一株南海海洋真菌Sporothrix sp.#4335的甾体类代谢产物研究

目的研究分离自香港近海岸的红树林样品Kandelia candel的树皮中,种属已鉴定为Sporothrixsp的海洋真菌Sporothrixsp.#4335的代谢产物。方法用硅胶柱层析法分离代谢产物。结果分离得到3种甾体类化合物。结论化合物的结构通过解析波谱数据分别鉴定为麦角甾醇,过氧化麦角甾醇,麦角甾-7,22(E)-二烯-3β,5α,6β-三醇。     

应用特异性引物鉴定人申克孢子丝菌感染

目的研究人申克孢子丝菌感染的分子生物学鉴定方法,为建立检测人申克孢子丝菌感染的快速、敏感、特异方法提供参考资料。方法从疑似申克孢子丝菌感染患者的感染部位表面刮取物、腐烂组织或活检组织中提取DNA,应用合成的申克孢子丝菌特异性引物ITS3-SSP进行核糖体DNA内1332靶序列PCR扩增,将检测结果与真菌培养鉴定结果或组织病理学检查对比,观察种特异性引物PCR扩增结果的敏感性和特异性。结果在17例经形态学检查和／或真菌培养检查证实的组织标本中,有12例扩增出191bp的片段,其中7例组织中发现染成紫红色的圆形或卵圆形的孢子;而在11例经真菌培养证实为申克孢子丝菌的标本中,有4例样品巢式PCR结果为阴性。结论应用所设计的特异性引物ITS3-SSP结合巢式PCR方法鉴定申克孢子丝菌特异、敏感,是一种非常有发展潜力的检查手段。     

Influence of TLR-2 in the immune response in the infection induced by fungus Sporothrix schenckii

Toll-like receptors (TLRs) play an important role in immunity, since they bind to pathogen surface antigens and initiate the immune response. However, little is known about the role of TLR-2 in the recognition of S. schenckii and in the subsequent immune response. Therefore, the aim of this study was to evaluate the involvement of TLR-2 in the immune response induced by S. schenckii. C57BL/6 mice (WT) and C57BL/6 TLR-2 knockout (TLR-2^?/?) were used to evaluate, over a period of 10 weeks of sporotrichotic infection, the influence of TLR-2 over macrophages production of IL-1β, IL-12 and TNF-α, their stimulation level by NO release and the production of IFN -γ, IL-6, IL-17 and TGF-β by spleen cells. The results showed that the production of pro-inflammatory mediators and NO, TLR-2 interference is striking, since its absence completely inhibited it. IL-17 production was independent of TLR-2. The absence of Th1 response in TLR2^?/? animals was concomitant with IL-17 production. Therefore, it can be suggested that TLR-2 absence interferes with the course of the infection induced by the fungus S. schenckii.     

申克孢子丝菌接种的无毛小鼠皮损处蛋白酶抑制剂的作用

Chymostatin和 Pepstatin能分别抑制申克孢子丝菌产生的蛋白酶 和 ,为了证明在孢子丝菌病中 Chymostatin和 Pepstatin能通过抑制这些酶的活性而抑制真菌的生长 ,我们研究了在小鼠的皮损上外用蛋白酶抑制剂的效应 ,用 0 .1% Chymostatin,0 .1% pepstatin和 0 .1% chymostatin+0 .1%pepstatin软膏涂于无毛小鼠的申克孢子丝菌接种部位与对照组相比 ,单用一种成分时 ,皮损处结节大小的变化曲线表现分轻度抑制 ,而用含 chymostatin和 pepstatin的软膏 ,则强烈地抑制了结节的形成。本研究不仅证明了两种蛋白酶对申克孢子丝菌在体内生长的作用 ,而且还发展了一种新的局部药物治疗方法     

Ritual Samoan body tattooing and associated sporotrichosis

A 36 year old Samoan male who developed cutaneous sporotrichosis as a result of traditional body tattooing is reported. It is considered that the tattooing procedure was responsible for introducing the infection. He was treated with itraconazole, which produced resolution of the lesions.     

申克孢子丝菌4Hppd基因结构功能的生物信息学分析

 目的:初步预测及分析申克孢子丝菌(Sporothrix schenckii，Ss.)对羟苯基丙酮酸双加氧酶基因（4Hppd）编码蛋白的结构和生物学功能特征。方法：应用生物信息学在线工具预测分析Ss.4Hppd编码蛋白的理化性质及功能特征。结果：Ss.4Hppd基因编码蛋白含对羟苯基丙酮酸双加氧酶保守域，由477个氨基酸构成，理论分子量为52.0 kD，等电点为5.98，半衰期较长，该蛋白为非分泌蛋白，定位于线粒体中。其二级结构以无规则卷曲及α-螺旋为主，另外还存在乙二醛酶功能域及丰富的磷酸化位点，与其他致病真菌同源蛋白有较高的相似度。结论：预测Ss.4HppD位于线粒体中，具有乙二醛酶功能域结构功能特点及丰富修饰位点，可为其表达、晶体结构分析及小分子抑制剂开发提供参考。     

Human sporotrichosis beyond the epidemic front reveals classical transmission types in Espírito Santo,Brazil

Sporotrichosis has emerged as the main subcutaneous mycosis of humans and animals around the world. With particular differences in frequency, the major species includes Sporothrix brasiliensis, S. schenckii, S. globosa and S. luriei. In Brazil, the main aspect of this epidemic is based on the zoonotic transmission through the scratches and bites of diseased cats contaminated with S. brasiliensis. Areas free of feline sporotrichosis are poorly characterised in Brazil. We investigated by molecular tools the epidemiology of human sporotrichosis in the Espírito Santo (ES) state, an area adjacent to Rio de Janeiro where is the epicentre of the long‐lasting outbreak of cat‐transmitted sporotrichosis. The human cases in the ES state reveal the prevalence of classical transmission types where subjects are mainly infected by accidental traumatic inoculation during manipulation of contaminated plant material. In agreement with an environmental source, Sporothrix schenckii was the major aetiological agent in the classical transmission. Unlike Rio de Janeiro, this study shows that cat‐transmitted epidemic in Espírito Santo is still scanty, although the geographic proximity and similar climatic features. Sporothrix brasiliensis was the agent in the feline‐transmitted cases. Sporothrix globosa was isolated from a patient with fixed cutaneous lesions that did not report any contact with diseased animals. In conclusion, beyond the borders of Rio de Janeiro epidemic, agents of sporotrichosis in Espírito Santo show a scattered occurrence with high species diversity.     

Genotyping species of the Sporothrix schenckii complex by PCR-RFLP of calmodulin

Sporotrichosis is one of the most common subcutaneous mycosis in Latin America and is caused by 4 pathogenic thermodimorphic fungi in the genus Sporothrix. From both therapeutic and epidemiological perspectives, it is essential to identify the causative agents down to the species level. Traditional parameters may overlap among closely related species, and we propose polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) as an alternative approach. In the present study, the calmodulin gene was amplified and digested with HhaI to yield 5 different electrophoretic patterns representing all medically important Sporothrix species: Sporothrix brasiliensis, Sporothrix schenckii sensu stricto, Sporothrix globosa, and Sporothrix luriei. The PCR-RFLP protocol described here is a simple and inexpensive method and is highly suitable for accurate routine genotyping of relevant Sporothrix species.     

Sporothrix schenckii isolated from a cat in Japan

Summary. A yeast-like fungus isolated from a granulomatous and ulcerative skin lesion in a mongrel cat was mycologically examined. The isolate was identified as Sporothrix schenckii from the morphological findings and its pathogenicity in a mouse, confirming the first case of feline sporo-trichosis in Japan. Fortunately, no transmission to humans occurred in this case, howeyer the risk of humans contracting Sporothrix schenckii infection increases with the increase in the number of animals with sporotrichosis. Consequently when handling such animals protective clothing such as gloves should be worn to reduce the risk of transmission of the fungus through open wounds.
Zusammenfassung. Ein hefeähnlicher Pilz, isoliert aus der granulomatösen, ulzerierenden Hautläsion einer Bastardkatze, wurde anhand morphologischer Mermale und der Mäusepathogenität als Sporothrix schenckii identifiziert. Damit wird der erste Fall einer felinen Sporotrichose in Japan beschrieben. Übertragungen auf den Menschen wurden hierbei nicht beobachtet. Es muß jedoch durch die steigende Zahl animaler Sporotrichose-Fälle mit einem solchen Übertragungsrisiko gerechnet werden. Im Umgang mit animaler Sporotrichose sollte daher durch Gebrauch von Schutzhandschuhen das Übertragungsrisiko über offene Wunden minimiert werden.