Periodontitis is one of the most prevalent bacterial diseases affecting man with up to 90% of the global population affected. Its severe form can lead to the tooth loss in 10-15% of the population worldwide. The disease is caused by a dysbiosis of the local microbiota and one organism that contributes to this alteration in the bacterial population is Prophyromonas gingivalis. This organism possesses a range of virulence factors that appear to contribute to its growth and survival at a periodontal site amongst which is its ability to invade oral epithelial cells. Such an invasion strategy provides a means of evasion of host defence mechanisms, persistence at a site and the opportunity for dissemination to other sites in the mouth. However, previous studies have demonstrated that invasion of the mammalian cells in a population by P. gingivalis is heterogenous, with some cells becoming heavily invaded while others harbour no or only a few bacteria. An understanding of this heterogeneity may throw light on the mechanisms involved and we hypothesised that the phase of the host cell cycle may explain this phenomenon. In an attempt to study the factors influencing P. gingivalis invasion and the cell response to that invasion, a standard antibiotic protection assay was employed and an oral keratinocyte cell line, H357. The results showed that P. gingivalis NCTC 11834 invasion was significantly increased with increasing time of exposure to the cells and the cell density. This may reflect an increased host cell surface area available for bacterial attachment. No effect on invasion of P. gingivalis invasion was observed by the bacterial growth phase, H357 cell passage number or whether cells were pre-incubated with P. gingivalis lipopolysaccharide. Epithelial cells did, however, respond to the presence of P. gingivalis in a number of ways. For example, the mRNA expression of endothelin-1 and urokinase receptor were upregulated with increasing P. gingivalis infection time, suggesting that these proteins could act as inflammatory mediators and possibly as useful markers of the severity of periodontal disease or in the diagnosis and treatment of periodontitis. iii Secondly, in an attempt to investigate the reason for the observed heterogeneous P. gingivalis invasion of H357 cell populations, the effect of cell cycle phase on P. gingivalis invasion was investigated. H357 cells were synchronized by serum starvation. On re-introduction of serum, characterisation of cell cycle phase distribution was performed by flow cytometry following staining with propidium idodide (PI) or by immunofluorescence using bromodeoxyuridine (BrdU), which specifically identifies cells in S-phase. The effect of cell cycle phases on P. gingivalis invasion was measured using the antibiotic protection assay, immunofluorescence and flow cytometry and these were correlated with gene and surface expression of the urokinase receptor and the α5-integrin subunit, which is thought to mediate P. gingivalis invasion. Results showed that the percentage invasion was enhanced with increasing serum re-introduction time, and positively correlated with the number of cells in S-phase. In addition, flow cytometry data showed that the highest association of fluorescent P. gingivalis was with PI positive S-phase cells. Moreover, BrdU positive S-phase cells were 3 times more likely to be invaded and contained 10 times more P. gingivalis than cells in other phases. Also, α5-integrin was more highly expressed in cells in S-phase than other phases, which could explain the mechanism underlying this enhanced invasion. Data presented here have suggested that P. gingivalis targeting of cells in S- phase could, in vivo, allow preferential invasion of the junctional epithelial cells which turns over rapidly. The data presented in this thesis suggest that P. gingivalis invasion is greatly dependent on several factors attributed to the host, the bacteria itself, and to the environment which the bacteria reside in. The invasion occurs within a population of host cells in a heterogeneous fashion, and is dependent on the cell cycle phase, specifically S-phase. This novel finding, in addition to the previously reported mechanisms of P. gingivalis invasion, increases our understanding of this virulence trait and suggests that such a strategy is a highly organised process which the bacteria can follow to ensure its survival within the host. Furthermore, knowledge of these mechanisms could provide novel approaches to treatment of periodontal diseases.
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Background: The anticancer impact of Epigallocatechin gallate (EGCG) the highly active polyphenol of green tea was abundantly studied. Though, the exact mechanism of its cytotoxicity is still under investigation. Objectives: Hence, the current study designed to investigate the molecular target of EGCG in HepG2 cells on thirteen autophagy- and/or apoptosis- related genes. Methods: The apoptosis detection analyses such as flow cytometry and dual apoptosis assay were used. The genes expression profile was explored by the real-time quantitative-PCR. Results: EGCG increases G0/G1 cell cycle arrest and the real-time apoptosis markers proteins leading to stimulate apoptos
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The reaction of 2, 4, 6-trihydroxyactophenonemonohydrate with hydrazine monohydrate was realized under reflux in methanol and a few drops of glacial acetic acid were added to give the (intermediate) 2-(1-hydrazono-ethyl)-benzene-1, 3, 5-triol, which reacted with salicylaldehyde in methanol to give a new type (NO) ligand [HL][(2-1-[(2-hydroxy-benzyliidene)-hydrazono]-ethyl) benzene-1, 3, 5-triol. The ligand was reacted with Mcl.(where M-Co, Ni, Cu, and Zn) under reflux in methanol with (l: 1) ratio to give complexes of the general formula [M (HL)]. All compounds have been characterized by spectroscopic methods I" H NMR, IR. UV-Vis, HPLC, atomic absorption] microanalysis along with conductivity measurement. From the above data the proposed mo
... Show MoreBackground: Diabetes mellitus is a major risk factor for chronic periodontitis (CP) and hyperglycemia has an important role in the enhancement of the severity of the periodontitis. It has been reported that the progression of CP causes shifting of the balance between bone formation and resorption toward osteoclastic resorption, and this will lead to the release of collagenous bone breakdown products into the local tissues and the systemic circulation. Cross-linked N-telopeptide of type I collagen (NTx) is the amino-terminal peptides of type I collagen which is released during the process of bone resorption. This study was conducted to determine the effects of nonsurgical periodontal therapy on serum level of NTx in type 2 diabetic patients
... Show MoreWe have investigated the photoemission and electronic properties at the PTCDI molecules interface on TiO2 and ZnO semiconductor by means of charge transition. A simple donor acceptor scenario used to calculate the rate for electron transfer of delocalized electronics in a non-degenerately TiO2 and ZnO electrodes to redox localized acceptors in an electrolytic. The dependent of electronic transition rate on the potential at contact of PTCDI with TiO2 and ZnO semiconductors, it has been discussion using TiO2 and ZnO electrodes in aqueous solutions. The charge transfer rate is determining by the overlapping electronic coupling to the TiO2 and ZnO electrodes, the transition energy, potential and polarity media within the theoretical scenario of
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Background/Objectives: Nonsurgical periodontal treatment (NSPT) is the gold-standard technique for treating periodontitis. However, an individual’s susceptibility or the inadequate removal of subgingival biofilms could lead to unfavorable responses to NSPT. This study aimed to assess the potential of salivary and microbiological biomarkers in predicting the site-specific and whole-mouth outcomes of NSPT. Methods: A total of 68 periodontitis patients exhibiting 1111 periodontal pockets 4 to 6 mm in depth completed the active phase of periodontal treatment. Clinical periodontal parameters, saliva, and subgingival biofilm samples were collected from each patient at baseline and three months after NSPT. A quantitative PCR assay was us
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The study aims to examine the emotional innovation, constructive thinking, foresight, and meditation among university students. Besides, attempts to explore the relationship between emotional innovation and constructive thinking, the relationship between emotional innovation and foresight, and meditation, the relationship between constructive thinking and foresight, and meditation, finally, the relationship among emotional innovation, constructive thinking, foresight, and meditation of university students regarding the variables of gender and major. A total of (400) students were selected as a sample for the study. The results revealed that university students have a high level of emotional innovation that induce them to take positive ro
... Show MoreThe current research aimed to identify psychological stability and its relationship to university integration and spiritual intelligence among university students. The research sample consisted of (158) students from the College of Education - Al-Mustansiriya University.
A scale was applied: psychological stability, university integration, and spiritual intelligence, and by using the (Pearson) correlation coefficient, and the t-test, the results showed: the sample members enjoy psychological stability, university integration, and spiritual intelligence, and there is a positive, statistically significant correlation between the research variables, and the results resulted in some recommendations and proposals.