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Comparison of Giemsa counts and ELISA for evaluation of in vitro P. carinii drug susceptibility testsDurkin, Michelle Marie January 1992 (has links)
This document only includes an excerpt of the corresponding thesis or dissertation. To request a digital scan of the full text, please contact the Ruth Lilly Medical Library's Interlibrary Loan Department (rlmlill@iu.edu).
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Sterol biosynthesis and sterol uptake in the fungal pathogen Pneumocystis cariniiJoffrion, Tiffany Michelle 12 April 2010 (has links)
No description available.
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Genetic investigations of pneumocystis jirovecii : detection, cotrimoxazole resistance and population structureRobberts, Frans Jacob Lourens 12 1900 (has links)
Thesis (PhD (Pathology. Medical Microbiology))--University of Stellenbosch, 2005. / Pneumocystis jirovecii is a significant contributor to the burden of disease in
immunocompromised patients. The polymerase chain reaction (PCR) is more
sensitive and specific than microscopy. Cotrimoxazole prophylactic breakthrough and
treatment failures have been reported, and associated with mutations at codons 55
and 57 of P. jirovecii dihydropteroate synthase (DHPS). No phylogenetic or
population genetic models have been successful in elucidating P. jirovecii
intraspecies strain relatedness.
Aims: 1) Compare detection rates of nine PCR techniques and immunofluorescence
microscopy (IF); 2) Determine the extent of co-infecting pathogens associated with
Pneumocystis Pneumonia (PcP); 3) Determine local P. jirovecii ITS1-5.8S-ITS2 rDNA
strain types, and model lineage evolution employing a coalescent-theory based
statistical parsimony network analysis; 4) Investigate the possible emergence of
cotrimoxazole-resistant strains
Methods: PCR was evaluated on clinical specimens employing: ITS nested; DHPS
single and nested; DHFR nested; major surface glycoprotein (MSG) heminested;
mitochondrial large subunit rRNA (mtLSUrRNA) single and nested; 18S rRNA onetube
nested, and real-time 5S rRNA PCR. Retrospective analysis of co-infecting
pathogens seen in PcP patients was conducted. ITS regions were amplified, cloned
and sequenced. Statistical parsimony was applied for coalescence based network
genotype analysis. DHPS genome walking was attempted and DHPS and DHFR
primer annealing sites explored. Amplified DHPS and DHFR genes were cloned and
sequenced.
Results: Most sensitive PCR technique was mtLSUrRNA nested followed by 5S realtime
PCR. A poor correlation exist between mtLSUrRNA PCR and IF. Review of
clinical records suggested a high rate of false-positive IF results. P. jirovecii was
detected in 4.3% M. tuberculosis-positive HIV-positive, and 2.5% M. tuberculosispositive
HIV-negative patients. P. jirovecii was detected in 45% HIV-negative patients. The most prevalent ITS type was Eg. Four new combinations: Eo, Je, Ge,
No; 11 new ITS1 and 13 new ITS2 sequences were identified. A new ITS2 type was
detected in three patients and designated u. More than one strain type was detected
in 15/19 patients. Analysis of 5.8SrDNA region revealed 13 clones containing 1-2
nucleotide polymorphisms. Of 85 mtLSUrRNA PCR-positive specimens, currently
employed primers amplified DHPS and DHFR genes from 53 and 27 specimens,
respectively. Newly designed DHPS primers increased detection in 3 / 28 previously
DHPS-negative mtLSUrRNA-positive specimens. Of 56 DHPS genes amplified and
sequenced, one contained the double mutation (Thr55Aa; Pro57Ser). DHFR
Ala67Val was detected in three specimens and a new DHFR genotype (Arg59Gly;
C278T) was demonstrated.
Conclusions: The study emphasises the need to evaluate PCR primers against local
strains. It is recommended that mtLSUrRNA PCR be performed in parallel to IF and
discordant results resolved with clinical evaluation. Co-infection with P. jirovecii and
M. tuberculosis occurs in South Africa, and treatment for both pathogens is
recommended when demonstrated by the laboratory. ITS genotyping employing
statistical parsimony network analysis suggests type Eg as major ancestral
haplotype, and supports recombination contributing to strain diversity worldwide.
DHPS mutations may signal emergence of resistance to cotrimoxazole in South
Africa, however, low sensitivity of primers limits surveillance efforts.
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Community acquired pneumonia in HIV and non-HIV infected patients presenting to a teaching hospital in KwaZulu-Natal : aetiology, distribution, and determinants of morbidity and mortality.Nyamande, Kennedy. January 2004 (has links)
No abstract available. / Thesis (M.D.)-University of KwaZulu-Natal, 2004.
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GROUP I INTRON-DERIVED RIBOZYME REACTIONSJohnson, Ashley Kirtley 01 January 2005 (has links)
Group I introns are catalytic RNAs capable of self-splicing out of RNA transcripts. Ribozymes derived from these group I introns are used to explore the molecular recognition properties involved in intron catalysis. New ribozyme reactions are designed based on the inherent ability of these ribozymes to perform site-specific nucleophilic attacks. This study explores the molecular recognition properties of group I intron-derived ribozyme reactions and describe a new ribozyme reaction involving molecular recognition properties previously not seen.We report the development, analysis, and use of a new combinatorial approach to analyze the substrate sequence dependence of suicide inhibition, cyclization, and reverse cyclization reactions catalyzed by a group I intron from the opportunistic pathogen Pneumocystis carinii. We demonstrate that the sequence specificity of these Internal Guide Sequence (IGS) mediated reactions is not high, suggesting that RNA targeting strategies which exploit tertiary interactions could have low specificity due to the tolerance of mismatched base pairs.A group I intron-derived ribozyme from P. carinii has been previously shown to bind an exogenous RNA substrate, splice-out an internal segment, and then ligate the two ends back together (the trans excision-splicing reaction). We now report that a group I intron derived ribozyme from the ciliate Tetrahymena thermophila can also perform the trans excision-splicing reaction, although not nearly as well as the P. carinii ribozyme.In addition, we discovered a new ribozyme reaction called trans insertion-splicing where the P. carinii ribozyme binds two exogenous RNA substrates and inserts one directly into the other. Although this reaction gives the reverse products of the trans excision-splicing reaction, the trans insertion-splicing reaction is not simply the reverse reaction. The ribozyme recognizes two exogenous substrates through more complex molecular recognition interactions than what has been previously seen in group I intron-derived ribozyme reactions. We give evidence for this new reaction mechanism composed of three steps, with intermediates attached to the ribozyme.
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Bioactivation of diacetyldapsone in cultured lung cellsNimbalkar, Dipali 01 January 2000 (has links)
Dapsone has been shown to be an effective agent against Pneumocystis carinii pneumonia, an opportunistic infection in AIDS patients. Oral administration of dapsone is associated with several adverse effects, including methemoglobinemia, hemolytic anemia and photosensitivity reactions. To reduce the adverse effects associated with oral dapsone, an alternative would be to administer the prodrug diacetyldapsone (DADDS) into the lung, which may be hydrolyzed to monoacetyldapsone and the active metabolite dapsone. The purpose of this investigation was to determine the effect of cyclodextrindiacetyldapsone (CD-DADDS) complex upon the cultured lung cells and whether or not cultured lung cells could activate DADDS into dapsone, the active metabolite. The effect of the CD-DADDS complex upon the growth of cultured CRL 7272 lung cells was assessed by the trypan blue dye exclusion technique. There was no significant reduction in cell number as compared to the control for incubations with three different concentrations of CD-DADDS complex. The amount of arylamine produced by hydrolysis was initially monitored by the Bratton-Marshall diazotization technique.
Only incubation with 0.01% DADDS in 1% CD showed a significant time dependent hydrolysis of DADDS over a period of 72 hours due to insensitivity of the assay method. Over the same period, cultured lung cells produced 1.65 μmoles of metabolite/106 cells. However interfering substances could contribute to this value. To provide additional evidence for hydrolysis and to quantitatively estimate the amount of dapsone, a more sensitive HPLC method was used. The results obtained from HPLC analysis demonstrated a significant concentration and time dependent increase in the amount of dapsone with 0.001% DADDS, 0.005% DADDS and 0.01% DADDS incubations, respectively. Over a period of 48 hours, 255ng of dapsone/ 106 cells was formed in an incubation containing 0.01% DADDS.
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Determination of Pentamidine Transfer in the in Vitro Perfused Human Cotyledon With High-Performance Liquid ChromatographyFortunato, Stephen J., Bawdon, Roger E. 01 January 1989 (has links)
Pentamidine is used to treat Pneumocystis carinii pneumonia. The incidence of this infection in pregnancy has paralleled the increasing incidence of acquired immunodeficiency syndrome in pregnancy. Using the in vitro bidirectionally perfused human placenta, we studied the transfer of pentamidine across the placenta. Pentamidine was added to the maternal circulation at therapeutic concentrations (2 wg/ml). No transfer of pentamidine was detectable with a newly devised high-performance liquid chromatography method sensitive to 0.05 wg/ml of pentamidine. Increasing the pentamidine concentration tenfold produced a low level of transfer to the fetal circuit. Fetal concentrations were far below maternal perfusate concentrations. Placental tissue levels were higher than media levels. These data are suggestive of minimal drug transfer to the fetus and significant concentration of the drug in placental tissue. (Am J Obstet Gynecol 1989;160:759-61.)
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Polyamines and Alveolar Macrophage Apoptosis during Pneumocystis PneumoniaLiao, Chung-Ping 01 October 2009 (has links)
Indiana University-Purdue University Indianapolis (IUPUI) / Pneumocystis pneumonia (PCP) is the leading opportunistic disease in immunocompromised individuals, particularly in AIDS patients. The alveolar macrophage (AM) is the major type of cell responsible for the clearance of Pneumocystis organisms; however, they undergo a high rate of apoptosis during PCP due to increased intracellular polyamine levels. This study examined the mechanism of this polyamine mediated apoptosis and investigated an alternative therapy for PCP by targeting this mechanism. The elevated polyamine levels were determined to be caused by increased polyamine synthesis and uptake. Increased polyamine uptake was found to be AM-specific, and recruited inflammatory cells including monocytes, B cells, and CD8+ T cells were found to be a potential source of polyamines. The expression of the antizyme inhibitor (AZI), which regulates both polyamine synthesis and uptake, was found to be greatly up-regulated in AMs during PCP. AZI overexpression was confirmed to be the cause of increased polyamine synthesis and uptake and apoptosis of AMs during PCP by gene knockdown assays. Pneumocystis organisms and zymosan were found to induce AZI overexpression in AMs, suggesting that the β-glucan of the Pneumocystis cell wall is responsible for this AZI up-regulation. In addition, levels of mRNA, protein, and activity of polyamine oxidase (PAO) were also found to be increased in AMs during PCP, and its substrates N1-acetylspermidine and N1-acetylspermine were found to induce its up-regulation. These results indicate that the H2O2 generated during PAO-mediated polyamine catabolism caused AMs to undergo apoptosis. Since increased polyamine uptake was demonstrated to be a pathogenic mechanism of PCP in this study, the potential therapeutic activity of five putative polyamine transport inhibitors against PCP was tested. Results showed that compound 44-Ant-44 significantly decreased pulmonary inflammation, organism burden, and macrophage apoptosis, and prolonged the survival of rats with PCP. In summary, this study demonstrated that Pneumocystis organisms induce AZI overexpression, leading to increased polyamine synthesis, uptake, and apoptosis rate in AMs and that targeting polyamine transport is a viable therapeutic approach against PCP.
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A pentecostal response to the challenges of HIV/AIDS in TumaholeSkhosana, Thabang Johannes 11 1900 (has links)
This dissertation is a challenge to the Pentecostal churches, particularly, the Apostolic Faith Mission Church in Tumahole, to take an action in meeting the challenges posed by HIV/AIDS. This disease, HIV/AIDS, is the latest enemy to human life that the nations are faced with. In the newspapers like Sowetan, there is an article almost daily about HIV and AIDS. In this dissertation, I have tried to show shocking figures of how this disease is spreading in Africa. The seriousness of the disease, unlike other diseases, is its in curability. The secular organisations are far ahead of the churches in as far as the relevant programmes on
combating HIV/AIDS are concerned. Despite these massive programmes, the disease is spreading like the wild fire. Deducing from this background, it is no longer the question of whether the Pentecostal churches have any role to play, but what specific role should the
church play in this challenge. In this challenging times, many people look at the church as one of the most important institute that would play a positive role in bringing hope to the hopeless. / Christian Spirituality, Church History and Missiology / M. Th. (Missiology (Urban Ministry))
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A pentecostal response to the challenges of HIV/AIDS in TumaholeSkhosana, Thabang Johannes 11 1900 (has links)
This dissertation is a challenge to the Pentecostal churches, particularly, the Apostolic Faith Mission Church in Tumahole, to take an action in meeting the challenges posed by HIV/AIDS. This disease, HIV/AIDS, is the latest enemy to human life that the nations are faced with. In the newspapers like Sowetan, there is an article almost daily about HIV and AIDS. In this dissertation, I have tried to show shocking figures of how this disease is spreading in Africa. The seriousness of the disease, unlike other diseases, is its in curability. The secular organisations are far ahead of the churches in as far as the relevant programmes on
combating HIV/AIDS are concerned. Despite these massive programmes, the disease is spreading like the wild fire. Deducing from this background, it is no longer the question of whether the Pentecostal churches have any role to play, but what specific role should the
church play in this challenge. In this challenging times, many people look at the church as one of the most important institute that would play a positive role in bringing hope to the hopeless. / Christian Spirituality, Church History and Missiology / M. Th. (Missiology (Urban Ministry))
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