Orientador: José Gonçalves Franco Júnior / Banca: Anice Maria Vieira Camargo Martins / Banca: Edson Borges / Banca: Jorge Nahas Filho / Banca: Ana Lúcia Kalinin / Resumo: Nas últimas décadas, alguns parâmetros laboratoriais têm sido propostos para identificar o potencial de saúde oocitária em Reprodução Assistida: 1. Maturidade nuclear e citoplasmática (pesquisa); 2. Dismorfismo (anomalias intracitoplasmáticas); 3. Fertilizaçao. Habitualmente, esses parâmetros (exceto a maturação citoplasmática) são avaliados pelo simples emprego de sistemas de microscopia de luz convencional. Nos últimos oito anos, foram relatados diversos estudos em oócitos humanos sobre a birrefringência da zona pelúcida (BF-ZP), a espessura da ZP, e a visualização do fuso meiótico (FM). Entretanto, ainda é discutível o poder desses novos parâmetros na seleção de óvulos com o pontecial de originar embriões com taxas de implantação elevadas, além disso, seu sistema de visualização e quantificação é complexo e dispendioso. O objetivo deste estudo foi investigar uma possível correlação entre esses parâmetros tradicionais com a BF-ZP, espessura da ZP e o FM. Dessa forma, um total de 73 pacientes que participaram do programa de injeção intracitoplasmática de espermatozóide (ICSI) no Centro de Reprodução Humana "Prof Franco Júnior" tiveram seus óvulos avaliados quanto a maturação nuclear (n=83), maturação citoplasmática (n=29), dismorfismo (n=280) e fertilização (n=216) sendo os resultados correlacionados com a imagem do FM (presença ou ausência), a intensidade da BF-ZP (positiva ou negativa) e a espessura da ZP. Os dados finais revelaram uma fraca correlação entre a maturação, o dismorfismo e a fertilização oocitária com a imagem do FM, a intensidade da BF-ZP, e a espessura da ZP. Entretanto, esses parâmetros poderiam ser responsáveis por identificar pontos biológicos diferentes do potencial de qualidade oocitária. Assim sendo, seria uma medida coerente, a utilização desses parâmetros, em forma conjunta, através de um escore de qualidade oocitária. / Abstract: During the last decades, three laboratorial parameters have been proposed to identify the potential of the oocyte quality in ART: 1- Nuclear and cytoplasmic maturation; 2-Dysmorphims (intracytoplasmic abnormalities); 3 - Fertilization. These parameters (except the cytoplasmic maturation) are assessed by a simple system of conventional light microscopy. In the last eight years, numerous studies about zona pellucida birefringence (BF-ZP), zona pellucida thickness and meiotic spindle visualization have been reported. However, it is still controversial the power of these parameters in selecting oocyte with potential to develop into implantation-competent embryos. Furthermore, the microscopy system is complex and expensive. The present study aims to evaluate whether there is any correlation between the classical parameters with BF-ZP, ZP thickness and spindle image. A total of 73 patients who participated in an ICSI program had their oocytes evaluated such nuclear maturation (n=83), cytoplasmic maturation (n=29), dysmorphism (n=280) and fertilization (n=216) being its results correlated to the spindle image (presence or absence), BF-ZP intensity (positive or negative) and ZP thickness. The final results showed a low correlation between maturation, dysmorphism and fertilization compared to spindle image and ZP characteristics (BF-ZP, ZP thickness). However, these factors can be responsible to identify specific biological differ from the oocyte potential. Concluding, the use of these factors in association, as a score (oocyte quality score), could be the correct evaluation. / Doutor

Made available in DSpace on 2014-06-12T16:24:46Z (GMT). No. of bitstreams: 2 arquivo1166_1.pdf: 4907678 bytes, checksum: 022eb5169fb973f57dfb3c991e5484ec (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2010 / Peperomia pellucida L. (H.B.K.) é conhecida popularmente como erva-de-jabuti e é usada como antimicrobiano, anti-hipertensivo e anti-inflamatório. Este trabalho teve por finalidade desenvolver tecnologicamente formas farmacêuticas à base de P. pellucida para o tratamento antimicrobiano e cardiovascular, realizando análises anatômica e microquímica das folhas, caracterização físico-química para o pó e para a solução extrativa da planta inteira, análises por espectrofotometria e cromatografia das frações flavonoídica e do extrato bruto, avaliação da sazonalidade e do processo de extração, a avaliação das atividades antimicrobiana e cardiovascular do extrato etanólico, clonagem, avaliação dos diferentes processos de secagem utilizando microscopia eletrônica de varredura e análise térmica, otimização do processo e caracterização físico-química do extrato seco por spray dryer e o desenvolvimento de cápsulas utilizando extrato seco por aspersão de P. pellucida. As características anatômicas e microquímicas foram fundamentais na identificação e caracterização da espécie. A avaliação da sazonalidade indicou que não há influência dos valores de precipitação das chuvas sobre o teor do marcador utilizado para P. pellucida, a 3 ,4 ,7-tri-O-metoxiflavona, nas amostras coletadas nos meses março, maio e julho de 2010. Na variação da concentração do solvente da maceração (50, 70 e 90%), o marcador apresentou um maior valor de teor na concentração de 70% de álcool etílico. O extrato etanólico demonstrou atividade antibacteriana contra Staphylococcus aureus e Pseudomonas aeruginosa. A micropropagação de P. pellucida foi possível utilizando-se gemas axilares jovens. O extrato seco obtido por spray dryer apresentou menor aderência ao equipamento utilizado e partículas esféricas, possibilitando uma melhor fluidez durante o desenvolvimento farmacotécnico-industrial de formas farmacêuticas sólidas. Os estudos de pré-formulação conduziram à definição da forma farmacêutica cápsula. Os resultados obtidos contribuem para a determinação de especificações de uma futura monografia farmacopeica e fornecerão parâmetros para que empreendedores da indústria farmacêutica baseiem-se para a produção de insumos, bem como para o controle de qualidade de intermediários e do produto acabado à base de P. pellucida

No description available.

Reproduction is an essential feature of all animals and a fundamental step to produce new generations. Study of sperm proteins is crucial for understanding of the sperm-egg recognition. We searched out sperm surface proteins involving in the zona pellucida (ZP) binding and studied whether these proteins are preserved throughout mammalian species. Indirect immunofluorescent technique was used to test a panel of monoclonal antibodies prepared against boar sperm surface proteins on spermatozoa of bull and mice. We found a cross-reactivity of some antibodies against boar sperm with bull ejaculated and mouse epididymal spermatozoa. Further, we isolated sperm proteins from different mammalian species, such as pig, bull, dog, cat, mouse and human. Proteins were separated by SDS- electrophoresis and protein/glycoprotein profiles from epididymal, ejaculated and in vitro capacitated sperm were compared. The interaction of sperm with ZP was studied on electrophoretically-separated sperm surface proteins from pig and bull with biotin-labeled ZP glycoproteins. Antibodies, which reacted with boar sperm surface proteins with ZP- binding activity, therefore could be potential egg-binding receptors, were used for monitoring of the sperm protein origin in reproductive fluids and tissues. (In Czech) Keywords: sperm...

In mammals, before fertilisation can occur, sperm have to bind to, and penetrate, the extracellular coat of the oocyte, the zona pellucida (ZP). In the laboratory mouse, which has been used as a model system for fertilization studies, sperm-ZP binding has been found to be mediated by a region near the carboxy terminal, encoded by exon 7 of the Zp3 gene. This region shows considerable interspecific sequence diversity in North American cricetid rodents, with some evidence of adaptive evolution, suggesting that this may contribute to species specific sperm-ZP binding. However, by contrast, in a preliminary study of three species of Australian murine rodents an identical protein sequence of the region encoded by exon 7 of Zp3 was found to be present. The aim of this present study was to determine the pattern of sequence diversity of this region in the most speciose subfamily of mammals, the murine rodents, and to obtain insight into the selective pressures involved in its evolution. For this, DNA was extracted from murine rodents of Africa, Eurasia, South-east Asia, New Guinea and Australia. The nucleotide and predicted amino acid sequence of exons 6 and 7 of Zp3 in 96 murine species from 14 divisions, as recently defined by Musser and Carleton (2005), was determined and compared. Generally, it was found that closely related species shared a highly similar ZP3 sequence. Maximum likelihood analyses of codon substitution models using representatives from 14 murine divisions, suggested that positive selection had occurred within only a few lineages at several different codon sites adjacent to, or within, the putative combining-site for sperm of ZP3. Positive selection was not evident when the analysis was restricted to the Australian taxa which showed low levels of both intra- and inter-generic sequence divergence. There was no good evidence that this region contributes to species specificity of sperm-ZP binding in these species. These findings thus suggest that the selective forces acting on the Zp3 exon 7 region during the evolution of the murine rodents have varied possibly due to a range of selective pressures not necessarily restricted to the prevention of hybridization. It seems unlikely, therefore, that the amino acid sequence of the exon 7 coding region contributes to species specificity of sperm-ZP binding within most of the lineages from this most speciose subfamily of eutherian mammals. / http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1294654 / Thesis(Ph.D.)-- School of Medical Sciences, 2007

Binding of sperm to the oocyte is mediated by complementary molecules on the surface of both gametes and involves the interaction of sperm protein receptors with the zona pellucida (ZP) saccharide structures. It has been shown that many proteins receptors are involved in the sperm-ZP interaction, and potential primary sperm receptors for ZP glycoproteins have been investigated in various mammals. The majority of proteins with identified sperm-ZP binding activity belong to the plasma membrane proteins. However, the exact methods for isolation of sperm membrane proteins are still to be standardized. This study is focused on investigating how employment of various isolation protocols leads to acquisition of various protein mixtures. Further in the work, two possible approaches towards identification of potential ZP-binding partners are implemented, in order to disclose novel primary ZP-binding receptor candidates. Sperm proteins of ejaculated and in vitro capacitated boar sperms were isolated by: Triton X-100, Triton X-114, acetic acid, sodium dodecyl sulphate (SDS), N-octyl-β-D- glucopyranoside (OBG), rehydration buffer (RHB), and finally by freezing-thawing extraction and they were characterized by 1-D, 2-D protein profiles, glycoprotein staining and substrate zymographic methods. The results have...

Abstract Despite the development of assisted reproduction technologies and significant advances in reproductive biology and medicine over the years the cause of infertility remains unexplained in 10–20% of cases. The cause of infertility in these cases may be connected to problems in fertilization or implantation and genetic factors may play a part in this. The zona pellucida (ZP) is an extracellular matrix surrounding the oocyte and early-stage embryos. It is important for folliculogenesis, fertilization and implantation. In humans, it is composed of four known ZP glycoproteins that all show varying degrees of structural and functional roles in reproduction. The aim of the present study was to examine the role of zona pellucida genes in cases of total fertilization failure and zona anomalies, and to study their expression in human fetal and adult ovaries. A total of 34 sequence variations were detected in genes expressing the four human ZP proteins (ZP1–ZP4) among women with fertilization failure and those with varying degrees of zona anomalies in their oocytes. Most of the variations were known single nucleotide polymorphisms, while three were novel findings. Women with fertilization failure had a higher mean number of sequence variations in ZP1 and ZP3 when compared with controls. Some of the most frequent zona anomalies may be at least partly explained by sequence variations in ZP1–ZP4 genes. In fetal life, the expression of ZP3 protein and mRNA could already be detected as early as at the 11th week of gestation and it peaked at the 20th week, the time of primordial follicle formation. This suggests that components needed for zona matrix are already present well before the formation of the zona pellucida and may have a role in the development of primordial follicles. Expression of the transcription factor FIGLA (factor in the germline alpha) was increased at around the 20th week of gestation, supporting previous findings of its critical role in the initiation of folliculogenesis and primordial follicle formation. The present study adds to our knowledge on the currently still incomplete picture of formation of the ZP and fertilization in humans. Understanding the genetic background of infertile patients may help us to develop new tools not only to evaluate but also to improve their fertilization potential, and to choose the optimal treatment to achieve pregnancy. / Tiivistelmä Diagnostiikan kehityksestä huolimatta hedelmättömyyden syy jää edelleen epäselväksi 10–20 %:ssa tapauksista. Niissä hedelmättömyyden taustalla voivat olla munasolun hedelmöittymiseen ja kohtuun kiinnittymiseen liittyvät ongelmat, jotka voivat osittain johtua geneettisistä syistä. Alkiokuori on munasolua ja varhaista alkiota ympäröivä rakenne, joka osallistuu munarakkulan kehittymiseen, munasolun hedelmöittymiseen ja alkion tarttumiseen kohdun limakalvolle. Ihmisellä alkiokuori muodostuu neljästä tunnetusta alkiokuoriproteiinista (ZP1–ZP4). Tutkimuksessa selvitettiin alkiokuoriproteiineja koodittavien geenien vaikutusta hedelmällisyyteen potilailla, joilla koeputkihedelmöitys ei ollut tuottanut yhtään hedelmöittynyttä munasolua (engl. total fertilization failure, TFF) tai joiden munasoluissa havaittiin alkiokuoren rakennemuutoksia (engl. zona anomalies, ZA). Lisäksi selvitettiin alkiokuoriproteiinien ja niiden lähetti-RNA:n esiintymistä sikiöiden ja aikuisten munasarjoissa. TFF- ja ZA-potilaiden alkiokuoriproteiineja koodittavista geeneistä löytyi yhteensä 34 nukleotidimuutosta. Muutoksista kolme oli uusia löydöksiä, mutta suurin osa oli ennalta tunnettuja yhden nukleotidin polymorfioita eli geneettisiä monimuotoisuuskohtia. TFF-potilailla havaittiin ZP1- ja ZP3-geeneissä keskimäärin enemmän polymorfioita kuin verrokeilla. Myös osa yleisimmistä alkiokuoren rakennemuutoksista voidaan mahdollisesti selittää ZP1–ZP4-geeneistä löytyneillä polymorfioilla. Sikiöllä ZP3:n ilmentyminen oli havaittavissa jo 11. raskausviikolla, mutta voimakkainta se oli primordiaalivaiheen munarakkuloiden muodostumisen aikaan 20. raskausviikolla. Tämä voi viitata siihen, että ZP3 saattaa osallistua primordiaalivaiheen munarakkulan kehittymiseen ennen varsinaisen alkiokuoren muodostumista. ZP-geenien säätelytekijän FIGLA:n esiintyminen lisääntyi 20. raskausviikolla, mikä tukee aikaisempia havaintoja FIGLA:n merkityksestä munarakkulan kehittymisen aktivaatiossa ja primordiaalivaiheen munarakkuloiden muodostumisessa. Tämä tutkimus tuo lisätietoa alkiokuoren merkityksestä munasolun hedelmöittymisessä ja syventää tietämystämme alkiokuoren muodostumisesta ihmisellä. Hedelmättömyyden taustalla olevien geneettisten tekijöiden tunteminen voi parantaa lapsettomuuspotilaiden hedelmällisyyden arviointia ja auttaa löytämään heille parhaiten sopivan hoidon.

Thesis (PhD)--Stellenbosch University, 2002. / ENGLISH ABSTRACT: In this study the author presents new data that will shed light on the fairly nebulous knowledge of intracellular pathways involved in the physiologically induced acrosome reaction and the subsequent events leading to fertilization. The zona pellucidainduced acrosome reaction, sperm-zona interaction as well as various sperm motion characteristics were investigated. The first part of the study focussed on the role of extracellular signal regulated kinase (ERK), a member of the family of mitogen activated protein kinases, during the zona pellucida-induced acrosome reaction and sperm-oocyte interaction. It was shown that the inhibition of ERK significantly reduced the zona pellucida-induced acrosome reaction as measured by fluorescent microscopy. This suggests that ERKs are directly or indirectly involved in the signal transduction pathway through which the human sperm acrosome reaction is induced by the zona pellucida. In an attempt to provide further proof that ERK was involved in human acrosome signalling hemizona assays were employed to test sperm-oocyte binding. From these sperm-oocytebinding experiments it was clear that the inhibition of ERK leads to increased binding. These results support the idea that the zona pellucida-induced acrosome reaction, and therefore the physiologically relevant exocytotic event, is regulated by an ERKmediated signal transduction process. In the second part of the study the significance of phosphatidylinositol 3-kinase (PI3K) in the process of human sperm motility, acrosome reaction and sperm-oocyte binding, was investigated by employing the specific PI3K, LY294002. PI3K inhibition increased the percentage motility and percentage progressive motility in asthenozoospermia patients. Under the present laboratory conditions PI3K inhibition furthermore did not influence the acrosome reaction, whilst it enhanced sperm-oocyte binding. These results therefore imply that PI3K negatively affect sperm motility and zona-binding. In the last part of the study the possible effects of intracellular cGMP accumulation via acute in vivo sildenafil citrate (ViagraTM) administration was investigated on seminal parameters, induction of the acrosome reaction, sperm-oocyte binding and sperm motility. As was expected no changes in the macro- and microscopically seminal parameters were caused by sildenafil citrate when compared to placebo. Furthermore the acrosome reaction was also not initiated or potentiated by sildenafil citrate at concentrations of 50mg orally. Sperm-oocyte binding, smooth path velocity, straight line velocity and the percentage rapid cells all increased after sildenafil citrate treatment. From these results it appear that there are various role players in the zona pellucidainduced acrosome reaction intracellular signalling system. A thorough understanding of such signal transduction systems and the crosstalk in-between will ultimately yield information regarding the nature of receptors to which these signal transduction pathways are coupled in human spermatozoa as well as the intracellular effectors that ultimately regulate sperm function. Moreover, an understanding of these regulatory pathways will be essential for the future development of clinical approaches designed to enhance or preclude fertilization. / AFRIKAANSE OPSOMMING: Die outeur lê in hierdie studie nuwe data voor ten einde meer lig te werp op die relatief vae veld van intrasellulêre seintransduksie paaie betrokke by die fisiologiesgeïnduseerde akrosoomreaksie en die daaropvolgende gebeure wat aanleiding gee tot bevrugting. Die zona pellucida-geïnduseerde akrosoomreaksie, sperm-zona interaksie sowel as spermmotiliteitseienskappe is ondersoek. Die eerste gedeelte van die studie fokus op die rol van ekstrasellulêreseingereguleerde- kinase (ERK), 'n lid van die familie van mitogeen-geaktiveerde proteïenkinases, tydens die zona pellucida-geïnduseerde akrosoomreaksie en sperm-oosiet interaksie. Daar word aangetoon dat die inhibisie van ERK die zona pellucida geïnduseerde akrosoomreaksie, soos gemeet met behulp van fluorosensie mikroskopie, betekenisvol verminder. Dit suggereer dat ERK direk of indirek betrokke is by die seintransduksie paaie waardeur die akrosoomreaksie van die menslike sperm deur die zona pellucida geïnduseer word. In 'n poging om onomwonde te bewys dat ERK betrokke is by menslike akrosoom-seintransduksie, is hemizona essais gebruik om sperm-oesiet binding te bepaal. Van hierdie sperm-oosiet bindingeksperimente is dit duidelik dat die inhibisie van ERK aanleiding gee tot verhoogde binding. Hierdie resultate ondersteun dus die idee dat die zona pellucidageïnduseerde akrosoomreaksie en dus die fisiologies relevante eksositotiese gebeurtenis gereguleer word deur 'n ERK-gemediëerde seintransduksie proses. In die tweede gedeelte van die studie is die belang van fosfatidielinositol 3-kinase (PI3K) in die proses van menslike spermmotiliteit, akrosoomreaksie en sperm-oesiet binding ondersoek deur van die spesifieke PI3K inhibitor LY294002, gebruik te maak. Pl3K-inhibisie het die persentasie motiliteit en progressiewe motiliteit by astenozoospermiese pasiënte verhoog. Onder hierdie laboratoriumtoestande het Pl3K-inhibisie nie die akrosoomreaksie beïnvloed nie, terwyl sperm-oosiet binding verhoog is. Hierdie resultate beteken dus dat PI3K spermmotiliteit en zona-binding negatief beïnvloed. In die laaste gedeelte van die studie is die effekte van intrasllulêre cGMP akkumulasie deur akute in vivo sildenafil sitraat (ViagraTM) toediening op seminale parameters, induksie van die akrosoomreaksie, sperm-oesiet binding en spermmotiliteit ondersoek. Soos verwag is geen veranderinge in die makro- en mikroskopiese seminale parameters deur sildenafil sitraat in vergelyking met plasebo veroorsaak nie. Verder is die akrosoomreaksies ook nie deur 50mg orale sildenafil sitraat geïnisieer of potensieer nie. Sperm-oosiet binding, geplaneerde snelheid, reguitlyn snelheid en persentasie vinnigbewegende selle was almal vehoog na sildenafil sitraat behandeling. Uit hierdie resultate blyk dit dat daar verskeie rolspelers in die zona pellucidageïnduseerde akrosoomreaksie is. 'n Deeglike insig van al hierdie seintransduksiepaaie en die onderlinge kruiskontak tussen mekaar sal uiteindelik die nodige inligting rakende die reseptore waaraan hierdie seintransduksie paaie gekoppel is, verskaf sowel as die intrasellulêre effektore wat uiteindelik spermfunksie beheer. Nog te meer sal die begrip van hierdie regulatoriese paaie verder noodsaaklik wees vir die toekomstige ontwikkeling van kliniese benaderings om bevrugting te bevorder of te voorkom.

Made available in DSpace on 2014-06-11T19:35:39Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-02-17Bitstream added on 2014-06-13T19:05:47Z : No. of bitstreams: 1 costa_cgpr_dr_botfm.pdf: 1744933 bytes, checksum: d90abe2ebd4b5e05246936b2f56031cd (MD5) / Nas últimas décadas, alguns parâmetros laboratoriais têm sido propostos para identificar o potencial de saúde oocitária em Reprodução Assistida: 1. Maturidade nuclear e citoplasmática (pesquisa); 2. Dismorfismo (anomalias intracitoplasmáticas); 3. Fertilizaçao. Habitualmente, esses parâmetros (exceto a maturação citoplasmática) são avaliados pelo simples emprego de sistemas de microscopia de luz convencional. Nos últimos oito anos, foram relatados diversos estudos em oócitos humanos sobre a birrefringência da zona pelúcida (BF-ZP), a espessura da ZP, e a visualização do fuso meiótico (FM). Entretanto, ainda é discutível o poder desses novos parâmetros na seleção de óvulos com o pontecial de originar embriões com taxas de implantação elevadas, além disso, seu sistema de visualização e quantificação é complexo e dispendioso. O objetivo deste estudo foi investigar uma possível correlação entre esses parâmetros tradicionais com a BF-ZP, espessura da ZP e o FM. Dessa forma, um total de 73 pacientes que participaram do programa de injeção intracitoplasmática de espermatozóide (ICSI) no Centro de Reprodução Humana “Prof Franco Júnior” tiveram seus óvulos avaliados quanto a maturação nuclear (n=83), maturação citoplasmática (n=29), dismorfismo (n=280) e fertilização (n=216) sendo os resultados correlacionados com a imagem do FM (presença ou ausência), a intensidade da BF-ZP (positiva ou negativa) e a espessura da ZP. Os dados finais revelaram uma fraca correlação entre a maturação, o dismorfismo e a fertilização oocitária com a imagem do FM, a intensidade da BF-ZP, e a espessura da ZP. Entretanto, esses parâmetros poderiam ser responsáveis por identificar pontos biológicos diferentes do potencial de qualidade oocitária. Assim sendo, seria uma medida coerente, a utilização desses parâmetros, em forma conjunta, através de um escore de qualidade oocitária. / During the last decades, three laboratorial parameters have been proposed to identify the potential of the oocyte quality in ART: 1- Nuclear and cytoplasmic maturation; 2-Dysmorphims (intracytoplasmic abnormalities); 3 - Fertilization. These parameters (except the cytoplasmic maturation) are assessed by a simple system of conventional light microscopy. In the last eight years, numerous studies about zona pellucida birefringence (BF-ZP), zona pellucida thickness and meiotic spindle visualization have been reported. However, it is still controversial the power of these parameters in selecting oocyte with potential to develop into implantation-competent embryos. Furthermore, the microscopy system is complex and expensive. The present study aims to evaluate whether there is any correlation between the classical parameters with BF-ZP, ZP thickness and spindle image. A total of 73 patients who participated in an ICSI program had their oocytes evaluated such nuclear maturation (n=83), cytoplasmic maturation (n=29), dysmorphism (n=280) and fertilization (n=216) being its results correlated to the spindle image (presence or absence), BF-ZP intensity (positive or negative) and ZP thickness. The final results showed a low correlation between maturation, dysmorphism and fertilization compared to spindle image and ZP characteristics (BF-ZP, ZP thickness). However, these factors can be responsible to identify specific biological differ from the oocyte potential. Concluding, the use of these factors in association, as a score (oocyte quality score), could be the correct evaluation.

Made available in DSpace on 2014-06-12T16:30:28Z (GMT). No. of bitstreams: 2 arquivo3052_1.pdf: 2447872 bytes, checksum: eb78f6d485668be053ba08a81af9b89d (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2011 / Peperomia pellucida L. (H.B.K.) é uma planta da família Piperaceae que apresenta ações anti-inflamatória, analgésica e antimicrobiana, comprovadas cientificamente. Popularmente é utilizada como anti-hipertensiva. Este trabalho tem por finalidade a caracterização físico-química do pó e dos extratos fluido e seco de P. pellucida e o desenvolvimento e validação de metodologia para doseamento do pó de P. pellucida. Os estudos realizados consistiram na caracterização físico-química do pó e da solução extrativa utilizando metodologia famacopeica, com exceção teor de extrativos e do resíduo seco; nas avaliações das técnicas de secagem (spray dryer, leito de jorro, evaporação rotativa e liofilização) utilizando microscopia eletrônica de varredura e análise térmica; no desenvolvimento e validação da metodologia para determinação quantitativa de flavonóides por espectrofotometria no UV visível. Os extratos secos por evaporação rotativa e liofilização se apresentaram com aspectos resinosos e pastosos. Na microscopia eletrônica de varredura, foi observado que as amostras obtidas por esses processos de secagem não ficaram totalmente secas. Os extratos secos obtidos por spray dryer e leito de jorro, devido à adição de aerosil® à solução extrativa, apresentaram uma melhora significativa no rendimento e nas características farmacotécnicas do pó obtido. O método de quantificação por espectrofotometria a 397 nm mostrou-se preciso, exato e reprodutível, aliado à acessibilidade e facilidade de execução. Os resultados obtidos contribuem para a determinação de especificações de uma futura monografia farmacopeica e fornece parâmetros para que empreendedores da indústria farmacêutica se baseiem para a produção de insumos bem como para o controle de qualidade de intermediários à base de P. pellucida