In July 2007, information on the current status of mycological examinations in Japanese university clinics and requests for a university network system was gathered by a questionnaire completed by 98 of 112 professors or...In July 2007, information on the current status of mycological examinations in Japanese university clinics and requests for a university network system was gathered by a questionnaire completed by 98 of 112 professors or directors of those clinics. A summary of the findings follows: only 9% of the hospitals performed fungal culture studies for all or most cases, indicating a drop of 7% from 2000 (16% in 2000, reported by Kasai) . Also, just 55% of relevant departments maintained the ability to identify most or all clinical isolates in-house, which down 27% from 2000 (83% in 2000, reported by Kasai). These findings indicate that mycological diagnoses by many departments have been rapidly decreasing. Eighty-nine percent of respondents indicated a desire for some help from the university network including: basic training in medical mycology of young doctors (senior residents in university hospitals); aid in diagnosing rare fungal infections; and support in obtaining knowledge and new technologies. The basic training of senior residents in each department is not possible in the network system so each department is required to take responsibility for training their own. However, a project of an advanced course at the leader level, which educates the person in charge of each department, may be possible in the network system. Employing this would make it realistic for the educated leaders to train the senior residents in their departments.
Fungal keratitis is one of the most challenging types of microbial keratitis for the ophthalmologist to diagnose and treat. Fungi causing human keratitis take the form of either yeasts or mold. Candida, the major pathoge...Fungal keratitis is one of the most challenging types of microbial keratitis for the ophthalmologist to diagnose and treat. Fungi causing human keratitis take the form of either yeasts or mold. Candida, the major pathogenic yeasts, can be detected in the normal ocular surface flora. Preceding ocular surface disorder, the wearing of contact lenses and the use of antibiotic/steroid eye drops may lead to candida keratitis. Infectious focus caused by Candida tends to melt the corneal stroma. Keratitis caused by mold often develops after an injury caused by soil and/or a plant. Mold can reach the anterior chamber without destroying the stromal layer of the cornea, which results in distinctive clinical features such as endothelial plaque and hyphate ulcer. Fungal keratitis needs to be managed by antifungal agents, most of which must be prepared by ourselves to apply to the ocular surface. Candida keratitis should be managed with azoles. If the infection seems to be caused by mold, several antifungal drugs including pimaricin, which is the only agent officially applicable to the eye, should be used. Some cases of mold keratitis need to have therapeutic penetrating keratoplasty because of their lack of response to intensive medication. Mold causing keratitis is variegated. Fusarium and Aspergillus can reach the intraocular space rapidly. Alternaria and some other unclassified molds remain in the superficial layer of the cornea for a long time. Our experiments indicate that the progress of focus in the cornea is regulated by the receptiveness of mold against temperature.
We provide foot and nail care services to outpatients at our dermatology clinic in Saiseikai Kawaguchi General Hospital. Patients with dermatological foot problems such as onychogryphosis, onychomycosis, and ingrown nail...We provide foot and nail care services to outpatients at our dermatology clinic in Saiseikai Kawaguchi General Hospital. Patients with dermatological foot problems such as onychogryphosis, onychomycosis, and ingrown nails are recommended by dermatologists to obtain foot and nail care services performed by a specially trained nurse. These services include a footbath, foot massage, nail clipping, and corn and nail filing. If ingrown nails exist, a small piece of cloth is placed under the nail edge.
In recent years fungi have been flourishing in immunocompromised patients of tertiary care centers. The data on the burden of opportunistic mycoses in India is not clear though the climate in this country is well suited...In recent years fungi have been flourishing in immunocompromised patients of tertiary care centers. The data on the burden of opportunistic mycoses in India is not clear though the climate in this country is well suited for a wide variety of fungal infections. There are very few good diagnostic mycology laboratories and clinicians are still not aware of the emerging trends. Within the limited data available, an increased incidence of invasive candidiasis, aspergillosis, and zygomycosis are reported. The emergence of fungal rhinosinusitis, penicilliosis marneffei and zygomycosis due to Apophysomyces elegans is unique in the Indian scenario. Invasive candidiasis is the most common opportunistic mycosis. The global change in spectrum of Candida species is also observed in India; however, the higher prevalence of candidemia due to Candida tropicalis instead of C. glabrata or C. parapsilosis is interesting. Invasive aspergillosis is the second contender. Though due to difficulty in antemortem diagnosis the exact prevalence of this disease is not known, high prevalence is expected in Indian hospitals where construction activities continue in the hospital vicinity without a proper impervious barrier. The other opportunistic mycosis, invasive zygomycosis is an important concern as the world's highest number of cases of this disease is reported from India. The infection is commonly observed in patients with uncontrolled diabetes mellitus. Though antiretroviral therapy in AIDS patients has been introduced in most Indian hospitals, no decline in the incidence of cryptococcosis and penicilliosis has yet been observed. Thus there is need of good diagnostic mycology laboratories, rapid diagnosis, and refinement of antifungal strategies in India.
The incidence of invasive aspergillosis (IA) , which is commonly caused by Aspergillus fumigatus, has increased recently in immunocompromised patients and has become the common cause of death in these patients. Antifunga...The incidence of invasive aspergillosis (IA) , which is commonly caused by Aspergillus fumigatus, has increased recently in immunocompromised patients and has become the common cause of death in these patients. Antifungal resistance is one of the reasons for treatment failure. Since the first itraconazole-resistant A. fumigatus was reported in 1997, the reports on clinical strains of triazole-resistant A. fumigatus have increased, as well as studies on the resistant mechanisms. In this paper, the known molecular mechanisms of antifungal resistance in Aspergillus, especially in A. fumigatus, are reviewed.
Sequence information of 25S rRNA gene was useful for the genotype determination in Candida albicans. Genotypes of 301 C. albicans strains by this single PCR method were determined, and out of them, two strains of new gen...Sequence information of 25S rRNA gene was useful for the genotype determination in Candida albicans. Genotypes of 301 C. albicans strains by this single PCR method were determined, and out of them, two strains of new genotype (genotype E) which are closely related to C. dubliniensis in its intron structure were discovered. Analysis of internal transcribed spacer (ITS) region sequence including 5.8S rRNA region in three varieties of C. neoformans was found to be an useful method for genotype determination, and a new genotype (Africa genotype) of var. gattii was discovered. In comparison with other traditional taxonomic methods in pathogenic fungi, usefulness of these genotype determination methods in their epidemiological studies was discussed. New PCR identification systems which were developed based on RAPD band pattern analyses for Histoplasma capsulatum, Paracoccidioides brasiliensis and Penicillium marneffei were also introduced. These genetic studies lead to a development of new DNA microarray identification method, and their usefulness was discussed.
Dimethyl sulfoxide (DMSO) is commonly used as a solvent for antifungal drugs. Earlier the author has reported the inhibitory effect of DMSO on the growth of many strains of dermatophytes' colonies in dermasel agar and pr...Dimethyl sulfoxide (DMSO) is commonly used as a solvent for antifungal drugs. Earlier the author has reported the inhibitory effect of DMSO on the growth of many strains of dermatophytes' colonies in dermasel agar and proposed that this could cause the variations between results of different studies for the evaluation of the activities of antifungal drugs. In studies regarding the determination of the effect of antifungal drugs on the germination of arthrospores of dermatophytes it was observed that relatively higher concentrations of DMSO were being used as a solvent for the antifungal drugs, the final concentration in the media being 5%. Therefore, the present study was aimed at determining the effect of different concentrations of DMSO (1.25 to 10%) on the growth of germ tubes of arthrospores of Trichophyton mentagrophytes and Candida albicans, in glucose peptone broth. With DMSO 10% there was a negligible growth of germ tubes of both the arthrospores and yeast; between 2.5 and 7.5% there was a rather linear dose-related inhibitory effect; whereas 1.25% had insignificant effect from controls. The present study shows that besides other factors, variations in the results of the susceptibility tests of antifungal drugs might occur due to the effect of DMSO on the growth of fungi and differences in the final concentration of DMSO in the medium.
Pseudallescheria boydii is a ubiquitous filamentous fungus. We report a case of cutaneous P. boydii infection of the left knee in a 79-year-old Japanese man who was receiving oral predonisolone (25 mg/day) for radiation...Pseudallescheria boydii is a ubiquitous filamentous fungus. We report a case of cutaneous P. boydii infection of the left knee in a 79-year-old Japanese man who was receiving oral predonisolone (25 mg/day) for radiation pneumonitis after radiation therapy on left breast cancer. He presented with a 2-week-history of a lesion on the left knee. A biopsy specimen from the skin lesion revealed granulomatous inflammation with hyphae. Culture of the pus from the skin specimen confirmed the diagnosis of cutaneous P. boydii infection. rDNA ITS sequence was analyzed to confirm the mycological diagnosis. The patient was treated orally with 200 mg/day of itraconazole. The lesion was gradually cured and left a hypertrophic scar. Cutaneous injury may be responsible for an incidence of localized infection. Such rare fungus infection in immunocompromised patients who have a persistent traumatic skin ulcer needs to be ruled out. An opportunistic infection in immunocompromised patients can be life-threatening and prompt treatment based on accurate diagnosis is important.
As there is not yet a standardized in vitro susceptibility test of micafungin (MCFG), we evaluated the methods of such testing, focusing on the judgment method of MIC, based on the National Committee for Clinical Laborat...As there is not yet a standardized in vitro susceptibility test of micafungin (MCFG), we evaluated the methods of such testing, focusing on the judgment method of MIC, based on the National Committee for Clinical Laboratory Standards (NCCLS) M27-A2, M38-A, the proposed standards of The Japanese Society for Medical Mycology (JSMM) for yeast (JSMM-Y) and for filamentous fungi (JSMM-F) against Candida spp. and Aspergillus spp. The judgment of MIC value was performed spectrophotometrically and visually in both (NCCLS and JSMM) assays. Only the spectrophotometric MIC judgment against Aspergillus spp. in the NCCLS assay used two end points: 80% inhibitory concentration (IC80) of the growth control and 50% inhibitory concentration (IC50). The end point for the visual judgment against Aspergillus spp. in the NCCLS assay was determined to be no growth from the small clumps of altered hyphae in the microtiter plate. The other MIC judgments used an IC80 end point. The MICs of MCFG for Candida spp. were </= 0.0039-1mug/ml in both judgments by the two assays. In contrast, there was a large difference of the MIC values against Aspergillus spp. between the NCCLS assay results using the IC80 end point and the visual judgment (> 4mug/ml and 0.0078-0.0313mug/ml). However, the MICs using the IC50 end point and those by JSMM assay agreed with the result of the visual assessment. Therefore, we recommend the JSMM assay, the NCCLS assay using the IC50 end point or the novel visual judgment for the susceptibility testing of MCFG against Aspergillus spp.
The genotypes and phenotypes of 77 isolates derived from ascospores produced from two genetically different Arthroderma benhamiae were studied. Specifically, mitochondrial DNA (mtDNA) type, nuclear DNA (nDNA) type, matin...The genotypes and phenotypes of 77 isolates derived from ascospores produced from two genetically different Arthroderma benhamiae were studied. Specifically, mitochondrial DNA (mtDNA) type, nuclear DNA (nDNA) type, mating type, colony texture, growth rate, urease activity, red pigmentation and hair perforation were examined. The nDNA types based on the restriction fragment length polymorphism (RFLP) in the internal transcribed spacer (ITS) region of ribosomal RNA (rRNA) genes and mating types were inherited from parents independently. mtDNA type was inherited from only one parent. All the phenotypes, except hair perforation and mating type, showed great variations. Those seemed not to be a conclusive factor for species identification. Additionally, these characteristics appeared in variable combinations suggesting that they are not interrelated. The intensity of red pigmentation varied even within a colony, implying that it is not a strain-specific characteristic. Hair perforation was observed in isolates of all but one atypical strain, and therefore could be one characteristic of this species.
An experimental study was conducted between January 2002 and April 2003 for the detection of delayed hypersensitivity to Fonsecaea pedrosoi metabolic antigen (chromomycin) in skin tests. A total of 194 subjects were atte...An experimental study was conducted between January 2002 and April 2003 for the detection of delayed hypersensitivity to Fonsecaea pedrosoi metabolic antigen (chromomycin) in skin tests. A total of 194 subjects were attended by spontaneous demand at the Infectious and Parasitic Diseases outpatient clinic of the Federal University of Maranhão-UFMA and at the Department of Microbiology, Federal University of Minas Gerais-UFMG and classified into three groups: patients with chromoblastomycosis caused by F. pedrosoi (n=20), healthy subjects (n=86) and patients with other diseases (n=88). For the skin test, 0.1 ml of the antigen was applied to the anterior side of the right forearm and 0.1 ml Smith medium was applied to the anterior side of the left forearm as control. The results were analyzed 48 h after inoculation of the antigen and an induration >/= 5 mm was considered to indicate a positive test. A cellular immune response to chromomycin was detected in 18 (90.0%) of the 20 patients with chromoblastomycosis caused by F. pedrosoi, and one of the patients with a negative test had reactional leprosy. Eighty-five (98.8%) of the 86 healthy subjects presented a negative reaction and only one reacted positively to the antigen. The skin test was negative in all 88 (100%) patients with other diseases, such as dermatophytosis, paracoccidioidomycosis, pulmonary aspergilloma, candidiasis, pityriasis versicolor, tuberculosis, leprosy, tegumentary leishmaniasis and syphilis, and one case of chromoblastomycosis caused by Rhinocladiella aquaspersa. Chromomycin was effective in detecting delayed hypersensitivity in patients with chromoblastomycosis caused by F. pedrosoi, with a sensitivity and specificity of 90.0% and 98.8%, respectively. These results suggest that this antigen can be used in the auxiliary diagnosis of the disease and also in epidemiological studies for determination of the prevalence of chromoblastomycosis infection in endemic areas.
The biological properties of elastase inhibitor from Aspergillus flavus (AFLEI) were investigated. AFLEI was produced at the highest rate when casamino acid was used as the nitrogen source. When a mixture of AFLEI (appro...The biological properties of elastase inhibitor from Aspergillus flavus (AFLEI) were investigated. AFLEI was produced at the highest rate when casamino acid was used as the nitrogen source. When a mixture of AFLEI (approx. molecular weight, 7,500) and elastase from A. flavus (approx. molecular weight, 40,000) was detected using anti-AFLEI antibody, molecular weight of the detected mixture was approximately 48,000, indicating that AFLEI and elastase bound at a proportion of 1 : 1. When immunocompromised mice administrered of immunosuppressive (cyclophosphamide) were infected by inhalation of A. flavus and administered amphotericin B (AMB) alone or in combination with AFLEI, survival rate tended to be higher with combination treatment than with AMB alone. Moreover, although extensive bleeding was seen in pathology sections taken from rat lung resected 24 hr after purified elastase was administered to the lung via the bronchus, this bleeding was inhibited by AFLEI. These findings indicate that for the treatment of aspergillosis, combination of an existing antifungal agent with AFLEI can be expected to provide greater therapeutic benefits than administration of an antifungal agent alone.
A 72 year-old man was referred to our department with white curd-like material on the surface of his tongue as well as the mucosal surface of the lower lip, after unsuccessful treatment with itraconazole for 3 weeks. He...A 72 year-old man was referred to our department with white curd-like material on the surface of his tongue as well as the mucosal surface of the lower lip, after unsuccessful treatment with itraconazole for 3 weeks. He also had a history of depression and had received topical steroid and/or antibiotics treatment for persistent oral aphtha and irritation of his upper lip for 4 years. A diagnosis of oral candidiasis was made through positive KOH direct microscopic examination and he was instructed to rinse his oral mucosal lesion with amphotericin B syrup. Although no significant eruption was observed on his upper lip at his first visit, he applied the steroid ointment for 4 weeks and came back to our clinic with his upper lip red and swollen. It was also covered with yellow crusty material mixed with a pustule. Histological examination of the lips revealed non-specific chronic inflammation in the mid to lower dermis. Hyphae in the cornea detected by PAS and Grocott staining. KOH direct microscopic examination from the pustule and crust showed positive pseudohyphae although no sign of parasitism to the hair was seen. Candida albicans and Candida parapsilosis were detected by culture from the crust and a biopsy sample. He was successfully treated with 2 courses of pulse therapy of oral itraconazole for sycosis candidiasis, accompanied by 2% miconazole gel for oral candidiasis.
Measurement of (1-->3)-beta-D-glucan for invasive fungal infection is used practically in Japan. The problem of false positive results due to the frequent occurrence of non-specific reaction in alkaline treatment, chromo...Measurement of (1-->3)-beta-D-glucan for invasive fungal infection is used practically in Japan. The problem of false positive results due to the frequent occurrence of non-specific reaction in alkaline treatment, chromogenic automated kinetic assay to measure (1-->3)-beta-D-glucan had been recognized in Japan. But this important problem was resolved in July 2005 by improvement made in the pretreatment reagent in a (1-->3)-beta-D-glucan measurement kit. In this manuscript, we describe the process of improvement of this kit and its clinical usefulness.
Comparative studies of random amplified polymorphic DNA (RAPD) band patterns of Candida tropicalis with those of clinically important Candida species have shown the presence of specific RAPD bands for C. tropicalis. A ba...Comparative studies of random amplified polymorphic DNA (RAPD) band patterns of Candida tropicalis with those of clinically important Candida species have shown the presence of specific RAPD bands for C. tropicalis. A band specific to C. tropicalis strains (ca. 400 bp) was extracted and sequenced. It was found to belong to a fragment of the Trf4 gene, which is essential for growth of these strains and has a characteristic sequence of C. tropicalis. A PCR primer was designed specifically for C. tropicalis which amplifies the 324 bp band. The PCR primer amplified DNA products for all C. tropicalis strains tested, but did not amplify any PCR bands from C. albicans, C. dubliniensis, C. glabrata, C. guilliermondii, C. kefer, C. krusei, C. parapsilosis, or C. zeylanoides. Usefulness of the PCR primer in differentiating from clinical isolates of other fungal species is discussed.
Ability for growth support and species differentiation by colony features were compared on two commercial chromogenic agars, CHROMagar Candida and newly developed Pourmedia Vi Candida. Eleven strains (ten species) of sta...Ability for growth support and species differentiation by colony features were compared on two commercial chromogenic agars, CHROMagar Candida and newly developed Pourmedia Vi Candida. Eleven strains (ten species) of standard strains and twenty-four isolates (five species) of clinical strains were tested. All isolates were grown on both agar plates in 48 hours at 35 degrees C, however, several species had not matured in 22 hours. Color of the colonies for each strain were stable on both agars. The results show that Pourmedia Vi Candida is equivalent to CHROMagar Candida in its ability to differentiate species as a primary culture plate.
BACKGROUND: Infection with the anthropophilic fungus Trichophyton tonsurans has spread among members of combat sports clubs and has become a serious public health problem in Japan and other countries. Infection usually p...BACKGROUND: Infection with the anthropophilic fungus Trichophyton tonsurans has spread among members of combat sports clubs and has become a serious public health problem in Japan and other countries. Infection usually provokes only a weak inflammatory response, and treatment compliance tends to be poor. OBJECTIVE: To evaluate the hairbrush method and the treatment protocol described in the guidelines for T. tonsurans infection. METHOD: The study subjects were 69 individuals with positive hairbrush culture from among 327 members of 12 judo clubs participating in the survey. (a) Subjects with no more than 4 colonies by the hairbrush method were treated with miconazole nitrate shampoo. (b) Subjects with 5 or more colonies were treated with (1) itraconazole at a dose of 100 mg/day for 6 weeks or at a dose of 400 mg/day for 1 week, or (2) terbinafine at a dose of 125 mg/day for 6 weeks or at a dose of 500 mg/day for 1 week. Treatment efficacy was monitored by the hairbrush method at 1.5 and 3 months after treatment. RESULTS: Of the 46 subjects with 5 or more colonies isolated by the hairbrush method, 32 (69.6%) took itraconazole or terbinafine in compliance with their treatment schedules and were negative for T. tonsurans after treatment. Of the 23 subjects with 4 or fewer colonies, 15 (65.2%) were negative for T. tonsurans after treatment with miconazole nitrate shampoo. CONCLUSION: The treatment protocol seems promising, but poor compliance is a problem with the oral treatment regimens. The shampoo therapy is only partially effective, with 35% of subjects remaining positive for T. tonsurans after this therapy. In order to eradicate this disease, we have renewed the guidelines for T. tonsurans infection.
This paper reports on medical history from the end of the Edo period to the present and development of studies on infectious diseases, especially medical mycology including systemic fungal diseases. With the inflow of Du...This paper reports on medical history from the end of the Edo period to the present and development of studies on infectious diseases, especially medical mycology including systemic fungal diseases. With the inflow of Dutch studies at the end of the Edo period and the adoption of European, mainly German, medicine in the Meiji Restoration, Japanese medical studies gradually developed. However, evolution in the medical field as well as other scientific fields was prevented during the 2nd World War. After the War, there was marked progress in scientific fields and medical research made strong advances. In the past 20 years, basic fungal studies and clinical fungal diseases, especially clinical analysis, clinical diagnosis and treatment of systemic fungal infections have progressed. The level in this field is now equivalent to or higher than that in European countries. Further development is necessary, however, to relieve patients suffering from systemic fungal infections. Members of the Japanese Association of Medical Mycology must be leaders among international medical mycologists.