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.
This work represents the preparation of the starting material, 3-chloro-2-oxo-1,4-dithiacyclohexane (S) using a new method. This material was reacted with, 4-phenylthiosemicarbazide to give (H3NS3) as a tetradentate ligand H3L. New complex of rhenium (V) with this ligand of the formula [ReO(L)] was prepared. New complexes of the general formula [M(HL)] of this ligand when reacted with some metal ions where: M = Ni(II), Cu(II), Cd(II), Zn(II), Hg(II) have been reported. The ligand and the complexes were characterized by infrared, ultraviolet–visible, mass, 1H nuclear magnetic resonance and atomic absorption spectroscopic techniques and by (HPLC), elemental analysis, and electrical conductivity. The proposed structure for H3L with Re (V) i
... Show MoreThe reaction of 2-amino benzoic acid with 1,2-dichloroethane under reflux in methanol and KOH as a base to gave the precursor [H4L]. The precursor under reflux and drops of CH3COOH which reacted with (2mole) from salicycaldehyde in methanol to gave a new type N2O4 ligand [H2L], this ligand was reacted with (MCl2) Where [M= Co (II), Ni(II), Cu(II) and Zn(II)] in (1:1) ratio at reflux in methanol using KOH as a base, to give complexes of the general formula [M(L)]. All compounds have been characterized by spectroscopic methods [1H NMR ( just to the ligand), FTIR, uv-vis, atomic absorption], melting point, conductivity, chloride content, as well as magnetic susceptibility measurements. From the above data, the proposed molecular structu
... Show MoreA specific, sensitive and simple method was used for the determination of: vitamin B9 (Folic acid) in pure and pharmaceutical formulations using continuous flow injection analysis. The method is based on formation of ion pair compound between folic acid and ammonium molybdate in an aqueous medium to obtain a gray precipitate complex, using homemade; Ayah-6SX1-ST-2D solar cell CFI Analyzer. Optimum parameters was studied to increase the sensitivity for developed method. The linear range for the calibration graph was 0.01-0.6 mMol.L-1 of vitamin B9 and LOD was 131.994 ng/sample with correlation coefficient ( r ) of 0.9810, RSD% was lower than 0.1%, (n=9) for the determination of vitamin B9 at concentration (0.07and 0.5) mMol.L-1 respectiv
... Show MoreDisease responses of eight wheat cultivars , Saber Beg. , Abu-Ghraib 3, Mexipak , Tamoz 2,Tamoz 3 , IPA 95 ,IPA 99 and Tahadi which were grown in four different sowing date , 25 th October , 19th September , 14th December and 8 January , to leaf rust disease caused by Puccinia recondita were investigated under natural infection conditions at the experimental farm , College of Agriculture , Abu-Ghraib, during the growing season of 1997-1998.Results of this study revealed that IPA 95, IPA 99 and Tahadi showed moderate resistant reaction, while Tamuz 3 was moderateley susceptable . Abu-Ghraib , Saber Beg, Tamuz 2 and Mexipak showed susceptible yeaction to the causal agent . The first sowing date was not suitable for disease progress in compars
... Show MoreThe aim of the research is to know the effect of a training program based on interactive teaching strategies on achievement and creative problem solving among fourth-grade students in chemistry of the directorate of education Rusafa first, the sample was divided into two groups, one experimental and numbering (29) students and the other control group numbering (30) students. The experimental group underwent the training program in the first semester of the year (2021-2022) and the control one studied according to the usual method. Two tools were built, the first being an academic achievement test consisting of (40) multiple-choice items, and the second a test of creative problem-solving skills in a chemistry subject and consisting o
... Show MoreThe industrial sector of the important drivers of economic activity sectors due to having the capabilities and the potential to qualify biggest contribution to the economic development of the country, which requires the availability of a higher degree of safety in industrial organizations, and the preservation of the lives of employees and workers in emergency situations, and that all the work is not without its dangers, which produces in a year of uncertainty in the possible events get in the future, therefore, reduce the dangers that could be exposed to industrial organizations and management are at the heart of the strategic direction which works to stimulate thinking about the future and respond to the changes that occur in theextern
... Show MoreBACKGROUND: Acute appendicitis (AA) remains a complex case even for experienced surgeons. Rate of negative appendectomy is 5–40% and delayed intervention result in perforated appendicitis in 5–30% of cases. AIM: The aim of the study was to evaluate NPT as a marker for the diagnosis of AA concerning its severity. And compare the diagnostic value of it with the ALV scoring system. METHODS: One hundred twenty patients presented with signs and symptoms of AA and underwent appendectomy, only 84 patients proved to be AA by histopathological examination, were included in the study. Blood samples for neopterin (NPT) estimation and Alvarado (ALV) score was calculated. Control group consists of 45 healthy individual. RESULTS: NPT levels were s
... Show MoreBackground/Objectives: The purpose of this study was to classify Alzheimer’s disease (AD) patients from Normal Control (NC) patients using Magnetic Resonance Imaging (MRI). Methods/Statistical analysis: The performance evolution is carried out for 346 MR images from Alzheimer's Neuroimaging Initiative (ADNI) dataset. The classifier Deep Belief Network (DBN) is used for the function of classification. The network is trained using a sample training set, and the weights produced are then used to check the system's recognition capability. Findings: As a result, this paper presented a novel method of automated classification system for AD determination. The suggested method offers good performance of the experiments carried out show that the
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