Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 15 de 15
Filter
Add more filters










Publication year range
2.
Eye (Lond) ; 21(6): 752-9, 2007 Jun.
Article in English | MEDLINE | ID: mdl-16575415

ABSTRACT

PURPOSE: Most uveal melanoma patients (UMP) do not show evidence of metastases upon diagnosis. However, despite local tumour control, approximately 50% of them will develop metastases. These findings suggest that malignant cells may have already disseminated by the time of initial diagnosis. The purpose of the study was to detect circulating malignant cells (CMCs) in UMP and to correlate them with prognostic factors and therapy. METHODS: Nested reverse transcriptase-polymerase chain reaction (RT-PCR) was used to detect CMCs. In each UMP, blood was collected every 3 months. In each visit, 20 RT-PCR tests were performed. The date of diagnosis, largest tumour dimension, type, and date of treatment were obtained. RESULTS: A total of 30 UMP were enrolled. Five patients were enrolled at the time of diagnosis and 25 patients between 1 and 17 years following diagnosis. No UMP showed clinical evidence of metastasis. A total of 136 visits were registered, 1360 samples collected, and 2720 RT-PCRs performed. CMCs were identified in 29 patients in 119 visits (87.5%). However, in each visit, a low number of positive tests were recorded. CMCs were found in newly diagnosed, irradiated, enucleated, and observed patients regardless of tumour size and time period following treatment. CONCLUSIONS: Uveal melanoma (UM) is not a localized ocular disease. CMCs were recorded at initial diagnosis confirming the early metastatic nature of UM. CMCs were present following treatment, including enucleation, demonstrating that CMCs are capable of disseminating and surviving, possibly as micrometastasis, which would contribute to the pool of CMCs at a later stage. Systemic therapy should be evaluated.


Subject(s)
Melanoma/secondary , Neoplastic Cells, Circulating/pathology , Uveal Neoplasms/pathology , Aged , Female , Follow-Up Studies , Humans , Male , Melanoma/diagnosis , Melanoma/pathology , Melanoma/therapy , Middle Aged , Neoplasm, Residual/pathology , Prognosis , Prospective Studies , Reverse Transcriptase Polymerase Chain Reaction/methods , Uveal Neoplasms/diagnosis , Uveal Neoplasms/therapy
3.
Dev Biol ; 297(2): 402-16, 2006 Sep 15.
Article in English | MEDLINE | ID: mdl-16916506

ABSTRACT

During early stages of brain development, neuroepithelial stem cells undergo intense proliferation as neurogenesis begins. Fibroblast growth factor 2 (FGF2) has been involved in the regulation of these processes, and although it has been suggested that they work in an autocrine-paracrine mode, there is no general agreement on this because the behavior of neuroepithelial cells is not self-sufficient in explants cultured in vitro. In this work, we show that during early stages of development in chick embryos there is another source of FGF2, besides that of the neuroepithelium, which affects the brain primordium, since the cerebrospinal fluid (E-CSF) contains several isoforms of this factor. We also demonstrate, both in vitro and in vivo, that the FGF2 from the E-CSF has an effect on the regulation of neuroepithelial cell behavior, including cell proliferation and neurogenesis. In order to clarify putative sources of FGF2 in embryonic tissues, we detected by in situ hybridization high levels of mRNA expression in notochord, mesonephros and hepatic primordia, and low levels in brain neuroectoderm, corroborated by semiquantitative PCR analysis. Furthermore, we show that the notochord segregates several FGF2 isoforms which modify the behavior of the neuroepithelial cells in vitro. In addition, we show that the FGF2 ligand is present in the embryonic serum; and, by means of labeled FGF2, we prove that this factor passes via the neuroepithelium from the embryonic serum to the E-CSF in vivo. Considering all these results, we propose that, in chick embryos, the behavior of brain neuroepithelial stem cells at the earliest stages of development is influenced by the action of the FGF2 contained within the E-CSF which could have an extraneural origin, thus suggesting a new and complementary way of regulating brain development.


Subject(s)
Cerebrospinal Fluid/metabolism , Fibroblast Growth Factor 2/physiology , Neuroepithelial Cells/cytology , Stem Cells/cytology , Animals , Cell Differentiation , Cell Proliferation , Cerebrospinal Fluid Proteins/physiology , Chick Embryo , Embryonic Development , Fibroblast Growth Factor 2/metabolism , Neurons/metabolism , PC12 Cells , Rats
4.
Eye (Lond) ; 18(2): 135-8, 2004 Feb.
Article in English | MEDLINE | ID: mdl-14762403

ABSTRACT

PURPOSE: A chalazion, localized lipogranulomatous inflammation of the eyelid, may simulate various eyelid lesions. This study was conducted to determine the accuracy of the clinical diagnosis of chalazion and demonstrate the importance of histopathological confirmation of the diagnosis. METHODS: Histopathological diagnoses of 1060 cases with the clinical diagnosis of chalazion, submitted to the Henry C Witelson Ophthalmic Pathology Laboratory and Registry between September 1993 and December 2001, were retrospectively evaluated. Discrepancies between clinical and histopathological diagnoses were classified. RESULTS: A total of 1033 (97.4%) of the 1060 cases were clinically diagnosed as primary and the remaining 27 (2.6%) as recurrent chalazions. Agreement was noted between clinical and histopathological diagnoses in 992 (93.6%) cases. Of the 68 (6.4%) clinically misdiagnosed cases, 15 (1.4%) were found to be malignant, two (0.2%) premalignant, and 51 (4.8%) benign conditions. Sebaceous cell carcinoma was the most commonly missed malignancy (12 cases, 1.1%) followed by basal cell carcinoma (three cases, 0.3%). Premalignant lesions, which masqueraded as chalazion, were chronic inflammation with cellular atypia and mitotic figures (two cases, 0.2%). Of these 17 cases with premalignant and malignant histopathologies, only six (35.3%) had a clinical diagnosis of recurrent chalazion, whereas the others (64.7%) were primary cases. Of the various benign conditions that were misdiagnosed as chalazion, different types of chronic inflammation (24 cases, 2.2%) were the most frequent. CONCLUSIONS: A number of different benign, premalignant, and malignant conditions may clinically masquerade as a chalazion. Delayed diagnosis and treatment of sebaceous cell carcinoma, which is the most frequently missed malignancy, may be life threatening for the patient. Therefore, all chalazion specimens, primary or recurrent, should be submitted for histopathological examination.


Subject(s)
Chalazion/diagnosis , Eyelid Neoplasms/diagnosis , Adenocarcinoma, Sebaceous/diagnosis , Adenocarcinoma, Sebaceous/pathology , Adult , Aged , Carcinoma, Basal Cell/diagnosis , Carcinoma, Basal Cell/pathology , Chalazion/pathology , Diagnosis, Differential , Eyelid Neoplasms/pathology , Humans , Middle Aged , Precancerous Conditions/diagnosis , Precancerous Conditions/pathology , Recurrence , Retrospective Studies
6.
Arch Soc Esp Oftalmol ; 76(10): 613-19, 2001 Oct.
Article in Spanish | MEDLINE | ID: mdl-11685679

ABSTRACT

PURPOSE: To analyse the putative toxic effect of three commercially available non-preserved artificial tear formulations on in vitro human conjunctival cells. MATERIAL AND METHOD: A conjunctival human epithelial cell line was exposed to Cellufresh, Oculotect and Acuolens formulations during 1, 3 and 24 hours. Cytotoxicity was measured by calculating the percentage of cell viability examination and scanning electron microscopy (SEM). Controls underwent exposure to supplement free DMEM-F12 (negative control) and exposure to 0.005% benzalkonium chloride solution (positive control). RESULTS: Cell viability after 1 or 3 hours incubation with Cellufresh and Oculotect was similar to that obtained for negative controls. With Acuolens incubation however, cell viability showed significant reduction after 3 and 24 hours compared to control. SEM showed that Cellufresh and Oculotect exposed cells presented similar behavior to control cells. All three cell lines presented evidence of cellular surface alteration after incubation for 1 or 3 hours compared to controls, Acuolens showing the highest rate of alterations in exposed cells and an additional increment in cell loss was observed. CONCLUSION: In the present study, non preserved artificial tears formulations showed a different degree in their in vitro toxicity, Acuolens being more toxic than Cellufresh or Oculotect.


Subject(s)
Conjunctiva/drug effects , Epithelial Cells/drug effects , Ophthalmic Solutions/adverse effects , Cell Survival/drug effects , Cells, Cultured , Conjunctiva/cytology , Drug Evaluation, Preclinical , Epithelial Cells/ultrastructure , Humans , Microscopy, Electron, Scanning , Ophthalmic Solutions/chemistry
7.
Arch. Soc. Esp. Oftalmol ; 76(10): 613-620, oct. 2001.
Article in Es | IBECS | ID: ibc-9059

ABSTRACT

Objetivo: Analizar los posibles efectos tóxicos de tres preparados comerciales de lágrimas artificiales sin conservantes sobre células conjuntivales humanas in vitro. Métodos: Cultivos de células epiteliales de conjuntiva humana fueron expuestos a la acción de los preparados comerciales Cellufresh, Oculotect y Acuolens durante 1, 3 y 24 horas. Transcurrido dicho tiempo se estudió su posible efecto tóxico, expresado como porcentaje de Viabilidad Celular, y se analizó la presencia de alteraciones en la superficie de las células mediante microscopia electrónica de barrido (MEB). El estudio incluyó como control negativo de toxicidad células expuestas a medio de cultivo DMEM-F12 sin suplementos y como control positivo, células expuestas a una solución de cloruro de benzalconio al 0,005 por ciento en dicho medio de cultivo. Resultados: La Viabilidad Celular obtenida tras incubar 1 ó 3 horas con Cellufresh y Oculotect fue similar a la del control negativo, si bien disminuyó algo cuando el tiempo de incubación fue de 24 horas. Sin embargo, la Viabilidad Celular tras incubar con Acuolens fue significativamente menor para todos los tiempos. La MEB mostró que con Cellufresh y Oculotect el aspecto general de las células era muy similar al observado en las células control. Sin embargo, con Acuolens se observaron alteraciones acusadas, tanto tras 1 hora como tras 3 horas de incubación, y una notable pérdida celular. Conclusión: De las lágrimas artificiales sin conservantes en estudio, Acuolens resultó ser la más tóxica in vitro, tanto a tiempos cortos como largos (AU)


Subject(s)
Humans , Microscopy, Electron, Scanning , Ophthalmic Solutions , Cells, Cultured , Cell Survival , Conjunctiva , Drug Evaluation, Preclinical , Epithelial Cells , Microscopy, Electron, Scanning
8.
Arch Ophthalmol ; 118(9): 1287-9, 2000 Sep.
Article in English | MEDLINE | ID: mdl-10980777

ABSTRACT

A "napkin-ring" subretinal membrane is an unusual expression of subretinal proliferation associated with retinal detachment. An 80-year-old man with a total funnel-shaped retinal detachment underwent pars plana vitrectomy, 360 degrees relaxing retinotomy, excision of a subretinal napkin-ring membrane, and silicone oil injection. Histopathologic examination of the removed napkin-ring subretinal membrane revealed the presence of retinal pigment epithelium (RPE) as the major source of cells within the membrane. Myofibroblasts were the most common cellular constituents; the total number of these cells may have correlated with the degree of clinical contraction, causing a funnel-shaped retinal detachment. Arch Ophthalmol. 2000;118:1287-1289


Subject(s)
Retinal Detachment/etiology , Vitreoretinopathy, Proliferative/complications , Actins/metabolism , Aged , Aged, 80 and over , Fibroblasts/pathology , Glial Fibrillary Acidic Protein/metabolism , Humans , Immunoenzyme Techniques , Injections , Male , Pigment Epithelium of Eye/pathology , Retinal Detachment/pathology , Retinal Detachment/surgery , S100 Proteins/metabolism , Silicone Oils/administration & dosage , Vitrectomy , Vitreoretinopathy, Proliferative/metabolism , Vitreoretinopathy, Proliferative/pathology , Vitreoretinopathy, Proliferative/surgery
9.
Ophthalmology ; 107(3): 550-4, 2000 Mar.
Article in English | MEDLINE | ID: mdl-10711894

ABSTRACT

OBJECTIVE: Malignant granular cell tumor is a rare type of soft tissue sarcoma. To our knowledge, ocular (eyelid) involvement has been described in only two cases. Herein, we report the clinicopathologic features of an unusual case of malignant granular cell tumor metastatic to the orbit. DESIGN: Observational case report. METHODS: Retrospective review of the medical record and the histopathologic and electron microscopic findings and review of the literature. RESULTS: A 72-year-old man with biopsy-proven granular cell tumor in the cervical region was initially seen with proptosis and motility disturbance. A magnetic resonance imaging scan showed a large intraconal mass, and biopsy of the orbital mass revealed granular cell tumor. Histopathologic examination of the primary neck tumor and the orbital mass revealed increased nuclear atypia and pleomorphism in the consecutive lesions. The morphologic impression of granular cell tumor was also supported by the immunohistochemical demonstration of S-100 protein expression and ultrastructural findings typical of granular cell tumor. Six months after the orbital involvement, systemic workup revealed multiple apparent bony and lung metastases. CONCLUSIONS: We report the first malignant granular cell tumor metastatic to the orbit and suggest the inclusion of this tumor in the differential diagnosis of metastatic orbital lesions.


Subject(s)
Granular Cell Tumor/secondary , Head and Neck Neoplasms/pathology , Orbital Neoplasms/secondary , Aged , Diphosphonates , Fatal Outcome , Granular Cell Tumor/diagnosis , Granular Cell Tumor/radiotherapy , Humans , Lung Neoplasms/diagnostic imaging , Lung Neoplasms/secondary , Magnetic Resonance Imaging , Male , Orbital Neoplasms/diagnosis , Orbital Neoplasms/radiotherapy , Radionuclide Imaging , Retrospective Studies , Spinal Neoplasms/diagnostic imaging , Spinal Neoplasms/secondary , Technetium Compounds
10.
Curr Eye Res ; 19(2): 95-105, 1999 Aug.
Article in English | MEDLINE | ID: mdl-10420178

ABSTRACT

PURPOSE: To demonstrate by ultrastructural techniques that human conjunctival epithelium cells in vitro can produce mucin-like secretion. METHODS: Primary cultures of human conjunctival epithelial cells were grown in different culture media. Cultures were allowed to grow and were processed after 5 days and 1, 2, 3, 4, or 5 weeks for transmission and scanning electron microscopy, according to the method of Nichols et al. modified in our laboratory. RESULTS: Marked differences were seen between primary cultures grown with or without hydrocortisone. A thick tannic acid-stained layer was observed when hydrocortisone was present in the culture medium; however, that layer was virtually absent in cultures grown with hydrocortisone-free media. Scanning electron microscopy revealed a dense deposit showing a network-like structure. Moreover, the age of the cultures clearly influenced the thickness of the tannic acid-stained deposit, which thickened as the cultures aged. CONCLUSIONS: These results strongly suggest that the layer growing in the presence of hydrocortisone is mucus. The fact that this material became more abundant as the cultures aged indicates that mucus is actively produced and secreted by conjunctival epithelial cells in vitro. This study might contribute to the knowledge of mucus-deficient pathologies of the ocular surface.


Subject(s)
Conjunctiva/metabolism , Mucus/metabolism , Animals , Cattle/blood , Cattle/embryology , Cells, Cultured , Conjunctiva/cytology , Conjunctiva/drug effects , Conjunctiva/ultrastructure , Epithelial Cells/drug effects , Epithelial Cells/metabolism , Epithelial Cells/ultrastructure , Fetal Blood/physiology , Humans , Hydrocortisone/pharmacology , Microscopy, Electron , Microscopy, Electron, Scanning
11.
Cornea ; 17(4): 433-40, 1998 Jul.
Article in English | MEDLINE | ID: mdl-9676917

ABSTRACT

PURPOSE: Frequent instillation of artificial tears is the primary disadvantage of the treatment for dry-eye syndrome. Recently gel formulations have been proposed as an alternative to classic cellulose formulations. The higher viscosity of these gels presumably prolongs tear-retention time in the eye and results in fewer daily applications. However, no toxicologic studies with gel formulations have been performed. Our aim was to study the toxic effects of these formulations on corneal cells. METHODS: SIRC cells from rabbit cornea were exposed for 30 min, and 1, 3, and 6 h to five commercially available artificial tears, three of them carbomer gel formulations (Lacrivisc, Lacrivisc unit-doses, and Viscotears) and two carboxymethylcellulose (CMC) formulations (Celluvisc and Cellufresh). A cytotoxicity assay and a scanning electron microscopy (SEM) study were used to analyze the putative toxic effects of the formulations. The preservatives of the gel formulations also were tested. RESULTS: Carbomer gel formulations, both with and without preservatives, caused more in vitro toxic effects in the corneal cells than did CMC formulations and caused severe damage even after 30 min of exposure. SEM revealed dramatic cell-surface alterations. Preservatives added to Lacrivisc and Viscotears also had toxic effects on cells, whose effects were not significantly different from those of the commercial preparations. CONCLUSION: These results demonstrate that in the in vitro study, CMC artificial tears are less toxic than carbomer gel formulations. Questions about the benefits of using high-viscosity gels in the treatment of dry-eye syndrome still remain.


Subject(s)
Carboxymethylcellulose Sodium/toxicity , Cornea/drug effects , Ophthalmic Solutions/toxicity , Polymers/toxicity , Animals , Cell Line , Cell Survival/drug effects , Cells, Cultured , Cornea/ultrastructure , Gels , Microscopy, Electron, Scanning , Rabbits
12.
Graefes Arch Clin Exp Ophthalmol ; 235(5): 268-76, 1997 May.
Article in English | MEDLINE | ID: mdl-9176674

ABSTRACT

Primary cultures of human epithelial cells from normal conjunctiva were developed and characterized to determine whether they retained epithelial characteristics. Conjunctival explants were obtained from the upper fornix of healthy donors and cultured in supplemented DMEM/F-12 medium for 5 days. The epithelial outgrowth was maintained for an additional 10 days. Primary cultures were then processed for light microscopy, transmission and scanning electron microscopy (TEM, SEM), and immunocytochemistry. They exhibited typical features of conjunctival epithelium on light microscopy (polygonal morphology, intimate cohesion, production of mucins), TEM (abundant desmosomes, keratin bundles, granules, microvilli), SEM (polygonal shape, microvilli, intimate cohesion), and immunocytochemistry (positivity for the receptor of epidermal growth factor, desmosomal proteins, and cytokeratins). In conclusion, primary cultures developed from normal human conjunctiva maintained the epithelial characteristics in vitro. Because the conjunctiva is a major component of the anterior ocular surface, we propose this in vitro system as suitable for physiopathologic and toxicologic studies.


Subject(s)
Conjunctiva/cytology , Antibodies, Monoclonal , Cells, Cultured , Conjunctiva/metabolism , Conjunctiva/ultrastructure , Culture Media , Cytoplasm/ultrastructure , Cytoskeletal Proteins/metabolism , Endoplasmic Reticulum/ultrastructure , Epithelial Cells , Epithelium/metabolism , Epithelium/ultrastructure , Golgi Apparatus/ultrastructure , Humans , Immunohistochemistry , Microscopy, Electron, Scanning Transmission , Microscopy, Phase-Contrast , Mitochondria/ultrastructure
13.
Biochem J ; 327 ( Pt 3): 917-23, 1997 Nov 01.
Article in English | MEDLINE | ID: mdl-9581574

ABSTRACT

The expression of cell-surface adhesion proteins and the release of extracellular-matrix degradative enzymes constitute crucial processes for the attachment of neutrophils to the endothelium and for the subsequent extravasation of these cells through the endothelial layer. We have analysed in resting human neutrophils the subcellular localization of heparanase, a heparan-sulphate-degrading endoglycosidase that can degrade basement-membrane components, thereby facilitating neutrophil passage into the tissue during an inflammatory reaction. By subcellular fractionation of postnuclear supernatants from resting human neutrophils on continuous sucrose gradients, we have found that heparanase activity was mainly located in gelatinase-containing tertiary granules. Using a specific antibody, the 96-kDa heparanase protein was further located in the gelatinase-rich subcellular fractions. Following immunoblotting and immunoprecipitation analysis in the distinct subcellular fractions, we also found co-localization of heparanase and Mo1 (CD11b/CD18), a leucocyte integrin involved in the attachment of neutrophils to the endothelium, in the fractions enriched in gelatinase-containing tertiary granules. Treatment of human neutrophils with tumour necrosis factor or granulocyte/macrophage colony-stimulating factor induced an increase in the CD11b/CD18 cell-surface expression, as well as the release of both gelatinase (matrix metalloproteinase-9) and heparanase, but not of other granule markers, indicating a major co-localization of gelatinase, heparanase and CD11b/CD18 in the same organelle. Furthermore, confocal laser scanning microscopy using specific antibodies against gelatinase and heparanase revealed a major co-localization of both enzymes in intracellular cytoplasmic granules. The major localization of heparanase and CD11b/CD18 in the gelatinase-containing tertiary granule supports the notion that mobilization of this organelle can regulate extravasation of human neutrophils.


Subject(s)
Cytoplasmic Granules/enzymology , Extracellular Matrix/enzymology , Gelatinases/metabolism , Glucuronidase , Glycoside Hydrolases/metabolism , Neutrophils/enzymology , CD18 Antigens/immunology , Cell Fractionation , Cytoplasmic Granules/immunology , Granulocyte-Macrophage Colony-Stimulating Factor/metabolism , Humans , Immunoblotting , Macrophage-1 Antigen/immunology , Microscopy, Confocal , Neutrophils/cytology , Neutrophils/immunology , Precipitin Tests , Tumor Necrosis Factor-alpha/metabolism
SELECTION OF CITATIONS
SEARCH DETAIL
...