Abstract Candida albicans is a commensal fungal pathogen that grows in yeast and hyphal forms in the human gut. C. albicans causes mucosal and cutaneous diseases that can result in significant mortality following systematic infections and it also exhibits drug resistance. Zebrafish have been an excellent model to investigate C. albicans infections because of their transparency and the availability of many transgenic lines. However, there is a limitation in using zebrafish as a model because the fish embryos cannot survive at 37°C therefore it is not suitable for studying Candida infections at physiological relevant human body temperature. In this thesis, the normal embryonic development of Arabian killifish (A. dispar) is investigated, revealing that embryogenesis was divided into 32 stages based on diagnostic patterns of development. A. dispar can also found to tolerate a wide range of temperatures and salinities. This suggests that A. dispar could be developed as a novel model to investigate host-pathogen interactions. The tolerance of A. dispar to high temperatures may in part be attributable to brown pigment cells with a highly fluorescent character that may have developed to allow the fish to adapt to live within extreme environmental conditions with strong sunlight and a wide range of temperatures (Chapter 3). In terms of Candida infections, this study examined A. dispar as a model to test C. albicans pathogenicity. The survival of A. dispar embryos following Candida infection showed a dose dependent relationship. We also found that A. dispar can survive longer than zebrafish after infection. Furthermore, C. albicans cells were observed to undergo a transition from yeast to hyphae at 37°C. An investigation of the ability of mutant strains of C. albicans with defects in cell wall mannosylation revealed a significant impact on virulence, host mortality, and the fishes’ immune response. The present study found that although the deletion of O- and N-mannan from the cell wall of C. albicans, affected fungal burden (attenuation), and the survival of the infected embryos per se was significantly decreased in the infections of the mutant strains compared to the WT. This data confirms the importance of the mannosylation state of the cell wall in triggering an immune recognition event (Chapter 4). A. dispar is also shown to be suitable for studying the effectiveness of 3 | P a g e antifungals. Fluconazole treatment of infected embryos and eggs promoted greater rates of survival at high doses, alongside a significant reduction of C. albicans CFUs (Chapter 4). When looking at the Candida-host interaction, we directly observed phagocytosed yeast cells within macrophages. Various detection methods were used to follow macrophages and neutrophils including Western blotting, immunostaining and histological staining (Sudan black and FITC-tyramide) allowing the monitoring of the time course of the immune cells. A biphasic response of macrophages was detected by L-plastin Western blotting, suggesting activation of two different type of macrophage: activated macrophage (M1) and alternative macrophage (M2). We also assayed reactive oxygen species (ROS) within infected embryos using a fluorescent probe (H2DCFDA), revealing the accumulation of the fluorescent probe at the sites of infection. Quantitative and qualitative analyses of the oxidative and immune response using the H2DCFDA and qPCR were also accomplished within A. dispar embryos after infection with both the WT and mutant strains of Candida albicans (WT, pmr1∆, mnt1-mnt2∆, and och1∆). The results confirmed that the mutant strains did not activate a host oxidative stress response nor immune cell accumulation when compared to WT, suggesting that the immune response is less activated against these mutants. Finally, a new transgenic line of A. dispar fish was developed using Betaactin-DsR-LoxP-GFP. The new transgenic A. dispar is suggested to be an ideal model for real time observation of host-pathogen interactions and for investigation of molecular functions of the immune response. Overall these results improve our understanding of the use of a new transparent fish model to study fungal pathogenesis and demonstrates the potential advantages of using this species in future studies of bacterial, fungal and viral pathogens at a physiologically relevant temperature for human infection. Such a model could lead us to investigate in more depth the key interactions between pathogens and their host and permit the screening and development of new antifungal therapies (that might target the pathogens directly or target the host immune system). View full metadata
Bioavailability is the objective for an optimum formulation. The target of the analysis is to maximize both the fluidity and disintegration profile of class II weakly compounds that are water-soluble. Anti-dyslipidemia drug rosuvastatin calcium (RC) (bioavailability 20%) through formulating as nanofibers (NFs) using electrospinning (ES) technology. Twenty formulas were prepared, and different polymers and polymer combinations with various concentrations were used such as polyethylene oxide (PEO) polyvinyl pyrrolidine (PVPK-30), and hydroxypropyl methylcellulose (HPMC). Three distinct groups of maximum parameters, including polymeric solution, electrospinning method, and ambient parameter, are capable of influencing the creation alon
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Numerous blood biomarkers are altered in COVID-19 patients; however, no early biochemical markers are currently being used in clinical practice to predict COVID-19 severity. COVID-19, the most recent pandemic, is caused by the SRS-CoV-2 coronavirus. The study was aimed to identify patient groups with a high and low risk of developing COVID-19 using a cluster analysis of several biomarkers. 137 women with confirmed SARS CoV-2 RNA testing were collected and analyzed for biochemical profiles. Two-dimensional automated hierarchy clustering of all biomarkers was applied, and patients were sorted into classes. Biochemistry marker variations (Ferritin, lactate dehydrogenase LDH, D-dimer, and C- reactive protein CRP) have split COVID-19 patien
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This study involved preparation of Graphene oxide (GO) and reduced graphene oxide (RGO) using Hummer method and chemical method respectively. These carbon nanomaterials were used as starting material to make novel functionalize with thiocarbohydrazide (TCH) which was prepared by reacting CS2 with hydrazine to form GO or RGO- 4-amino,5-substituted 1H,1,2,4 Triazole 5(4H) thion (ASTT) ,(GOT) and( RGOT) respectively via cyclocondensation reaction. Also MnO2 nanorod was prepared to form hybridized with GOT and RGOT. A commercial multiwall carbon nanotube (MWCNT) and functionalization with carboxylic groups' (f-MWCNT) and its nanocomposite with GOT were also prepared. All carbon nanomaterials were characterized with different techniques such as
... Show MoreAbstract: The development of highly sensitive sensors has become an efficient field of research. In this work, an ArF Excimer laser of 193 nm with a maximum pulse energy of 275 mJ, 15 ns pulse duration and a repetition rate of 1 Hz is utilized to form a Laser Induced Periodic Surface Structures (LIPSS) of three different morphologies (nanochains, contours, grooves) on surface of CR39 polymer at a fluence range above the ablation threshold (250 mJ/cm2). The laser ablated polymer surface is then Surface Enhanced Raman Scattering (SERS) activated by deposition of a gold layer of 30 nm thickness. The capability of the produced substrate for surface enhanced Raman scattering is evaluated through thiophenol as an analyte molecule. It is observ
... Show MoreCertain cyclic amine derivatives of metronidazole via acetate spacer were prepared. Cyclic amines used are piperidine and piperazine to improve the physicochemical properties and reduce some of metronidazole side effects. This is believed to be done by modification of its structural features to get prodrugs with improved properties over that of metronidazole. The present work includes esterification of metronidazole with choroacetic acid, N-alkylation of the cyclic amines by the halogenated ester and characterization of their structures by spectral(UV and IR) and elemental(CHN)analysis.The melting points, degree of solubilities and partition coefficients were also determined. Both metronid
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Bladder cancer (BC) is the predominant malignant neoplasm in the urinary system and ranks as the tenth most prevalent malignant tumor worldwide. Compared with females, males displayed a four-fold more common incidence of bladder cancer. It mainly affects men. Bladder cancer is the fourth most prevalent neoplasm in males. The most important protein that makes up high density lipoprotein (HDL), ApoA-I apolipoprotein A1 is essential in regulating the right amount of cholesterol. Multiple inquiries have demonstrated that APOA1 plays a pivotal role in the progression, infiltration, and spread of tumors. Objectives. The objective of this study was to measure the level of urine to serum apolipoprotein A1 in patients suffering from bladder
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Background: Pelvic masses are common in women & can present at any age of woman life, it could be benign or malignant mass and may originate from gynecological organs like cervix, uterus, uterine adnexia, or from other pelvic organs like intestine, bladder, ureters, skeletal muscle, and bone.Objective: We attempted to determine the increasing of platelet counts(> 450.000 /micro liter) and CA125serum level (> 35 U/mL) as useful tools for predicting and confirming malignancy in gynecological pelvic mass.Patients and methods: A prospective unmatched hospital based case-control study carried out at Baghdad Teaching Hospital, about 126 women were enrolled in our study, divided into two groups 60 women were control group (free o
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Darifenacin hydrobromide (DH) is the more recent uroselective M3 receptor antagonist for treating uncomplicated overactive bladder (OAB). This study was aimed to formulate DH as fast dissolving buccal films (FDBFs) using a solvent casting method to enhance patient’s compliance.
Films were prepared by using polyvinyl alcohol (PVA) as a film forming polymer. Different types and concentrations of superdisintegrants (croscarmellose sodium, sodium starch glycolate, indion 414) were used to select the best formula by studying the physicochemical properties of the films, disintegration time (DT) and percent drug release.
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In this work preparation of antireflection coating with single layer of MgO using pulsed laser deposition (PLD) method which deposit on glass substrate with different thicknesses (90 and 100) nm annealed at temperature 500 K was done.
The optical and structural properties (X-ray diffraction) have been determined. The optical reflectance was computed with the aid of MATLAB over the visible and near infrared region. Results shows that the best result obtained for optical performance of AR'Cs at 700 shots with thickness 90 nm nanostructure single layer AR'Cs and low reflection at wavelength 550 nm.
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