Sebastian Radej

Effect of polyamidoamine dendrimer G3 and G4 on skin permeation of 8-methoxypsoralene--in vivo study.

In the present study we have assessed the ability of (PAMAM) dendrimers G3 and G4 to facilitate transdermal delivery of 8-methoxypsoralen (8-MOP) in vivo. In vitro study using Franz diffusion cell revealed an enhanced transdermal flux for 8-MOP in complex with G3 and G4 dendrimer in relation to standard 8-MOP solution. In present study in vivo skin permeation potential of 8-MOP complex with G3 and G4 PAMAM dendrimer was assessed using confocal laser scanning microscopy (CLSM), which revealed an enhanced permeation of the 8-MOP to the deeper layers of the skin and significantly higher concentration in comparison with standard 8-MOP solution. Skin tissue 8-MOP concentration, evaluated by HPLC indicates that G3 and G4 PAMAM application significantly increase 8-MOP skin deposition in comparison with standard 8-MOP solutions after 1 and 2h. G4 appeared to be a more effective 8-MOP penetration enhancer than G3 PAMAM. Our results suggest the feasibility of G3 and G4 PAMAM dendrimers for transdermal delivery of 8-MOP resulting in better skin permeation and higher concentration of 8-MOP in epidermis and dermis of the drug that could help to improve effectiveness and safety of PUVA therapy.

Transdermal delivery of 8-methoxypsoralene mediated by polyamidoamine dendrimer G2.5 and G3.5--in vitro and in vivo study.

In this work, we have focused on 8-methoxypsoralene (8-MOP) complexed with G2.5 and G3.5 poly(amido amine) (PAMAM) dendrimers. The purpose of this study was to investigate the efficacy of half-generation G2.5 and G3.5 PAMAM dendrimers conjugated with 8-MOP for delivery of 8-MOP in vitro study through polivinyldifluoride membrane (PVDE) and prepared pig ear skin (PES) using Franz diffusion and in vivo study through the skin of experimental animals (hairless rat skin). The tissue concentration of 8-MOP in hairless rat skin was analyzed by high performance liquid chromatography (HPLC) after 1 and 2 h. Detailed distribution of 8-MOP in skin layers and cellular structures were analyzed using laser scanning microscopy (CLSM). In vitro and in vivo studies showed that half-generation G2.5 and G3.5 PAMAM dendrimers are able to facilitate transdermal delivery of 8-MOP. G2.5 PAMAM dendrimer appeared to be more effective 8-MOP penetration enhancer than G3.5 PAMAM dendrimer, but in vivo the differences are not statistically significant. The concept of using G2.5 and G3.5 PAMAM dendrimers as carriers seems to be a promising method for the delivery of 8-MOP for PUVA (psoralen-UV-A) therapy.

Biochemical markers of psoriasis as a metabolic disease

Psoriasis is a chronic immune mediated inflammatory skin disease with a population prevalence of 2-3%. In recent years, psoriasis has been recognized as a systemic disease associated with metabolic syndrome or its components such as: obesity, insulin resistance, hypertension and atherogenic dyslipidemia. Many bioactive substances have appeared to be related to metabolic syndrome. Based on current literature, we here discuss the possible role of adiponectin, leptin, ghrelin, resistin, inflammatory cytokines, plasminogen activator inhibitor 1, uric acid, C-reactive protein and lipid abnormalities in psoriasis and in metabolic syndrome.

Highly increased maspin expression corresponds with up-regulation of miR-21 in endometrial cancer: a preliminary report.

Background: Maspin and programmed cell death 4 (Pdcd4) are tumor suppressor genes, and miR-21 is overexpressed in many solid tumors and was proven to negatively regulate a number of tumor suppressor genes including maspin and Pdcd4. The purpose of this study was to investigate the expression of maspin, Pdcd4, and miR-21 and their interrelations with clinicopathologic features in endometrial cancer using a quantitative approach. Methods: Maspin, Pdcd4, and miR-21 expressions were evaluated by a real-time polymerase chain reaction in 20 endometrial cancer and 10 normal endometrium samples. Results: Maspin showed a significantly increased expression in endometrial cancer samples compared with the control group and was up-regulated by a mean factor of 46.54 (SE range, 2.367-1160.26; 95% confidence interval, 0.515-15001, P < 0.0001). Expression of miR-21 was found significantly up-regulated in the sample group in comparison to control group by a mean factor of 2.312 (SE range, 0.741-7.778; 95% confidence interval 0.191-15.0, P = 0.028). No significant differences were present in the expression level of Pdcd4 between endometrial cancer and control groups. Comparison between IA and more advanced International Federation of Gynecology and Obstetrics stages of endometrial cancer in regard to expression levels of maspin, Pdcd4, and miR-21 did not reveal any significant differences. Similarly, no differences were encountered when histopathologic grading, myometrial invasion, age, body mass index, and parity were taken into consideration. Conclusions: Association between increased maspin expression and up-regulation of miR-21 in endometrial cancer suggests distinct and tissue-specific relationships of the 2 molecules in this type of malignancy and requires further studies that would reveal its clinical relevance.

Immunomodulatory Effects of IFN-β and Lovastatin on Immunophenotype of Monocyte-Derived Dendritic Cells in Multiple Sclerosis

Multiple sclerosis (MS) is an autoimmune disease of the central nervous system and current MS treatment is only partially effective. Recent data suggest that statins may be potent immunomodulatory agents. In order to evaluate their role in MS, we analyzed the in vitro effects of interferon (IFN)-β and lovastatin on the differentiation and maturation of monocyte-derived dendritic cells (DCs) of MS patients. Twenty-seven patients with relapsing–remitting MS were recruited for the study. DC differentiation and maturation were evaluated based on surface phenotypic changes and the expressions of CD14, CD83, CD1a, CD80, CD86, CD206, and C209 were analyzed by flow cytometry. The results showed that IFN-β and lovastatin affect DC phenotype. Both agents decrease the expression of CD1a, which indicates a weakened presentation of glycolipid antigens. IFN-β causes up-regulated and lovastatin down-regulated expression of CD86, which results in a biased Th-cell responses in MS. Furthermore, high doses of lovastatin cause a decrease in CD209 expression on the surface of DCs and can limit their migration to various tissues. One of the mechanisms of the beneficial action of IFN-β and statins may be associated with their influence on DCs.

Activated and inactivated PPARs-gamma modulate experimentally induced colitis in rats

Summary Background This study sought to define the mechanism by which PPAR-γ ligands affect the course of experimentally induced colitis in rats. Material/Methods Inflammation was induced in Wistar rats by a single rectal administration of 2,4,6,-trinitrobenzene sulfonic acid (TNBS). The antagonist of PPARγ antagonist, bisphenol A diglycidyl ether (BADGE), was administrated intraperitoneally 120 mg/kg 4 times every other day. Rosiglitazone 8 mg/kg was administrated by gastric tube 4 times. Body weight was measured daily. After killing, the large intestinal tissue was weighed and collected for histopathologic and immunoenzymatic tests. Levels of IL-6, IL-10, and myeloperoxidase (MPO) were determined in serum and in intestinal homogenates. Results Rats receiving rosiglitazone had higher body weight, whereas large intestine weight/length ratio was lower; histology showed fewer inflammatory markers. Rats receiving TNBS and TNBS along with BADGE had more intensive inflammatory changes. Rosiglitazone alone decreased expression of IL-6; used with TNBS it decreased expression of MPO in intestinal tissue, yet did not increase the expression of IL-10. Decreased levels of MPO indicate reduced neutrophil-dependent immune response. The antagonist of PPAR-γ increased IL-6 in serum and decreased IL-10 in intestinal homogenates. Bisphenol A diglycidyl ether administrated to healthy animals increases serum IL-6 levels. Conclusions Rosiglitazone inhibits experimental inflammation; administration of its selective antagonist abolishes this protective influence. Rosiglitazone inhibits expression of proinflammatory IL-6 and does not affect IL-10. Agonists of PPARs-γ are possibilities for inflammatory bowel disease prevention. Exogenous substances blocking PPARs-γ may contribute to development or relapse of nonspecific inflammatory bowel diseases.

Evaluation of immature monocyte-derived dendritic cells generated from patients with colorectal cancer

UNLABELLED Dendritic cells are heterogeneous population of the leukocytes and most potent APC in activation of naive T lymphocytes. Therefore the DCs generated in vitro are under research for their application in anti-tumor immunotherapy. The aim of the study was generation of the immature dendritic cells from peripheral blood monocytes collected from colorectal cancer patients and comparison of their ability to endocytosis, cytokine production and immunophenotype to DCs generated from healthy donors. MATERIAL AND METHODS 16 adenocarcinoma stage II patients were included in the study. Dendritic cells were generated in the presence of rhGM-CSF and IL-4. PBMC were isolated from the blood of patients and 16 healthy donors - control group. Immunophenotype, ability of endocytosis of Dextran- FITC as well as intracellular IL-12 expression of the generated dendritic cells was measured using flow cytometry. The cytokines (IL-6, IL-10, IL-12p70, IFN-γ) concentration in the supernatants of DCs culture was measured by ELISA. RESULTS The percentage of the immature dendritic cells and expression of CD206 and CD209 antigens was significantly higher in patients group (p <0.05 and p <0.001 respectively). Significantly (p <0.001) higher expression of the antigens which initiate the Th2 immune response (CD80-/CD86 + and B7-H2 + / CD209 +) was in the patients group. There were no differences in endocytosis ability and the cytokines (IL-6, IL-10, IL-12p70, IFN-γ) concentration between investigated groups. CONCLUSIONS High immature markers expression on the generated dendritic cells together with identical endocytosis ability in patients group is advantageous in antitumor autologous cells immunotherapy planning. However there is one troubling fact--high expression of markers, which may induce tolerance to particular antigen. It seems to be more reasonable to use the autologous DCs in the antitumor immunotherapy, especially due to the incompatibility in allogenic cells in the context of HLA complex.

Immune system disturbances in Clouston syndrome

Clouston syndrome belongs to the family of ectodermal dysplasias. So far, a defective immune response has not been reported in Clouston syndrome. We report, for the first time, immunological particularities of a large multigenerational Polish family with Clouston syndrome.

Ocena apoptozy limfocytów krwi obwodowej u chorych leczonych z powodu raka krtani

Summary The laryngeal cancer is the most often cancer among others in head and neck region. It occurs mostly among 55 and 69. Its development depends on immunological state of the body. Vitality of the immunological system cells was considered due to growth, treatment sensitivity and prognosis of some neoplasms. The aim of this work were estimation and comparison the phenomenon of lymphocytes T and B apoptosis in laryngeal cancer patients treated with surgery and radiotherapy. Material and methods The material were 30 patients hospitalized in The Department of Otolaryngology Medical University of Lublin. They all were treated with surgery or surgery and radiotherapy. Apoptosis was estimated on different stages of the treatment process. All samples were examined with the flow cytometry method. The control group were 21 patients hospitalized because of the suspicion of the apnea syndrome, which wasn’t confirmed with polysomnographic examination. Results Results of this study show significantly increasing percentage of peripheral blood apoptotic B (CD19+) cells caused by surgical treatment. The results considering radiotherapy showed different influence on the phenomenon of immunological cells apoptosis, still those results weren’t significant. Conclusions The surgical treatment causes increased amount of apoptotic peripheral blood lymphocytes.

Selected metabolites of neutrophils in patients with 2-type diabetes complicated and non complicated with diabetic foot syndrome during colonization of E. coli toxin.

Neutrophils play an important role in the pathogenesis of complications of diabetes mellitus. The aim of the study was to evaluate the metabolism of neutrophiles activation markers during the colonization of E. coli endotoxin in order to determine their potential role in the treatment of 2-type diabetes complicated and non-complicated with the diabetic foot syndrome and to evaluate production of peroxide anions by stimulated and non-stimulated neutrophils depending on the exposition time. 54 patients were divided into 3 groups (15 healthy volunteers--control group (1), group 2 - 17 patients with 2-type non-complicated diabetes group 3 - 22 patients with diabetes and diabetic foot syndrome). Blood samples from all subjects were examined. Results show significant differences of E.coli endotoxin influence on metabolism of neutrophiles in study groups. Production of peroxide anions by non-stimulated neutrophils in 20th minute of the experiment was 15 times higher in the group with no diabetic foot and 18 times higher in the group with diabetic foot as compared to the control group. Production of peroxide anions produced by neutrophils increased significantly with the exposure time. The results correspond to data in the literature, that suggest, that type, time of exposition and concentration of pathogens may significantly interfere with neutrophiles activity in the course of diabetes.