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1.
J Dairy Sci ; 105(12): 9367-9386, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-36241439

RESUMO

A growing number of companies within the cheese-making industry are now using high-protein (e.g., 4-5%) milks to increase cheese yield. Previous studies have suggested that cheeses made from high-protein (both casein and whey protein; WP) milks may ripen more slowly; one suggested explanation is inhibition of residual rennet activity due to elevated WP levels. We explored the use of microfiltration (MF) to concentrate milk for cheese-making, as that would allow us to concentrate the casein while varying the WP content. Our objective was to determine if reducing the level of WP in concentrated cheese milk had any impact on cheese characteristics, including ripening, texture, and nutritional profile. Three types of 5% casein standardized and pasteurized cheese milks were prepared that had various casein:true protein (CN:TP) ratios: (a) control with CN:TP 83:100, (b) 35% WP reduced, 89:100 CN:TP, and (c) 70% WP reduced, 95:100 CN:TP. Standardized milks were preacidified to pH 6.2 with dilute lactic acid during cheese-making. Composition, proteolysis, textural, rheological, and sensory properties of cheeses were monitored over a 9-mo ripening period. The lactose, total solids, total protein, and WP contents in the 5% casein concentrated milks were reduced with increasing levels of WP removal. All milks had similar casein and total calcium levels. Cheeses had similar compositions, but, as expected, lower WP levels were observed in the cheeses where WP depletion by MF was performed on the cheese milks. Cheese yield and nitrogen recoveries were highest in cheese made with the 95:100 CN:TP milk. These enhanced recoveries were due to the higher fraction of nitrogen being casein-based solids. Microfiltration depletion of WP did not affect pH, sensory attributes, or insoluble calcium content of cheese. Proteolysis (the amount of pH 4.6 soluble nitrogen) was lower in control cheeses compared with WP-reduced cheeses. During ripening, the hardness values and the temperature of the crossover point, an indicator of the melting point of the cheese, were higher in the control cheese. It was thus likely that the higher residual WP content in the control cheese inhibited proteolysis during ripening, and the lower breakdown rate resulted in its higher hardness and melting point. There were no major differences in the concentrations of key nutrients with this WP depletion method. Cheese milk concentration by MF provides the benefit of more typical ripening rates.


Assuntos
Queijo , Animais , Queijo/análise , Leite/química , Proteínas do Soro do Leite/análise , Caseínas/análise , Proteólise , Cálcio/análise , Manipulação de Alimentos/métodos , Nitrogênio/análise , Concentração de Íons de Hidrogênio
2.
Regul Toxicol Pharmacol ; 117: 104752, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32791089

RESUMO

At the 8th conference of Occupational and Environmental Exposure of the Skin to Chemicals (OEESC) (16-18 September 2019) in Dublin, Ireland, several researchers performing skin permeation assays convened to discuss in vitro skin permeability experiments. We, along with other colleagues, all of us hands-on skin permeation researchers, present here the results from our discussions on the available OECD guidelines. The discussions were especially focused on three OECD skin absorption documents, including a recent revision of one: i) OECD Guidance Document 28 (GD28) for the conduct of skin absorption studies (OECD, 2004), ii) Test Guideline 428 (TGD428) for measuring skin absorption of chemical in vitro (OECD, 2004), and iii) OECD Guidance Notes 156 (GN156) on dermal absorption issued in 2011 (OECD, 2011). GN156 (OECD, 2019) is currently under review but not finalized. A mutual concern was that these guidance documents do not comprehensively address methodological issues or the performance of the test, which might be partially due to the years needed to finalize and update OECD documents with new skin research evidence. Here, we summarize the numerous factors that can influence skin permeation and its measurement, and where guidance on several of these are omitted and often not discussed in published articles. We propose several improvements of these guidelines, which would contribute in harmonizing future in vitro skin permeation experiments.


Assuntos
Congressos como Assunto/normas , Exposição Ambiental/normas , Fidelidade a Diretrizes/normas , Exposição Ocupacional/normas , Organização para a Cooperação e Desenvolvimento Econômico/normas , Absorção Cutânea/efeitos dos fármacos , Exposição Ambiental/prevenção & controle , Substâncias Perigosas/metabolismo , Substâncias Perigosas/toxicidade , Humanos , Irlanda , Exposição Ocupacional/prevenção & controle , Absorção Cutânea/fisiologia
3.
J Dairy Sci ; 103(11): 9906-9922, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32921459

RESUMO

Some European dairies use low concentration factor microfiltration (MF) in their cheese plants. Removal of whey protein (WP) from milk before cheesemaking using microfiltration without concentration provides the opportunity to produce a value-added by-product, milk-derived whey. However, few studies have focused on the effects on cheese properties caused by the depletion of WP from cheese milk. Most studies have concentrated cheese milk using MF in addition to depletion of WP. In our approach, cheese milk was not concentrated during WP depletion using MF. We wanted to quantify residual WP levels in cheese made from MF milk and to explore whether WP depletion from milk would influence functionality, nutritional profile, and cheese quality during ripening. Casein (CN) contents for all milks were kept at ∼2.5%, to eliminate the confounding factor of concentration of CN, which was observed in some previous MF studies. Cheese milks had similar ratios of CN to fat. Three standardized milks were produced with various CN:true protein (TP) ratios: (a) control with a CN:TP ratio of 83:100, (b) 35% WP depletion, 89:100 CN:TP, and (c) 70% WP depletion, 95:100 CN:TP. Cheddar cheeses were made from MF milk with various WP depletion levels and aged for 9 mo, and their functionality was evaluated during ripening. We found no major differences in cheese composition or pH values between samples. Cheese yield, solids recovery, and nitrogen recovery were slightly higher in the 95:100 CN:TP cheeses compared with the control. These enhanced recoveries reflect that MF-treated milk started with a higher fraction of CN-based protein solids, rather than WP solids. The standardized milk from the 95:100 CN:TP treatment also had a slightly higher fat content compared with the control, likely helping to increase cheese yield. Rheological properties of cheeses during heating were similar between treatments. Hardness initially decreased with age for all cheeses due to proteolysis or solubilization, or both, of calcium phosphate. Maximum loss tangent (LT), an index of cheese meltability, was slightly lower for the control cheese until 30 d of ripening, but after 30 d, all treatments exhibited similar maximum LT values. The temperature where LT = 1 (crossover temperature), an index of softening point during heating, was slightly lower for MF cheese compared with the control cheeses during ripening. Microfiltration treatment had no significant influence on proteolysis. Sensory properties were similar between the cheeses, except for bitterness. Bitterness intensity was slightly lower in the MF cheeses than in the control cheeses and increased in all cheeses during ripening. We detected no major differences in the concentrations of key nutrients or vitamins between the various cheeses. Depletion of WP in cheese milk by MF did not negatively affect cheese quality, or its nutritional profile, and resulted in similar cheesemaking yields.


Assuntos
Queijo/análise , Leite/química , Proteínas do Soro do Leite/análise , Animais , Caseínas/análise , Queijo/normas , Filtração , Manipulação de Alimentos , Nitrogênio/análise , Reologia , Paladar , Temperatura
4.
Fish Physiol Biochem ; 42(3): 995-1004, 2016 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-26746846

RESUMO

Renin immunoreactivity was localized at the light and electron microscopic level in two elasmobranch fish species, the Atlantic stingray, Dasyatis sabina, and river ray, Potamotrygon humerosa. At the light microscopic level, the peroxidase-anti-peroxidase method showed a positive immunoreactivity in modified smooth muscle cells in kidney afferent arterioles as well as in arterioles of several organs: rectal gland, inter-renal gland, conus arteriosus, and gill. Electron microscopic renin-positive immunogold localization was confined to the contents of membrane bound granules in the modified smooth muscle cells of these arterioles. The presence of renin-containing granules in the modified smooth muscle, "granular cells," of the renal glomerular afferent arteriole of these two stingray species adds support to earlier studies which showed the structural components of a complete juxtaglomerular apparatus and some of the biochemical and molecular components of a renin-angiotensin system (RAS) as found in teleost fish, reptiles, birds, and mammals. A notable result, however, was the renin-positive immunoreaction in the arteriolar wall of all other organs studied here. The presence of this "diffuse renin system" in the connective tissue of various organs suggests that in these two stingray species in addition to local organ-specific functions, the RAS may act as a systemic mechanism to regulate blood pressure and blood flow in the body.


Assuntos
Elasmobrânquios/metabolismo , Miócitos de Músculo Liso/metabolismo , Renina/metabolismo , Animais , Arteríolas/anatomia & histologia , Arteríolas/metabolismo , Arteríolas/ultraestrutura , Elasmobrânquios/anatomia & histologia , Mucosa Gástrica/metabolismo , Imuno-Histoquímica , Rim/anatomia & histologia , Rim/metabolismo , Microscopia Eletrônica , Miócitos de Músculo Liso/ultraestrutura , Reto/metabolismo , Reto/ultraestrutura , Estômago/anatomia & histologia
5.
Sci Total Environ ; 886: 163767, 2023 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-37156387

RESUMO

Although organic solvents have been associated with CNS toxicity, neurotoxicity testing is rarely a regulatory requirement. We propose a strategy to assess the potential neurotoxicity of organic solvents and predict solvent air concentrations that will not likely produce neurotoxicity in exposed individuals. The strategy integrated an in vitro neurotoxicity, an in vitro blood-brain barrier (BBB), and an in silico toxicokinetic (TK) model. We illustrated the concept with propylene glycol methyl ether (PGME), widely used in industrial and consumer products. The positive control was ethylene glycol methyl ether (EGME) and negative control propylene glycol butyl ether (PGBE), a supposedly non-neurotoxic glycol ether. PGME, PGBE, and EGME had high passive permeation across the BBB (permeability coefficients (Pe) 11.0 × 10-3, 9.0 × 10-3, and 6.0 × 10-3 cm/min, respectively). PGBE was the most potent in in vitro repeated neurotoxicity assays. EGME's main metabolite, methoxyacetic acid (MAA) may be responsible for the neurotoxic effects reported in humans. No-observed adverse effect concentrations (NOAECs) for the neuronal biomarker were for PGME, PGBE, and EGME 10.2, 0.07, and 79.2 mM, respectively. All tested substances elicited a concentration-dependent increase in pro-inflammatory cytokine expressions. The TK model was used for in vitro-to-in vivo extrapolation from PGME NOAEC to corresponding air concentrations (684 ppm). In conclusion, we were able to predict air concentrations that would not likely result in neurotoxicity using our strategy. We confirmed that the Swiss PGME occupational exposure limit (100 ppm) will not likely produce immediate adverse effects on brain cells. However, we cannot exclude possible long-term neurodegenerative effects because inflammation was observed in vitro. Our simple TK model can be parameterized for other glycol ethers and used in parallel with in vitro data for systematically screening for neurotoxicity. If further developed, this approach could be adapted to predict brain neurotoxicity from exposure to organic solvents.


Assuntos
Éter , Propilenoglicóis , Humanos , Toxicocinética , Propilenoglicóis/metabolismo , Propilenoglicóis/toxicidade , Éteres/toxicidade , Etilenoglicóis/toxicidade , Etilenoglicóis/metabolismo , Solventes
6.
Toxicol In Vitro ; 73: 105129, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33662515

RESUMO

Bisphenol A (BPA) in vitro skin permeation studies have shown inconsistent results, which could be due to experimental conditions. We studied the impact of in vitro parameters on BPA skin permeation using flow-through diffusion cells with ex-vivo human skin (12 donors, 3-12 replicates). We varied skin status (viable or frozen skin) and thickness (200, 400, 800 µm), BPA concentrations (18, 250 mg/l) and vehicle volumes (10, 100 and 1000 µl/cm2). These conditions led to a wide range of BPA absorption (2%-24% after 24 h exposure), peak permeation rates (J = 0.02-1.31 µg/cm2/h), and permeability coefficients (Kp = 1.6-5.2 × 10-3 cm/h). This is the first time steady state conditions were reached for BPA aqueous solutions in vitro (1000 µl/cm2 applied at concentration 250 mg/l). A reduction of the skin thickness from 800 and 400 µm to 200 µm led to a 3-fold increase of J (P < 0.05). A reduction of the vehicle volume from 1000 to 100 led to a 2-fold decrease in J (P > 0.05). Previously frozen skin led to a 3-fold increase in J compared to viable skin (P < 0.001). We found that results from published studies were consistent when adjusting J according to experimental parameters. We propose appropriate J values for different exposure scenarios to calculate BPA internal exposures for use in risk assessment.


Assuntos
Compostos Benzidrílicos/farmacologia , Fenóis/farmacologia , Absorção Cutânea , Pele/metabolismo , Administração Cutânea , Humanos , Técnicas In Vitro , Pele/anatomia & histologia
7.
Aliment Pharmacol Ther ; 22 Suppl 2: 71-3, 2005 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-16225478

RESUMO

The mechanisms responsible for the progression of nonalcoholic fatty liver disease (NAFLD) to more severe liver injury are still poorly understood. Data from animal models suggest that oxidative stress contributes to steatohepatitis and an increase of lipid peroxidation has been documented in human NAFLD. By measuring the titers of circulating antibodies against lipid peroxidation products as markers of oxidative stress we have observed that NAFLD patients have titers of these antibodies significantly higher than in controls. Moreover, the titers of lipid peroxidation-related antibodies are associated with a 3-fold increase in the risk of developing advanced fibrosis/cirrhosis. Although the mechanisms causing oxidative stress in NAFLD have not been elucidated, these results support the involvement of lipid peroxidation in the processes leading to liver fibrosis associated with NAFLD.


Assuntos
Fígado Gorduroso/metabolismo , Estresse Oxidativo/fisiologia , Anticorpos/sangue , Progressão da Doença , Fígado Gorduroso/complicações , Fígado Gorduroso/imunologia , Humanos , Peroxidação de Lipídeos/imunologia , Peroxidação de Lipídeos/fisiologia , Cirrose Hepática/etiologia , Cirrose Hepática/metabolismo
8.
Matrix Biol ; 20(1): 37-51, 2001 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-11246002

RESUMO

The organization of the aggregates occurring in the stroma: (1) of the murine and human cornea after incubation in an ATP acidic solution; (2) of surgically excised epiretinal membranes (ERM); and (3) of the trabecular meshwork of monkey eyes was investigated morphologically and immunocytochemically on thin section electron microscopy. Morphology. The aggregates in the cornea appeared as cross-banded fibrils. The bands were uniformly electron dense (single banded form); they were separated from each other by interbands consisting of a bundle of filaments emerging in cross section as small areas of randomly assembled dot-like structures. In the ERM, most of the aggregates stood out as heteromorphic cross-banded bodies showing dense bands with electron denser borders (double banded form) and interbands composed of longitudinally oriented, parallel sheets or laminae of amorphous material enclosing thin, similarly oriented filaments. These extended, thinner and double in number (since interlacing with similar components of the opposite sheet), into the pale central zone of the dense band. The aggregates of the trabecular meshwork were heteromorphic, had uniformly dense bands (single banded form as in the cornea), but their interbands displayed longitudinal sheets (as the ERM aggregates). Immunocytochemistry revealed type VI collagen in the three eye aggregates with gold particles preferentially localized at the interbands. The specificity of the antibodies used was tested by Western blot analysis of type VI collagen samples extracted from human placenta and on homogenates of human cornea. In conclusion, the results indicate that the tetramers of type VI collagen may aggregate differently into structures with distinct supramolecular arrangements. These are illustrated in schematic drawings.


Assuntos
Colágeno/metabolismo , Olho/metabolismo , Idoso , Animais , Olho/patologia , Feminino , Humanos , Masculino , Camundongos , Microscopia Imunoeletrônica/métodos , Pessoa de Meia-Idade , Coelhos , Retina/metabolismo , Retina/patologia , Inclusão do Tecido , Malha Trabecular/metabolismo , Malha Trabecular/patologia
9.
Matrix Biol ; 14(1): 101-11, 1994 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-8061916

RESUMO

In the female German black-faced sheep the posterior cruciate ligament was replaced by a free patellar tendon autograft and after 2, 6, 16, 26 and 52 weeks tissue samples of the graft's center (axial region far from bones) were removed for histochemistry and electron microscopy. To localize the proteoglycans Alcian Blue and 0.3 M MgCl2 were added to the fixative solution. The distribution of the proteoglycans in the graft was compared to that of a normal patellar tendon and of a normal posterior cruciate ligament. In the patellar tendon spindle-shaped cells predominated and proteoglycans appeared as short filaments at regular intervals between the collagen fibrils. In the posterior cruciate ligament chondroid cells and long filaments in a net-work-like arrangement were seen. In the patellar tendon autografts short interfibrillar filaments prevailed after 2, 6 and 16 weeks. After 26 weeks and particularly after 52 weeks long filaments also appeared. Digestion with Chondroitinase ABC, AC and Hyaluronidase suggested that the short filaments were PGs containing dermatan sulfate. In grafts, in the early phases the fibroblasts predominated, while in the late phases mainly chondroid cells were observed. The grafts showed aspects of the normal posterior cruciate ligament. However, differences remained, for example the thin collagen fibrils, which could represent one of the reasons for a secondary graft failure.


Assuntos
Matriz Extracelular/metabolismo , Ligamentos/cirurgia , Proteoglicanas/biossíntese , Tendões/metabolismo , Tendões/transplante , Citoesqueleto de Actina/ultraestrutura , Animais , Colágeno/análise , Colágeno/metabolismo , Matriz Extracelular/ultraestrutura , Feminino , Microscopia Eletrônica , Patela , Proteoglicanas/análise , Ovinos , Tendões/citologia , Fatores de Tempo , Transplante Autólogo
10.
Neurology ; 35(5): 746-9, 1985 May.
Artigo em Inglês | MEDLINE | ID: mdl-2581176

RESUMO

In a patient with hyperkalemic periodic paralysis, metaproterenol prevented muscular weakness and hyperkalemia in periods of rest after exercise. During a severe attack, the drug rapidly corrected hyperkalemia and seemed to enhance the return of strength. The action of metaproterenol may involve a beta-adrenergic-mediated increase of potassium transport via the sodium-potassium pump.


Assuntos
Hiperpotassemia/tratamento farmacológico , Metaproterenol/uso terapêutico , Paralisias Periódicas Familiares/tratamento farmacológico , Adolescente , Humanos , Hiperpotassemia/metabolismo , Hiperpotassemia/fisiopatologia , Canais Iônicos/metabolismo , Masculino , Contração Muscular , Paralisias Periódicas Familiares/metabolismo , Paralisias Periódicas Familiares/fisiopatologia , Esforço Físico , Potássio/metabolismo , Descanso
11.
J Histochem Cytochem ; 33(4): 360-6, 1985 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-3980985

RESUMO

After fixation in the presence of filipin, the membrane of the goblet cell granules of the distal colonic mucosa (guinea pig) displays, in freeze-fracture replicas, a rhombic pattern composed of intersecting "lines" of unknown nature. The filipin-cholesterol complexes of the granule membrane are ordered according to the same geometry. This arrangement--but not the "lines"--has been found for the filipin-cholesterol complexes of other membranes of the same goblet cells as well as of other colonic cells. These observations indicate that in some biological membranes cholesterol has an ordered distribution, comparable to that suggested by investigations on artificial membranes.


Assuntos
Colesterol , Colo/citologia , Filipina , Mucosa Intestinal/citologia , Polienos , Animais , Grânulos Citoplasmáticos , Feminino , Técnica de Fratura por Congelamento , Cobaias , Técnicas In Vitro , Membranas Intracelulares , Masculino
12.
J Histochem Cytochem ; 49(3): 397-406, 2001 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-11181742

RESUMO

Epoxy resins provide optimal tissue morphology at both the light and the electron microscopic level and therefore enable correlative studies on semithin and thin sections from the same tissue block. Here we report on an approach to retain these advantages for immunolabeling studies by adapting and combining well-known techniques, i.e., surface etching with sodium ethoxide and heat-mediated antigen retrieval. We propose a simple procedure for immunostaining semithin and thin epoxy resin sections. To check its applicability, well characterized, commercially available antibodies (against E-cadherin, alpha-catenin, and beta-catenin) were used on sections of human small intestine. By light microscopy, the immunostaining efficiency was compared on cryo-, paraffin, and epoxy semithin sections processed in parallel. The most detailed results were obtained on semithin sections, where the labeling precisely delineated the lateral plasma membrane of the enterocytes. At the electron microscopic level the procedure did not damage the structures and allowed an efficient, reproducible immunogold labeling extending homogeneously over exceptionally wide tissue areas. The three antibodies specifically labeled the zonula adherens of the junctional complex between epithelial cells and, in agreement with light microscopic observations, the lateral plasma membrane.


Assuntos
Resinas Epóxi , Microscopia/métodos , Inclusão em Plástico , Transativadores , Caderinas/metabolismo , Proteínas do Citoesqueleto/metabolismo , Humanos , Imuno-Histoquímica , Jejuno/metabolismo , Luz , Microscopia Imunoeletrônica/métodos , alfa Catenina , beta Catenina
13.
J Histochem Cytochem ; 31(5): 662-8, 1983 May.
Artigo em Inglês | MEDLINE | ID: mdl-6188781

RESUMO

Rat kidneys were perfused with fixative solutions containing either a) a polycationic dye (Alcian blue 8 GX, Astra blue 6 GLL, cuprolinic blue, ruthenium red), b) a monocationic dye (safranine 0), or c) Alcian blue in the presence of a 0.3 M MgCl2 concentration. Whereas solutions of a revealed the glomerular basement membrane proteoglycans as particles or threads 60 nm apart and arranged in a reticular pattern, solutions of b and c demonstrated new morphological aspects of these molecules. They appeared as tiny filamentous structures, about 100 to 160 nm long, ordered in a network-like pattern with a mesh of about 60-nm width. The filaments displayed lateral branches about 20 nm apart and about 25 nm long, projecting within the meshes. We suggest that the filamentous structures are the protein core, and the branches are the glycosaminoglycans of proteoglycan molecules. Because of this arrangement the negatively charged sites of the glomerular basement membrane would lie closer to each other than previously assumed.


Assuntos
Glomérulos Renais/ultraestrutura , Proteoglicanas/análise , Animais , Membrana Basal/ultraestrutura , Histocitoquímica , Magnésio , Cloreto de Magnésio , Masculino , Ratos , Ratos Endogâmicos , Coloração e Rotulagem
14.
J Histochem Cytochem ; 42(10): 1323-32, 1994 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-7930514

RESUMO

Virtually all vertebrates acidify their gastric contents to a pH between 0.8 and 2.0. In mammals, acid secretion is mediated by a K-stimulated proton-translocating adenosine triphosphatase (H,K-ATPase), which establishes a million-fold gradient of protons across the apical membrane of the gastric parietal cell. The earliest phylogenetic appearance of gastric acid secretion is in cartilaginous fish, and we sought to verify in this class (Chondrichthyes) the presence and distribution of H,K-ATPase in gastric epithelial cells. An antibody against a synthetic peptide based on the C-terminus of pig H,K-ATPase alpha-subunit was localized in the gastric glands of the Atlantic stingray Dasyatis sabina. The C-terminal antibody stained all cells with tubulovesicles and the apical membrane domain of mucous neck cells. In proximal stomach, gastric glands showed the strongest immunoreactivity in cells close to the isthmus; in the distal stomach, strongest immunoreactivity was found in cells at the base of the glands. Oxyntic cells were more intensely immunoreactive than oxynticopeptic cells. This antibody labeled a single band of M(r) 100,600 on immunoblots of D. sabina gastric microsomes. These results show the earliest phylogenetic occurrence of a gastric ATPase in putative acid-secreting cells and suggest that this enzyme shares structural features with mammalian H,K-ATPase.


Assuntos
ATPase Trocadora de Hidrogênio-Potássio/metabolismo , Rajidae/metabolismo , Estômago/enzimologia , Animais , Técnicas Imunoenzimáticas , Fatores de Tempo
15.
J Histochem Cytochem ; 37(9): 1421-5, 1989 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-2768810

RESUMO

Freeze-fracture replicas of filipin-treated samples of guinea pig colon mucosa reveal areas in the membrane of the goblet cell granules labeled by filipin-cholesterol complexes (FCC) intermingled with regions patterned by "lines." The FCC and "lines" are arranged in an approximately rhombic pattern. Other membranes of the same cell or of other cells display either FCC only, aligned and occasionally ordered in "rhombs," "lines" only, with a similar pattern, or randomly distributed FCC. Optical diffraction was used to analyze and compare replicas of membranes with ordered FCC and "lines", as well as randomly distributed FCC. The results demonstrate that all these structures are reciprocally related through a common distribution pattern in the membrane. This observation supports the assumption that cholesterol has a preferential ordered distribution within the membrane bilayer.


Assuntos
Colesterol/análise , Colo/citologia , Mucosa Intestinal/citologia , Animais , Membrana Celular/análise , Membrana Celular/ultraestrutura , Colesterol/metabolismo , Colo/análise , Colo/ultraestrutura , Filipina/análise , Filipina/metabolismo , Técnica de Fratura por Congelamento , Cobaias , Mucosa Intestinal/metabolismo , Mucosa Intestinal/ultraestrutura , Microscopia Eletrônica , Mucosa/metabolismo , Mucosa/ultraestrutura , Difração de Raios X
16.
Invest Ophthalmol Vis Sci ; 35(2): 711-9, 1994 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-8113022

RESUMO

PURPOSE: In patients with several disorders of the ocular surface, in wearers of contact lenses, and occasionally in patients considered "normal," the epithelial cells of the bulbar conjunctiva present in their nucleus peculiar alterations of the heterochromatin arrangement that, because of the shape this assumes, are named "snakelike chromatin," or "snakes." To obtain some information about the yet unknown etiology of these snakes, the authors investigated them by electron microscopy. METHODS: Identical conjunctival epithelial cells, collected by impression cytology from long-time contact lens wearers, were first identified by light microscopy and then observed by scanning and transmission electron microscopy. RESULTS: In scanning electron microscopy, cytoplasmic and nuclear components of air-dried cells were seen collapsed on the snake, which became prominent at the surface, proving its high degree of compactness and showing its axial position and characteristic shape in the elongated nucleus. In transmission electron microscopy, the marginal heterochromatin of the affected nuclei was detached peripherally, forming thin chromatin strands directed toward the main nuclear axis and accumulating there into the snake structure. An important component of the nuclear skeleton, the fibrous lamina, was altered or lost, whereas the nuclear envelope itself did not move and remained intact. Stages of the snake alteration considered as advanced showed an almost completely reversed eu- and heterochromatin distribution, and nucleoli were usually no longer seen. In cases with increased epithelial alteration, there occurred various stages of segmentation of nuclei, induced by an atypical accumulation of cytoplasmic filaments, rolling up around the nucleus and constricting it like a cuff. CONCLUSIONS: The presence of a mechanical stimulus is shown in the ultrastructural findings, and these strongly suggest that it is altering the nuclear and cytoplasmic skeleton, producing snakes and their segmentation. Therefore, snakelike chromatin is suggested as an indicator of mechanical stress on the ocular surface.


Assuntos
Cromatina/ultraestrutura , Túnica Conjuntiva/ultraestrutura , Núcleo Celular/ultraestrutura , Túnica Conjuntiva/citologia , Lentes de Contato/efeitos adversos , Citoplasma/ultraestrutura , Citoesqueleto/ultraestrutura , Células Epiteliais , Epitélio/ultraestrutura , Humanos , Microscopia Eletrônica de Varredura , Estresse Mecânico
17.
Placenta ; 1(3): 245-58, 1980.
Artigo em Inglês | MEDLINE | ID: mdl-7443643

RESUMO

Terminant chorionic villi of the human placenta at 8, 9, 12, 14 weeks' gestation and at full term have been studied in freeze-fracture replicas and in thin sections. In replicas, maculae occludentes of different size have been detected in all placental samples on the cleaved plasma membrane of the free (maternal) surface of the syncytiotrophoblast. In thin sections they are located in narrow invaginations of the syncytium luminal plasma membrane. These invaginations are in turn similar to flattened vesicles (also associated with junctions) located in the underlying syncytioplasm. Possible origin and function of the junctions on the syncytium free surface are discussed.


Assuntos
Vilosidades Coriônicas/ultraestrutura , Placenta/ultraestrutura , Membrana Celular/ultraestrutura , Feminino , Técnica de Fratura por Congelamento , Humanos , Junções Intercelulares/ultraestrutura , Gravidez , Primeiro Trimestre da Gravidez , Segundo Trimestre da Gravidez , Terceiro Trimestre da Gravidez
18.
Virchows Arch ; 429(2-3): 165-72, 1996 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-8917718

RESUMO

The effect of externally applied mechanical stress was investigated by thin section electron microscopy of the microvessels in the unaffected palmar fascia in the carpal tunnel syndrome and in patients with Dupuytren's contracture before and after application of a continuous elongation device. In the unaffected palmar fascia the microfilaments of the endothelial cells were connected to a few adherens junctions and focal contacts; stress fibres were absent. In the cord of Dupuytren's disease the microfilaments were increased in quantity. The length ratios of the connections with the lateral and basal cell membrane were significantly higher than in the control group and increased to an even greater extent in the continuously extended fascia. Stress fibres appeared in the endothelial cells of postcapillary venules in the nonextended cord and in the endothelium of both arterioles and venules after extension elongation. the numerous intermediate filaments and the rare microtubules remained unchanged in the endothelial cells of all palmar fasciae analysed. In the endothelial cells of the microvessels the mechanical stress applied from outside mainly affected the contractile component of the cytoskeleton.


Assuntos
Citoesqueleto de Actina/ultraestrutura , Síndrome do Túnel Carpal/patologia , Contratura de Dupuytren/patologia , Endotélio Vascular/patologia , Fáscia/irrigação sanguínea , Mãos/irrigação sanguínea , Idoso , Feminino , Humanos , Masculino , Microcirculação , Microscopia Eletrônica , Pessoa de Meia-Idade , Estresse Mecânico
19.
Microsc Res Tech ; 38(6): 598-608, 1997 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-9330348

RESUMO

The brush cells (BC) are the second most frequent cellular component of the epithelium of the mouse gallbladder. They have a topographical distribution, being present in large numbers toward the neck and in the fundic regions of the organ and are scattered in the body. Serial section studies demonstrate that BC have a characteristic shape consisting of a narrow apical portion, bulky body and basal cytoplasmic projections. BC are located obliquely among the principal cells. Scanning electron microscopy demonstrates that the microvilli forming the prominent brush border, after which the cell was named, have a triangular arrangement. Due to their size and stiffness, the microvilli of BC have more similarity with stereocilia of sensory cells than with conventional microvilli. Furthermore freeze-fracture replicas demonstrate that, like stereocilia, the P face of the microvilli plasma membrane of BC is smoother than the E face but several intramembranous particles form small aggregates on the microvillus tip of both P and E faces. Numerous intramembranous particles are scattered on the lateral plasma membrane. An unusual, spatially organized cytoskeleton characterizes the apical cytoplasm of BC. The use of the appropriate fixative reveals that it consists of bundles of actin filaments originating from the axis of the apical microvilli and stretching continuously up to the supranuclear region of the cell. Microtubuli, also assembled in bundles, flank in alternating manner the actin filaments over their whole course. Due to the strong parallel arrangement of both cytoskeletal structures, the apical cytoplasm of the BC assumes a typical stiffness, observable in both thin sections and freeze-fracture replicas. A variable number of vesicles of different size are aligned between the bundles of actin filaments and microtubuli; their shape is highly influenced by the fixative used. Intraluminal injection of horseradish peroxidase demonstrates that these vesicles are not resorptive as they are not filled by the tracer. The BC possess a large number of lateral microvilli. These, whether single or in pairs, are rigid cytoplasmic protrusions that leave the lateral surface of the cell in all directions and penetrate deeply into the cytoplasm of the adjacent principal cells. The bundle of actin filaments emanating from each lateral microvillus extends at different angles into the cytoplasm. A conspicuous amount of bundles of 10 nm filaments is intertwined around the nucleus and extends toward the desmosomes of the lateral plasma membrane and into the basal cellular body. Arguments are considered in support of the view that interactions between the plasma membrane with its differentiations on the one hand and the cytoskeleton elements on the other hand, play a key role in the function of BC as a receptor (sensory) cell.


Assuntos
Vesícula Biliar/ultraestrutura , Animais , Núcleo Celular/ultraestrutura , Tamanho Celular/fisiologia , Citoplasma/ultraestrutura , Camundongos , Microvilosidades/ultraestrutura
20.
J Neurol ; 237(6): 382-4, 1990 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-2277275

RESUMO

A case of a primary intracerebral sarcoma is described in a 5-year-old girl. Histology and immunohistochemistry excluded the diagnosis of a leiomyosarcoma, a malignant haemangiopericytoma or a fibrosarcoma; electron-microscopical findings indicated that the origin of the sarcoma was in the pia mater.


Assuntos
Neoplasias Meníngeas/ultraestrutura , Pia-Máter/ultraestrutura , Sarcoma/ultraestrutura , Pré-Escolar , Diagnóstico Diferencial , Feminino , Hemangiopericitoma/diagnóstico , Hemangiopericitoma/ultraestrutura , Humanos , Imuno-Histoquímica , Leiomiossarcoma/diagnóstico , Leiomiossarcoma/ultraestrutura , Neoplasias Meníngeas/diagnóstico , Microscopia Eletrônica , Sarcoma/diagnóstico
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