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Infections with pneumocystis aspects of the intriguing relationship with its host /Beckers, Pieter Jozef Antonius, January 1984 (has links)
Thesis (doctoral)--Nijmegen, 1984.
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Pneumocystis jiroveci and respiratorey bacterial pathogens in cases of pneumonia at hospitals in Port ElizabethDu Plessis, Sarah Jane January 2008 (has links)
Pneumocystis jiroveci, Mycoplasma pneumoniae and Mycobacterium tuberculosis are respiratory pathogens associated with pneumonia, with increasing prevalence of Pneumocystis pneumonia (PcP) and tuberculosis (TB) in AIDS patients. Increased resistance of M. tuberculosis has emphasized the need for rapid susceptibility testing, such as flow cytometry. Sputum specimens (102) were assessed by PCR employing primers directed at the following genes: P. jiroveci: mitochondrial large subunit ribosomal RNA (mtLSUrRNA), dihydropteroate synthase (DHPS) and dihydrofolate reductase (DHFR), and for M. pneumoniae: 16S rRNA and P1 adhesin. Positive P. jiroveci samples were genotyped by PCR-SSCP (single-strand conformation polymorphism) targeting the: internal transcribed spacer region (ITS), intron of the nuclear 26S rRNA gene (26S), variable region of the mitochondrial 26S rRNA gene (mt26S) and β-tubulin gene (β-tub). Multi-drug resistant (MDR-TB) cultures grown in the presence and absence of four antibiotics (rifampicin, isoniazid, ethambutol and ofloxacin) were heat killed, stained with SYTO16 and Propidium Iodide and analysed using flow cytometry. Rifampicin resistance gene mutations were screened by PCR and DNA sequencing. Details of patient’s gender, age, HIV and M. tuberculosis status were provided by the hospitals. Women were seen to be at high risk for community-acquired P. jiroveci colonisation. Overall, prevalence of P. jiroveci was 55.1 percent (54/102 patients). P. jiroveci was mainly associated with HIV (25/102 P. jiroveci positive patients for which clinical data was available) and co-colonisation with M. tuberculosis was observed in 11 cases. Sequence analysis of DHPS and DHFR products found no resistance associated mutations. M. pneumoniae was detected in one patient. Four simple SSCP patterns were identified and there were no co-infections with other P. jiroveci strains. Nine M. tuberculosis samples [8 MDR-TB isolates (NHLS) and M. tuberculosis ATCC® 27294TM] were tested. There was a 53 percent (19 out of 36 tests) agreement of flow cytometry with the BACTEC MGIT 960. Mutations (at two specific codons, namely 516 and 531) in the rifampicin resistance-determining region (RRDR) of the rpoB gene were observed in eight M. tuberculosis isolates. Evaluation of methods for genotyping and drug susceptibility testing of PcP and TB are imperative for epidemiology and drug resistance studies, and impact on treatment protocols.
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Invasive yeast infections: understanding the current scene. / CUHK electronic theses & dissertations collectionJanuary 2011 (has links)
Abstract not available. / Hui, Mamie. / "Dec 2010." / Source: Dissertation Abstracts International, Volume: 73-04, Section: B, page: . / Thesis (M.D.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 209-236). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [201-] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
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Genetic investigations of pneumocystis jirovecii : detection, cotrimoxazole resistance and population structure /Robberts, Frans Jacob Lourens. January 2005 (has links)
Thesis (PhD)--University of Stellenbosch, 2005. / Bibliography. Also available via the Internet.
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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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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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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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Development of the direct fluorescent antibody method for identification of Pneumocystis carinii and diagnosis of pneumocystis carinii pneumonitisLim, Sook Kyung. January 1972 (has links)
Thesis (D.P.H.)--University of Michigan.
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Development of the direct fluorescent antibody method for identification of Pneumocystis carinii and diagnosis of pneumocystis carinii pneumonitisLim, Sook Kyung. January 1972 (has links)
Thesis (D.P.H.)--University of Michigan.
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