Gingival crevicular fluid and serum levels of APRIL,BAFF and TNF-alpha in rheumatoid arthritis and osteoporosis patients with periodontal disease

Background

This study was performed to evaluate gingival crevicular fluid (GCF) and serum levels of a proliferation inducing ligand (APRIL) and B cell activating factor (BAFF) and compare this to differences between TNF-alpha levels in rheumatoid arthritis (RA), osteoporosis (OPR) and systemically healthy women with periodontal disease (SH).

Design

Gingival crevicular fluid (GCF) and serum samples were obtained before any periodontal intervention from 17 RA, 19 OPR patients and 13 SH women with periodontitis. Full-mouth clinical periodontal measurements were recorded. APRIL, BAFF and TNF-α levels were determined by ELISA. Statistical analysis was performed using multivariate analysis, ANOVA and Spearman correlation.

Results

Pocket depths differed in site-specific comparisons, but otherwise clinical measurements were similar in the three study groups. Multivariate least squares regression ANOVA adjusted for age and for plaque index indicated that total amounts of TNF-α and concentrations of TNF-α, BAFF and APRIL were significantly greater in the RA patients than in the SH group (p < 0.05), and GCF concentrations of BAFF were greater in OPR patients than in SH. Serum TNF-α and BAFF were significantly higher in the RA group compared to SH (p < 0.05) and serum TNF-α was greater in RA than in OPR (p < 0.05). APRIL and BAFF correlated with RANKL levels in GCF and serum (p < 0.05).

Conclusion

Despite long-term usage of anti-inflammatory drugs in the RA and OPR patients, increased TNF-family cytokines, might suggest that these patients have a propensity to overproduce these inflammatory mediators but whether this results from greater disease activity or contribute to greater disease activity remains moot.     

Leakage evaluation of three different root canal obturation techniques using electrochemical evaluation and dye penetration evaluation methods

The aim of this study was to compare the apical sealing ability of three different obturation techniques using an electrochemical evaluation and evaluating dye penetration. One hundred and thirty-two maxillary anterior teeth were randomly divided into six groups. There were 20 teeth in each group. The teeth were obturated as follows: Groups 1 and 4 with Thermafil, Groups 2 and 5 with System B, and Groups 3 and 6 with cold lateral condensation (CLC). The apical leakage in these groups was evaluated using an electrochemical method for Groups 1, 2, 3 and a dye penetration method for Groups 4, 5 and 6. In both methods, the lowest mean leakage values were observed for Thermafil and the highest were observed for the CLC groups. The difference between Thermafil and CLC was statistically significant (P < 0.05). In both groups, System B fillings showed moderate leakage and the difference was not significant with Thermafil and CLC groups.     

The role of lymphocyte subsets and adhesion molecules in T cell-dependent cytotoxicity mediated by CD3 and CD28 bispecific monoclonal antibodies

The cure of human Hodgkin's tumors heterotransplanted into SCID mice can be achieved by two bispecific monoclonal antibodies (Bi-mAb) directed against the tumor-associated CD30 antigen and CD3 and CD28, respectively, and normal peripheral human blood T cells. We investigated the role of lymphocyte subsets and adhesion molecules in this Bi-mAb-mediated cytolysis. CD4⁺ lymphocytes were the most rapidly expanding subpopulation, but Bi-mAb-directed cytotoxicity was mediated preferentially by CD8⁺ lymphocytes and effector cells belonging to the CD45RO⁺ “memory” pool. Blocking of the LFA-1/ICAM-1 or CD2/LFA-3 adhesion pathways by mAb decreased Bi-mAb-mediated cytotoxicity. This was not due to inhibition of aggregate formation between Bi-mAb-coated T lymphocytes and target cells. Cross-linking of LFA-1 or CD2 molecules on lymphocytes prestimulated with Bi-mAb bound to CD3 and CD28 antigen lead to a more pronounced and prolonged rise in the intracellular concentration of free Ca²⁺. Additional CD2 cross-linking resulted in the tyrosine phosphorylation of distinct proteins. These findings indicate that adhesion molecules play a critical role and function as co-stimulatory signals rather than as cellular contact mediators in CD3 and CD28 Bi-mAb-stimulated T lymphocytes.     

Early Contribution of Pericytes to Angiogenic Sprouting and Tube Formation

Immunostaining with endothelial and pericyte markers was used to evaluate the cellular composition of angiogenic sprouts in several types of tumors and in the developing retina. Confocal microscopy revealed that, in addition to conventional endothelial tubes heavily invested by pericytes, all tissues contained small populations of endothelium-free pericyte tubes in which nerve/glial antigen 2 (NG2) positive, platelet-derived growth factor beta (PDGF beta ) receptor-positive perivascular cells formed the lumen of the microvessel. Perfusion of tumor-bearing mice with FITC-dextran, followed by immunohistochemical staining of tumor vasculature, demonstrated direct apposition of pericytes to FITC-dextran in the lumen, confirming functional connection of the pericyte tube to the circulation. Transplantation of prostate and mammary tumor fragments into NG2-null mice led to the formation of tumor microvasculature that was invariably NG2-negative, demonstrating that pericytes associated with tumor microvessels are derived from the host rather than from the conversion of tumor cells to a pericyte phenotype. The existence of pericyte tubes reflects the early participation of pericytes in the process of angiogenic sprouting. The ability to study these precocious contributions of pericytes to neovascularization depends heavily on the use of NG2 and PDGF beta -receptor as reliable early markers for activated pericytes.     

DNA end-independent activation of DNA-PK mediated via association with the DNA-binding protein C1D

DNA-dependent protein kinase (DNA-PK), which is involved in DNA double-strand break repair and V(D)J recombination, is comprised of a DNA-targeting component termed Ku and an ~465-kD catalytic subunit, DNA-PK_cs. Although DNA-PK phosphorylates proteins in the presence of DSBs or other discontinuities in the DNA double helix in vitro, the possibility exists that it is also activated in other circumstances via its association with additional proteins. Here, through use of the yeast two-hybrid screen, we discover that the recently identified high affinity DNA binding protein C1D interacts with the putative leucine zipper region of DNA-PK_cs. Furthermore, we show that C1D can interact with DNA-PK in mammalian cells and that C1D is a very effective DNA-PK substrate in vitro. Finally, we establish that C1D directs the activation of DNA-PK in a manner that does not require DNA termini. Therefore, these studies provide a function for C1D and suggest novel mechanisms for DNA-PK activation in vivo.     

Correlation between putative indicators of primary restless legs syndrome severity

BACKGROUND AND PURPOSE: Several methods of assessing disease severity in restless legs syndrome (RLS) have been suggested. The purpose of this study was to examine the relationship between the suggested immobilization test (SIT), the International RLS Study Group rating scale (IRLS), sleep efficiency, and periodic leg movements of sleep index (PLMI). PATIENTS AND METHODS: Forty primary RLS patients with periodic leg movements of sleep were included in this prospective study. Study procedures were all performed during the same night, beginning with IRLS administration and following with SIT and polysomnography (PSG) evaluations, in that order. SIT was composed of two parameters: SIT mean discomfort score (SIT-MDS) and SIT periodic leg movements of wakefulness index (SIT-PLMW). PSG target measures were PLMI and sleep efficiency. Pearson's correlation was used for analysis at a P<0.01 significance level. RESULTS: PSG-PLMI correlated with IRLS (r=0.462; P=0.003) and with SIT-PLMW (r=0.681; P=0.0004). A correlation was also found between IRLS and SIT-MDS (r=0.447; P=0.004), even though SIT-PLMW and IRLS did not correlate with each other (P=0.286). A negative correlation was found between PSG-PLMI and sleep efficiency (r=-0.435; P=0.005). Neither SIT nor IRLS correlated with sleep efficiency. Only SIT discomfort scores from the second half of SIT correlated with SIT-PLMW (r=0.457, P=0.004), and they had a stronger correlation with IRLS (P=0.003). CONCLUSIONS: This study attempted a much needed comprehensive evaluation of the relationship between various RLS severity indicators. Our findings support a strong role of motor dysfunction on sleep quality in RLS, as well as the potential use of SIT-PLMW as a sensitive indicator of RLS severity.