ABSTRACT
yielded an average particle size of 120 nm with 70% encapsulation-efficiency. In vitro release profile of NP-OP showed sustained release of OP for 21 days. In vivo anti-fertility studies were conducted in marmosets. Results indicated that control animals conceived in the same cycle while two of three treated animals failed to conceive in treatment cycle. The in vivo studies thus corroborate with in vitro release of OP, demonstrating its anti-fertility activity in 66% of animals.
Subject(s)
Animals , Callithrix/physiology , Carrier Proteins/administration & dosage , Carrier Proteins/chemistry , Contraception , Female , Humans , Nanoparticles/administration & dosage , Nanoparticles/chemistry , Ovarian Follicle/chemistry , Particle Size , Peptide Fragments/administration & dosage , Peptide Fragments/chemistry , Polymers/administration & dosage , Polymers/chemistryABSTRACT
O órgão do Bidder e o ovário do sapo Bufo ictericus foram analisados por meio de microscopia de luz, utilizando a coloração pela hematoxilina-eosina (HE) e o método do ácido periódico de Schiff (PAS). A expressão e a distribuição de carboidratos foram verificadas por meio da histoquímica com lectinas, tendo sido utilizadas 8 lectinas com diferentes especificidades para carboidratos (Ulex europaeus (UEA I), Lens culinaris (LCA), Erythrina cristagalli (ECA), Arachis hypogaea (PNA), Ricinus communis (RCA I), Aleuria aurantia (AAA), Triticum vulgaris (WGA) e Glycine maximum (SBA). Os resultados mostraram que a zona pelúcida Bidderiana apresenta resíduos de a-mannose, a-L-fucose, b-D-galactose, N-acetilDglicosamine e a/b-N-acetil-galactosamina. As células foliculares Bidderianas mostraram a presença de b-D-galactose e N-acetil-D-glicosamina. Na matriz de extracelular foram detectados resíduos de a-mannose e a/b-N-acetil-galactosamina. Resíduos de a-L-fucose, N-acetyl-D-glicosamina e a/b-N-acetil-galactosamina foram evidenciados na zona pelúcida ovariana, enquanto na célula folicular foi detectado o resíduo de a-mannose e de N-acetil-D-glicosamina. Assim, a zona pelúcida, em ambos os órgãos, contém resíduos de N-acetil-D-glicosamina e a/b-N-acetil-galactosamina. O resíduo de a-L-fucose foi detectado na zona pelúcida de ambos os órgãos, mas utilizando-se diferentes lectinas. Considerando que o resíduo de a-D-galactose é ausente no ovário, mas presente no órgão de Bidder, a a-D-galactose pode ter um papel importante no controle do desenvolvimento folicular, bloqueando o desenvolvimento dos folículos Bidderianos e impedindo que o órgão de Bidder se transforme em um ovário funcional.
Subject(s)
Animals , Male , Female , Bufonidae , Glycoconjugates/analysis , Ovary/chemistry , Hematoxylin , Histocytochemistry , Lectins , Ovarian Follicle/chemistry , Zona Pellucida/chemistryABSTRACT
Human follicular fluid (HFF) includes various biologically active proteins which can affect follicle growth and oocyte fertilization. Thus far, these proteins from mature follicles in human follicular fluid have been poorly characterized. Here, two-dimensional polyacrylamide gel electrophoresis (2-DE) with matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) was used to identify new proteins in HFF. Mature follicular fluids were obtained from five females after oocyte collection during in vitro fertilization (IVF). We directly rehydrated HFF samples, obtained high-resolution 2-DE maps, and processed them for 2-DE and MALDI-MS. One hundred eighty spots were detected and 10 of these spots were identified. By the 2-DE database, six of them had been reported, as proteins already existing in HFF. Hormone sensitive lipase (HSL), Unnamed protein product 1 (UPP1), Unnamed protein product 2 (UPP2), and apolipoprotein A-IV precursor were newly detected. HSL and apolipoprotein A-IV participate in lipid metabolism. UPP1 has a homology with selenocysteine lyase. We found by RT-PCR that these genes are expressed from human primary granulosa cells. The proteins identified here may emerge as potential candidates for specific functions during folliculogenesis, hormone secretion regulation, or oocyte maturation. Further functional analysis of these proteins is necessitated to determine their biological implications.
Subject(s)
Adult , Female , Humans , Electrophoresis, Gel, Two-Dimensional/methods , Follicular Fluid/chemistry , Gene Expression , Granulosa Cells/metabolism , Ovarian Follicle/chemistry , Proteins/analysis , RNA, Messenger/genetics , Reverse Transcriptase Polymerase Chain Reaction , Spectrometry, Mass, Matrix-Assisted Laser Desorption-IonizationABSTRACT
The ability of Triatoma infestans ovarian follicles to synthesize a very high-density lipoprotein (VHDL) has been examined by immunohistochemical methods. This kind of lipoprotein can be envisaged as a storage hexameric protein present in the hemolymph of some insect species. VHDL immunoreactivity is observed in oocytes at different stages of maturation. The antigen is present in the oocyte cytoplasm as well as in the follicular epithelial cells. The immunopositive reaction in the apical surface of follicle cells suggests both a VHDL synthesis and a secretion process. Furthermore, VHDL seems to be stored into oocyte in yolk granules. On the contrary, no immunopositive reaction is observed in the intracellular spaces between follicle cells, suggesting that VHDL is not incorporated from hemolymph into the oocyte.