Die Besthmung von hefeähnlichen Pilzen aus klinischem Untersuchungsmaterial mit der Uni-Yeast-Tek-Platte1,2

Summary: The Uni-Yeast-Tek system was studied in its ability to differentiate the clinical significant yeasts and yeastlike organisms. Evaluation was based on the examination of 268 routine clinical isloates and 53 stock cultures. The Uni-Yeast-Tek system gives reliable results with morphologic features. It turned out that Candida parapsilosis didn't react to the assimilation test trehalose. Zusammenfassung: Das Uni-Yeast-Tek-System ermöglicht die Bestimmung der wichtigsten Hefen und hefeähnlichen Pilze aus klinischem Material. Es wurden insgesamt 321 Stämme von 22 verschiedenen Spezies untersucht. Die Uni-Yeast-Tek-Platte hat sich bei genauer Kenntnis morphologischer Merkmale als ein schnelles und zuverlässiges Hilfsmittel erwiesen. Schwierig-keiten traten nur bei Candida parapsilosis ad, bei der eine Zucker-Assimilations-Reaktion (Trehalose) versagte. Sproßpilze     

In vitro activity of terpenes against Candida biofilms

The antibiofilm activity of 10 terpenes was tested in vitro against three Candida species by 24-h treatment of biofilms aged 1-5 days. Treatment of 24-h-old Candida albicans biofilms with carvacrol, geraniol or thymol (0.06%) resulted in >80% inhibition. Carvacrol (0.03%) inhibition was > or =75% independent of the age of the C. albicans biofilm. Carvacrol (0.125%) inhibition was >75% against Candida glabrata and Candida parapsilosis biofilms. Geraniol (> or =0.125%) and thymol (0.06% or 0.125%) inhibition was >75% against C. parapsilosis biofilms whatever their age. This study demonstrates the antibiofilm activity of terpenes and points out the exceptional efficiency of carvacrol, geraniol and thymol, which could represent candidates in the treatment of candidiasis associated with medical devices.     

氟康唑纸片扩散法和浓度梯度法检测酵母菌耐药性的研究

目的 对比研究酵母菌氟康唑纸片扩散法和浓度梯度法的相关性。方法 分别应用纸片扩散法和浓度梯度法检测142株临床分离的酵母菌对氟康唑的敏感性；采用BIOMIC仪自动读取培养板上的抑菌环直径、记录试验结果并对质控数据进行核实；用WHONET-5．3软件分析结果，比较两者的相关性。结果 两种体外药敏试验方法的一致率可达82．4％，极重要误差率为0．7％，重大误差率为2．8％，次要误差率为14．7％。对于光滑念珠菌两种方法的检测结果有较大差异。结论 应用WHONET-5．3软件分析得出氟康唑纸片扩散法和浓度梯度法对于检测大多数酵母菌具有非常好的一致性，但对于一部分菌株需要进一步应用常量肉汤稀释法确定其最低抑菌浓度值。     

Three-dimensional structure of tubular networks,presumably Golgi in nature,in various yeast strains: A comparative study

Background: In the yeast Saccharomyces cerevisiae, the Golgi apparatus consists of discrete units distributed throughout the cytoplasm. When such units are examined in three dimensions, in relatively thick sections prepared for the electron microscope, they usually appear as small tubular networks with a stained material accumulating in dilations located at the junctions of membranous tubules. To see whether such tubular networks are observed in other yeast species, the three-dimensional structure of organelles in eight additional yeast strains, endowed with diverse biological properties, are examined. Methods: Yeast strains were grown at 24°C in YPD medium (2% Bactopeptone, 1% Bactoyeast extract, and 2% glucose). Cells that were examined by electron microscopy came from exponentially growing cultures grown in a shaking water bath and maintained at a OD 600 (optical density at 600 nm) of 0.5. Cells were fixed in a fixative containing 2% glutaraldehyde in 0.1 M cacodylate buffer pH 7.4 and 0.8 M sorbitol. They were then treated for 15 min in 1% sodium metaperiodate and postfixed for 1 hr in potassium ferrocyanide-osmic acid. They were preembedded in agarose prior to dehydration and finally embedded in Epon. In these conditions, the preservation of cell organelles was improved and the cytoplasmic retraction from the cell wall was minimized. Photographs of sections tilted at ± 15° from the 0° position of the goniometric stage were used to prepare stereopairs from which the three-dimensional configuration of the organelles was visualized. Results: In all yeast strains, tubular networks appeared as separate elements or units dispersed throughout the cytoplasm. Each unit consisted of anastomosed membranous tubules. In some strains such as Saccharomyces cerevisiae, Zygosaccharomyces rouxii, or Saccharomyces pombe, such units appeared mainly as polygonal networks of intensely stained membranous tubules. Along these networks, distensions filled with stained material were similar in size to nearby secretory granules, suggesting that the latter formed by fragmentation of the tubular networks. In Hansenula polymorpha, Pichia pastoris, and Debaryomyces hansenii, networks of anastomosed tubules were closely superposed to each other and formed parallel arrays reminiscent of the stacks of Golgi saccules seen in mammalian cells. However, in contrast to what is usually found in the latter, the layers making up the parallel arrays in yeasts, were clearly continuous to each other. In other strains, i.e., Kluyveromyces lactis, Candida albicans, and Candida parapsilosis, the situation was intermediate and their cytoplasm contained only arrays of small size with two or at most three superposed layers of membranous tubules. Small vesicles in the 30–50 nm range were rarely encountered in most yeast strains. Conclusions: It is therefore concluded that tubular networks, presumably Golgi in nature, are present in all yeasts examined so far. Yet, in some strains, these tubular networks may be arranged in parallel arrays or stacks. © 1995 Wiley-Liss, Inc.     

Wechselwirkungen zwischen 5-Fluorcytosin und Polyen-Antibiotica bzw. Imidazol-Derivaten

Zusammenfassung: Mittels Warburg-Technik wurde an Myzeten (Saccharomyces cerevisiae und Candida albicans) untersucht, ob 5-Fluorcytosin die antimyzetische Aktivität von Polyen-Antibiotika (Amphotericin B und Nystatin) bzw. von Imidazol-Derivaten (Econazolnitrat und Ketoconazol) verstärkt, oder ob ein Antagonismus vorliegt.
Im Falle der Polyen-Antibiotica ließ sich ein statistisch signifikanter additiver Effekt mit 5-Fluorcytosin nach weisen. Bei den Imidazol-Derivaten hingegen fehlte jegliche additive Wirkung des 5-Fluorcytosins; eine wechselseitige Abschwächung war aber auch nicht nachzuweisen. Die Ergebnisse werden im Hinblick auf ihre klinisch therapeutische Bedeutung kurz diskutiert.
Summary: On resting yeasts (Candida albicans, Saccharomyces cerevisiae), investigations were carried out to find out whether there exist interactions between 5-fluorocytosine and polyene antibiotics, respectively imidazole derivatives. Among the polyenes, amphotericine B and nystatin, among the imidazoles, ketoconazole and econazole nitrate were tested.
With the polyene antibiotics, a significant additive action with 5-fluorocytosine could be demonstrated. Such additive effect lacked in the case of the imidazole antifungals. However, no decrease in activity could be shown neither.
For practical purposes in human systemic antifungal therapy, these results permit the simultaneous application of 5-fluorocytosine together with polyene antibiotics (amphotericine B deoxycholate) or imidazole derivatives (ketoconazole). In the first case, an additive therapeutic effect may be expected, in the latter case at least no interactions causing a decrease in antifungal activity will occur.     

Current approaches to diagnosis and treatment of fungal infections in children infected with human immuno deficiency virus

The prolonged survival of profoundly immunocompromised patients has revealed mucosal and invasive fungal infections to be major causes of morbidity and mortality in advanced HIV disease in children of all age groups. Antifungal resistance has become a clinically relevant problem. Paediatricians caring for HIV-infected children need to be aware of these increasingly frequent and often life-threatening infectious complications. This article reviews what is currently known about epidemiology, clinical presentation, diagnosis and treatment of mucosal and invasive fungal infections in children and adolescents with HIV disease. Candida spp. have become a leading bloodstream isolate in hospitalised patients; mucosal candidiasis is the most prevalent opportunistic infection in HIV-infected patients, and in both invasive and superficial infections, non Candidaalbicans spp. are on the increase. Invasive pulmonary aspergillosis has surfaced as an HIV-associated complication and previously uncommon fungi are more frequently encountered. HIV-infected individuals are particularly susceptible to Peumocystis carinii, Cryptococcus neoformans and infections by endemic fungi, such as Histoplasma capsulatum, Coccidioides immitis, Penicillium marneffei, and others. Newer immunological and molecular-based methods provide early and rapid diagnosis and monitoring. Potent and broad-spectrum third generation triazoles and novel fungicidal lipopeptides of the echinocandin class of antifungal antibiotics have entered clinical trials. Immunmodulation by recombinant cytokines and antifungal vaccines are very actively pursued inroads to adjunctive and preventive immunotherapy. Conclusion Mucosal and invasive fungal infections will remain important complications in HIV-infected children of all age groups. Interventional studies and well documented case series are needed to improve the molecular diagnosis, treatment and prevention of invasive fungal infections in the paediatric HIV-infected population. Received: 13 June 1998 / Accepted: 14 July 1998     

丙型肝炎病毒NS3基因酵母表达载体构建及表达

目的　为探讨丙型肝炎病毒 (HCV)非结构蛋白NS3的功能 ,在真核生物酵母细胞中表达HCVNS3基因。方法　用聚合酶链反应 (PCR)的方法以HCV全长质粒pBRTM/HCV 1为模板扩增HCVNS3基因 ,克隆到 pGEM T载体中 ,双酶切后回收连接到酵母表达质粒 pGBKT7中表达。提取酵母蛋白质 ,进行十二烷基磺酸钠 聚丙烯酰胺凝胶电泳 (SDS PAGE)和Western免疫印迹分析。结果　成功构建HCVNS3基因酵母表达载体 ,Western免疫印迹显示了HCVNS3在酵母细胞中表达。表达产物在胞内存在 ,相对分子质量为 70 0 0 0。结论　HCVNS3蛋白在酵母中表达成功。     

难辨梭状芽孢杆菌相关性腹泻研究进展

难辨梭状芽孢杆菌是一种革兰阳性肠道病原体。难辨梭状芽孢杆菌相关性腹泻（CDAD）及其引起的假膜性肠炎为消化道多发病。难辨梭状芽孢杆菌可产生毒素A和毒素B，侵入肠黏膜后引起细胞病变，导致一系列感染相关临床表现。免疫学机制在难辨梭状芽孢杆菌感染相关疾病的发病中起重要作用。本文就CDAD的发病机制、临床表现及其检测和治疗作一综述。     

泌尿系感染假丝酵母菌的临床特点及耐药性分析

目的 了解泌尿系感染假丝酵母菌菌株分布及耐药现状,为临床合理选择抗菌药物提供依据.方法 回顾性调查2011年10月～ 2013年12月本院住院患者泌尿系发生假丝酵母菌感染的相关资料及药敏结果.结果 从送检的1688份泌尿系感染患者的尿培养标本中共分离268株假丝酵母菌,检出率为15.87％,菌种以热带假丝酵母菌为主,占52.6％,其次为白色假丝酵母菌61株,占22.76％,克柔假丝酵母菌26株,占9.70％,光滑假丝酵母菌为19株,占7.09％.所有菌株体外药敏实验对两性曲霉B100％敏感,对5-氟胞嘧啶、氟康唑、伊曲康唑、酮康唑的耐药率分别是38.43％、76.49％、73.88％、74.25％.结论 假丝酵母菌是泌尿系发生感染最常见的真菌,耐药性有增长趋势,应对分离菌株进行药敏试验,根据实验室的药敏试验结果指导临床合理.