Photo-induced destruction of giant vesicles in methylene blue solutions

We study the photodecomposition of phospholipid bilayers in aqueous solutions of methylene blue. Observation of giant unilamellar vesicles under an optical microscope reveals a consistent pattern of membrane disruption as a function of methylene blue concentration and photon density for different substrates supporting the vesicles.

Pharmacological and local toxicity studies of a liposomal formulation for the novel local anaesthetic ropivacaine

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Ultramorphology and histochemistry of fat body cells from last Instar larval of the Pachycondyla (=Neoponera) villosa (Fabricius) (Formicidae: Ponerinae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Structural and ultrastructural characteristics of interrenal gland and chromaffin cell of matrinxa, Brycon cephalus Gunther 1869 (Teleostei-Characidae)

This work presents the structure and ultrastructure of the interrenal gland and chromaffin cells, as well as the morphology of the head kidney of Brycon cephalus, the head kidney is composed of fused bilateral lobes located anterior to the swim bladder and ventrolateral to the spinal column, the parenchyma revealed lympho-haematopoietic tissue, melano-macrophage centres, interrenal gland and chromaffin cells. The interrenal gland consisted of cords or strands of cells grouped around the posterior cardinal vein and their branches. Chromaffin cells are found in small groups, closely associated with the interrenal gland and/or under the endothelium of the posterior cardinal vein. So far, the ultrastructural analysis has revealed only one interrenal cell type which contained abundant smooth endoplasmic reticulum and numerous mitochondria with tubulo-vesicular cristae, characteristic of steroid-producing cells. Two types of chromaffin cells were observed. The first type was characterized by the presence of vesicles with round, strongly electron-dense granules, which were eccentrically located, Such cells were interpreted as noradrenaline cells, Meanwhile, cells which contained smaller vesicles and electron-lucent granules, with a small halo separating the granule from the vesicular limiting membrane, were identified as adrenaline cells.

Isolamento e caracterização ultraestrutural de folículos pré-antrais de vacas da raça Nelore (Bos taurus indicus)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Ultrastructural morphology and morphometry of the normal corneal endothelium of adult crossbred pig

O endotélio corneal é uma monocamada de células poligonais. A integridade e saúde dessa camada são essenciais para a manutenção da transparência corneal normal. Este estudo reportou pela primeira vez, de forma detalhada, a morfologia ultra-estrutural e a morfometria do endotélio corneal de suínos adultos mestiços à microscopia eletrônica de varredura (MEV). A superfície endothelial corneal apresentou um padrão regular de células poligonais, com predomínio da forma hexagonal e de bordas celulares nítidas. O núcleo foi observado como protuberância arredondada no centro da célula. Também foram observados os cílios (2-4) em apenas algumas células da região periférica da córnea, as aberturas das vesículas pinocitóticas na proximidade dos cílios, as microvilosidades, as varas da borda e as bordas celulares em formato de zigzag. A área celular média foi significativamente maior (P<0,05) no centro da córnea do que na periferia, com um coeficiente de variação menor no centro da córnea. A densidade celular média foi significativamente maior na periferia (P<0,05) e 43,9% maior que os dados reportados por outros autores na microscopia especular, o que demonstra o efeito da retração celular durante o processamento das amostras. O valor médio do número de lados das células (pleomorfismo) foi de 5,9, o que evidencia um predomínio do formato hexagonal. A percentagem de células hexagonais foi significativamente maior no centro (P<0,001). Os parâmetros obtidos nesta pesquisa servirão de base para estudos futuros sobre o efeito de medicamentos, cirurgias intracamerulares ou soluções para armazenamento de córneas para transplantes no endotélio corneal do suíno.

Aspectos morfológicos do saco vitelino em roedores da subordem Hystricomorpha: paca (Agouti paca) e cutia (Dasyprocta aguti)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Variabilidade sazonal no ducto epididimário de codorna doméstica: observações morfológicas

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Early development of Astronotus ocellatus under stereomicroscopy and scanning electron microscopy

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Construction of an alkaline phosphatase-liposome system: a tool for biomineralization study

Alkaline phosphatase is required for the mineralization of bone and cartilage. This enzyme is localized in the matrix vesicle, which plays a role key in calcifying cartilage. In this paper. we standardize a method for construction an alkaline phosphatase liposome system to mimic matrix vesicles and examine a some kinetic behavior of the incorporated enzyme. Polidocanol-solubilized alkaline phosphatase, free of detergent, was incorporated into liposomes constituted from dimyristoylphosphatidylcholine (DMPC), dilaurilphosphatidylcholine (DLPC) or dipalmitoylphosphatidylcholine (DPPC). This process was time-dependent and >95% of the enzyme was incorporated into the liposome after 4 h of incubation at 25 degreesC. Although, incorporation was more rapid when vesicles constituted from DPPC were used, the incorporation was more efficient using vesicles constituted from DMPC. The 395 nm diameter of the alkaline phosphatase-liposome system was relatively homogeneous and more stable when stored at 4 degreesC.Alkaline phosphatase was completely released from liposome system only using purified phosphatidylinositol-specific phospholipase C (PIPLC). These experiments confirm that the interaction between alkaline phosphatase and lipid bilayer of liposome is via GPI anchor of the enzyme, alone. An important point shown is that an enzyme bound to liposome does not lose the ability to hydrolyze ATP, pyrophosphate and p-nitrophenyl phosphate (PNPP), but a liposome environment affects its kinetic properties, specifically for pyrophosphate.The standardization of such system allows the study of the effect of phospholipids and the enzyme in in vitro and in vivo mineralization, since it reproduces many essential features of the matrix vesicle. (C) 2002 Elsevier B.V. Ltd. All rights reserved.

Erythrocyte ghost cell-alkaline phosphatase: construction and characterization of a vesicular system for use in biomineralization studies

Alkaline phosphatase is required for the mineralization of bone and cartilage. This enzyme is localized in the matrix vesicle, which plays a role key in calcifying cartilage. In this paper we standardize a method to construction a resealed ghost cell-alkaline phosphatase system to mimic matrix vesicles and examine the kinetic behavior of the incorporated enzyme. Polidocanol-solubilized alkaline phosphatase, free of detergent, was incorporated into resealed ghost cells. This process was time-dependent and practically 50% of the enzyme was incorporated into the vesicles in 40 h of incubation, at 25 degreesC. Alkaline phosphatase-ghost cell systems were relatively homogeneous with diameters of about 300 nm and were more stable when stored at -20 degreesC.Alkaline phosphatase was completely released from the resealed ghost cell-system using only phospholipase C. These experiments confirm that the interaction between alkaline phosphatase and the lipid bilayer of resealed ghost cell is exclusively via glycosylphosphatidylinositol (GPI) anchor of the enzyme.An important point shown is that an enzyme bound to resealed ghost cell does not lose the ability to hydrolyze ATP, pyrophosphate and p-nitrophenyl phosphate (PNPP), but the presence of a ghost membrane, as a support of the enzyme, affects its kinetic properties. Moreover, calcium ions stimulate and phosphate ions inhibit the PNPPase activity of alkaline phosphatase present in resealed ghost cells. (C) 2002 Elsevier B.V. B.V. All rights reserved.

Cellular and molecular characterization of an embryonic cell line (BME26) from the tick Rhipicephalus (Boophilus) microplus

The cellular and molecular characteristics of a cell line (BME26) derived from embryos of the cattle tick Rhipicephalus (Boophilus) microplus were studied. The cells contained glycogen inclusions, numerous mitochondria, and vesicles with heterogeneous electron densities dispersed throughout the cytoplasm. Vesicles contained lipids and sequestered palladium meso-porphyrin (Pd-mP) and rhodamine-hemoglobin, suggesting their involvement in the autophagic and endocytic pathways. The cells phagocytosed yeast and expressed genes encoding the antimicrobial peptides (microplusin and defensin). A cDNA library was made and 898 unique mRNA sequences were obtained. Among them, 556 sequences were not significantly similar to any sequence found in public databases. Annotation using Gene Ontology revealed transcripts related to several different functional classes. We identified transcripts involved in immune response such as ferritin, serine proteases, protease inhibitors,. antimicrobial peptides, heat shock protein, glutathione S-transferase, peroxidase, and NADPH oxidase. BME26 cells transfected with a plasmid carrying a red fluorescent protein reporter gene (DsRed2) transiently expressed DsRed2 for up to 5 weeks. We conclude that BME26 can be used to experimentally analyze diverse biological processes that occur in R. (B.) microplus such as the innate immune response to tick-borne pathogens. (C) 2008 Elsevier Ltd. All rights reserved.

Caracterização morfofisiológica de isolados do gênero Cylindrocladium

Com o objetivo de caracterizar o complexo gênero anamórfico Cylindrocladium, dezoito isolados foram cultivados em meios de cultura distintos, mantidos sob diferentes temperaturas, além de ter conídios e vesículas terminais analisados morfologicamente para identificação correta das culturas. Inoculações em diferentes hospedeiros foram, também, realizadas para avaliação do comportamento patogênico dos isolados estudados. Com relação aos caracteres morfológicos, pôde-se observar que ocorrem alterações nas dimensões de conídios e na morfologia da vesícula terminal devido, provavelmente, à mudança do substrato de cultivo. Porém, a variabilidade natural nas características dessas estruturas é tão elevada que dificulta a identificação correta dos isolados. em uma mesma cultura, por exemplo, foram observadas vesículas terminais de diferentes morfologias. Pôde-se constatar que existem diferenças patogênicas e fisiológicas entre os isolados, uma vez que houve a formação de grupos distintos quando tais características foram consideradas. Estas diferenças, provavelmente, sejam devidas à constituição genética distinta existente entre os isolados.

Ultrastructural Study of the Neuromuscular Junctions of Extensor Digitorum Longus Muscle (M. extensor digitorum longus) of the Mice

The neuromuscular junction of the extensor digitorum longus muscle of fingers was analyzed in 21 young (three months) and old (from six to 25 months) mice, from both genders. Morphologic changes were found throughout the mouse life, being more frequent and visible with aging. According with the data described in the literature consulted and the observations taken in this research, it becomes clear that a continuous process of morphological remodeling occurs in all neuromuscular ultrastructural junctions of the extensor digitorum longus muscle of fingers, during the life of the animal. Theses changes are characterized by figures of myelin in the cytoplasm of Schwann cells, pleomorphic and multivesiclar bodies, mitochondrias with morphologically altered crests in the axon terminal and degenerated junction folders. Coated vesicles are common in older animals and rare in young animals.

Taking Advantage of Electrostatic Interactions To Grow Langmuir-Blodgett Films Containing Multilayers of the Phospholipid Dipalmitoylphosphatidylglycerol

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)