Eva Dionyssopoulou

Serum concentrations of growth factors in women with and without endometriosis: the action of anti-endometriosis medicines

Endometriosis is a common gynecologic syndrome of unknown etiology and pathogenesis. Growth factors and inflammatory mediators produced by peritoneal leukocytes have recently been postulated to participate in the pathogenesis of endometriosis. Angiogenic factors released from peritoneal macrophages may also play a role in the development of this disease. In the present study, we investigate the soluble levels of vascular endothelial growth factor (VEGF), epidermal growth factor-receptor (EGF-R), granulocyte/macrophage-colony stimulating factor (GM-CSF), Insulin-like growth factor-1 (IGF-1) and interferon-gamma (IFN-gamma) in the serum of 28 women with and 20 without endometriosis. We also compared these levels before, during and after treatment with danazol and leuprorelin acetate depot, the two therapeutic regiments of choice concerning this disease. We found that only sVEGF levels were higher in women with endometriosis in comparison to controls (P < 0.001) while sEGF-R is not present. GM-CSF, IGF-1 and IFN-gamma soluble levels are not affected in either healthy or endometriotic subjects. The 6-month treatment with danazol decreased sVEGF levels (P < 0.02) and increased sEGF-R levels (P < 0.001). These observations support the view that VEGF may be associated with the disease process and that danazol may bring sVEGF levels to a normal threshold. However, future studies will be focused on the anti-angiogenic control of the action of VEGF in patients with endometriosis.

Carnitine deficiency in children and adolescents with type 1 diabetes

Carnitine is essential for the lipid and carbohydrate metabolism, and proper metabolic control in type 1 diabetes has potential impact on long-term complications. The plasma total, free, and acylcarnitine levels in 47 children and adolescents with type 1 diabetes were determined by a radioisotopic assay and compared to the values of a series of anthropometric measurements and metabolic parameters, including blood glycosylated hemoglobin Alc, serum cholesterol and triglycerides, and urine microalbumin levels. Plasma values for total, free, and acylcarnitine were 30.1+/-7.26, 20.0+/-4.50, and 10.2+/-6.47 micromol/l, respectively. Acyl/free carnitine ratio was 0.544+/-0.369. Individuals with type 1 diabetes had significantly lower total and free carnitine levels and significantly higher acyl/free carnitine ratios than controls (P<.001). Plasma total and free carnitine levels were inversely correlated to the duration of diabetes (P=.036 and P=.071, respectively). No statistical relationship was documented between carnitine levels and the remaining anthropometric and metabolic variables. In conclusion, total and free carnitine levels are decreased in children and adolescents with type 1 diabetes. This reduction is time related and may have potential interactions with the long-term complications of type 1 diabetes. Larger studies are required for final conclusions to be drawn on the precise role of carnitine and the possible benefit, if any, of carnitine supplementation in diabetic patients.

Expression of serum human leukocyte antigen and growth factor levels in a Greek family with familial endometriosis.

Objective: An increased incidence of endometriosis in the first-degree relatives of patients with endometriosis has been reported, suggesting a familial predisposition and possible genetic influence. In this study, we present a family with four members who have histologically proven endometriosis (mother and three daughters) in two generations and one member with suspected endometriosis in the third generation. The aim of this study was to investigate the presence of serum-soluble class I and class II human leukocyte antigen (sHLA) levels, because they have been shown to be reduced in women with endometriosis. We also studied the levels of vascular endothelial growth factor (VEGF) and epidermal growth factor-receptor (EGF-Rc) whose function in angiogenesis implies an active role in endometriosis. Methods: Apart from the family members under study, the control groups consisted of 38 women with endometriosis and 30 without any pelvic disease. All the soluble factors under investigation were measured by an enzyme-linked immunosorbent assay technique using a specific immunoassay. Results: All the affected family members and the 38 women with endometriosis had very low levels of serum-soluble class I and class II HLA levels compared with healthy subjects. The circulating levels of VEGF were higher in the endometriosis group than the healthy control group, a pattern in accordance with the family members. On the contrary, EGF-Rc was negative in controls and women with endometriosis, with the exception of certain family members in specific stage of endometriosis. Conclusion: We studied the association of endometriosis with circulating levels of human leukocyte antigens and VEGF in two generations of a single family (mother and three daughters). These markers were expressed distinctly in women with familial endometriosis.

Early events of the exogenously provided L-Carnitine in murine macrophages, T- and B-lymphocytes: modulation of prostaglandin E1 and E2 production in response to arachidonic acid.

L-carnitine is an essential energy-providing compound to the cell since it transports long chain fatty acids through the mitochondrial membrane and delivers them to the beta-oxidation pathway for catabolism and/or entrance to biosynthetic pathways. Some of the early events taking place in immune cells after L-carnitine inoculation in vitro are defined in this report. Using arachidonic acid as a fatty acid source, we determined the utilization rate of L-carnitine by murine T-, B-lymphocytes and macrophages within two hours of cell culture, its effect on prostaglandin E1 and E2 production and the levels of beta-hydroxy-butyrate. The results show that although all immune cells consume a small portion of L-carnitine, beta-hydroxy-butyrate decreases upon addition of arachidonic acid and/or L-carnitine indicating that active biosynthetic pathways are induced. L-carnitine is shown to increase the arachidonic acid-induced production of prostaglandins E1 and E2 in macrophages, while their secretion from T- and B-lymphocytes is decreased. These findings indicate the L-carnitine may very rapidly alter the activation state of immune cells and lead to the development of various reactions, beneficial or not to the organism.

Characterization of intracellular HLA-DR, DM and DO profile in K562 and HL-60 leukemic cells.

Surface class-II antigen expression fires-up the antigen presentation process and development of immune response. The absence of surface HLA-DR is used in various systems to avoid immune recognition. Most leukemic cells use such mechanism to escape immune surveillance. Here, K562 and HL-60 leukemic cells were examined as to intracellular HLA-DR, -DM and -DO expression, if any. Immunofluorescence scored by UV-microscopy, flow cytometry or confocal microscope analysis detected intracellular pools of HLA-DR, -DO and to a lesser degree HLA-DM, whereas sub-cellular fractionation localized these molecules within endosomes. RT-PCR experiments revealed the presence of HLA-DRalphabeta, HLA-DMalphabeta and HLA-DObeta but not HLA-DOalpha transcripts. Despite the absence of the HLA-DOalpha chain, stable transfectants of K562 with a full length HLA-DObeta-EGFP construct showed that DObeta chain could be translocated to endosomes and form stable complexes with HLA-DR. Such complexes could be responsible for arresting HLA-DR molecules within endosomes, maintaining their surface class-II negative state.

Role of prostaglandin E2 in the experimental L-carnitine-induced endometriosis-like inflammatory model

Purpose Ectopic implantation of endometrium leading to endometriosis has been correlated with various immune and non-immune parameters. Inflammatory cytokines including IL-2, IFN-γ, TNF-α, increase in serum and peritoneal fluid of endometriotic patients, while elevated levels of prostaglandins PGE1 and PGE2 have been accused of facilitating the inflammation process. Using the experimental model of L-carnitine (L-Cn)-induced endometriosis, the aim of this study was to explore the role of PGE1 and PGE2 in endometriosis development and define whether L-Cn had a direct effect on endometrial cells. Methods During treatment with L-Cn, experimental mice were injected with indomethacin or specific PGE1 and PGE2 analogs and their effect on the L-Cn-induced inflammatory state was examined. Primary uterine cells or human endometrial adenocarcinoma cell lines were used to assess the direct or indirect effect of L-Cn. Results The seven-day treatment of virgin females with L-Cn induced obvious endometrial abnormalities with accumulation of CD90 and CD11 cells. Administration of indomethacin alleviated some of the L-Cn-induced effects, but could not inverse the phenotype. Interestingly, 11-deoxy-PGE1 aggravated the L-Cn-induced effects, whereas 17-phenyl-trimor-PGE2 reversed them. In vitro experiments showed that the effect of L-Cn depended on the nature of endometrioma cells. L-Cn could aggravate the inflammatory state of Ishikawa-inoculated mice, but could alleviate symptoms from MFE-319 inoculated females. Conculsions Although PGE2 appeared to play a key role in the L-Cn-induced uterus inflammation, the actual role of L-Cn depends on the nature of endometrioma cells and can aggravate or protect from inflammation according to the induced regulatory pathways.