Increased number of circulating endothelial cells (CECs) in patients with psoriasis--preliminary report.

BACKGROUND: Numerous studies have demonstrated increased cardiovascular risk in psoriasis. Circulating endothelial cells (CECs) have been proposed as a new marker of endothelial dysfunction that plays an important role in pathogenesis of atherosclerosis. OBJECTIVE: The aim of this study was to compare the number of CECs in psoriatic patients to a control group and to analyze possible correlations between the numbers of CECs and the plasma levels of classical markers of endothelial dysfunction, such as: sICAM-1, sE-selectin and von Willebrand factor (vWF). METHODS: The number of CECs, identified as CD146 + / CD45- cells, were determined in peripheral blood with using flow cytometry in psoriatic patients (n = 63) and controls (n = 31). The plasma levels of: sICAM-1, sE-selectin, vWF were measured with ELISA. The severity of psoriasis was assessed with PASI. RESULTS: The number of CECs was significantly increased in psoriatic patients compared with controls (P < 0.00001) and positively correlated with disease severity (R = 0.360; P = 0.0037). The levels of sICAM-1, sE-selectin and vWF were significantly elevated in psoriatic patients (P < 0.00001; P < 0.00001; P = 0.00072, respectively). The number of CECs was significantly, positively correlated with the levels of sICAM-1 (R = 0.393; P = 0.0014) and vWF (R = 0.314; P = 0.012) in psoriatic patients. The levels of sICAM-1 and sE-selectin were positively correlated with disease severity (R = 0.356; P = 0.0041 and R = 0.407; P = 0.0009, respectively). CONCLUSION: The increased number of CECs that correlates with disease severity and plasma levels of sICAM-1 and vWF may indicate endothelial dysfunction or injury in patients with psoriasis.

Fas expression in primary breast cancer is related to neoplastic infiltration of perilymphatic fat.

PURPOSE: Various studies have revealed that both Fas and its ligand play an important role in cancer biology. The aim of our study was to determine if there is a relationship between the expression of Fas or Fas-ligand in breast cancer and the presence of malignant cells in perilymphatic fat. MATERIAL/METHODS: Tumor samples from 147 consecutive breast cancer patients, aged 35-81 (median, 59), were subjected to analysis. The expressions of Fas and Fas-ligand were determined immunohistochemically. RESULTS: The expression of Fas, but not Fas-ligand, was significantly less frequent in breast cancer patients in whom malignant cells infiltrated through the perilymphatic fat (p=0.042). The infiltration of paranodal fatty tissue occurred more often in cases of ductal carcinomas (p=0.008), larger primary tumors (pT>or=2, p=0.030) and regional lymph node involvement (pN>or=1, p=0.021). Univariate analysis revealed that perilymphatic fat infiltration shortened overall survivals in breast cancer patients (p=0.05), similarly to postmenopausal status (p=0.034), age >60 years (p=0.05) and regional lymph node involvement (p=0.05). None of the aforementioned factors, however, was revealed as an independent predictor of survival in multivariate analysis. CONCLUSIONS: The study showed that lack of Fas in primary breast cancer is associated with perilymphatic fat infiltration. Consequently, both the absence of Fas in the primary tumor and the occurrence of neoplatic cells in paranodal fatty tissue should be considered in the prognosis, complementing existing conventional factors.

Calcium inhibits muscle FBPase and affects its intracellular localization in cardiomyocytes.

As our recent investigation revealed, in mammalian heart muscle, fructose 1,6-bisphosphatase (FBPase)--a key enzyme of glyconeogenesis--is located around the Z-line, inside cells' nuclei and, as we demonstrate here for the first time, it associates with intercalated discs. Since the degree of association of numerous enzymes with subcellular structures depends on the metabolic state of the cell, we studied the effect of elevated Ca2+ concentration on localization of FBPase in cardiomyocytes. In such conditions, FBPase dissociated from the Z-line, but no visible effect on FBPase associated with intercalated discs or on the nuclear localization of the enzyme was observed. Additionally, Ca2+ appeared to be a strong inhibitor of muscle FBPase.

Loss of tumorigenicity of murine colon carcinoma MC38/0 cell line after transduction with a retroviral vector carrying murine IL-12 genes.

Cells of transplantable MC38 colon carcinoma of C57BL/6 mice were adapted to growth in vitro as the MC38/0 cell line. Along the establishing process, MC38/0 cells preserved their tumorigenicity. After transduction with a retroviral vector carrying murine interleukin 12 (mIL-12) genes and further selection, stable MC38/IL-12 transductant cells were obtained. These cells produced IL-12 (approx. 2500 ng/ml/5x10(5) cells/48 h) as evaluated in the optimized bioassay. After subcutaneous inoculation into syngeneic mice, the IL-12-modified cells demonstrated reduced tumorigenicity as compared to parental MC38/0 cells. Mice that rejected the MC38/IL-12 tumour became protected against subsequent challenge with MC38/0 cells. The obtained data indicate that the IL-12-transduced murine colon carcinoma cells could be used both as a model tumour for the study of mechanisms of anticancer immunity and/or as an adjuvant to cancer vaccines.

Bcl-Xl protein expression in laryngeal squamous cell carcinoma.

The Bcl-2 family of proteins regulate one of the steps in an evolutionary conserved apoptotic pathway. The long splice variant of Bcl-X (Bcl-Xl) is a potent antagonist of apoptosis. The aim of the study was to evaluate the relation between the presence of immunohistochemically detectable Bcl-Xl protein in laryngeal squamous cell carcinomas (LSCCs) and clinicopathological data, as well as DNA ploidy status and proliferative activity. In 50 specimens of LSCC, Bcl-Xl protein expression was evaluated immunohistochemically. Proliferative activity (SG2M-phase index) and DNA ploidy were measured by flow cytometry. In our study, Bcl-Xl protein expression decreased with decreasing tumour differentiation (P = 0.04). The majority of patients with Bcl-Xl protein immunoreactivity had no metastatic lymph node involvement (P = 0.01). Other factors such as age, gender, primary tumour size (pT) and type of cancer (keratinizing/non-keratinizing) were not associated with Bcl-Xl protein level. There was no correlation between Bcl-Xl protein and SG2M-phase index or DNA ploidy status. Our findings show that expression of Bcl-Xl protein is increased in a great fraction of laryngeal cancers. Further studies, however, are needed to clarify association between Bcl-Xl protein expression and clinical course of patients.

Trophoblasts and soluble adhesion molecules in peripheral blood of women with pregnancy-induced hypertension.

PROBLEM: The current hypothesis on the pathogenesis of pregnancy-induced hypertension (PIH) considers it as an endothelial disorder that is first local but with the potential of becoming general. The aim of the work was to investigate the relation of the number of trophoblast cells in maternal peripheral blood against the serum levels of soluble vascular and intercellular cell adhesion molecule (sVCAM-1 and sICAM-1) in PIH. METHOD OF STUDY: Women with PIH were at 28th to 40th week of gestation. Control group were normotensive (NT) pregnant women at 28th to 41st week of gestation. Flow cytometry was used to assess the relative number of the trophoblasts in the peripheral blood. Trophoblasts were labeled with monoclonal anti-human trophoblast protein antibody MCA 277. The presence of sVCAM-1 and sICAM-1 was determined using the enzyme-linked immunosorbent assay method. RESULTS: Women with PIH had significantly higher trophoblasts number than NT women (median 19.0, range 5.0-57.0/400 microL versus median 7.0, range 0.0-18.0/400 microL; P = 0.000011) as well as plasma level of sVCAM-1 when compared with NT women (median 730.0, range 325.0-1525.0 ng/mL versus median 493, range 310-1075 ng/mL; P = 0.02). ICAM-1 level in the PIH group was slightly elevated (median 280.0, range 174.0-524.0 ng/mL) when compared with NT women (median 260.0, range 190.0-464.0 ng/mL, P = 0.322). Eight of 21 women with PIH had proteinuria but no correlation was found between this symptom and the laboratory findings. CONCLUSION: The increased number of trophoblast cells in maternal peripheral blood and higher levels of sVCAM-1 correlate with the presence of PIH. The differences of sVCAM levels were significantly higher than those observed for sICAM. The results indicate an association between circulating trophoblasts and vascular endothelium activation, during PIH.

Colocalization of muscle FBPase and muscle aldolase on both sides of the Z-line.

Previously we have reported that in vitro muscle aldolase binds to muscle FBPase [Biochem. Biophys. Res. Commun. 275 (2000) 611-616] which results in the changes of regulatory properties of the latter enzyme. In the present paper, the evidence that aldolase binds to FBPase in living cell is presented. The colocalization experiment, in which aldolase was diffused into skinned fibres that had been pre-incubated with FBPase, has shown that aldolase in the presence of FBPase binds predominantly to the Z-line. The existence of a triple aldolase-FBPase-alpha-actinin complex was confirmed through a real-time interaction analysis using the BIAcore biosensor. The colocalization of FBPase and aldolase on alpha-actinin of the Z-line indicates the existence of glyconeogenic metabolon in vertebrates' myocytes.

New human microvascular endothelial cell lines with specific adhesion molecules phenotypes.

Vascular endothelial cells recognize blood-borne circulating cells and allow them to extravasate in a tissue-specific manner. Because this property determines the selectivity of lymphocyte homing, it is fundamental in physiological as well as pathological processes (inflammation, autoimmune diseases, metastasis). As a tool to assess the molecular basis of endothelium selectivity, microvascular endothelial cell lines of distinct tissue origin were established. Endothelial cells, isolated from lymphoid tissues (lymph nodes and appendix) and from nonlymphoid immune sites--intestine, lung, and skin--were immortalized in vitro. Their general endothelial characteristics, such as the presence of von Willebrand factor (wWf), angiotensin-converting enzyme (ACE), VE-cadherin, and the intracellular E-selectin, were preserved. This article shows that these cell lines display phenotypic characteristics related to their tissue origin. Hence, endothelial cells from lymph nodes expressed peripheral lymph node addressins (PNAds). Endothelial cells from nonlymphoid tissues were ICAM-1 (intercellular adhesion molecule-1) and CD49e positive, whereas P-selectin was not equally distributed among the cell lines. Endothelial cells from mucosal sites reacted with antibody against human MAdCAM-1 (mucosal addressin cell adhesion molecule). In the adhesion test, lymphoid and myeloid cells adhere to endothelial cell lines in a distinct manner. These lines could be useful to study molecular mechanisms involved in tissue-specific cell-cell interaction.

KDR promoter can transcriptionally target cytosine deaminase suicide gene to cancer cells of nonendothelial origin.

The KDR/flk-1 gene promoter is considered to be endothelial cell-specific. We show its activity in two cancer cell lines of non-endothelial origin: in murine L1 sarcoma and OVP-10 human ovarian carcinoma cell lines. KDR promoter-driven cytosine deaminase gene can be efficiently expressed in these cells leading to sensitization to 5-fluorocytosine, as demonstrated both in vitro and in vivo. Our results indicated that KDR promoter activity is not endothelial cell-exclusive and that this promoter can also be used to obtain specific expression of therapeutic genes in certain cancer cells.

PIH is associated with an increase of trophoblasts circulating in maternal blood.

One of the reason of PIH problems may be due to exposition to placental trophoblast. The objective of the work was to evaluate the number of trophoblast cells deported into maternal peripheral blood of patients with PIH (pregnancy induced hypertension) as compared to normal pregnancy. Trophoblasts have been detected, by cytofluorimetry, in peripheral maternal venous blood of hypertensive woman (15 cases) and normotensive pregnancy (16 cases). Women with PIH had statistically significant (p < 0.005) higher trophoblasts number than that found in normotensive pregnant women without PIH (16 cases). Our results indicate that the increased trophoblasts deportation into peripheral blood could be a marker of the maternal syndrome of PIH.

Increased maternal plasma levels of soluble vascular cell adhesion molecule-1 (VCAM-1) in pregnancy induced hypertension (PIH).

One of the reason of PIH problems may be due to the presence of increased circulating levels of cell adhesion molecules, markers of endothelial damage and leukocyte activation. The objective was to evaluate the plasma levels of soluble vascular cell adhesion molecule in maternal peripheral blood of patients with PIH (pregnancy induced hypertension) and compared to those of normal healthy women with uncomplicated pregnancy. Maternal plasma samples were prepared from peripheral venous blood collected from 10 patients with PIH and 10 matched normotensive patients with uncomplicated pregnancies. Samples were assayed for soluble VCAM-1 by specific enzyme-linked immunosorbent assay (ELISA). Women with PIH had significantly higher plasma level of soluble VCAM-1 as compared with healthy pregnant women without PIH (653.50 vs. 456.39 ng/mL, respectively, p < 0.005). Our results on the increased plasma levels of soluble VCAM-1 in patients with PIH provide evidence for endothelial activation of PIH. It suggest that increased plasma level of soluble VCAM-1 could be an early marker of the maternal syndrome of PIH.

Colorectal cancer-associated nuclear antigen.

By using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting assays in the presence of polyclonal antiserum raised against electrophoretically specific polypeptides of colorectal cancer nuclear polypeptides with M(r) of 35-40 kDa, we have identified p36 protein whose expression accompanies tumorigenesis of large intestine. Immunological analysis of 35 nuclear protein preparations has indicated expression of p36 antigen in nine of 11 right-sided (81.8%) and 21 of 24 (87.5%) left-sided colorectal tumor cases, but not in any control tissue samples. In this study, we have identified p36 antigen in two colon tumor cell lines, i.e., SW620 and HT29 as well. Fractionation experiments based on selective extraction of nuclei isolated from cancerous specimens, which enables their separation into chromatin, nuclear matrix and its subfraction, i.e., internal and peripheral matrix have revealed the concentration of this particular antigen in the internal matrix.

[Animal lectins--structure and function]. / Lektyny zwierzece--struktura i funkcja.

Lectins are structurally and functionally diverse group of proteins or protein domains capable of specific binding of oligosaccharide structures present on cell surfaces, the extracellular matrix, and secreted glycoproteins. Animals lectins are classified into six groups: C-type lectins, S-type lectins (galectins), I-type lectins, P-type lectins, pentraxins, and others. In this review, the basic knowledge regarding structure and function of animal lectins is presented.

Human in vitro cell lines verification by minisatellite DNA restriction fragment length polymorphism.

Four families of human in vitro cell lines were tested for minisatellite restriction fragment length polymorphism (RFLP) using multilocus probes MZ1.3 and/or 33.15 after digestion of DNA with restriction enzymes HinfI or HaeIII. These results confirmed that (i) the RFLP pattern is relatively stable in established cell lines and, therefore, could be used as a specific marker of a cell line identity, (ii) the use of MZ1.3 and 33.15 probes permits the identification of hybridomas and (iii) one of the cell lines tested, a lymphoblastoid cell line HAJ, may possess a hot spot of mutation.

Synthesis and cytostatic properties of 5-substituted derivatives of 3-methylisoxazolo[5,4-d]1,2,3-triazine-4-ones and 3-methyl-5-triazene 4-isoxazolecarboxylic acid ethyl esters.

Six new 5-substituted derivatives of 3-methyl-isoxazolo[5,4-d]1,2,3-triazine-4-one and 3-methyl-5-triazene-4-isoxazolecarboxylic acid ethyl ester have been synthesized from 5-amino-3-methylisoxazole-4-carboxylic acid amides, and ethyl ester. They were examined for cytostatic activity in comparison with Dacarbazine. The 3-methyl-5-(4-chlorophenyl)isoxazolo[5,4-d]1,2,3-triazine-4-one showed better activity than Dacarbazine.

[Hemocarboperfusion in the treatment of patients with an opiate abstinence syndrome]. / Gemokarboperfuziia v lechenii bol'nykh s opiinym abstinentnym sindromom.

The authors analyze of the results of applying hemocarboperfusion in treatment of opiate addiction. All the patients were treated in the Belorussian center of sorption methods of detoxication and plasmapheresis. It is shown that 4-5 procedures of hemocarboperfusion in combination with infusion and drug therapy allow to correct of withdrawal mental and somatic disorders.

Effect of lactoferrin on proliferation and differentiation of the Jurkat human lymphoblastic T cell line.

The effect of human lactoferrin on the human lymphoblastic T cell line (Jurkat) was tested with regard to proliferation and differentiation. Lactoferrin enhanced cell proliferation in a serum-reduced (1% fetal calf serum) culture. The stimulatory effect of lactoferrin on proliferation depended on the degree of iron saturation but the amplitude of the effect was low, similar to that obtained in the presence of serum transferrin. The proliferation stimulatory effect of lactoferrin was not observed in the presence of 10% fetal calf serum (FCS) in the culture medium. These results suggest that Fe-lactoferrin can substitute for Fe-transferrin during the prolonged culture of cells in a low serum concentration. Iron-saturated lactoferrin was also shown to promote T cell differentiation. Jurkat cells, when exposed to iron-saturated lactoferrin in the presence of 10% FCS, gradually exhibited a decrease in the cell volume, cell surface density of CD71 antigen, the nuclear incorporation of [methyl-3H]thymidine, but an increase of the percentage of cell population in the G0/1 phase of the cell cycle. These modifications were accompanied by the appearance of CD4 antigen at the cell surface. Therefore, in the continuous presence of lactoferrin, proliferating cells slowly enter into quiescence state, undergoing cell differentiation.

Conjugation of the monoclonal antibody 17-1A with the nitroacridine compound C921 with the poly-L-lysine as an intermediate agent.

Monoclonal antibody 17-1A specific for human gastric carcinoma was coupled directly or indirectly, using poly-L-lysine as an intermediate, with nitroacridine compound C921. The aim of the study was to obtain selectively cytotoxic immunoconjugates for experimental therapy. Directly coupled conjugates retained antibody specificity towards cells of several human adenocarcinoma lines but were non cytotoxic in vitro, whereas conjugates obtained with the use of intermediate poly-L-lysine expressed only low, non-specific cytotoxicity, probably exerted by the poly-L-lysine content.

Adhesion of human uroepithelial cells to E-selectin: possible involvement of sialosyl LewisA-ganglioside.

In a previous study we showed that tumorigenic and invasive human uroepithelial cell lines are characterized by the presence of sialosyl Le(a) (sLe(a)) ganglioside. Our data suggested that expression of this glycolipid correlated with acquisition of the malignant phenotype by human urothelial cells. To evaluate the postulated adhesion function of sLe(a) antigen, we studied the adherence of 6 human urothelial cell lines with different expressions of this carbohydrate structure to E-selectin-expressing CHO cells. The only cell line that bound specifically to E-selectin was Hu 1703He, which expressed the highest level of sLe(a) antigen. The involvement of carbohydrate-E-selectin interaction in the adhesion of Hu 1703He cells was indicated by the following facts: (i) anti-E-selectin monoclonal antibody (MAb) completely abolished binding to E-selectin-expressing CHO cells; (ii) removal of sialic acid from Hu 1703He cells highly decreased the adhesion. Adhesion correlated with the presence of several sLe(a)-carrying glycoproteins, which was shown by immunoblotting of Hu 1703He cell lysate with anti-sLe(a) MAb 19-9. The binding of antibody was abolished when cell lysate was treated with O-sialoglycoprotein endopeptidase, suggesting that sLe(a) is present on O-linked oligosaccharides. However, incubation of Hu 1703He cells with O-sialoglycoprotease had no effect on adhesion to E-selectin or on binding of 19-9 MAb to the cell surface. Our data suggest that (i) protein-bound sLe(a) oligosaccharides represent only a minor portion of whole sLe(a) antigen produced by uroepithelial cells; (ii) effective binding to E-selectin occurs when sLe(a) oligosaccharide present on cell-surface glycosphingolipids is expressed in high density since the cell lines with moderate expression of sLe(a) ganglioside did not bind to E-selectin-transfected CHO cells.