Imaging of cellular aging in human retinal blood vessels.

To date two main aging vascular lesions have been reported in elderly human retinas: acellular capillaries and microaneurysms. However, their exact mechanism of formation remains unclear. Using high resolution microscopy techniques we revise cellular alterations observed in aged human retinal vessels, such as lipofuscin accumulation, caveolae malfunction, blood basement membrane disruption and enhanced apoptosis that could trigger the development of these aging vascular lesions. Moreover, we have generated a set of original images comparing retinal vasculature between middle and old aged healthy humans to show in a comprehensive manner the main structural and ultrastructural alterations occurred during age in retinal blood vessels.

Miconazole-loaded nanostructured lipid carriers (NLC) for local delivery to the oral mucosa: improving antifungal activity.

Miconazole is a widely used antifungal agent with poor aqueous solubility, which requires the development of drug delivery systems able to improve its therapeutic activity. For this purpose, a miconazole-loaded nanostructured lipid carriers (NLC) dispersion was prepared and characterized. Further, the dispersion was used to prepare a NLC-based hydrogel formulation proposed as an alternative system to improve the local delivery of miconazole to the oral mucosa. NLC dispersion showed particles in the nanometer range (≈ 200 nm) with low polidispersity index (<0.3), good physical stability and high encapsulation efficiency (>87%). A controlled miconazole release was observed from NLC and NLC-based hydrogel formulations, in contrast to a commercial oral gel formulation, which demonstrated a faster release. Additionally, it was observed that the encapsulation of miconazole in the NLC improved its antifungal activity against Candida albicans. Therefore, it was demonstrated that the encapsulation of miconazole in NLC allows for obtaining the same therapeutic effect of a commercial oral gel formulation, using a 17-fold lower dose of miconazole.

Monosegmented flow-analysis of serum cholesterol.

A monosegmented flow system is designed for enzymatic spectrophotometric determination of cholesterol in blood serum. The sample (4.5 microliters), enzymatic reagent (150 microliters) and an air plug (100 microliters) are simultaneously inserted into a carrier stream buffered to pH 7.4 (potassium dihydrogenphosphate). In order to avoid the step of air removal, a relocating detector was used. This system handles about 42 samples per hour, yielding precise results (R.S.D. usually < 3.0%). Sensitivity is 46 mAU 1/mmol (mAU stands for milliabsorbance units), being the method linear up to 10.3 mmol/l cholesterol. Accuracy was assessed by running 30 samples already analysed by a conventional procedure: no statistical difference between methods was found at the 95% confidence level.

Molecular cloning of metaphase chromosome protein 1 (MCP1), a novel human autoantigen that associates with condensed chromosomes during mitosis.

Systemic lupus erythematosus autoantibodies were used to identify and to characterize new human chromosome-associated proteins. Previous immunolocalization studies in human and murine tissue culture cells showed that some of these monoclonal antibodies recognize nuclear antigens that associate with condensed chromosomes during mitosis. One antibody was selected for screening a human HeLa S3 cDNA expression library, and cDNAs that code for an antigen of 31-33 kDa were isolated. Immunological, biochemical and cell fractionation data indicate that the 31- to 33-kDa antigen corresponds to the chromosome-associated protein recognized by the original monoclonal antibody. Sequence analysis shows that we isolated a novel human gene. Immunolocalization to human tissue culture cells shows that during interphase the antigen is dispersed in the nucleus and that during mitosis it associates exclusively with condensed chromosomes. A similar pattern of localization was also observed in mouse fibroblasts, suggesting that the antigen is conserved among different species. Finally, we show that part of the antigen remains bound to the scaffold/matrix component, even after high salt extraction.

Ewing's sarcoma in children: twenty-five years of experience at the Instituto Portugês de Oncologia de Francisco Gentil (I.P.O.F.G.).

Fifty children with Ewing's sarcoma were consecutively treated from 1962 to 1987 and retrospectively analyzed at the I.P.O.F.G. of Lisbon. At first diagnosis, 10 cases had distant metastases. The remaining 40 patients had clinically localized disease, and different protocols were followed over the years. The best results were obtained with chemotherapy and radiotherapy with or without surgery; and for these children the two-year survival rate was 42.8% vs. 8.3% for the group of patients submitted to local treatment alone. Besides the treatment modality, other factors influenced the prognosis, such as inflammatory signs, sex, tumor volume, and tumor site as well as evidence of distant metastases.

Palliative telecobalt-60 with concentrated dose in TNM classification T4 of the breast.

A method is presented of rapid, palliative radiotherapy for carcinoma of the breast, composed of two sessions of 6.5 or 8.5 Gy delivered at a 48-hour interval. The radiobiological equivalence of this unconventional technique is presented. Our preliminary results in a series of 80 patients were published in 1978. The present report is based on the analysis of 112 patients with T4 of the breast submitted to the "flash" dose, from March 1978 to December 1981. The method was used not only for relief of symptoms such as pain and hemorrhage, but also as part of intensive radiotherapy in all patients with good response and with no manifestation of distant dissemination so that they were able to resume treatment 2.5 or 3 weeks later, with conventional fractionation. Some of these patients became operable. About half of the cases had castration and/or hormone therapy and/or chemotherapy. In the group of patients with T4M0, 59.8% were alive after more than 1 year, and of them, 43.7% had no evidence of disease. These encouraging results suggest that this method be advocated in T4 of the breast because of its rapidity and good tolerance with no significant complications.