BACKGROUNDThe prevalence of dermatophytoses and the development of new antifungal agents has focused interest on susceptibility tests of dermatophytes. The method used universally for susceptibility tests of dermatophytes was published as document (M38-A) in 2002 by the Clinical and Laboratory Standards Institute (CLSI), dealing with the standardization of susceptibility tests in filamentous fungi, though not including dermatophytes especially. However, it is not a very practical method for the clinical laboratory in routine susceptibility testing. In this test, we developed a novel rapid susceptibility assay-glucose consumption method (GCM) for dermatophytes.

METHODSIn this study, we investigated the antifungal susceptibilities of dermatophytes to itraconazole (ITC), voriconazole (VOC), econazole nitrate (ECN) and terbinafine (TBF) by glucose consumption method (GCM), in comparison to the Clinical and Laboratory Standards Institute (CLSI) M38-A method. Twenty-eight dermatophyte isolates, including Trichophyton rubrum (T. rubrum) (n = 14) and Trichophyton mentagrophytes (T. mentagrophytes) (n = 14), were tested. In the GCM, the minimum inhibitory concentrations (MICs) were determined spectrophotometrically at 490 nm after addition of enzyme substrate color mix. For the CLSI method, the MICs were determined visually.

RESULTSComparison revealed best agreement for TBF against T. mentagrophytes and T. rubrum, since MIC range, MIC50, and MIC90 were identical from two methods. However, for ITC and VOC, GCM showed wider MIC ranges and higher MICs than CLSI methods in most isolates. For ECN against T. rubrum, high MICs were tested by GCM (0.125-16 microg/ml) but not M38-A method (0.5-1 microg/ml). The overall agreements for all isolates between the two methods within one dilution and two dilutions for ITC, VOC, ECN and TBF was 53.6% and 75.0%, 57.1% and 75.0%, 82.1% and 89.3%, and 85.7 and 85.7%, respectively.

CONCLUSIONMeasurement of glucose uptake can predict the susceptibility of T. rubrum and T. mentagrophytes to ECN and TBF.

Antifungal Agents ; pharmacology ; Econazole ; pharmacology ; Glucose ; metabolism ; Itraconazole ; pharmacology ; Microbial Sensitivity Tests ; Naphthalenes ; pharmacology ; Pyrimidines ; pharmacology ; Triazoles ; pharmacology ; Trichophyton ; drug effects ; metabolism ; Voriconazole

OBJECTIVETo establish a murine transplant model for bone marrow purging of metastatic breast cancer and to explore the efficiency of Econazole (Ec) as a purging agent.

METHODSMixtures of TSA /Neo breast cancer cells and murine bone marrow cells were transplanted into lethally irradiated mice following purging with Econazole or saline in vitro. The recipient mice were monitored for hematopoietic engraftment, appearance of metastatic nodules in lungs and the overall survival.

RESULTSAll the mice receiving i.v. injection of TSA cells developed metastatic lung nodules. The hematological recovery was not delayed in mice transplanted with Ec purged bone marrow. More importantly, metastatic lung nodules were not seen in Ec treated group and the overall survival was improved.

CONCLUSIONThe purged metastatic breast cancer cell bone marrow transplant model was easily established and reproducible. Ec could be used to purge the bone marrow grafts contaminated with breast cancer cells.

Animals ; Antineoplastic Agents ; pharmacology ; Bone Marrow Purging ; Bone Marrow Transplantation ; Cell Line ; Econazole ; pharmacology ; Female ; Mammary Neoplasms, Experimental ; pathology ; Mice ; Mice, Inbred BALB C

Diverse signaling pathways have been proposed to regulate store-operated calcium entry (SOCE) in a wide variety of cell types. However, it still needs to be determined if all of these known pathways operate in a single cell type. In this study, we examined involvement of various signaling molecules in SOCE using human fibroblast cells (HSWP). Bradykinin (BK)-stimulated Ca2+ entry, previously shown to be via SOCE, is enhanced by the addition of vanadate, an inhibitor of tyrosine phosphatases. Furthermore, SOCE is regulated by cytochrome P-450, as demonstrated by the fact that the products of cytochrome P-450 activity (14,15 EET) stimulated SOCE while econazole, an inhibitor of cytochrome P450, suppressed BK-stimulated Ca2+ entry. In contrast, Ca2+ entry was unaffected by the guanylate cyclase inhibitor LY83583, or the membrane permeant cyclic GMP analog 8-bromo-cyclic GMP (8-Br-cGMP). Neither nitric oxide donors nor phorbol esters affected BK-stimulated Ca2+ entry. SOCE in HSWP cells is primarily regulated by tyrosine phosphorylation and the cytochrome P-450 pathway, but not by cyclic GMP, nitric oxide, or protein kinase C. Thus, multiple pathways do operate in a single cell type leading to the activation of Ca2+ entry and some of these signaling pathways are more prominently involved in regulating calcium entry in different cell types.
Vanadates/pharmacology ; Tetradecanoylphorbol Acetate/pharmacology ; Protein-Tyrosine-Phosphatase/*metabolism ; Phosphotyrosine/metabolism ; Phosphorylation/drug effects ; Nitric Oxide/metabolism ; Humans ; Fibroblasts ; Epidermal Growth Factor/pharmacology ; Enzyme Inhibitors/pharmacology ; Econazole/pharmacology ; Cytochrome P-450 Enzyme System/antagonists & inhibitors/*metabolism ; Cyclic GMP/analogs & derivatives/metabolism ; Cells, Cultured ; Calcium Channels/*metabolism ; Calcium/metabolism ; Bradykinin/pharmacology

The development of selective and safe antifungal agents are relatively delayed, compared to that of other antibiotics. The reasons are the relatively lesser interest of pharmaceutical companies because of the fewer occurrence of fungal disease and the apparent lack of a highly selective fungal target, not present in other eukaryotic (including mammalian) cells. Until the 1940s, fungal skin infection was treated by keratinolytics, antiseptics, and antibacterial chemicals. The first selective antifungal agent was polylene compounds in the early 1950s, which were topical nystatin and fungizone (amphotericin-B). In 1958, the first oral fungal agents, 'griseofulvin', as developed and have been used effectively to tinea capitis and other dermatophytes. Between the late 1960s and early 1970s, the azole compound, 'the real broad spectrum antifungal agents' was introduced, and clotrimazole was the first topical azole compound followed, by miconazole and econazole. Ketoconazole was released in early 1980s and it was the first real oral antifungal agent for systemic and superficial fungal infections. However, because of serious side effects of symptomatic hepatic injury, its use was replaced by triazole antifungal agents such as itraconazole and fluconazole. Triazole was more safe and effective, and caused advancement in the treatment of onychomycosis. In addition, terbinafine 'belonging to the allylamine compounds and developed in 1984', has been approved as a very potent antifungal agent for dermatophytes and also is being used widely to cutaneous infection by candidia species and some molds.
Allylamine ; Amphotericin B ; Anti-Bacterial Agents ; Anti-Infective Agents, Local ; Antifungal Agents* ; Arthrodermataceae ; Clotrimazole ; Danazol ; Dermatomycoses* ; Econazole ; Fluconazole ; Fungi ; Itraconazole ; Ketoconazole ; Miconazole ; Nystatin ; Onychomycosis ; Skin ; Tinea Capitis

To investigate apoptosis of mouse leukemia cell (WEHI-3) induced by econazole and its mechanism, apoptosis induced by econazole was examined by flow cytometry, while free calcium ([Ca(2+)]i) was determined by Fura-2 fluorescein load technique. The protein was isolated from endoplasmic reticulum of WEHI-3 cells, and then the expression of caspase-12 and caspase-7 was evaluated by Western blot. The results showed that WEHI-3 exhibited typical change of apoptosis when it was treated by econazole, [Ca(2+)]i was significantly higher in comparison with the control. The expression of caspase-12 and caspase-7 enhanced as the econazole concentration increased. In conclusion, econazole can induce WEHI-3 cell apoptosis and the caspase-12 plays a key role in this process.
Animals ; Apoptosis ; drug effects ; Blotting, Western ; Calcium ; metabolism ; Caspase 12 ; metabolism ; Caspase 7 ; metabolism ; Cell Line, Tumor ; Dose-Response Relationship, Drug ; Econazole ; pharmacology ; Flow Cytometry ; Leukemia ; metabolism ; pathology

BACKGROUND: The epidemiology of Candida species isolated from nonsterile as well as normally sterile sites is important because colonization of the former may precede invasive Candida infections. METHODS: We investigated the epidemiology and antifungal susceptibility of Candida species recovered in Busan Paik Hospital during the past 6 years and compared these results according to the type of specimens. RESULTS: Among the 2364 strains, C. albicans (53.8%) was the most frequently isolated, followed by C. tropicalis (17.5%), and C. guilliermondii (10.0%). Non-albicans Candida species were more prevalent in normally sterile sites (P<0.001); the prevalence of C. tropicalis and C. parapsilosis was significantly higher in normally sterile than in nonsterile sites (P<0.001). The prevalence of C. parapsilosis was higher in blood, intravenous catheter tips, and ear discharge, whereas C. tropicalis was more frequently isolated from urine. C. guilliermondii was the most frequently isolated from bronchial washings. The susceptibilities of Candida species to 5-flucytosine, amphotericin B, nystatin, miconazole, econazole, and ketoconazole were 98.3, 99.3, 99.7, 94.9, 86.3, and 94.5%, respectively. The susceptibilities of the organisms from normally sterile sites were lower than those from nonsterile sites. CONCLUSION: The distribution of Candida species differed among various types of specimens, especially those from normally sterile versus nonsterile sites. We assume that the frequency of infections of exogenous origin is high. We presume that the candidemia of C. parapsilosis is associated with the use of central venous catheter and that C. parapsilosis is acquired from exogenous sources.
Amphotericin B ; Busan ; Candida* ; Candidemia ; Catheters ; Central Venous Catheters ; Colon ; Ear ; Econazole ; Epidemiology ; Ketoconazole ; Miconazole ; Nystatin ; Prevalence

BACKGROUND: The frequency of nosocomial bloodstream infections by Candida species has risen dramatically in the past two decades, and a noticeable shift in the species of Candida causing bloodstream infection toward non-albicans species has occurred. Also, the isolation frequency of Candida species are influenced by patient type, nation and region, study period, and investigators. The aim of this study is to investigate the isolation rates and antifungal susceptibility of Candida species isolated from blood cultures at Wonju Christian Hospital during the recent four years (1997~2000). METHODS: For one-hundred twenty-seven isolates of Candida species from blood cultures, we analyzed the isolation frequency by year, age/sex, and department. Identification of yeasts was done by germ tube test and ATB ID 32 C kit. Antifungal susceptibilities to flucytosine, amphotericin B, nystatin, miconazole, econazole, and ketoconazole were determined by ATB FUNGUS. RESULTS: The isolation rates of Candida species in decreasing order were C. albicans (44.9%), C. parapsilosis (21.3%), C. glabrata (14.2%), and C. tropicalis (9.5%). The isolation rates of Candida species by year were as follows; C. albicans decreased from 61.5% in 1997 to 33.3% in 2000; C. tropicalis decreased from 23.1% in 1997 to 5.0% in 2000; C. parapsilosis increased from 0% in 1997 to 30.8% in 2000; and C. glabrata increased from 7.7% in 1997 to 18.0% in 2000. Of 127 Candida species, all but one isolates were susceptible to amphotericin B. CONCLUSION: This data showed that the candidemia by C. albicans and C. tropicalis are decreasing trend, and candidemia by C. parapsilosis and C. glabrata are increasing trend in recent four years.
Amphotericin B ; Candida* ; Candidemia ; Econazole ; Flucytosine ; Fungi ; Gangwon-do ; Humans ; Ketoconazole ; Miconazole ; Nystatin ; Research Personnel ; Yeasts

BACKGROUND: We thought that nitroimidazoles including metronidazole had been overused empirically for treatment of trichomoniasis in Korea. But there were not any reports about in vitro-drug susceptibility and distribution of resistant strains of Trichomonas vaginalis up to date. Therefore, we made an experiment in order to observe the susceptibility of clinical isolates of T. vaginalis to a variety of antiprotozoal agents. METHODS: Twenty-six strains of T. vaginalis isolated from 217 patients afflicted with the increased vaginal discharge were tested by Meingassner's microtiter plate method in newly devised anaerobic box, in which anaerobic and microaerobic conditions were more easily manipulated. The agents used in this study for testing the minimal lethal concentration (MLC) to the clinical isolates were as follows; nitroimidazoles, doxycycline, Zinc sulfate and gentian violet as chemotherapeutic agents and povidone-iodine as vaginal cleansing agents were studied. RESULTS: In anaerobic culture, according to anaerobic resistance cut-point (minimal lethal concentration >3.1 microgram/mL) proposed by M ller etc., metronidazole (MTZ)-, tinidazole (TNZ)-and ornidazole (ONZ)-resistant strains were four (4/26, 15.4%), two (2/26, 7.7 %) and two (2/26, 7.7%) strains, respectively. Among these resistant strains, two strains (G7 and G16) were resistant to two drugs and one strain (G20) resistant to three drugs concomitantly. Their resistance range was narrow as 6.25~12.5 microgram/mL. MLC of clotrimazole was > or = 2,000 microgram/mL in all strains, econazole was as high as 62.5~250 microgram/mL and miconazole was also high as 62.5~> or = 2,000 microgram/mL. In microaerobic culture (O2 concentration <5%), all strains showed lower MLC to MTZ, TNZ and ONZ than >100 microgram/ mL (aerobic resistance cut-point proposed by M ller etc.). MLC of doxycycline ranged 62.5 to 250 microgram/mL both in microaerobic and anaerobic conditions. All strains of T. vaginalis growed well in 3,000 microgram/mL of povidone-iodine. 22 strains (84.6%) among 26 T. vaginalis strains showed MLCs of 3.5 mM~7.0 mM to zinc sulfate. Gentian violet showed 15.6~62.5 microgram/mL of MLC. CONCLUSION: In absolute anaerobic culture, 4 strains (15.4%) among 26 T. vaginalis strains were resistant to metronidazole. But these 4 strains were not resistant in microaerobic culture depending on Miler's aerobic resistance cut-point (>50~100 microgram/mL), the value decided in normal O2 pressure. Vaginal PO2 is 0~28mm Hg (median 1 mmHg) at healthy or trichomonas-infected women. Therefore, we think that his aerobic resistance cut-point value is hard to be available in microaerobic condition and microaerobic resistance guide-line is to be established newly.
Anti-Infective Agents ; Antiprotozoal Agents* ; Clotrimazole ; Detergents ; Doxycycline ; Econazole ; Female ; Gentian Violet ; Humans ; Korea ; Metronidazole ; Miconazole ; Nitroimidazoles ; Ornidazole ; Povidone-Iodine ; Tinidazole ; Trichomonas vaginalis* ; Trichomonas* ; Vaginal Discharge ; Zinc Sulfate

Angiotensin II (ANG II) has a biphasic effect on Na+ transport in proximal tubule: low doses of ANG II increase the Na+ transport, whereas high doses of ANG II inhibit it. However, the mechanisms of high dose ANG II-induced inhibition on Na+ uptake are poorly understood. Thus the aim of the present study was to investigate signal transduction pathways involved in the ANG II-induced inhibition of Na+ uptake in the primary cultured rabbit renal proximal tubule cells (PTCs) in hormonally defined serum-free medium. ANG II (10-9 M)-induced inhibition of Na+ uptake was blocked by losartan (10-8 M, AT1 antagonist), but not by PD123319 (10-8 M, AT2 antagonist) (P < 0.05). ANG II-induced inhibition of Na+ uptake was also completely abolished by neomycin (10-4 M, PLC inhibitor), W-7 (10-4 M, calmodulin antagonist), and AACOCF3 (10-6 M, PLA2 inhibitor) (P < 0.05). ANG II significantly increased (3H)arachidonic acid (AA) release compared to control. The ANG II-induced (3H)AA release was blocked by losartan, AACOCF3, neomycin, and W-7, but not by PD123319. ANG II-induced (3H)AA release in the presence of extracellular Ca2+ was greater than in Ca2+-free medium, and it was partially blocked by TMB-8 (10-4 M, intracelluar Ca2+ mobilization blocker). However, in the absence of extracellular Ca2+, it was completely blocked by TMB-8. In addition, econazole (10-6 M, cytochrome P-450 monooxygenase inhibitor) and indomethacin (10-6 M, cyclooxygenase inhibitor) blocked ANG II-induced inhibition of Na+ uptake, but NGDA (10-6 M, lipoxygenase inhibitor) did not affect it. In conclusion, PLA2-mediated AA release is involved in ANG II-induced inhibition of Na+ uptake and is modulated by (Ca2+)i in the PTCs.
Angiotensin II ; Angiotensins* ; Arachidonic Acid* ; Calcium* ; Calmodulin ; Cytochrome P-450 Enzyme System ; Econazole ; Indomethacin ; Kidney ; Lipoxygenase ; Losartan ; Neomycin ; Prostaglandin-Endoperoxide Synthases ; Signal Transduction

Chronic dermatophyte infection rarely fails to respond to topical or systemic antifungal therapy. Such refractory condition relates to many factors and one of them is the decreased response of delayed type hypersensitivity. A plausible mechanism by which the delayed hypersensitivity response may cause dermatophyte inhibition has been proposed already. Our patient had skin rashes for 6 years. It was diagnosed as tinea corporis and treated with various systemic antifungal agents, such as griseofulvin, itraconazole, fluconazole, terbinafine and topical forms of econazole and terbinafine. But the skin lesions did not resolve completely and reaggravated frequently. Numerous verrucae planar were found on face, neck and both extremities. Trichophyton rubrum was identified by fungus culture study. Laboratory examination showed no response against multi-CMI test, DPCP sensitization and prick test for trichophytons. We challenged the therapy with the combined antifungal agents and immune stimulatory drugs. This case is thought to be a chronic dermatophyte infection due to the defects in the both cell mediated immunity and immediate type hypersensitivity which is crucial for the host defence mechanisms against fungal infection.
Antifungal Agents* ; Arthrodermataceae* ; Econazole ; Exanthema ; Extremities ; Fluconazole ; Fungi ; Griseofulvin ; Humans ; Hypersensitivity ; Hypersensitivity, Delayed ; Immunity, Cellular ; Itraconazole ; Neck ; Skin ; Tinea ; Trichophyton ; Warts