Bcl-2 and Fas expression in peripheral blood leukocytes of patients with alcoholic and autoimmune liver disorders.

BACKGROUND: This article is devoted to the inquiry of three diseases of the liver: alcoholic liver disease (ALD), primary biliary cirrhosis (PBC), and autoimmune hepatitis (AIH). The aim of the study was to assess the changes in populations of circulating lymphocytes expressing antiapoptotic bcl-2 molecule and proapoptotic Fas (cluster of differentiation 95(CD95)) receptor in patients with ALD, AIH, and PBC. MATERIALS AND METHODS: The study population consisted of 110 patients with ALD (n = 50), PBC (n = 30), and AIH (n = 30) and age-matched healthy volunteers (n = 25). Peripheral blood lymphocytes were isolated, stained with monoclonal antibodies against CD4, CD8, and CD19 antigen; intracellular bcl-2; and surface Fas receptor (CD95) antigens, and estimated using the flow cytometric method. RESULTS: Bcl-2 expression was the highest in CD4+ and CD19+ T lymphocytes in ALD; however, only the differences in median/mean fluorescence intensity values of CD4+bcl-2+ lymphocytes between ALD and PBC group and CD19+bcl-2+ between ALD and PBC groups were statistically significant, indicating the different role of B cells in pathology of ALD and PBC. In contrast to that, statistically significant higher percentage of CD4+, CD8+, and CD19+ bearing Fas receptor in all groups of patients with liver diseases in comparison with the control subjects were estimated. The highest expression of Fas in CD4+ lymphocytes in ALD and in CD8+ cells of PBC and AIH groups were detected. CONCLUSION: Low expression of bcl-2 molecule and high expression of Fas in peripheral blood lymphocytes indicate significant dysregulation of apoptotic mechanisms not only in the liver but also in peripheral blood lymphocytes in all examined groups, especially in ALD group.

Analysis of immunohistochemical expression of proinflammatory cytokines (IL-1α, IL-6, and TNF-α) in gallbladder mucosa: comparative study in acute and chronic calculous cholecystitis.

BACKGROUND: Several studies have shown increased serum levels of proinflammatory cytokines (IL-1α, IL-6, and TNF-α) in patients with cholelithiasis. The local expression of the proteins involved in pathogenesis of the disease is poorly recognised. MATERIALS AND METHODS: The authors examined immunohistochemically (IHC) the expression status of IL-1α, IL-6, and TNF-α in gallbladder mucosa of the patients with cholelithiasis as related to acute (ACC) and chronic (CCC) types of cholecystitis. Proinflammatory cytokines were quantitatively evaluated in gallbladder mucosa (epithelium and lamina propria) in ACC (n = 16) and CCC (n = 55) groups using modern spatial visualisation technique. RESULTS: Quantitative analysis of IHC signals showed no significant differences in IL-1α and IL-6, and immunoexpression in patients with ACC and CCC. A significantly greater IHC expression of TNF-α was detected in CCC as compared with ACC group. In either of the patient groups immunoexpression of IL-1α and of TNF-α was significantly higher than that of IL-6. Immunoexpression of TNF-α was significantly higher than that of IL-1α only in CCC group. A positive correlation was disclosed between IHC expression of IL-1α and body mass index in CCC group. IHC expression of TNF-α correlated positively with expression of CD68 molecule (histiocytic marker), number of leukocytes in blood and higher grading of gallbladder wall in ACC group. CONCLUSIONS: A more pronounced IHC expression of TNF-α and IL-1α than IL-6 in both types of cholecystitis may suggest the role of these cytokines in pathogenesis of cholelithiasis. IHC expression of TNF- α shows better correlation with clinical/laboratory data in acute cholecystitis, and its quantitative prevalence over the remaining cytokines points to the role of the TNF-α in maintenance of inflammation in the course of cholelithiasis.

A comparative study of the diagnostic accuracy of cone beam computed tomography and intraoral radiographic modalities for the detection of noncavitated caries.

OBJECTIVES: The aim of this study was to determine the diagnostic accuracy of cone beam computed tomography (CBCT) in the detection of approximal and occlusal noncavitated carious lesions. METHODS: A total of 135 extracted human posterior teeth were used. They were radiographed using the following: conventional film (Kodak Insight), a digital system (PSP plates Digora Optime, Soredex), and a CBCT system (NewTom 3G, Quantitative Radiology). All the images were assessed by two independent observers twice. Receiver operating characteristic analysis (Az) was used. RESULTS: NewTom 3G 9″ CBCT was statistically inferior to conventional film and a digital system for the detection of approximal caries. NewTom 3G 9″ had significantly higher Az values than PSP plate (p < 0.05), but there were no statistically significant differences between the Az values for CBCT and film (p > 0.33) for occlusal surface caries. CONCLUSION: The diagnostic accuracy of all three tested radiographic systems is low, and it is necessary to use other methods to improve early caries detection. CLINICAL RELEVANCE: CBCT has found a wide application in different fields of dentistry. The report from a CBCT examination performed for any of these reasons should include all abnormal findings, and the detection of noncavitated caries lesions is especially important because it facilitates the use of noninvasive treatment.

The influence of amalgam fillings on the detection of approximal caries by cone beam CT: in vitro study.

OBJECTIVES: The aim of this CBCT investigation on the detection of caries was to assess the influence of artefacts produced by the presence of amalgam fillings located in the vicinity. METHODS: 102 non-cavitated pre-molar and molar teeth were placed in blocks of silicone with approximal contacts consisting of 3 sound or carious teeth and 1 mesial-occlusal-distal amalgam-filled tooth in-between. Radiographs of all the teeth were recorded using the CBCT system (NewTom™ 3G; QR Srl, Verona, Italy; field of view, 9 inches). Data from the CBCT unit were reconstructed and sectioned in the mesiodistal tooth plane. Images were evaluated twice by two observers, using a five-step confidence scale. After the CBCT examination, the teeth were individually sectioned in the mesiodistal direction with a diamond saw. Using a light microscope at ×40 magnification, the true morphological status of all approximal surfaces was established. RESULTS: Sensitivity of the CBCT for the detection of caries on surfaces located proximally and distally to an amalgam filing ranged from 0.27 to 0.30 for enamel and from 0.47 to 0.56 for dentin. Specificity values for enamel proximal and distal lesions were 0.48 and 0.53, respectively, for enamel and 0.33 to 0.38, respectively, for proximal and distal dentin cases. Intra-observer reliability was 0.84, and interobserver reliability was 0.49. CONCLUSIONS: Owing to its low specificity, scans from a CBCT examination should not be used to determine the presence of demineralization of the tooth surface when amalgam fillings are present in the region of interest.

Role of high endothelial postcapillary venules and selected adhesion molecules in periodontal diseases: a review.

Periodontitis is accompanied by the proliferation of small blood vessels in the gingival lamina propria. Specialized postcapillary venules, termed periodontal high endothelial-like venules, are also present, and demonstrate morphological and functional traits similar to those of high endothelial venules (HEVs) in lymphatic organs. The suggested role of HEVs in the pathogenesis of chronic periodontitis involves participation in leukocyte transendothelial migration and therefore proinflammatory effects appear. Recent observations suggest that chronic periodontitis is an independent risk factor for systemic vascular disease and may result in stimulation of the synthesis of acute phase protein by cytokines released by periodontal high endothelial cells (HECs). However, tissue expression of HEV-linked adhesion molecules has not been evaluated in the gingiva of patients with chronic periodontitis. This is significant in relation to potential therapy targeting expression of the adhesion molecules. In this review, current knowledge of HEV structure and the related expression of four surface adhesion molecules of HECs [CD34, platelet endothelial cell adhesion molecule 1, endoglin and intercellular adhesion molecule 1 (ICAM-1)], involved in the key steps of the adhesion cascade in periodontal diseases, are discussed. Most studies on the expression of adhesion molecules in the development and progression of periodontal diseases pertain to ICAM-1 (CD54). Studies by the authors demonstrated quantitatively similar expression of three of four selected surface markers in gingival HEVs of patients with chronic periodontitis and in HEVs of reactive lymph nodes, confirming morphological and functional similarity of HEVs in pathologically altered tissues with those in lymphoid tissues.

Peripheral blood dendritic cells in alcoholic and autoimmune liver disorders.

Little is known about effects of alcohol consumption on dendritic cell (DC) function and resultant immune response. However, quantitative and qualitative disturbances of DCs are speculated to be involved in alcohol-related as well as in other liver pathology. The present study aimed to evaluate changes in circulating DC subsets in alcoholic liver disease (N = 43), autoimmune hepatitis (N = 26) and primary biliary cirrhosis (N = 20). DCs isolated from the peripheral blood of recruited participants were stained with monoclonal antibodies against blood dendritic cell antigens (BDCAs) and estimated using the flow cytometry. Myeloid DCs were defined as BDCA-1(+)/CD19(-) cells, and lymphoid DCs as BDCA-2(+)/CD123(+) cells. Total numbers of circulating DCs in subjects with some liver diseases were markedly lower than in the healthy participants (p = 0.03). There was a significantly lower percentage of circulating BDCA-2(+)/CD123(+) (p = 0.02), and a tendency for the percentage of circulating BDCA-1(+)/CD19(-) cells to decrease in patients with liver diseases compared to the controls (p = 0.09). These results may suggest that decreased numbers of DCs may be responsible for reduced adaptive immune responses and increased susceptibility to infections and cancer development observed in patients exposed to alcohol. Moreover, numerical abnormalities of DCs may contribute to the breakdown of self-tolerance, a feature of autoimmune diseases.

The expression and concentration of CD40 ligand in normal pregnancy and pre-eclampsia.

This study investigated the surface expression of the CD154 antigen (CD40L) on peripheral blood CD4+ T lymphocytes and the sera concentrations of soluble CD40L antigens (sCD40L) in non-pregnant women, normal pregnant women, and patients with pre-eclampsia. Twenty-five patients with pre-eclampsia, 18 healthy pregnant women, and 10 healthy non-pregnant women were included in the study. The expression of the CD154 antigen on CD4+ T lymphocytes was determined using flow cytometry. The serum concentration of sCD40L was measured using an ELISA. Statistical analysis was performed using the Mann-Whitney U and ANOVA tests. The expression of the CD154 antigen on CD4+ T lymphocytes and the serum concentration of soluble CD40L were significantly higher in pre-eclampsia than in normal pregnant women. In normal pregnancy the expression of the CD154 antigen on CD4+ T lymphocytes and the concentration of sCD40L were significantly lower than in non-pregnant women. We conclude that during normal pregnancy the levels of these inflammatory mediators are lower than in non-pregnant women. In pre-eclampsia the levels are higher than those in normal pregnancy, but stable compared with the non-pregnant state. These results suggest that in pre-eclampsia, there are disturbances in the mechanisms responsible for the decrease in innate immunity which occurs in normal pregnancy.

Evaluation of bone loss at single-stage and two-stage implant abutments of fixed partial dentures.

INTRODUCTION: Fixed partial dentures (FPDs) can be supported on implant abutments only or on single-stage and two-stage implants and teeth. PURPOSE: The purpose of this study was a comparative analysis of bone loss at the single-stage and two-stage implant abutments of fixed partial dentures used to restore missing teeth classified as Class I or Class II according to the Kennedy classification. MATERIAL AND METHODS: 32 patients were treated by using 49 FPDs supported on implants and teeth worn for 2-6 years. Bone loss at the implant abutments of FPDs was evaluated by one examiner using a special ruler with a measuring scale and images of implants. Measurements were conducted at 26 single-stage implants and 50 two-stage implant abutments based on panoramic radiographs. RESULTS: Statistical analysis showed that the mean bone loss at implants after 2 years was 0.70 mm +/- 0.50. The mean bone loss at implants after 6 years was 1.73 mm +/- 0.41. The bone loss of the alveolar ridge at the single-stage implants was greater than at the two-stage implants but it was not statistically significant. CONCLUSION: Prosthetic treatment of missing teeth classified as Class I or II according to the Kennedy classification with FPDs may result in bone loss less than 2 mm after 6 years. Both single-stage and two-stage intraosseous implants can be suitable for the implant-prosthetic treatment of patients with alar lack of teeth.

Influence of ethyl acetate extract and quercetin-3-methyl ether from Polygonum amphibium on activation lymphocytes from peripheral blood of healthy donor in vitro.

The influence of an ethyl acetate extract from Polygonum amphibium L. and quercetin-3-methyl ether isolated from them were examined on the human immune system. The investigations were made on peripheral blood mononuclear cells from healthy donors. The cells were stimulated by plant extract and quercetin-3-methyl ether in 24 h culture and analysed inflow cytometry. The following mice monoclonal antibody anti human activate markers were used: anti HLA-DR PE, anti CD25 FITC, anti CD69 FITC and anti CD71 FITC. The level of Interleukine-2 in blood serum and in culture supernatants was measured by ELISA method. Ethyl acetate extract from Polygonum amphibium caused the rise in the number of CD25 and HLA-DR positive lymphocytes and increased the expression of CD25 and CD71 antigens. The level of IL-2 was increasing for the duration of the culture, independently from the presence of stimulators. Besides quercetin-3-methyl ether, from herb of Polygonum amphibium L., trans-taxifolin, quercetin and kaempferol were isolated The structure of the isolated compounds was determined by spectroscopic (UV, (1)H NMR, (13)C NMR and CI-MS) methods.

Short Communication: Expression of Apoptosis Regulating Factors on T Lymphocytes in Multiple Myeloma Patients.

Multiple myeloma (MM) is a malignant proliferation of plasma cells in which dysregulation of programmed cell death (apoptosis) is responsible for tumor cell expansion. However some phenotypic and functional alterations of T cells in MM patients have been reported, that also can influence the plasma cell growth. The aim of the study was to assess some aspects of T lymphocyte apoptosis in MM to obtain a better understanding of the changes in the immune system in this disease. Flow cytometry was used to analyze the expression of two main regulators of apoptosis: the pro-apoptotic Fas antigen and the anti-apoptotic protein BCL-2 in patients with untreated MM and in healthy controls. ELISA was used to determine soluble Fas ligand (sFasL) serum levels in patients and control groups. We detected statistically significant higher Fas expression in patients than in controls both on CD4 and CD8 lymphocytes, but no differences in BCL-2 expression by these cells. The sFasL level was statistically significant lower in patients than in controls. Our results indicate that T cells in MM are controlled by up-regulation of Fas. The Fas/FasL system induces the killing of T cells expressing Fas antigen, what could account for the incapability of the immune system to protect host against tumor expansion.

Assessment of Apoptosis Regulating Factors BCL-2 and Fas Antigens on Malignant and Normal Plasma Cells.

Multiple myeloma (MM) is characterised by slow proliferation of malignant plasma cells and their accumulation within the bone marrow. The dysregulation of programmed cell death (apoptosis) is a very important mechanism in the pathogenesis of this tumour. It prompted us to investigate the apoptosis regulating factors such as the pro-apoptotic Fas antigen and the anti-apoptotic protein BCL-2 on bone marrow malignant plasma cells in untreated patients with newly diagnosed MM and to compare them with their normal counterparts-plasma cells isolated from bone marrow of healthy individuals. Twenty-nine MM patients and 16 healthy persons were studied. Bone marrow mononuclear cells were isolated, indicated by monoclonal antibodies and analysed using the flow cytometry method. There was no statistically significant difference in BCL-2 expression in plasma cells between patients and control groups. However the percentage of BCL-2 positive cells was significantly related to the clinical stage of the disease. We detected statistically significant lower percentage of Fas positive cells in the patient group than in control. We concluded that in MM at diagnosis the expression of BCL-2 in bone marrow malignant plasma cells was comparable to normal plasma cells but expression of Fas antigen on these cells was lower. It suggests that down regulation of Fas and normal regulation of BCL-2 may be implicated for myeloma cell survival and their escape from apoptosis in vivo.

Flow cytometric analysis in diagnosis of thymoma metastases.

Thymomas are relatively rare tumours of the anterior mediastinum, constituting approximately 10-15% of all mediastinal tumours. In contrast to other neoplasms, they rarely present distant metastases. We describe a case of thymoma with long survival and skin metastases diagnosed by two-colour flow cytometry.

[Ultrastructural pattern of enamel in initial caries]. / Ultrastrukturalny obraz szkliwa w próchnicy poczatkowej.

In beginning caries of particular interest are destructive changes in the enamel layers just below the surface. The studied material comprised about twenty enamel blocks obtained from 10 teeth with carietic spots removed for orthodontic reasons in children aged 12-14 years. The aim of the study was assessment of the initial enamel damage under electron microscope. In all electronograms of the enamel involved by the carietic spot various grades of demineralization of the enamel below its surface were seen. The changes in initial caries seemed well delineated from the remaining healthy enamel, although within the carietic focus demineralization was observed from very slight widening of the area around the prisms to massive destruction of prism structure. The changes in the carietic spot under electron microscope showed a wide variety of patterns of destruction of enamel prisms, although clinically the surface of the enamel involved by beginning caries was small and not damaged.

[Trials of remineralization of artificially produced enamel lesions]. / Próby remineralizacji sztucznie wywolanych uszkodzen szkliwa.

The purpose of the study was checking in vitro whether after application of calcifying preparations remineralization occurs of artificially damaged enamel prisms. The study was carried out on 32 enamel samples obtained from 8 human teeth. For remineralization saturated solutions were used of phosphate-calcium compounds in the form of fluid or gel. The degree of demineralization and remineralization of enamel was assessed on the basis of images of enamel breaks in JSM-50A scanning microscope. Partial remineralization of artificially damaged enamel was found. Detailed evaluation of the remineralization progress requires extension of investigation methods.

[Chemical composition of the saliva in children and adolescents with black tartar]. / Sklad chemiczny sliny dzieci i mlodziezy z czarnym osadem nazebnym.

The chemical composition of the saliva was assessed in children and adolescents with black tartar and children without these deposits. Group I (studied) included 60 children aged 4-16 years with black stain present on the teeth, group II (control) comprised 60 children in the same age range without black tartar. For the study stimulated mixed saliva was taken. In saliva samples determinations were done of sodium, potassium, chlorides, copper, zinc, iron, total calcium, inorganic phosphates, glucose and total protein. In the saliva of children with black tartar a significantly higher content was found of total calcium, inorganic phosphates, copper, sodium, total protein, and lower glucose content than in controls. The chemical composition of saliva in children and adolescents with black tartar is characteristic of subjects with low susceptibility to caries. Changes in the content of certain salivary components were noted in children with black stain at the time of permanent teeth eruption.

[Amount and pH of the saliva in children and adolescents with black tartar]. / Ilosc oraz pH sliny dzieci i mlodziezy z czarnym osadem nazebnym.

A comparative study was carried out of the volume of secreted saliva and its pH in children with black tartar and without it. Group I comprised 60 children with black tartar (aged 4-16 years) and group II (control) contained 60 children without black tartar in the same age range. Mixed stimulated saliva was obtained. Its volume was measured after 5, 10, 15 and 20 minutes, calculating then the minute index of saliva flow. The pH of the saliva was determined using indicator papers (in vivo) or pH-meter (in vitro). No significant differences were found in the amounts of saliva secreted in group I and group II. The pH of the saliva was significantly higher in children with black tartar than in those without it.

[Study of the initial enamel injuries in vitro]. / Badania poczatkowych uszkodzen szkliwa in vitro.

The purpose of the study was production under laboratory conditions of enamel changes corresponding to early caries. The study was carried out on 20 healthy human teeth in vitro, in two phases differing in the conditions of the experiment. Carietic changes were produced with lactate buffer (in concentrations from 0.025 to 1.0 M) as fluid without added inhibitor of the solubility of the superficial layer. The assessment of the degree of enamel demineralization was based on images obtained in JSM 50A scanning microscope. In the first stage of the experiment a minimal demineralization of the subsuperficial layer of the enamel was obtained. In the second stage besides damage to the subsuperficial layer enamel erosion was noted. In this connection further studies are necessary for determining of the proper conditions for obtaining initial enamel damage.

[Demineralization and remineralization of enamel in the light of the most recent literature reports]. / Demineralizacja i remineralizacja szkliwa w swietle najnowszego pismiennictwa.

In the light of a review of the most recent literature reports the present views on demineralization and remineralization of enamel in vivo and in vitro are presented. Modes and conditions of the development of artificial enamel damage of the character of incipient caries are discussed. The possibility in stressed of cure of initial enamel damage after application of calcification-inducing solutions and human saliva.