Detection of EWS/FLI-1 fusion in non-Ewing soft tissue tumors.

OBJECTIVES: EWS/FLI-1 fusion mainly appears in Ewing's sarcoma or the primitive neuroectodermal tumors and represents a genomic marker for these tumors. However, it can appear with lower frequency in other soft tissue tumors. The paper investigates the presence of EWS/FLI-1 fusion in clinically diagnosed sarcoma belonging to different non-Ewing connective tissue tumors in order to search for a possible new biomarker valuable for investigators. METHODOLOGY: 20 patients with soft tissue tumors, who underwent surgery, were tested. Intra-operative samples of normal and tumor tissue were collected for histopathological diagnosis and genetics determinations. The patients' RNA from tumor and normal peritumoral tissue was extracted and EWS/FLI-1 fusion screened by quantitative real-time PCR. The relative expression of the fusion in the tumor sample was compared to the similar expression in normal tissue. RESULTS: The amplification in the threshold zone was shown by 5 samples (25%): 2 clear cell sarcoma, 1 fibrosarcoma, 1 malignant tumor of nerve sheath, 1 metastatic adenocarcinoma. We differentiated between the unspecific amplification and concluded that these are weak positive results. CONCLUSIONS: Genomic investigation may establish the tumor malignancy and its possible affiliation earlier than histopathology. It can support the screening of EWS/FLI-1 fusion in a larger variety of clinically diagnosed soft tissue tumors.

Dynamics of telopodes (telocyte prolongations) in cell culture depends on extracellular matrix protein.

Telocytes (TC) are cells with telopodes (Tp), very long prolongations (up to 100 µm) with an uneven caliber ( www.telocytes.com ). Factors determining the dynamics of cellular prolongations are still unknown, although previous studies showed telopode motility in TC cultures. We comparatively investigated, by time-lapse videomicroscopy, the dynamics of Tp of mouse heart TC seeded on collagen, fibronectin, and laminin. Under our experimental conditions, TC and fibroblasts (cell line L929) behaved differently in terms of adherence, spreading, and prolongation extension. Fibroblasts showed lower spreading on the matrix proteins used. The time needed for spreading was 2-4 h for TC, versus 8-10 h for fibroblasts. The values for final cell surface area after spreading were between 200 and 400 µm(2) for fibroblasts and 800-2,000 µm(2) for TC. TC showed a more than three times higher ability to spread on the tested matrix proteins. An extremely low capacity to extend prolongations with lengths shorter than cell bodies was noted for fibroblasts, while TC extended prolongations longer than the cell body length, with a moniliform appearance. The stronger adherence and spreading were noted for TC seeded on fibronectin, while the lowest were on laminin. Collagen determined an intermediate adherence and spreading for TC, but the highest dynamics in Tp extensions. In conclusion, TC behave differently than fibroblasts in terms of adherence, spreading, and cell prolongation extension when seeded on various matrix proteins in cell culture.

Detection of EWS/FLI-1 fusion in non-Ewing soft tissue tumors.

OBJECTIVES: EWS/FLI-1 fusion mainly appears in Ewing's sarcoma or the primitive neuroectodermal tumors and represents a genomic marker for these tumors. However, it can appear with lower frequency in other soft tissue tumors. The paper investigates the presence of EWS/FLI-1 fusion in clinically diagnosed sarcoma belonging to different non-Ewing connective tissue tumors in order to search for a possible new biomarker valuable for investigators. METHODOLOGY: 20 patients with soft tissue tumors, who underwent surgery, were tested. Intra-operative samples of normal and tumor tissue were collected for histopathological diagnosis and genetics determinations. The patients' RNA from tumor and normal peritumoral tissue was extracted and EWS/FLI-1 fusion screened by quantitative real-time PCR. The relative expression of the fusion in the tumor sample was compared to the similar expression in normal tissue. RESULTS: The amplification in the threshold zone was shown by 5 samples (25%): 2 clear cell sarcoma, 1 fibrosarcoma, 1 malignant tumor of nerve sheath, 1 metastatic adenocarcinoma. We differentiated between the unspecific amplification and concluded that these are weak positive results. CONCLUSIONS: Genomic investigation may establish the tumor malignancy and its possible affiliation earlier than histopathology. It can support the screening of EWS/FLI-1 fusion in a larger variety of clinically diagnosed soft tissue tumors.

CD28 T-cell costimulatory molecule expression in pemphigus vulgaris.

BACKGROUND: CD28 superfamily of immune costimulatory molecules could play an important role in autotolerance control. CD28 costimulation seems to be necessary for regulatory T cell (Treg) activation and successive suppressive activities involved in autoimmunity protection. This study investigates CD28 expression, especially inducible costimulator fraction, on T lymphocytes in pemphigus vulgaris (PV) patients. METHODS: CD28 expression on T lymphocytes was assessed in 16 PV patients during acute attack. All patients and 10 healthy control subjects were tested for lymphocyte populations, T-cell subpopulations (T-CD4+, T-CD8+), Treg and CD28 expression on T-cell subpopulations. RESULTS: T, B and natural killer cells average values in PV patients were close to the control group values. Compared with control group, PV values showed lower Treg (2.2% compared with 4.7%), slightly decreased CD4+ CD28+ T cells (91% compared with 95%), higher CD4+ CD28- T cells (9% compared with 5%), decreased CD8+ CD28+ T cells (57% and 73%, respectively) and significantly enhanced CD8+ CD28- T cells (43% compared with 27%). CONCLUSIONS: These data suggest that Treg-mediated suppressor T-cell effects could be diminished in PV, together with an abnormal or ineffective subsequent helper T-cell suppression. CD28 high expression on helper T cells and low expression on suppressor T cells are arguments for a potential CD28 role in PV autoimmune response mechanism.

[TGF-beta2 involvements in open angle glaucoma]. / Implicatiile TGF-beta2 în glaucomul primitiv cu unghi deschis.

PURPOSE: To emphasize the correlation between TGF beta2 and primary open angle glaucoma (POAG). METHODS: Observational clinical study on two groups of patients: group I--17 patients with POAG who need surgical approach and group II--16 patients without glaucoma. The groups were homogeneous regarding age and sex. The exclusion criteria were any associated systemic pathology. The concentration of TGF beta2 in the aqueous humor of each patient of both groups was measured by ELISA method. RESULTS AND DISCUSSIONS: After the comparative analysis we observed that TGF beta2 was more increased in patients with POAG than in patients without the disease. The changes of trabecular extra cellular matrix are induced by many factors. TGF beta2 is a special cytokine witch is believed to increase the fibronectin concentration in the trabecular meshwork and alter the extra cellular matrix turn-over with a final profibrotic effect.

ICAM-1, ELAM-1, TNF-alpha and IL-6 in serum and blister liquid of pemphigus vulgaris patients.

The levels of ICAM-1, ELAM-1, TNF-alpha and IL-6 were determined in 12 patients with pemphigus vulgaris (PV) both in serum and the blister liquid. As a control, the same parameters were determined in 7 patients with herpes zoster (HZ). The patients with PV presented significantly higher values of ICAM-1 in the blister liquid, as compared to the serum values. The values of TNF-alpha and IL-6 were increased both in serum and the blister liquid. The ELAM-1 values did not show significant differences between serum and the blister liquid. In HZ patients, the blister liquid values did not significantly exceed the serum values both for ICAM-1 and ELAM-1. TNF-alpha and IL-6 presented high values both in serum and the blister liquid. We consider that the high values of ICAM-1 in the blister liquid from PV patients suggest the involvement of this adhesion molecule in the PV pathogenic features. The implication of ICAM-1 could be nonspecific and limited, and could possibly represent a reaction to the destruction of the desmosomal bonds within keratinocytes.

Photodynamic treatment of basal cell carcinoma and squamous cell carcinoma with hypericin.

Hypericin displays antiproliferative and cytotoxic effects on tumor cells. This effect depends on photodynamic activation with visible light and oxygen. Hence, we explored its potential use in treating skin cancer. Eight patients with squamous cell carcinoma (SCC) and eleven patients with basal cell carcinoma (BCC) were treated topically with hypericin. After intralesional injection, the hypericin was irradiated with visible light. Patients with SCC were given 40-100 micrograms hypericin intralesionally, 3-5 times per week for 2-4 weeks; patients with BCC 40-200 micrograms hypericin 3-5 times per week for 2-6 weeks. Hypericin displayed selective tumor-targeting: penetration in the surrounding tissues did not induce necrosis or cell loss and even the generation of a new epithelium at the surface of the malignancy was noticed. The effectiveness of the therapy depends on the concentration and total dose of hypericin, the frequency and duration of the therapy; clinical remissions can be expected after 6-8 weeks.

Tumoral infiltrate after local treatment with interferon in squamous cell carcinoma.

Using monoclonal antibodies UCHL-1 (T lymphocytes), MT-1 (pan T) and L-26 (B lymphocytes) in the study of the tumoral infiltrate after local treatment with alpha interferon (Roferon) in patients with squamous cell carcinoma of the lower lip, it was observed that: the proportion of UCHL-1 positive cells was between 30% and 80%, the proportion of MT-1 positive cells was of 85% and that of the L-26 positive cells was of 30% of all the cells in the infiltrate. In the area in which after treatment with interferon the tumoral structures had disappeared, the proportion of T lymphocytes was smaller than in the areas in which the tumoral structures were still present. The therapeutic effect of interferon is due both to the direct effects on the tumoral cell and also to the indirect effects, namely the activation of the cytotoxic T lymphocytes and of other cells in the tumoral infiltrate.

Interleukin-2 (IL-2) in the treatment of malignant brain tumors (glioblastomas).

The present study is aimed to provide a new possible approach to the treatment of glioblastomas, in an effort to control the extremely severe evolution of these tumors. Thus, from May 28, 1991, we started the administration of IL-2 (produced at the "Victor Babes" Institute, Bucharest) in 9 glioblastoma patients (operated on at the Neurosurgery department VII of the "G. Marinescu" Hospital, Bucharest) by repeated intraoperative and postoperative local injections. The survival period from the first IL-2 administration was 7 months (1 case), 8 months (2 cases), 11 months (2 cases), 14 months (1 case) and 17 months (1 case). There are only 2 survivors: one 4 months and the other 23 months following the first injection of IL-2. The survivor whose condition 23 months following introduction of IL-2 therapy is very good, demonstrates that IL-2 offers some chance to these hopeless patients.

[The action of the supernatants from human lymphocyte cultures on E-active rosette formation]. / Actiunea supernatantelor din culturi de limfocite umane asupra rozetarii E active.

Lymphocytes separated from 20 normal subjects were introduced in two rosetation systems: the first was modulated rosetation (REM) with phytohemaglutinina (PHA), concanavalina A (Con. A). Mycobacterium tuberculosis (Mbt), and tumoral extract from ocular malignant melanoma (ET-MMO): the second was E-active modulated rosetation with supernatants from human lymphocyte cultures stimulated with PHA, Con A, Mbf and ET-MMO (REM-S), he average values of percentages of rosettes forming cells in the two systems REM and REM-S are comparable. We may assume the existence of some modulating factors which operate in the same manner on the functionality of T-lymphocyte in the two systems (REM and REM-S).

[Functional E rosette study pre and post laser treatment in malignant ocular melanoma]. / Explorare prin rozetare E "functionala" pre si post tratament laser în melanomul malign ocular.

For 11 patients with ocular malignant melanoma (OMM) suspicion, before and after laser photocoagulation therapy, by E rosette test were studied. A higher activation and sensitisation to OMM antigen was obtained after laser therapy in 9 patients. These results suggest the influence of laser treatment on lymphocyte function by "revealing" the physiological reactivity. Our results point out the "functional" E rosette test is useful for OMM diagnosis and therapy, as an indicator for disease prognosis.