IN VITRO SUSCEPTIBILITY OF DERMATOPHYTES TO ANTIFUNGAL DRUGS: A COMPARISON OF TWO METHODS

Background. There is an evidence that some fungal strains are resistant to certain antimycotic drugs with resulting therapeutic failures. Unlike testing of antibacterial agents, the activity of these drugs against the given fungal strains is not tested routinely. The paper presented is an approach to such a procedure. Methods. The minimum inhibitory concentration (MIC) values obtained using the dilution method were compared with the diameters of growth inhibition zones using the disk-diffusion method. Seven antifungal drugs (graiseofulvin, natamycin, naftifine, clotrimazole, bifonazole, miconazole, ketoconazole) were tested with 80 fungal strains, including Trichophyton rubrum, T. mentagrophytes, Microsporum canis, and Epidermophyton floccosum. Results. The majority of fungal strains were inhibited by griseofulvin at concentrations not exceeding 10 mg/L. The least susceptible was T. mentagrophytes. The most effective drug was naftifine, and the least effective was natamycin. Naftifine inhibited 98.75% of the strains tested at a concentration not exceeding 1.0 mg/L. An increase of the MIC value was accompanied by a decrease in the growth inhibition diameter. The relation was statistically significant. Conclusions. T. rubrum strains are more susceptible to antifungal drugs than T. mentagrophytes. Naftifine proved to be the most effective drug in dermatophytoses. Natamycin is a useful drug against yeast-like fungi but not against dermatophytes. The disk-diffusion method of fungal susceptibility assessment yields data consistent with results obtained from the dilution method and is recommended for routine use because of its simplicity.     

Topical application of acidified nitrite to the nail renders it antifungal and causes nitrosation of cysteine groups in the nail plate

BACKGROUND: Topical treatment of nail diseases is hampered by the nail plate barrier, consisting of dense cross-linked keratin fibres held together by cysteine-rich proteins and disulphide bonds, which prevents penetration of antifungal agents to the focus of fungal infection. Acidified nitrite is an effective treatment for tinea pedis. It releases nitric oxide (NO) and other NO-related species. NO can react with thiol (-SH) groups to form nitrosothiols (-SNO). OBJECTIVES: To determine whether acidified nitrite can penetrate the nail barrier and cure onychomycosis, and to determine whether nitrosospecies can bind to the nail plate. METHODS: Nails were treated with a mixture of citric acid and sodium nitrite in a molar ratio of 0.54 at either low dose (0.75%/0.5%) or high dose (13.5%/9%). Immunohistochemistry, ultraviolet-visible absorbance spectroscopy and serial chemical reduction of nitrosospecies followed by chemiluminescent detection of NO were used to measure nitrosospecies. Acidified nitrite-treated nails and the nitrosothiols S-nitrosopenicillamine (SNAP) and S-nitrosoglutathione (GSNO) were added to Trichophyton rubrum and T. mentagrophytes cultures in liquid Sabouraud medium and growth measured 3 days later. Thirteen patients with positive mycological cultures for Trichophyton or Fusarium species were treated with topical acidified nitrite for 16 weeks. Repeat mycological examination was performed during this treatment time. RESULTS: S-nitrothiols were formed in the nail following a single treatment of low- or high-dose sodium nitrite and citric acid. Repeated exposure to high-dose acidified nitrite led to additional formation of N-nitrosated species. S-nitrosothiol formation caused the nail to become antifungal to T. rubrum and T. mentagrophytes. Antifungal activity was Cu(2+) sensitive. The nitrosothiols SNAP and GSNO were also found to be antifungal. Topical acidified nitrite treatment of patients with onychomycosis resulted in > 90% becoming culture negative for T. rubrum. CONCLUSIONS: Acidified nitrite cream results in the formation of S-nitrosocysteine throughout the treated nail. Acidified nitrite treatment makes a nail antifungal. S-nitrosothiols, formed by nitrosation of nail sulphur residues, are the active component. Acidified nitrite exploits the nature of the nail barrier and utilizes it as a means of delivery of NO/nitrosothiol-mediated antifungal activity. Thus the principal obstacle to therapy in the nail becomes an effective delivery mechanism.     

国产萘替芬体外抗真菌作用和对豚鼠皮肤真菌病的疗效观察

用国产萘替芬（Ｎａｆｔｉｆｉｎｅ）体外对２３株常见致病真菌及１２０株临床分离菌株的抑菌试验结果表明，萘替芬抑制红色毛癣菌、石膏样毛癣菌等皮肤真菌的效果较好、萘替芬的ＭＩＣ在０．０２～１．５６ｍｇ／Ｌ，咪康唑和克霉唑ＭＩＣ多在０．３９～３．１２ｍｇ／Ｌ，差异显著（Ｐ＜０．０１）；动物实验，１％萘替芬霜和１％萘替芬溶液剂在１０天内临床治愈率和平均治愈天数明显优于１％克霉唑软膏；与３％克霉唑软膏比较，除１０天内临床治愈率低于萘替芬外，其他则无明显差异。     

肟康唑抑菌试验及治疗271例股癣

肟康唑是一种新的广谱抗真菌药物,体外抑菌试验表明对红色毛癣菌、石膏样毛癣菌、石膏样小孢子菌、絮状表皮癣菌、白念珠菌、新型隐球菌、孢子丝菌等均有明显的抑菌作用.我们采用双盲法,用1%肟康唑霜剂与1%益康唑霜剂进行对照观察,结果表明肟康唑的二周治愈率为94.1%,益康唑为84.24%,二者经统计学处理有非常显著的意义.     

成人及儿童皮肤癣菌病患者红色毛癣菌的基因分型与药物敏感性分析

目的 探讨成人和儿童皮肤癣菌病临床分离菌株中红色毛癣菌基因型的差异，基因型与发病部位、基因型与药物敏感性的关系。方法 利用随机引物OPAA11 5′-ACCCGACCTC-3′，OPD18 5′-GAGAGCCAAC-3′分别对来自成人及儿童皮肤癣菌病的红色毛癣菌菌株进行任意引物PCR，根据产物电泳带型进行基因分型，采用微量液体稀释法对氟康唑、伊曲康唑、特比萘芬、酮康唑、利拉萘酯、布替萘芬、益康唑、联苯苄唑进行体外敏感性分析。结果 两组的主要致病菌都为红色毛癣菌，两种随机引物扩增出的红色毛癣菌不同株带型均稳定清楚，按随机引物 OPAA11的扩增结果，其基因型分为4型。成人中Ⅰa、Ⅲa型分别占41.94%，儿童中Ⅰa型为65.96%，基因型构成在两组人群中的差异有统计学意义（P < 0.05）；体癣、足癣中基因型的分布在两组间差异均有统计学意义（P < 0.01；P < 0.05）。甲癣和股癣中基因型的分布差异无统计学意义（P > 0.05）；各基因型对8种抗真菌药物的MIC值均较低，特比萘芬的MIC最低，氟康唑的MIC值相对较高，酮康唑和氟康唑对Ⅰa、联苯苄唑对Ⅱa的活性高于其他基因型，伊曲康唑对Ⅲa型的活性略低于其他基因型，差异有统计学意义（P < 0.01或P < 0.05）。结论 红色毛癣菌为南京及周边地区儿童及成人皮肤癣菌病的主要致病菌，成人感染以Ⅰa、Ⅲa型为主，儿童感染以Ⅰa型为主。8种抗真菌药物对各基因型均有较好的抗菌活性，但酮康唑、氟康唑、联苯苄唑、伊曲康唑、特比萘芬对各基因型的敏感性有差异。     

家兔饲养者两侧面颌部的难辨认癣1例

患者女,33岁。两侧面颌红斑瘙痒1年余。曾外用多种糖皮质激素霜治疗无效。刮取皮屑镜检见大量真菌菌丝,培养见棒形分隔大分生孢子、葡萄串状小分生孢子。尿素酶实验阳性、毛发穿孔实验阳性,鉴定为须癣毛癣菌。经内服特比萘芬胶囊和外用萘替芬酮康唑乳膏治疗28天后皮损消退。复查真菌阴性。     

Comparison of the in vitro antifungal activities of clotrimazole, miconazole, econazole and exalamide against clinical isolates of dermatophytes

The in vitro antifungal activities of clotrimazole, miconazole nitrate, econazole nitrate, and exalamide against 64 clinical isolates of various dermatophyte species obtained from out-patients seen at the Nippon Medical School Hospital were simultaneously compared using a liquid microculture method. The rank order of activity against the 64 isolates of dermatophytes was clotrimazole > econazole nitrate > miconazole nitrate > exalamide. For a given antimycotic, the minimal inhibitory concentration was affected by the dermatophyte species.     

Naftifine cream in the treatment of cutaneous candidiasis

Naftifine, a member of a new class of synthetic antifungal drugs, the allylamines, was evaluated for the treatment of cutaneous candidiasis. In a double-blind, parallel-group clinical trial, sixty patients with cutaneous candidiasis were randomly assigned to receive either naftifine cream 1 percent or its vehicle twice a day for three weeks. Two weeks after the end of therapy, 77 percent of the naftifine-treated patients were mycologically cured (negative results on potassium hydroxide preparations and culture) and had no clinically apparent disease, compared with 3 percent of the patients treated with vehicle (p less than 0.001). Side effects reported with naftifine cream were few and minor.     

Trichophyton species: use of volatile fingerprints for rapid identification and discrimination

BACKGROUND: Fungal infection of the skin is a common clinical problem, and laboratory confirmation of the diagnosis is important to ensure appropriate treatment. The identification of the species of fungus is also important, because different fungal species have different modes of transmission, and this may be of importance both in preventing re-infection and in avoidance of infection of others. OBJECTIVES: This study examined the potential of using volatile production patterns for the detection and discrimination between four Trichophyton species (T. mentagrophytes, T. rubrum, T. verrucosum and T. violaceum) in vitro on solid media and in broth culture. METHODS: Two different sensor array systems (conducting polymer and metal oxide sensors) were examined for comparing the qualitative volatile fingerprints produced in the headspace by these species over periods of 24-120 h. The relative sensitivity of detection of two of the species (T. mentagrophytes, T. rubrum) was determined for log 1 to log 7 inoculum levels over the same time period. RESULTS: The conducting polymer-based system was unable to differentiate between species based on volatile fingerprints over the experimental period. However, metal oxide-based sensor arrays were found to be able to differentiate between the four species within 96 h of growth using principal component analysis which accounted for approximately 94% of the data in principal components 1 and 2 based on the qualitative volatile production patterns. This differentiation was confirmed by cluster analysis of the data using Euclidean distance and Ward's linkage. Studies of the sensitivity of detection showed that for T. mentagrophytes and T. rubrum it was possible to differentiate between log 3, log 5 and log 7 inoculum levels within 96 h. CONCLUSIONS: This is the first detailed study of the use of qualitative volatile fingerprints for identification and discrimination of dermatophytes. This approach could have potential for rapid identification of patient samples, reducing significantly the time to treatment.     

A Clinical and Mycologic Study of Tinea Corporis and Pedis in Puerto Rico

A study of the causative agents of tinea corporis and pedis in Puerto Rico in 1982 disclosed four different dermatophytes from a total of 97 cases. Among the total of 49 fungus specimens collected in tinea corporis, the mycologic flora consisted of four different fungus species: Trichophyton rubrum, 42 isolates (85.7%); Epidermophyton floccosum, 4 (8.1%); Trichophyton mentagrophytes, 2 (4%); and Microsporum ferrugineum, 1 (2%). Among the total of 48 fungus specimens collected in tinea pedis, the mycologic flora consisted of 3 different fungus species as follows: T. rubrum, 35 isolates (72.9%); T. mentagrophytes 8 (16.6%); and E. floccosum, 5 (10.4%). Trichophyton rubrum caused 77 of 97 fungal isolates (79.3%). One surprising finding was the isolation of M. ferrugineum in a case of tinea corporis, documenting the first autochtonous case of this species in the island. This dermatophyte had previously been known only in Japan and adjacent areas of the Far East, Southern Europe, and Africa.     

Naftifine: a review

BACKGROUND: Naftifine is a topical allylamine that is effective and safe in the management of superficial dermatomycoses. Naftifine is fungicidal in vitro against a broad spectrum of dermatophyte fungi and provides good activity against Candida and Aspergillus species. It is also effective against gram-negative and gram-positive bacteria. OBJECTIVE: To provide a review of the pharmacologic properties and clinical efficacy of topical naftifine preparations. METHODS: A review of the medical literature was performed using PubMed (1965-2006) using the search term "naftifine." All available English-language articles discussing the pharmacology and clinical use of naftifine were reviewed for the article. RESULTS: Naftifine causes interruption of fungal ergosterol synthesis and accumulation of squalene in fungal organisms. Naftifine also has demonstrated anti-inflammatory properties such as a reduction in superoxide production and a reduction in polymorphonuclear leukocyte chemotaxis/endothelial adhesion. Naftifine has shown good efficacy and safety for a variety of conditions and is a useful treatment that provides both antifungal action and relief of inflammatory signs and symptoms. Few adverse events have been noted with naftifine use, the most frequent being mild and transient burning, stinging, or itching in the application area. CONCLUSION: Naftifine remains a reliable multifunctional agent for a variety of superficial infections.     

Treatment of tinea pedis with econazole nitrate cream

This study investigated the clinical effectiveness and the long-term relapse rate of the topical imidazole, econazole nitrate, in the treatment of tinea pedis. Econazole nitrate cream was applied by twenty-two patients with tinea pedis for twenty-eight consecutive days. Therapy was then discontinued and patients were evaluated to determine the long-term relapse rate. Nineteen (86 percent) of the twenty-two patients were considered cured at the end of twenty-eight days of treatment. At the follow-up examinations, 74 percent of the group considered to be clinically cured at the end of therapy remained clear for one month, 84 percent for two months, and 63 percent for three months after stopping active treatment. These data demonstrate that econazole nitrate is a highly effective topical agent in the treatment of tinea pedis and that clinical and mycologic cures in almost two thirds of the patients last for at least three months.     

Dermatophyte infections in hereditary palmo-plantar keratoderma. Frequency and therapy

The frequency of dermatophyte infections in hereditary palmo-plantar keratoderma ( HPPK ) of the Unna - Thost variety was investigated in 280 patients admitted to the Department of Dermatology, Central Hospital, Boden , during 1977-1981, and was found to be 35.0%. The distribution of fungi did not differ from that found for the total number of dermatophytes. An almost complete therapeutical resistance was found especially in Trichophyton rubrum infections, when patients were treated with micronized griseofulvin and topical econazole cream. Treatment of dermatophyte infections in HPPK with 50% propylene glycol in distilled water gave poor results but when 1% econazole nitrate was added negative cultures were found in 86.4% of the patients treated for 3 weeks.     

联苯苄唑乳膏体内外抗真菌活性及体内抗炎作用研究

目的 探讨1%联苯苄唑乳膏体内外抗真菌活性及体内抗炎作用。方法 采用豚鼠癣病模型、真菌体外药敏试验及小鼠二甲苯耳廓肿胀法评价1%联苯苄唑乳膏体内外抗真菌活性及体内抗炎作用。结果 1%联苯苄唑乳膏能有效治愈豚鼠须毛癣菌感染；体外抗皮肤癣菌和念珠菌最小抑菌浓度范围为 0.016 ～ 0.5 mg/L，最小杀菌浓度范围为0.063 ～ 2.0 mg/L；该药能显著抑制二甲苯所致的小鼠耳廓肿胀。结论 1%联苯苄唑乳膏具有明显抗皮肤癣菌、抗念珠菌和抗炎作用。     

茶皂甙体外抗真菌作用和治疗皮肤癣菌病的疗效观察

本文报告茶皂甙的体外抗真菌作用和茶皂甙酊治疗体癣、股癣的疗效观察。体外试验显示茶皂甙有较强的抑制真菌作用,对红色癣菌、石膏样癣菌、断发癣菌、黄癣菌、紫色癣菌、絮状表皮癣菌的MIC为0.125～1g/L,对白念珠菌为0.25～0.0625g/L,应用茶皂甙酊治疗皮肤癣菌病的结果是十分有效的。     

银屑病皮损鳞屑中两种微生物的检测及外用派瑞松效果

目的探讨银屑病发病与金黄色葡萄球菌(简称金葡菌)、糠秕孢子菌感染的关系及观察外用派瑞松治疗效果。方法对银屑病病人皮损进行细菌培养及真菌真接镜检,并外用派瑞松治疗。结果银屑病患者金葡菌和糠秕孢子菌带菌率均高于正常对照(P<0.01;P<0.05),治疗后带菌率明显下降;治愈率和有效率分别为18.0%,72.0%。结论金葡菌、糠秕孢子菌在银屑病的发病和发展中起一定作用,派瑞松是治疗银屑病较好的药物之一。     

曲安奈德益康唑乳膏长疗程间歇疗法治疗局限性慢性湿疹

目的 探讨外用糖皮质激素长疗程间歇疗法治疗局限性慢性湿疹的疗效、安全性及复发情况,以指导临床用药.方法 选取门诊局限性慢性湿疹患者,外用曲安奈德益康唑乳膏短程治疗.症状积分下降指数(SSRI)≥190%的患者进入长疗程,随机分两组,分别外用曲安奈德益康唑乳膏和润肤霜,间歇用药,共治疗8周.长疗程治疗4周、8周时及长疗程治疗停药12周时各随访1次.结果 长疗程治疗8周及停药12周时,曲安奈德益康唑组SCORAD评分均显著低于润肤霜组(t值分别为3.076和2.367,P值均<0.05).长疗程治疗4周、8周及停药12周时,曲安奈德益康唑组复发率均显著低于润肤霜组(x2值分别为4.426、7.683、8.199,P值均<0.05).治疗期间不良反应发生率3.1%,无严重不良事件.结论 外用糖皮质激素长疗程间歇治疗慢性湿疹,有防止症状加重、延缓湿疹复发的作用.     

曲安奈德益康唑乳膏长疗程间歇疗法治疗局限性慢性湿疹

目的 探讨外用糖皮质激素长疗程间歇疗法治疗局限性慢性湿疹的疗效、安全性及复发情况,以指导临床用药.方法 选取门诊局限性慢性湿疹患者,外用曲安奈德益康唑乳膏短程治疗.症状积分下降指数(SSRI)≥190%的患者进入长疗程,随机分两组,分别外用曲安奈德益康唑乳膏和润肤霜,间歇用药,共治疗8周.长疗程治疗4周、8周时及长疗程治疗停药12周时各随访1次.结果 长疗程治疗8周及停药12周时,曲安奈德益康唑组SCORAD评分均显著低于润肤霜组(t值分别为3.076和2.367,P值均<0.05).长疗程治疗4周、8周及停药12周时,曲安奈德益康唑组复发率均显著低于润肤霜组(x2值分别为4.426、7.683、8.199,P值均<0.05).治疗期间不良反应发生率3.1%,无严重不良事件.结论 外用糖皮质激素长疗程间歇治疗慢性湿疹,有防止症状加重、延缓湿疹复发的作用.     

曲安奈德益康唑乳膏长疗程间歇疗法治疗局限性慢性湿疹

目的 探讨外用糖皮质激素长疗程间歇疗法治疗局限性慢性湿疹的疗效、安全性及复发情况,以指导临床用药。方法 选取门诊局限性慢性湿疹患者,外用曲安奈德益康唑乳膏短程治疗。症状积分下降指数（SSRI）≥90％的患者进入长疗程,随机分两组,分别外用曲安奈德益康唑乳膏和润肤霜,间歇用药,共治疗8周。长疗程治疗4周、8周时及长疗程治疗停药12周时各随访1次。结果 长疗程治疗8周及停药12周时,曲安奈德益康唑组SCORAD评分均显著低于润肤霜组（t值分别为3.076和2.367,P值均 < 0.05）。长疗程治疗4周、8周及停药12周时,曲安奈德益康唑组复发率均显著低于润肤霜组（χ2值分别为4.426、7.683、8.199,P值均 < 0.05）。治疗期间不良反应发生率3.1％,无严重不良事件。结论外用糖皮质激素长疗程间歇治疗慢性湿疹,有防止症状加重、延缓湿疹复发的作用。     

Identification of several clinical isolates of dermatophytes based on the nucleotide sequence of internal transcribed spacer 1 (ITS 1) in nuclear ribosomal DNA.

Nucleotide sequences of internal transcribed spacer 1 (ITS 1) in nuclear ribosomal DNA from seven morphologically unidentified dermatophyte isolates were determined. The sequences were compared with those of typical isolates of Trichophyton (T.) mentagrophytes var. interdigitale, T. rubrum, and Epidermophyton floccosum. Two of the isolates were classified as T. rubrum and the other five as T. mentagrophytes var. interdigitale. The results did not conflict with identifications using other molecular techniques, including random amplification of polymorphic DNA (RAPD) analysis and restriction enzyme analysis of mitochondrial DNAs. Thus, the nuclotide sequence of ITS 1 is possibly a good molecular marker for identification of these major anthropophilic dermatophyte species.