Ursodeoxycholic and chenodeoxycholic bile acids attenuate systemic and liver inflammation induced by lipopolysaccharide in rats.

Bacterial lipopolysaccharide (LPS) induces general inflammation, by activating pathways involving cytokine production, blood coagulation, complement system activation, and acute phase protein release. The key cellular players are leukocytes and endothelial cells, that lead to tissue injury and organ failure. The aim of this study was to explore the anti-inflammatory, antioxidant, and cytoprotective properties of two bile acids, ursodeoxycholic acid (UDCA) and chenodeoxycholic acid (CDCA) in LPS-induced endotoxemia in rats. The experiment involved six distinct groups of Wistar rats, each subjected to different pretreatment conditions: control and LPS groups were pretreated with propylene glycol, as a bile acid solvent, while the other groups were pretreated with UDCA or CDCA for 10 days followed by an LPS injection on day 10. The results showed that both UDCA and CDCA reduced the production of pro-inflammatory cytokines: TNF-α, GM-CSF, IL-2, IFNγ, IL-6, and IL-1ß and expression of nuclear factor-κB (NF-κB) induced by LPS. In addition, pretreatment with these bile acids showed a positive impact on lipid profiles, a decrease in ICAM levels, an increase in antioxidant activity (SOD, |CAT, GSH), and a decrease in prooxidant markers (H2O2 and O2-). Furthermore, both bile acids alleviated LPS-induced liver injury. While UDCA and CDCA pretreatment attenuated homocysteine levels in LPS-treated rats, only UDCA pretreatment showed reductions in other serum biochemical markers, including creatine kinase, lactate dehydrogenase, and high-sensitivity troponin I. It can be concluded that both, UDCA and CDCA, although exerted slightly different effects, can prevent the inflammatory responses induced by LPS, improve oxidative stress status, and attenuate LPS-induced liver injury.

Anti-inflammatory and immunomodulatory effects of Biodentine on human periapical lesion cells in culture.

AIM: To evaluate the effect of Biodentine eluate on cytotoxicity and production of pro- and anti-inflammatory cytokines and osteodestructive/osteoprotective cytokines in cultures of human periapical lesion cells. METHODOLOGY: Conditioned Biodentine Medium (CBM) was prepared according to ISO 10993-12, by incubating Biodentine in RPMI medium (0.2 g mL-1 ) for 3 days at 37 °C. CBM contained both released microparticles and leachable soluble components. Inflammatory cells were isolated from 22 human periapical lesions after apicectomy or tooth extraction, by collagenase/DNase digestion, and cultured in several dilutions of CBM. The composition of periapical lesion cells was determined by morphological criteria, cytotoxicity was quantified by MTT and flow cytometric apoptosis/necrosis assays, whereas the levels of produced cytokines in cell culture supernatants were measured by flow cytometry and ELISA. Student t-test and Friedman test with Dunn's post-test were used for comparison of parametric and nonparametric variables, respectively. RESULTS: Undiluted (100%), 75% and 50% CBM were cytotoxic for periapical lesion cells due to induction of both necrosis (100% CBM) and apoptosis (75% and 50% CBM). Noncytotoxic concentrations of CBM (25%) inhibited the production of pro-inflammatory cytokines: TNF-α, (P < 0.005); IL-1ß (P < 0.01); IL-6 (P < 0.005) and chemokines IL-8: (P < 0.005); MCP-1 (P < 0.005), stimulated the production of anti-inflammatory cytokine (IL-10; P < 0.005), Th2 cytokines: IL-4, IL-5 and IL-33 (all P < 0.01), and IL-17A (P < 0.01). The concentration of CBM (12.5%) inhibited the production of IL-6 (P < 0.05), IL-8 (P < 0.01) and MCP-1 (P < 0.005) and augmented the production of IL-10 (P < 0.05). No significant effects on Th1-related cytokines (IFN-γ and IL-12) and IL-23 were detected with 25% and 12.5% CBM concentrations. Both CBM concentrations inhibited the production of osteolytic receptor activator of nuclear factor kappa-Β ligand (RANKL), dose dependently (P < 0.005 and P < 0.01, respectively). Higher CBM concentrations decreased RANKL/osteoprotegerin (OPG) ratio (P < 0.05), without significant influence on the levels of osteoprotective OPG. CONCLUSION: Biodentine possesses immunomodulatory properties by suppressing pro-inflammatory and augmenting anti-inflammatory cytokines. Together with the reduction of osteodestructive mediators, this novel root-end filling cement could be beneficial for healing and bone reparation after the surgical treatment of periapical lesions.

IL-17 and glutamate excitotoxicity in the pathogenesis of multiple sclerosis.

Immunoinflammatory-mediated demyelination, the main pathological feature of multiple sclerosis (MS), is regularly accompanied by neurodegenerative processes, mostly in the form of axonal degeneration, which could be initiated by glutamate excitotoxicity. In the current study, the relationship between Th17-mediated inflammatory and excitotoxic events was investigated during an active phase of MS. Cerebrospinal fluid (CSF) of patients with MS and control subjects was collected, and IL-17A and glutamate levels were determined. IL-17A level was significantly higher in patients with MS; whereas no statistically significant changes in glutamate concentrations were found. There was a direct correlation between IL-17A and glutamate levels; IL-17A levels were also associated with the neutrophil expansion in CSF and blood-brain barrier disruption. However, IL-17A level and the number of neutrophils tended to fall with disease duration. The results suggest that Th17 cells might enhance and use glutamate excitotoxicity as an effector mechanism in the MS pathogenesis. Furthermore, Th17 immune response, as well as neutrophils, could be more important for MS onset rather than further disease development and progression, what could explain why some MS clinical trials, targeting Th17 cells in the later stage of the disease, failed to provide any clinical benefit.

E-cadherin expression in acquired cholesteatoma.

OBJECTIVES: The epithelial cell adhesion molecule E-cadherin is important for maintenance normal tissue architecture and for cell-cell communication and immune cell migration. E-cadherin is also present in cholesteatomas. This study determined E-cadherin expression in acquired cholesteatomas and analyzed its expression according to cholesteatoma clinical and histological characteristics. METHODS: We investigated E-cadherin expression in 30 samples from operated patients with acquired middle ear cholesteatomas that were classified according to their clinical and histological characteristics. E-cadherin expression in cholesteatoma was determined immunohistochemically. A semi quantitative method was used to determine the index of expression of E-cadherin and t-tests, Mann-Whitney U tests and Spearman correlation analysis were used for statistical analysis. RESULTS: We found significant expression of E-cadherin on CD1, CD3 total, CD4 (p < 0.05), high expression of E-cadherin on CD8 total and CD19/CD38 lymphocytes (p < 0.01) and very high expression of E-cadherin on mast cells and antigen-presenting cells, including Langerhans cells (p < 0.005). We graduated results as no statistically significant (p>0.05), statistically significant (0.05 > p > 0.01), highly statistically significant (0.01 > p > 0.005) and very highly statistically significant (p < 0.005). CONCLUSION: E-cadherin expression was the same in the cholesteatoma matrix in all samples. There were no differences in expression according to the clinical and histological characteristics of the cholesteatomas.

Salivary interleukin-8 levels in children suffering from type 1 diabetes mellitus.

OBJECTIVE: The aim of this study was to investigate the differences between the salivary levels of IL-8 in patients with Type 1 diabetes mellitus (DM) with (DM+P) or without (DM-P) concomitant periodontitis and healthy subjects. The correlations between the levels of these cytokines and clinical periodontal parameters were also established. METHODS: Twenty children and adolescents with Type 1 DM (10 diagnosed with periodontitis, 10 presenting no signs of periodontitis) and a control group consisting of 20 healthy children and adolescents aged 7-18 years were recruited for this study. RESULTS: The Salivary IL-8 level was statistically significantly (p < 0.005) elevated in subjects with Type 1 DM (474.47 +/- 716.76) compared to non-diabetic control group (101.99 +/- 68.32). There was no difference (p > 0.05) in the salivary IL-8 level when subjects with Type 1 DM with concomitant periodontitis were compared to diabetics without periodontitis. When the salivary IL-8 level in subjects with Type 1 DM was correlated with the clinical parameters, no statistical significance was found. CONCLUSION: An elevated salivary IL-8 level in subjects with Type 1 DM without concomitant periodontitis plays a major role in the development of diabetic micro and macroangiopathy and pathogenesis of atherosclerosis. Consequently, this may offer a basis for the assessment of risk, prophylaxis and treatment of diabetic complications.

Response of monocyte-derived dendritic cells to rapidly solidified nickel-titanium ribbons with shape memory properties.

Ni-Ti Shape Memory Alloys (SMAs) have attracted considerable attention as biomaterials for medical devices. However, the biocompatibility of Ni-Ti SMAs is often unsatisfactory due to their poor surface structure. Here we prepared Rapidly Solidified (RS) Ni-Ti SMA ribbons by melt-spinning and their surface was characterised by Auger-electron spectroscopy, X-ray photoelectron spectrometry and scanning electron microscopy. The biocompatibility of the produced ribbons and their immunomodulatory properties were studied on human monocyte-derived dendritic cells (MoDCs). We showed that melt-spinning of Ni-Ti SMAs can form a thin homogenous oxide layer, which improves their corrosion resistance and subsequent toxicity to MoDCs. Ni-Ti RS ribbons stimulated the maturation of MoDCs, as detected by changes in the cells' morphology and increased expression of HLA-DR, CD86, CD40 and CD83 molecules. However, Ni-Ti RS ribbons enhanced the tolerogenic properties of immature MoDCs, which produced higher levels of IL-10 and IL-27, driving the differentiation of IL-10- and TGF-ß-producing CD4+T cells. On the other hand, in the presence of lipopolysaccharide, an important pro-inflammatory biomolecule, Ni-Ti RS ribbons enhanced the allostimulatory and Th1 polarising capacity of MoDCs, whereas the production of Th2 and Th17 cytokines was down-regulated. In conclusion, Ni-Ti RS ribbons possess substantial immunomodulatory properties on MoDCs. These findings might be clinically relevant, because implanted Ni-Ti SMA devices can induce both desired and adverse effects on the immune system, depending on the microenvironmental stimuli.

Cytotoxicity of gold nanoparticles prepared by ultrasonic spray pyrolysis.

The aim of this work was to study the cytotoxicity of different fractions of gold nanoparticles prepared by ultrasonic spray pyrolysis from gold scrap. The target cells were rat thymocytes, as a type of nonproliferating cells, and L929 mouse fibroblasts, as a type of continuous proliferating cells. Fractions 1 and 2, composed of pure gold nanoparticles, as determined by scanning electron microscopy with a combination of energy dispersive X-ray analysis, were nontoxic for thymocytes, but reduced moderately the proliferative activity of L929 cells. The inhibitory effect of fraction 2, containing particles smaller in size than fraction 1, was stronger. Fraction 3, composed of Au and up to 3% Cu was noncytotoxic for thymocytes, but was cytotoxic for L929 cells. Fraction 4, composed of Au and Ag nanoparticles, and fraction 5, composed of Au together with Cu, Ni, Zn, Fe, and In were cytotoxic for both thymocytes and L929 cells. These results suggest that USP enables the synthesis of pure gold nanoparticles with controlled size, even from gold scrap. However, microstructural analyses and biocompatibility testing are necessary for their proper selection from more cytotoxic gold nanoparticles, contaminated with other elements of gold alloys.

Effects of conventional versus biocompatible peritoneal dialysis solutions on peritoneal and systemic inflammation, malnutrition and atherosclerosis in CAPD patients.

BACKGROUND: Chronic inflammation, malnutrition and atherosclerosis (MIA syndrome) are important predictors of high mortality in continuous ambulatory peritoneal dialysis (CAPD) patients. We aimed to evaluate the effects of PD solutions (standard vs. biocompatible) on some parameters of MIA syndrome in patients undergoing CAPD. METHODS: 42 stable patients who were on CAPD at least 2.5 years participated in this cross-sectional study. Patients who had severe anemia (Hb < 10 g/l), immunomodulatory therapy, peritonitis or any inflammatory conditions for at least 3 months before the analysis, malignant disease and acute exacerbation of heart failure, were excluded. 21 (50%) patients were treated with standard PD solutions (CAPDP-1), while the remaining 21 (50% of patients) were treated with biocompatible PD solutions (neutral solutions with lower level of glucose degradation products and lower concentration of calcium, CAPDP-2). All patients underwent echocardiography and B-mode ultrasonography of common carotid arteries together with assessments of nutrition status and parameters of systemic and local inflammation. RESULTS: There were no significant differences between the groups concerning age, gender, underlying disease, residual renal function, peritoneal transport characteristics, comorbidity or therapy applied. Patients from group CAPDP-2 had a significantly lower serum level of hs-CRP (3.7 ± 2.6 mg/l vs. 6.3 ± 4.5 mg/l; p = 0.023) and significantly better nutritional status confirmed by mid-arm circumference (p = 0.015), mid-arm muscle circumference (p = 0.002) and subjective global assessment (14.28% of patients in CAPDP-2 vs. 71% of patients in CAPDP-1 were malnourished; p = 0.000). Group CAPD-2 had less frequent left ventricular hypertrophy (p = 0.039), thinner intima-media thickness (p = 0.005), smaller carotid narrowing (p = 0.000) and fewer calcified plaques of common carotide arteries (p = 0.003). No significant difference between the CAPDP groups was observed in serum and effluent levels of inflammatory cytokines (IL-1, IL-6 and TNF-α) and CA-125 effluent level. Logistic regression analysis did not confirm that biocompatibility of PD solutions was an independent predictor of any parameter of MIA syndrome. CONCLUSIONS: According to the present study and logistic regression analysis, the effect of biocompatible CAPD solutions on parameters of malnutrition, inflammation and atherosclerosis have to be confirmed by well-designed and controlled studies in a higher number of patients.

The impact of Trichinella spiralis excretory-secretory products on dendritic cells.

Parasitic nematode Trichinella spiralis exert immunomodulatory effect on the host immune response through excretory-secretory products (ES L1) released from the encysted muscle larvae. Rat bone-marrow derived dendritic cells (DCs) stimulated with ES L1 antigens acquire semi-matured status and induce Th2 and regulatory responses in vitro and in vivo. Priming naïve T cells in vitro with ES L1 pulsed DCs caused strong Th2 polarization, accompanied by elevated production of regulatory cytokines IL-10 and TGF-ß and no increase in the proportion of CD4+CD25+Foxp3+ among the effector T cell population. In vivo T cell priming resulted in mixed Th1/Th2 cytokine response, with the dominance of the Th2 type and elevated levels of regulatory cytokines. Significant increase in the proportion of CD4+CD25+Foxp3+ cells was found among recipient's spleen cells. We have achieved to create immune status characteristic for the live infection by in vivo application of DCs educated with ES L1 antigens.

Immunomodulatory activity of IL-27 in human periapical lesions.

IL-27, a cytokine with pro-inflammatory and anti-inflammatory properties, is a new member of the IL-6/IL-12 family, whose function in periapical lesions is unknown. We hypothesized that the production of IL-27 and its effect depend upon the type of immune/inflammatory response and clinical presentation of periapical lesions. We tested this hypothesis by studying the expression and function of IL-27 in human periapical lesions, both in situ and in culture. Immunohistochemistry demonstrated the strongest expression of IL-27 by endothelial cells and mononuclear phagocytes. Its production by periapical lesion mononuclear cells (PL-MNC), especially in symptomatic lesions, was significantly higher compared with that in peripheral blood MNC and correlated with the frequency of CD14(+) and CD3(+) cells. Exogenous IL-27 stimulated Th1 and down-regulated Th17 cytokine production by PL-MNC from symptomatic lesions, but down-regulated Th1 and Th2 responses in asymptomatic lesions. These findings suggest that IL-27 is an immunomodulatory cytokine in periapical lesions, with complex biological effects.

Regulatory T-cells in periapical lesions.

CD4(+)CD25(hi)Foxp3(+) regulatory T-cells (Tregs) are of crucial importance in regulating the immune response, including the control of any defense against infection. Their presence in periapical lesions has not been demonstrated, as yet. We hypothesized that Tregs infiltrate periapical lesions, where they inhibit T-cell proliferation. The aim of this study was to characterize Tregs in periapical lesions by confocal microscopy, flow cytometry, and functional assays. We showed that CD4(+)CD25(hi)Foxp3(+) cells in periapical lesions expressed IL-10 and TGF-beta. Their frequency was significantly higher than in peripheral blood and correlated with the levels of TGF-beta and IL-10 in culture supernatants of periapical lesion mononuclear cells. Tregs inhibited the proliferation of responder T-cells in vitro, at least in part, by stimulating the production of IL-10. These findings suggest that CD4(+)CD25(hi)Foxp3(+) cells in periapical lesions may play regulatory roles in controlling local immune/inflammatory processes.

[Strength required for mobilization of reconstructed flexor tendons in the front paw of the dog].

Adhesions of tendons with surrounding tissue, especially in zone II, are frequent complication at the reconstruction of flexor tendons of the hand. Through knowing of biomechanical incidents between the reconstructed flexor tendon and the surrounding tissue we get significant data for post-operative procedures, and that improves the opportunity for obtaining good functional results. By successive loading of operated flexor digitorum profundus in the experimental animals - dogs, medium values of force necessary for mobilization of tendons obtained on 3. post-operative day are 0 = +/- 3.36 N, 5. day 0 = +/- 6.85 N, 10. day 0 = +/- 9.58 N and 15. day 0 = +/- 10.19 N. It can be concluded that peritendon adhesions occur between 3 and 5 post-operative day. In time, the adhesions get stronger and block the movements of the tendon, and growing force is necessary for their mobilization.

Characterization of rat bone marrow dendritic cells initially primed by Trichinella spiralis antigens.

Pathogen-derived products have the capacity to induce maturation of bone marrow-derived dendritic cells (BMDCs)into populations of effectors cells that polarize Th cells toward Th1 or Th2 phenotype via different mechanisms. Since those mechanisms are not entirely clear for helminths, and almost completely unknown for Trichinella spiralis(TS), we started an investigation of the effects of TS antigens (four different antigens isolated from all three life-cycle stages of parasite)on maturation of BMDCs and their potential to present TS antigens. The expression of MHC class II, costimulatory molecules CD86, CD54, IL-10 and IL-12p70 cytokine production were measured after 2 days of BMDCs cultivation with TS antigens. While parasitic antigens did not significantly alter the expression of MHC II, most of them, except crude muscle larvae antigens, up-regulated the expression of costimulatory molecules. BMDCs, primed with all TS antigens, released increased amounts of IL-10 and decreased amounts of IL-12p70. BMDCs, primed with TS antigens, induced significant proliferation of syngeneic TS sensitized lymph nodes cells and also stimulated the production of IL-4 by T cells purified from of TS infected DA rats. The results indicate that TS stimulated BMDCs leads to the polarization of the immune response towards regulatory and Th2 type.

[Reconstruction of the alae nasi by folded nasolabial flap].

Reconstruction of the nose is very old surgical procedure and, in fact, represents the beginnings of plastic surgery. In reconstruction, an effort has to be made in order to achieve a normal look. A surgeon must choose carefully the method of reconstruction, taking into consideration the skin color, texture and nasal topography. Full-thickness nasal defects in alar region are very difficult for reconstruction due to anatomic characteristics and structures, which are very important for normal breathing and cosmetic result. Our study analyzed 16 patients with full-thickness defect of alae nasi. Folded nasolabial flaps were used for reconstruction of these defects. Good results were achieved in all cases. The flap was vital. No flap necrosis was reported in any of these patients. The lining of the nose was good. Postoperatively, the alar edge was thickened, but it became thinned after six months. The incision in donor's region was well placed in the natural line of nasolabial fold. It was concluded that folded nasolabial flap was extremely good one-step procedure for reconstruction of full-thickness defects of alar region.

Evaluation of the stimulatory effect of Epimedium alpinum L. methanolic extract on the immune response in vivo.

The effect of the methanolic extract of the underground parts of Epimedium alpinum L. (MEEA) on the immune response to Keyhole Limpet Hemocyanine (KLH) or alloantigens in vivo was studied in AO rats. Immunization of experimental animals with KLH or allogeneic lymphocytes together with MEEA was followed by an increase in cellularity of draining lymph nodes (LN) and enhanced proliferation of LN lymphocytes after their restimulation with specific antigens in vitro, compared to control rats immunized without MEEA. These effects correlated with an increase in relative values of B, MHC class II+, CD25+ and CD71+ cells, whereas percentages of T cells and both subsets of T cells (CD4+ and CD8+) were not significantly altered. As a consequence of higher LN cellularity, total numbers of all cell subsets in the MEEA-treated group of rats were significantly increased, compared to the corresponding control. The addition of MEEA together with KLH in vitro to LN lymphocytes of rats immunized with KLH or KLH and MEEA in vivo was manifested by significant increase (0.1 microg/ml of MEEA) and decrease (50 microg/ml and 100 microg/ml of MEEA) of cell proliferation, respectively. However, when LN lymphocytes from rats, immunized in vivo with KLH and MEEA, were stimulated in vitro with MEEA together with an anti-alphabeta T cell receptor monoclonal antibody (R73), their proliferation was siginificantly inhibited. Taken together, obtained results suggest that MEEA possesses immunostimulatory activity in vivo, but some components from the extract exert immunosuppressive effects in vitro on previously in vivo activated T cells.

[The tunneled supraclavicular island flap--a fasciocutaneous flap for head and neck reconstructions].

Reconstructive procedures in the head and neck region use a wide range of flaps for defect closure. The methods range from local, mostly myocutaneous flaps and skin grafts to free microsurgical flaps. To ensure a satisfactory functional and aesthetic result, good texture and color of the flap are always essential. Moreover, the donor-site defect needs to be reduced, with no resulting functional or aesthetic impairment. We have found that the shoulder is a region providing an optimum skin texture match to the neck and face. Fasciocutaneous island flap, nourished by the supraclavicular artery is used purely as a subcutaneously tunneled island flap. The tunneling maneuver significantly improves the donor site by reducing scarring. The flap is characterized by a long subcutaneous pedicle of up to 20 cm. The pivot point is in the supraclavicular region and allows the flap to be used in the upper chest, neck, chin, and cheek. In this article we present clinical cases in reconstructive procedures with supraclavicular flap and its expanded indications.

Production of interleukin-8 in vitro by mononuclear cells isolated from human periapical lesions.

INTRODUCTION: Interleukin-8 (IL-8) is an important mediator of inflammation. However, little is known about its production in chronic dental periapical lesions and this was the main aim of this work. METHODS: Inflammatory cells were isolated from clinically different periapical lesions and analyzed by morphological criteria. The mononuclear cells were isolated, phenotypically analyzed by immunocytochemistry and cultivated in vitro. IL-8 was measured in culture supernatants of these periapical lesion mononuclear cells (PL-MNC) using a microbeads fluorescence assay. RESULTS: We found a relatively high production of IL-8 in 19 out of 21 periapical lesions included in the study. The level of IL-8 and the proportion of neutrophil granulocytes were significantly higher in the group of symptomatic lesions, compared to the asymptomatic lesions, but there was no statistically significant correlation between these parameters. According to the predominance of CD3(+) T cells and Ig(+)/CD19(+) B cells and plasma cells, lesions were divided into T-type and B-type lesions, respectively. The levels of IL-8 were significantly higher in the culture supernatants of PL-MNC in the T-type lesions and were positively correlated with the proportion of macrophages/dendritic cells (CD11c(+) cells) and CD4(+) T cells. Such a correlation was not shown in B-type lesions. CONCLUSION: These results suggest that PL-MNC are a significant source of IL-8, which is probably an important chemokine for the migration and function of different cell types at the site of chronic inflammation.

Characterization of antigen-presenting cells in human apical periodontitis lesions by flow cytometry and immunocytochemistry.

AIM: To analyse phenotypic characteristics of antigen-presenting cells (APC), isolated from human periapical lesions by flow cytometry and immunocytochemistry. METHODOLOGY: Sixteen periapical lesions were digested for 15 min with 0.05% collagenase. Mononuclear cells, separated from other inflammatory cells by density centrifugation, were processed for flow cytometry and/or immunocytochemistry. Single and double immunostainings were performed using monoclonal antibodies specific for human CD45, CD3, CD19, CD14, HLA-DR, CD1a, CD83 and CD123. RESULTS: Antigen-presenting cells (HLA-DR(+) cells) represented 32.9 +/- 17.8% of total mononuclear cells. Amongst them, B cells (HLA-DR(+) CD19(+)) were the predominant APC population, followed by activated macrophages (HLA-DR(+) CD14(+)), dendritic cells (DC) (HLA-DR(+) CD14(-) CD19(-) CD3(-)) and activated T cells (HLA-DR(+) CD3(+)). Based on the predominance of T cells (CD3(+)) or B cells and plasma cells (CD19(+) and CD19(lo), respectively) amongst mononuclear cell infiltrates, lesions were divided into T- and B-types. The percentage of DC in T-type lesions (27.1 +/- 6.8% of total HLA-DR(+) cells) was higher, compared with B-type lesions (10.3 +/- 5.2%) (P < 0.01). Within the DC population, the percentages of CD1a (Langerhans cell type) and CD123 (probably plasmacytoid DC type) did not differ significantly between the groups (P > 0.05). However, the percentage of mature DC (CD83(+)) was significantly higher in T-type periapical lesions (P < 0.05). CONCLUSIONS: Flow cytometry and immunocytochemistry are suitable methods for phenotypic analysis of APC after their isolation from human periapical lesions. APC, that were phenotypically heterogeneous, constituted a significant component of infiltrating cells. Lesions with the predominance of T cells were characterized by a higher proportion of mature DC (HLA-DR(+)CD83(+) cells) than lesions with predominance of B cells/plasma cells.

[The excision width in surgical treatment of basal cell carcinoma].

Basal cell carcinoma originates from pluripotent cells of basal layer of epiderm, external covering of hair follicles, sebaceous glands or other skin adnexa. It is characterized by local infiltrating and sometimes destructive growth. There are several types of basal cell carcinomas that may be manifested in over 12 clinical forms. Surgical treatment depends to a large extent on the histological type, localization and its clinical manifestation. The analysis included 250 patients of both gender and different age, operated for basal cell carcinoma. Clinical characteristics of basal cell carcinoma and the width of the excision were described. It was concluded that the width of the excision of basal cell cancer was in relation to histological type.

[Fournier's gangrene and reconstruction of its defects].

Fournier's gangrene is remarkebly rare but severe, life threatening condition. Thanks to better earlz recongnition of disease in past few years, the percentage of patients which survive this condition is considerable, but consequence of this fact is large number of patients with defects of cutis and subcutis in perineal and scrotal reigions. Reconstruction of this defects is cimplicate and require helathy, well vascularised tissue from another anatomical region which is not involved in gangreneous process. Authors present two cases of reconstruciton of defects in perineal and scrotal regions. First case is large defect which is covered by transposition of musculus gracilis musculo-cutaneous flap. Second case is lesser defect which is enclosed by mobilisation of local skin flaps.