Partial cDNA cloning of a putative membrane transporter protein gene expressed in Sporothrix schenckii and DNA polymorphism of the isolated gene are described here. DNA fragments were isolated from S. schenckii, and the deduced amino acid sequence from one of the fragments contained a region homologous to the conserved sequence of the membrane transporter protein family. 188-bp fragments encoding the homologous region were amplified from many strains of S. schenckii, and were subjected to polymerase chain reaction single strand conformation polymorphism (PCR-SSCP) analysis. The results demonstrated that the strains of S. schenckii were divided into three groups, and several base substitutions among these groups were observed. This finding agreed with the classification of S. schenckii strains based on the mitochondrial DNA diversity, because the three groups were clearly located on the branches of the phylogenetic tree constructed by digestion profiles of mitochondrial DNA with restriction enzymes. The correlation of the results of PCR-SSCP analysis with the mitochondrial DNA diversity might indicate linkage of the mutation of the membrane transporter protein gene with the evolution of S. schenckii, suggesting the protein plays an essential role in S. schenckii.
A 38-year-old Japanese AIDS patient delveloped papular lesions which rapidly increased in number, eroded and crusted, and spread over not only skin but also the mucosal surface. High fever, sore throat, malaise and hepatosplenomegaly were also noted, and he died despite 2 months of intensive treatment. An autopsy revealed numerous histiocytes infected with Penicillium marneffei in the lymph nodes, liver, spleen, bone marrow, skin, and mucosal surface of the oral cavity to the pharynx. This case is thought to be the first Japanese case of penicilliosis marneffei.
A patient treated with itraconazole (ITCZ) under the diagnosis of Aspergillus flavus-induced chronic hypertrophic pachymeningitis is presented. The reason for the successful cure of this patient was investigated by the pharmacokinetic analysis of serum levels of ITCZ. Concurrently administered digoxin was also investigated for its drug-drug interaction. The patient (a 75-year-old male) developed ophthalmopathy, and was diagnosed as having A. flavus hypertrophic pachymeningitis by pachymeninx biopsy. After admission, he was treated with FLCZ, AMPH, 5-FC and MCZ. The infection tended to subside with the AMPH administration. Since renal insufficiency was induced by AMPH and the other antifungal drugs were ineffective, daily administration of 200mg of ITCZ was initiated, and the inflammatory signs and symptoms gradually subsided. The symptoms did not recur during the 36 months of itraconazole treatment after discharge, and it was concluded that ITCZ was effective for A. flavus hypertrophic pachymeningitis. Pharmacokinetic parameters of ITCZ and OH-ITCZ as follows: ITCZ: Cmax 93.2ng/ml, T1/2β 11 hours, AUC0-24 999ng·h/ml, OH-ITCZ: Cmax 159.4ng/ml, T1/2β 16.2 hours, AUC0-24 of 1391ng·h/ml. Both ITCZ and OH-ITCZ reached steady states seven days after administration began. The ITCZ and OH-ITCZ levels in serum collected 36 months after the initiation of administration were 452.9ng/ml and 1233.6ng/ml, respectively. Cmax and AUC0-24 of ITCZ and OH-ITCZ on the second day were markedly lower than those in healthy adults reported by Oguchi et al., and hypoalbuminemia observed at administration on that day was considered the most probable cause. It was assumed that the most plausible reason for a successful cure even at a low dose of ITCZ was the increase of distribution to tissue by the increase of the unbound form. Digoxin was concurrently given to this patient at 0.125mg/day, but the blood digoxin level was not elevated. Consideration of the blood level of albumin is believed to be important for evaluating the blood concentration of ITCZ.
Aspergillus fumigatus is an opportunistic pathogen, especially in an immunocompromised host. This fungus grows in a hyphal form in infected tissues; therefore, new tests to examine hyphal susceptibility are needed. In this study, we measured the mycotic activity of miconazole (MCZ) contained in human serum against A. fumigatus using the BioCell-Tracer method. Three serum samples were obtained from the same patient who was injected with 600mg b. i. d. MCZ daily for 2 days. The concentrations of MCZ in the serum sample were 8.8, 3.5, and 1.6μg/ml, respectively. The serum containing 8.8μg/ml of MCZ inhibited hyphal growth 90 minutes after administration, and the hypha stopped growing. The serum containing 3.5μg/ml MCZ stopped hypha growth 100 minutes after administration, but re-growth of the hypha was observed at this concentration of MCZ. Serum containing 1.6μg/ml did not inhibite hyphal growth, nor did control serum have any inhibitory activity foward hyphae. Based on these results, we conclude that the BioCell-Tracer is a useful method for determining the effects on filamentous fungi of antifungal agents in the serum.