgânica e gás. A componente inorgânica da lama extraída do cristalizador foi estudada por Difracção de Raios-X (DRX) e Fluorescência de Raios-X (FRX), enquanto a componente orgânica da mesma lama foi estudada por Cromatografia de Gás- Espectrometria de Massa (GC-MS). Os estudos efectuados incidiram sobre amostras representativas de lama, obtidas antes e após refinação e beneficiação a que foi submetida a lama tal-qual colhida nas salinas. Foram utilizados métodos geofísicos para caracterizar e distinguir as lamas depositadas na época de safra e no período de interregno. Para o efeito, foram cravados tubos amostradores no sedimento que reveste o fundo dos tanques, tubos que seguidamente foram transportados para o laboratório para medição da condutividade eléctrica do topo até à base da coluna de sedimento amostrado. A refinação foi efectuada por elutriação de suspensões aquosas de lama utilizando um equipamento desenvolvido para o efeito e que permitiu concentrar a lama fina no overflow. Após floculação, sifonagem da água sobrenadante, dessalinização e centrifugação a lama refinada e beneficiada pôde ser incorporada em formulações com objectivos terapêuticos e cosméticos. O estudo microbiológico efectuado nas amostras de lama com sal e sem sal e na água das salinas permitiu identificar diversos tipos de bactérias e colónias presentes na lama e avaliar também os processos de esterilização testados. Concluiu-se que a lama hipersalina ou dessalinizada resultante do processamento a que foram submetidas, não deve ser aplicada ou incorporada em formulações tal-qual, pelo facto de em ambas terem sido identificadas bactérias como é o caso de Clostridium perfringens. Não obstante, se submetidas a esterilização utilizando autoclave a lama salina refinada e beneficiada poderá ser aplicada como peloide extemporâneo. Assim sendo foram desenvolvidas formulações dermoterapêuticas e dermocosméticas contendo lama beneficiada e dessalinizada e esterilizada termicamente.

Limitações voluntárias dos direitos de personalidade no ordenamento jurídíco brasileiro:em especial, os direitos à vida e à integridade física

Tese de doutoramento, Ciências Jurídicas (Direito Civil), Universidade de Lisboa, Faculdade de Direito, 2014

Sperm utilization patterns in Gryllus integer (orthoptera: gryllidae

Sperm competition is the competition for fertilizations between ejaculates, within a female, following multiple mating. There are four sperm utilization or precedence patterns: first male precedence, where the first male to mate fertilizes most of the eggs laid by a female; last male precedence, where the last male to mate fertilizes most of the eggs laid by a female; "all-or-none" pattern, where sperm from either male fertilizes all the eggs laid by a female but which male's sperm that is used is random; or sperm mixing, where sperm from each male is used equally in fertilizing eggs laid by a female. Intermediate utilization patterns are also possible. Sperm competition occurs in a wide variety of insect species as well as other animals. This study was undertaken to study sperm competition in the field cricket, Gryllus integer. Four experiments were conducted: a radiation and sterilization experiment, a diapause experiment, and 2 competition experiments. It was found that 7,000 rad of gamma radiation sterilized adult ~ integer males. There was no diapause in the laboratory in ~ integer eggs. In the first competition experiment, three groups of females were used: females mated with a normal male, then with a second normal male (NN group); females mated with a normal male, and then with a sterile male (NR group); and females mated with a sterile male, and then with a normal male (RN group). The results obtained from this experiment showed that the mean proportion of eggs hatched was significantly different between 3 groups of females, with the proportion hatched much greater in the NN group than in either the NR or RN groups. The pattern for the proportion of eggs hatched following a double mating most closely resembled a pattern expected if sperm mixing is occurring. Results obtained in the replicate competition experiment showed that the mean proportion of eggs hatched for the females in the NR group was significantly lower than the proportion hatched in the other two groups. This also supports a model of sperm mixing as a precedence pattern. Values calculated following Boorman and Parker (1976), for the proportion of eggs fertilized by the second male to mate following a double mating, were 0.57 in competition experiment 1 and 0.62 in the replicate. These values indicate that sperm mixing occurs in~ integer.

Étude des mécanismes d’inactivation des microorganismes suite à un traitement à l’ozone

Une partie du travail a mené a un dépôt de brevet.

Les politiques de stérilisation sexuelle au Canada et aux États-Unis : une pratique à l'intersection de rapports de genre, de race et de classe

Dans ce mémoire, l’objectif poursuivi sera d’éclairer les dynamiques de genre, de race, de classe, de nation et de handicap à travers le phénomène du contrôle des capacités reproductives des femmes. Dans un premier temps, j’essaierai de comprendre comment les passés coloniaux du Canada et des États-Unis ont structuré leur rapport à la reproduction et comment celle-ci est devenue un enjeu politique de premier plan au sein de l’idéologie eugéniste. Dans un deuxième temps, j’explorerai quel a été le rôle de la science dans la mise en place, en Occident, de systèmes experts capables de guider la société vers le Progrès. Ces réflexions me permettront de retracer quel a été le contexte d’émergence des lois sur la stérilisation sexuelle et quels discours de légitimation ont été mis de l’avant afin de justifier l’appropriation des capacités reproductives de certaines populations jugées « indésirables ». Ainsi, je poserai l’hypothèse que les valeurs et présupposés « scientifiques » racistes, sexistes et classistes sous-jacents à l’élaboration de ces lois ont mené à des stérilisations forcées de certains groupes minorisés, c’est-à-dire les femmes autochtones au Canada et les femmes noires aux États-Unis. Je tenterai alors d’évaluer si, effectivement, les politiques de stérilisation aux Canada et aux États-Unis ont été discriminatoires dans leur formulation et dans leur mise en application à l’égard de ces populations. Finalement, je mobiliserai les figures de la welfare queen et de la squaw afin de comprendre comment ces identités assignées ont permis de légitimer un traitement différencié à leur égard et comment elles structurent encore aujourd’hui leur rapport à la sexualité et à la reproduction.

Évaluation in vitro de la stérilisation au peroxyde d'hydrogène sur les propriétés biologiques de prothèses utilisées lors de stabilisation du genou chez le chien

Objectif—Comparer les effets de la stérilisation au plasma de gaz de peroxyde d’hydrogène (HPGP) à l’oxyde d’éthylène (EO) et à la vapeur (ST) sur les propriétés physico-chimiques et d’adhésion bactérienne de fils de nylon et de polyéthylène. Design expérimental—Etude in vitro. Matériel—Des brins non stérilisés, stérilisés au HPGP, à l’EO et ST; de fil nylon leader (FNL), de fil de nylon pêche (FNP) et de fil de polyéthylène (PE) ont été utilisés. Méthodes—Une analyse de surface au spectroscope photo-électronique à rayons X (XPS), une mesure de l’angle de contact, une analyse par microscopie à force atomique (AFM) et l’adhésion bactérienne de Staphylococcus intermedius et d’Escherichia Coli ont été testés sur les brins. Résultats—Une oxydation de la surface de tous les échantillons stérilisés a été observée quelque soit la méthode de stérilisation. La stérilisation a augmenté significativement l’angle de contact pour tous les types de fil quelque soit la méthode. La rugosité n’a pas été affectée significativement par la méthode de stérilisation pour le FNL et FNP. L’adhésion bactérienne a été affectée significativement par la méthode de stérilisation. Le PE a un angle de contact, une rugosité et une adhésion bactérienne significativement plus élevée que le FNL et FNP, peu importe la méthode de stérilisation. Conclusion—La stérilisation au HPGP constitue une alternative intéressante à la vapeur et l’EO. Le PE n’est peut être pas un matériel idéal par sa capacité d’adhésion bactérienne. De futures études sont nécessaires pour déterminer la signification clinique de ces trouvailles.

Étude de l'influence de la réassociation en surface des atomes N et O sur l'inactivation des spores bactériennes dans une post-décharge N2-O2 basse pression en flux

Le recours au plasma pour stériliser des dispositifs médicaux (DM) est un domaine de recherche ne datant véritablement que de la fin des années 1990. Les plasmas permettent, dans les conditions adéquates, de réaliser la stérilisation à basse température (≤ 65°C), tel qu’exigé par la présence de polymères dans les DM et ce contrairement aux procédés par chaleur, et aussi de façon non toxique, contrairement aux procédés chimiques comme, par exemple, l’oxyde d’éthylène (OEt). Les laboratoires du Groupe de physique des plasmas à l’Université de Montréal travaillent à l’élaboration d’un stérilisateur consistant plus particulièrement à employer les effluents d’une décharge N2-%O2 basse pression (2-8 Torrs) en flux, formant ce que l’on appelle une post-décharge en flux. Ce sont les atomes N et O de cette décharge qui viendront, dans les conditions appropriées, entrer en collisions dans la chambre de stérilisation pour y créer des molécules excitées NO*, engendrant ainsi l’émission d’une quantité appréciable de photons UV. Ceux-ci constituent, dans le cas présent, l’agent biocide qui va s’attaquer directement au bagage génétique du micro-organisme (bactéries, virus) que l’on souhaite inactiver. L’utilisation d’une lointaine post-décharge évite du même coup la présence des agents érosifs de la décharge, comme les ions et les métastables. L’un des problèmes de cette méthode de stérilisation est la réduction du nombre de molécules NO* créées par suite de la perte des atomes N et O, qui sont des radicaux connus pour interagir avec les surfaces, sur les parois des matériaux des DM que l’on souhaite stériliser. L’objectif principal de notre travail est de déterminer l’influence d’une telle perte en surface, dite aussi réassociation en surface, par l’introduction de matériaux comme le Téflon, l’acier inoxydable, l’aluminium et le cuivre sur le taux d’inactivation des spores bactériennes. Nous nous attendons à ce que la réassociation en surface de ces atomes occasionne ainsi une diminution de l’intensité UV et subséquemment, une réduction du taux d’inactivation. Par spectroscopie optique d’émission (SOE), nous avons déterminé les concentrations perdues de N et de O par la présence des matériaux dans le stérilisateur, ainsi que la diminution de l’émission UV en découlant. Nous avons observé que cette diminution des concentrations atomiques est d’autant plus importante que les surfaces sont catalytiques. Au cours de l’étude du phénomène de pertes sur les parois pour un mélange N2-%O2 nous avons constaté l’existence d’une compétition en surface entre les atomes N et O, dans laquelle les atomes d’oxygènes semblent dominer largement. Cela implique qu’au-delà d’un certain %O2 ajouté à la décharge N2, seuls les atomes O se réassocient en surface. Par ailleurs, l’analyse des courbes de survie bi-phasiques des micro-organismes a permis d’établir une étroite corrélation, par lien de cause à effet, entre la consommation des atomes N et O en surface et la diminution du taux d’inactivation des spores dans la première phase. En revanche, nous avons constaté que notre principal agent biocide (le rayonnement ultraviolet) est moins efficace dans la deuxième phase et, par conséquent, il n’a pas été possible d’établir un lien entre la diminution des concentrations et le taux d’inactivation de cette phase-là.

STANDARDIZATION OF PROCESS PARAMETERS FOR READY TO EAT FISH PRODUCTS IN INDIGENOUS POLYMER COATED TIN FREE STEEL CANS

In the present work, Indigenous polymer coated Tin Free Steel cans were analyzed fortheir suitability for thermal processing and storage of fish and fish products following standard methods. The raw materials used for the development of ready to eat thermally processed fish products were found to be of fresh condition. The values for various biochemical and microbiological parameters of the raw materials were well within the limits. Based on the analysis of commercial sterility, instrumental colour, texture, WB-shear force and sensory parameters, squid masala processed to F0 value of 8 min with a total process time of 38.5 min and cook value of 92 min was chosen as the optimum for squid masala in tin free steel cans while shrimp curry processed to F0 7 min with total process time of 44.0 min and cook value of 91.1 min was found to be ideal and was selected for storage study. Squid masala and shrimp curry thermally processed in indigenous polymer coated TFS cans were found to be acceptable even after one year of storage at room temperaturebased on the analysis of various sensory and biochemical parameters. Analysis of the Commission Internationale d’ Eclirage L*, a* and b* color values showed that the duration of exposure to heat treatment influenced the color parameters: the lightness (L*) and yellowness (b*)decreased, and the redness (a*) significantly increased with the increase in processing time or reduction in processing temperature.Instrumental analysis of texture showed that hardness-1 & 2 decreased with reduction in retort temperature while cohesiveness value did not show any appreciable change with decrease in temperature of processing. Other texture profile parameters like gumminess, springiness and chewiness decreased significantly with increase of processing time. W-B shear force values of mackerel meat processed at 130 °C were significantly higher than those processed at 121.1 and 115 °C. HTST processing of mackerel in brine helped in reducing the process time and improving the quality.The study also indicated that indigenous polymer coated TFS cans with easy openends can be a viable alternative to the conventional tin and aluminium cans. The industry can utilize these cans for processing ready to eat fish and shell fish products for both domestic and export markets. This will help in reviving the canning industry in India.

The impact of sterilisation on family relationships

The major objective of the researcher was to assess the gains or losses in relationship between spouses after sterilization of the wife. The second objective was to gauge the impact of sterilisation upon parent—child relationships. Factors that were responsible in motivating women to accept sterilization were subjected to enquiry with the conviction that the information would give insight into the inadequacies of the official Family Planning Programme, offering prospects for feasible and salutary changes. The report is arranged under six chapters. The first chapter, Introduction, has three parts. The global and natural dimension of the problem of population is considered in Part I; The Importance of the topic studied namely sterilisation is discussed in the second Part: and Part III explains the background and methodology %f the study. The second chapter is a description of tubecto~ mized women who were the respondents. Their personal and family data, economic, social, educational and demographic profilesform its contents. The direct and indirect influences that were-responsible to make women decide in favour of sterilization are dealt with in the Third chapter. The Fourth Chapter is an analysis of the interspouse relations pre and post—operatively and an assessment of gains and losses in the area subsequent to sterilisation. How children have benefitted physically and emotionally as a consequence of improved care/attention from parents post~operatively constitutes the analysis attempted in the Fifth chapter.

Studies on arbuscular mycorrhiza (AM) in the Alentejo (Portugal) using pea mutants resistant to AM fungi as a control tool for field conditions

The utilization and management of arbuscular mycorrhiza (AM) symbiosis may improve production and sustainability of the cropping system. For this purpose, native AM fungi (AMF) were sought and tested for their efficiency to increase plant growth by enhanced P uptake and by alleviation of drought stress. Pot experiments with safflower (Carthamus tinctorius) and pea (Pisum sativum) in five soils (mostly sandy loamy Luvisols) and field experiments with peas were carried out during three years at four different sites. Host plants were grown in heated soils inoculated with AMF or the respective heat sterilized inoculum. In the case of peas, mutants resistant to AMF colonization were used as non-mycorrhizal controls. The mycorrhizal impact on yields and its components, transpiration, and P and N uptake was studied in several experiments, partly under varying P and N levels and water supply. Screening of native AMF by most probable number bioassays was not very meaningful. Soil monoliths were placed in the open to simulate field conditions. Inoculation with a native AMF mix improved grain yield, shoot and leaf growth variables as compared to control. Exposed to drought, higher soil water depletion of mycorrhizal plants resulted in a haying-off effect. The growth response to this inoculum could not be significantly reproduced in a subsequent open air pot experiment at two levels of irrigation and P fertilization, however, safflower grew better at higher P and water supply by multiples. The water use efficiency concerning biomass was improved by the AMF inoculum in the two experiments. Transpiration rates were not significantly affected by AM but as a tendency were higher in non-mycorrhizal safflower. A fundamental methodological problem in mycorrhiza field research is providing an appropriate (negative) control for the experimental factor arbuscular mycorrhiza. Soil sterilization or fungicide treatment have undesirable side effects in field and greenhouse settings. Furthermore, artificial rooting, temperature and light conditions in pot experiments may interfere with the interpretation of mycorrhiza effects. Therefore, the myc- pea mutant P2 was tested as a non-mycorrhizal control in a bioassay to evaluate AMF under field conditions in comparison to the symbiotic isogenetic wild type of var. FRISSON as a new integrative approach. However, mutant P2 is also of nod- phenotype and therefore unable to fix N2. A 3-factorial experiment was carried out in a climate chamber at high NPK fertilization to examine the two isolines under non-symbiotic and symbiotic conditions. P2 achieved the same (or higher) biomass as wild type both under good and poor water supply. However, inoculation with the AMF Glomus manihot did not improve plant growth. Differences of grain and straw yields in field trials were large (up to 80 per cent) between those isogenetic pea lines mainly due to higher P uptake under P and water limited conditions. The lacking N2 fixation in mutants was compensated for by high mineral N supply as indicated by the high N status of the pea mutant plants. This finding was corroborated by the results of a major field experiment at three sites with two levels of N fertilization. The higher N rate did not affect grain or straw yields of the non-fixing mutants. Very efficient AMF were detected in a Ferric Luvisol on pasture land as revealed by yield levels of the evaluation crop and by functional vital staining of highly colonized roots. Generally, levels of grain yield were low, at between 40 and 980 kg ha-1. An additional pot trial was carried out to elucidate the strong mycorrhizal effect in the Ferric Luvisol. A triplication of the plant equivalent field P fertilization was necessary to compensate for the mycorrhizal benefit which was with five times higher grain yield very similar to that found in the field experiment. However, the yield differences between the two isolines were not always plausible as the evaluation variable because they were also found in (small) field test trials with apparently sufficient P and N supply and in a soil of almost no AMF potential. This similarly occurred for pea lines of var. SPARKLE and its non-fixing mycorrhizal (E135) and non-symbiotic (R25) isomutants, which were tested in order to exclude experimentally undesirable benefits by N2 fixation. In contrast to var. FRISSON, SPARKLE was not a suitable variety for Mediterranean field conditions. This raises suspicion putative genetic defects other than symbiotic ones may be effective under field conditions, which would conflict with the concept of an appropriate control. It was concluded that AMF resistant plants may help to overcome fundamental problems of present research on arbuscular mycorrhiza, but may create new ones.

Exploratory Study on Occupational Health Exposure to Chemical Agents, in a Public Hospital in Valencia, Venezuela. Preliminar Assessment

Descriptive study that identified chemical agents (AQ) use and training on risk management and waste disposal techniques in a public Hospital in Valencia. A questionnaire was answered by 48 workers. Information obtained was: personal data, occupational history, AQ used; knowledge of risk management and waste disposal. There were 16 occupations from 12 “High Risk” areas. “Adult emergency” was the one with more workers (11 individuals), followed by “sterilization��� and “clinical laboratory” (7 each) and oncology (5). The remained areas had less than 8.3% workers. The most used anesthetic agents were: Halothane, Enfluorane and Isofluorane 4.17% each and main antineoplastics used were: Doxorubicin 16.67% and Paclitaxel, 5-Fluoracil and Etoposide, 8.33% each. The most mentioned substances were: alcohol (70.8%) and Chlorine (64.6%). None of the answers regarding knowledge of AQ’ risk management and waste disposal was satisfactory. Statistical associations between training and several variables such as age, time in their job and being or not a professional, resulted non-significant. The correlation between training and the knowledge of AQ’s management was significant (p < 0.001). Participants showed that their knowledge about chemical occupational risk factors they are exposed to is still insufficient. Therefore, this theme should be included in graduate course curricula. These results provide important data and will serve as a pilot research for the follow up Phase II study that will include clinical aspects and environmental and biological monitoring.

Estudio de cultivos celulares primarios derivados de Lucilia sericata (Diptera: Calliphoridae)

The main purpose of this study was to obtain primary cell cultures derived from Lucilia sericata (Diptera: Calliphoridae). Necrophagous this fly is used for determination of post-mortem interval and larval therapy. Since explants embryonated eggs were performed in various culture media (Grace Schneider, MM/VP12, DMEM, Grace/L-15 and L-15), supplemented with 20% fetal serum. Sterilization of the biological material was carried out by immersing it in formaldehyde and sodium hypochlorite solutions. The cell growth was initiated in the L-15, MM/VP12, and Schneider Grace/L-15 in an average time of 10 days after completion of planting by the proliferation of groups of colonies scattered on the surface of the boxes crops and also from the endings of larval fragments. The evolution of cell growth to the formation of monolayer semi-confluent was relatively fast, reaching at 3 weeks post-explant. Cellular morphology in cultured cells was heterogeneous, especially epithelioid forms, similar to nerve, giant and irregular. Comparison of the growth characteristics of these cell cultures with those obtained from other species of flies was more favorable in the evolution of those obtained from L. sericata, on the grounds that the cells are better adapted to the physical-chemical conditions of several culture media. This is the first report of a cell culture-fly family Calliphoridae.

Controlo microbiológico de formulações farmacêuticas parentéricas

O controlo microbiológico de formulações injectáveis visa garantir a isenção de um produto farmacêutico em termos microrganismos, possivelmente patogénicos, que possam induzir sintomas patológicos após a sua administração. Assim, o estudo das técnicas que permitam a eliminação de agentes patogénicos e a sua prevenção é de extrema importância. Nesta área são utilizadas diversas técnicas clássicas para avaliar determinados medicamentos e estão a ser desenvolvidas outras novas técnicas. Estas últimas revelam-se mais vantajosas comparativamente às clássicas uma vez que apresentam maior eficácia e rigor. Este artigo de revisão apresenta uma descrição simples das principais técnicas disponíveis (clássicas e as mais recentes) para o controlo microbiológico de medicamentos injectáveis podendo ser visto como uma ferramenta prática útil para os profissionais que trabalham ou poderão vir a trabalhar nesta área.

Flavour characterization of fresh and processed pennywort (Centella asiatica L.) juices

The flavour characteristics of fresh and processed pennywort juices treated by pasteurization, sterilization and high pressure processing (HPP) were investigated by using solid-phase micro-extraction combined with gas chromatography-mass spectrometry. Sesquiterpene hydrocarbons comprised the major class of volatile components present and the juices had a characteristic smell due to the presence of volatile compounds including β-caryophyllene, humulene, E-β-farnesene, α-copaene, alloaromadendrene and β-elemene. All processing operations caused a reduction in the total volatile concentration, but HPP caused more volatile acyclic alcohols, aldehydes and oxygenated monoterpenoids to be retained than pasteurization and sterilization. Ketones were not present in fresh pennywort juice, but 2-butanone and 3-nonen-2-one were generated in all processed juices, and 2-nonanone and 2-hexanone were present in pasteurized and sterilized juices. Other chemical changes including isomerization were also reduced by HPP compared to pasteurization, and sterilization.