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.
Experimental tests were conducted to study the behavior of skirted foundations rested on dry medium sandy soil subjected to vertical and inclined loads. To achieve this goal, a small-scale physical model was designed and performed which contained an aluminum circular footing (100 mm) in diameter and (10 mm) in thickness and skirts with different heights, local medium poorly graded dry sand is placed in a steel soil container (2 mm) thick with internal dimensions (1000 mm x 1000 mm in cross section and 800 mm in height). The main objective of this study was to evaluate the response of skirt attached to the foundation at different skirt (L/D) ratios (0.0, 0.5, 1.0 and 1.5) and is subjected to point load at different angles of inclinat
... Show MoreManagement of Foreign Exchange Rate Exposure by Using Financial Hedging An Analytical Empirical Study The main purpose of this Research is to investigate the ability to reduce the effect of exchange rate fluctuation on firm value , by usage appropriate hedging strategies to provide the firms force to adopted with complex and highly uncertainty conditions , characteristic of the most of the financial markets . The field of this study is the giant five Multinational on the world. Nokia, Toyota Motor, Intel , Coca Cola, Microsoft. practical analysis is provide the truth of all study's hypothesis , and it is reach to many of conclusion, the most important of them is Stem from unexpected fluctuation on nominal ex
... Show MoreIn this study, the CR-39 detector technique was used, to estimate the uranium concentration from the soil in midland refineries Company (Doura refine (, Baghdad, Iraq. Uranium concentrations in soil samples have been measured using solid state nuclear track detector type CR-39. Nine soil samples were collected from different areas within the Doura refinery and other soil samples were collected form Abu Tayara Street and ALshortaa District outside the refinery for comparison. The results showed variable values for uranium concentrations. The average value of uranium concentration was found to be 0.37 ppm in doura refinery. For areas outside the refinery, the concentration of uranium was 0.008 ppm. Thes
... Show MoreThis paper presents an experimental study of cooling photovoltaic (PV) panels using evaporative cooling. Underground (geothermal energy) water used to extract heat from it during cooling and cleaning of PV panels. An experimental test rig was constructed and tested under hot and dusty climate conditions in Baghdad. An active cooling system was used with auxiliary an underground water tank to provide cold water as a coolant over both PV surfaces to reduce its temperature. The cellulose pad has been arranged on the back surface and sprays cooling on the front side. Two identical PV panels modules used: without cooling and evaporative water cooling. The experiments are comprised of four cases: Case (I): backside cooling, Ca
... Show MoreSynthesis three organic inhibitors for carbon steel corrosion: 2-(propylthio)-1H-benzo[d]imidazole (PTBI), 2-(allylthio)- 1H-benzo[d]imidazole (ATBI) and 2-(prop-2-ynylthio)-1H-benzo[d]imidazole (YTBI) were prepared from reaction of 2-mercapto benzimidazole with different alkyl halide. The melting point and TLC were used to confirm the purity of the inhibitors as well as using the [FTIR, 1H-NMR and 13C-NMR] for the identify structures. The synthesized inhibitors were examined by potentiostatic polarization measurement as corrosion inhibitors of carbon steel in acidic media [1M H2SO4 ].The polarization measurement results showed that the mixed type inhibitors. In addition, the efficiency of inhibitors (YTBI) were studied at different con
... Show MoreResults of the current study demonstratedthat out of eighty-three isolatesof Pseudomonas aeruginosa,only twenty-five isolateswere resistant to five different antibiotics (of different classes) that were consequentlyconsideredmultidrug resistant isolates.These isolates developed variable susceptibility toward Eucalyptuscamaldulensisleavesoil (ECO). GC-MS analysis of ECOrevealed that the aromatic oil eugenol is the major constituent.However, the most frequent MIC was 0.39 µg/ml, while the lowest frequent MIC was 3.125 µg/ml.Moreover, this oil at ½ MIC (0.195µg/ml) increased the gene expression of exoU. Itis concluded from the outcomes of the studythat ECOmay cause severe damagewhen used to treat infections caused by P. aeruginosa.
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