Molecular detection of dermatophytes and nondermatophytes in onychomycosis by nested polymerase chain reaction based on 28S ribosomal RNA gene sequences

Background  Onychomycosis is often caused by dermatophytes, but the role of nondermatophytes is underestimated due to the difficulty of identifying them by conventional direct microscopy and culture.
Objectives  This study aims to detect nondermatophytes, as well as dermatophytes, in the nail samples of patients with onychomycosis using a polymerase chain reaction (PCR)-based culture-independent method.
Materials and methods  The nested PCR assay targeting the sequence of the 28S ribosomal RNA gene was used to amplify fungal DNAs from 50 microscopy-positive nail specimens. Newly designed primer sets for dermatophyte universal, Trichophyton rubrum , T. mentagrophytes , Aspergillus spp., Scopulariopsis brevicaulis , Fusarium solani , F. oxysporum , F. verticillioides , Candida albicans and C. tropicalis were used after confirmation of their specificity.
Results  Forty-seven cases (94%) were positive for fungal DNA, among which dermatophytes were detected in 39 cases (83·0%): T. rubrum in 35 cases (74·5%) and T. mentagrophytes in eight cases (17·0%). Surprisingly, nondermatophytes were detected in 18 cases (38·3%), both dermatophytes and nondermatophytes in 10 cases (21·3%) and nondermatophytes alone in eight cases (17·0%). Aspergillus spp. alone was observed in five cases (10·6%).
Conclusions  This study indicates that most of the affected nail plates of patients with onychomycosis were positive for specific fungal DNAs, and suggests that nondermatophytes detected at high rates may be involved in the pathogenesis of onychomycosis.     

根癌农杆菌介导的尿嘧啶缺陷烟曲霉转化是基因敲除的有效方法

目的:探讨用嘧啶营养缺陷作为筛选标记、根癌农杆菌介导的转化方法敲除烟曲霉基因.方法:以pyrG作为筛选标记,在双元载体pDHt/sk上分别构建烟曲霉FAPI,和SHOI基因的打靶序列.构建好的质粒分别转化入根癌农杆菌.24℃条件下,含打靶质粒的根癌农杆菌与烟曲霉在不含尿嘧啶和尿苷的诱导培养基上共培养48h;然后将培养物转移到37℃,筛选转化子.结果:根癌农杆菌介导的烟曲霉转化同源重组率较高,FAPI基因为44%(7/16)、SHOI基因为35%(7/20).结论:以pyrG作为筛选标记、根癌农杆菌介导的转化方法能高效敲除烟曲霉基因,为烟曲霉基因功能研究提供了有效方法.     

Renal Aspergilloma--A Case Illustrating the Problems of Medical Therapy

The second documented case of renal aspergilloma due to Aspergillusflavus is presented. The merits of the medical therapy thatfailed are discussed. Pathological examination showed a nidusof aspergillus around suture material persisting from a pyelolithotomyoperation 2 years before in India. We argue that this was thereason for the failure of the medical therapy. This is the firstcase of its kind reported.     

Mycotic Dermatitis in Domestic Fowl. A Case Report./Pilzdermatitis am Haushuhn. Ein Fallbericht

Summary: Two cases of mycotic dermatitis, probably caused by Aspergillus spec, are presented.
Zusammenfassung: Es werden zwei Fälle von Pilzdermatitis, wahrscheinlich hervorgerufen durch Aspergillus, vorgestellt     

EFFICACY OF MICONAZOLE IN EXPERIMENTAL KERATOMYCOSIS

For the first time comparative efficacy of 1 % miconazole in ointment base and drops has been evaluated against Candida, Aspergillus and Fusarium organisms In experimental heratomycosis. The findings of the present study reveal that miconazole in ointment drug delivery system has the advantage of being more effective in healing the fungal ulcers and associated lesions. It requires much less frequent application than drops, a parameter of great importance in improving patient compliance. Miconazole ointment was well tolerated by the rabbit eye and produced no ocular or systemic toxicity. The present study confirms that miconazole is a broad-spectrum antifungal agent and it could be a better alternative to presently available drugs to treat human heratomycosis .     

A strategy for managing fungal infections in haematopoietic stem cell transplantation

As more indications continue to be found for allogeneic haematopoietic transplantation, more patients are at risk for invasive fungal infectious diseases (IFID), particularly candidiasis and aspergillosis. Risk factors for disease are becoming better defined and diagnostic methods have improved considerably. In addition, there is now international agreement that three elements form the basis for defining IFID (host factors, clinical evidence, and mycological results), that imaging is acceptable for diagnosing disease, and that indirect tests such as antigen detection are also adequate mycological proof of cause. There are also more drugs available and still more to come, offering the potential for selective prophylaxis as well as preemptive and specific therapy, as well as for flexible administration. Hence, all the elements are in place for designing and testing an effective and economically sound strategy for dealing with IFID.     

Cloning and expression of the cDNA encoding an alternative oxidase gene from Aspergillus niger WU-2223L

A cDNA fragment encoding the mitochondrial alternative oxidase, the enzyme responsible for cyanide-insensitive and salicylhydroxamic acid (SHAM)-sensitive respiration, from the citric acid-producing fungus Aspergillus niger WU-2223L was cloned and expressed in Escherichia coli as a host strain. Synthetic primers were designed from the conserved nucleotide sequences of the alternative oxidase genes from higher plants and a yeast. The 210-bp DNA fragment was amplified by PCR with these primers using chromosomal DNA of WU-2223L as a template, and was employed to screen a cDNA library of A. niger. One full-length cDNA clone of 1.2 kb was obtained, and was sequenced to reveal that the clone contained an open reading frame (ORF-AOX1) encoding a polypeptide of 351 amino acids. The predicted amino-acid sequence exhibited 50%, 55%, and 52% homology to the alternative oxidases of Hansenula anomala, Neurospora crassa and Sauromatum guttatum, respectively. In the 5′-terminus region of the ORF-AOX1, a mitochondrial targeting motif was found. The whole open reading frame of ORF-AOX1 was ligated to plasmid pKK223-3 to construct the expression vector pKAOX1. The E. coli transformant harboring pKAOX1 showed cyanide-insensitive and SHAM-sensitive respiration, and expression was increased approximately two-fold by the addition of IPTG. These results indicated that the ORF-AOX1 encodes an alternative oxidase of A. niger. Received: 10 August / 13 October 1998     

Melanin as a virulence factor in pathogenic fungi

The pigment melanin is found universally in nature and is attributed to a variety of functions. In some fungi it is thought to play a decisive role in the determination of virulence. This review examines the experimental evidence which has led to an understanding of the mechanisms by which melanin functions in pathogenic fungi, particularly in plant pathogens, in Cryptococcus neoformans and Wangiella dermatitidis.     

Aspergillus Iris Granuloma: A Case Report with Review of Literature

We report the case of a 25-year-old male patient who presented with complaints of redness, photophobia, and decreased vision in the right eye of a week's duration. Slit-lamp biomicroscopic examination revealed a cream-colored, irregular elevated inferior iris mass, extending on to the anterior lens surface. Differential diagnoses of a fungal granuloma, a medulloepithelioma, and an amelanotic melanoma were considered. An excisional biopsy of the mass was performed through a superior clear corneal incision. Polymerase chain reaction analysis of the aqueous humor showed a positive pan fungal genome. Histopathology of the biopsied mass showed a giant cell granuloma with surrounding numerous branching, septate hyphae. Culture growth revealed Aspergillus fumigatus We report this case because of the rarity of Aspergillus iris granuloma as a primary presentation of endogenous Aspergillosis and review the relevant literature. Absence of a significant systemic history compounded the diagnostic dilemma in our patient. Definitive differentiation of this rare entity from a foreign body, amelanotic melanoma, and other inflammatory conditions such as sarcoidosis and tuberculosis, may be possible only on microbiological and histo-pathological evaluation.     

菊芋原料同步糖化发酵生产丁二酸

对菊芋原料发酵生产丁二酸进行了研究,用 Actinobacillus succinogenes 和 Aspergillus niger 同步糖化发酵,发现同步糖化发酵效果优于糖化后再发酵,在同步糖化发酵过程中还原糖质量浓度始终保持在10～40 g/L,可以避免高浓度的还原糖对 A.succinogenes 的抑制.5 L搅拌罐中同步糖化补料分批发酵96 h产丁二酸98.2 g/L,对消耗糖产率95.4%,生产强度1.02 g/(L·h) .