BACKGROUND & AIMS: :Lectins play a critical role in host protection against infection. The galectin family of lectins recognizes saccharide ligands on a variety of microbial pathogens, including viruses, bacteria, and parasites. Galectin-3, a galectin expressed by macrophages, dendritic cells, and epithelial cells, binds bacterial and parasitic pathogens including Leishmania major, Trypanosoma cruzi, and Neisseria gonorrhoeae. However, there have been no reports of galectins having direct effects on microbial viability. We found that galectin-3 bound only to Candida albicans species that bear beta-1,2-linked oligomannans on the cell surface, but did not bind Saccharomyces cerevisiae that lacks beta-1,2-linked oligomannans. Surprisingly, binding directly induced death of Candida species containing specific beta-1,2-linked oligomannosides. Thus, galectin-3 can act as a pattern recognition receptor that recognizes a unique pathogen-specific oligosaccharide sequence. This is the first description of antimicrobial activity for a member of the galectin family of mammalian lectins; unlike other lectins of the innate immune system that promote opsonization and phagocytosis, galectin-3 has direct fungicidal activity against opportunistic fungal pathogens.

背景与目标： ：凝集素在宿主预防感染中起着至关重要的作用。凝集素的半乳凝素家族可识别多种微生物病原体（包括病毒，细菌和寄生虫）上的糖配体。 Galectin-3是一种由巨噬细胞，树突状细胞和上皮细胞表达的半乳糖凝集素，结合细菌和寄生性病原体，包括大利什曼原虫，克氏锥虫和淋病奈瑟氏球菌。但是，尚未有关于半乳凝集素对微生物生存能力有直接影响的报道。我们发现，galectin-3仅与在细胞表面带有β-1,2-连接的寡甘露聚糖的白色念珠菌结合，但不与缺乏β-1,2-连接的寡甘露聚糖的酿酒酵母结合。出人意料的是，结合直接导致含有特定的β-1,2-连接的寡甘露糖苷的念珠菌死亡。因此，galectin-3可用作识别独特病原体特异性寡糖序列的模式识别受体。这是对哺乳动物凝集素半乳凝素家族成员的抗菌活性的首次描述；与先天免疫系统中促进调理作用和吞噬作用的其他凝集素不同，galectin-3具有直接针对机会性真菌病原体的杀真菌活性。

BACKGROUND & AIMS: :Candida albicans is an important human pathogen, causing opportunistic infections. The adhesion of planktonic cells to a substrate is the first step for biofilm development. The antimicrobial peptide (AMP) Psd1 is a defensin isolated from Pisum sativum seeds. We tested the effects of this AMP on C. albicans biofilms and planktonic cells, comparing its activity with amphotericin B and fluconazole. Three C. albicans variants were studied, one of them a mutant deficient in glucosylceramide synthase, conferring resistance to Psd1 antifungal action. Atomic force microscopy (AFM) was used to assess morphological and biomechanical changes on fungal cells. Surface alterations, with membrane disruption and leakage of cellular contents, were observed. Cytometry assays and confocal microscopy imaging showed that Psd1 causes cell death, in a time and concentration-dependent manner. These results demonstrate Psd1 pleiotropic action against a relevant fungal human pathogen, suggesting its use as natural antimycotic agent.

背景与目标： 白念珠菌是重要的人类病原体，引起机会性感染。浮游细胞与基质的粘附是生物膜发展的第一步。抗菌肽（AMP）Psd1是从豌豆（Pisum sativum）种子中分离出来的防御素。我们测试了这种AMP对白色念珠菌生物膜和浮游细胞的作用，并将其与两性霉素B和氟康唑的活性进行了比较。研究了三个白色念珠菌变体，其中一个是葡萄糖基神经酰胺合酶缺陷的突变体，赋予了对Psd1抗真菌作用的抗性。原子力显微镜（AFM）用于评估真菌细胞的形态和生物力学变化。观察到具有膜破坏和细胞内容物泄漏的表面变化。细胞计数法和共聚焦显微镜成像显示，Psd1以时间和浓度依赖性方式引起细胞死亡。这些结果证明了Psd1对相关真菌人类病原体的多效作用，表明其可作为天然抗真菌剂使用。

BACKGROUND & AIMS: :In this study, we investigated the yeasts colonization of genus Candida, including C. dubliniensis, isolated of HIV-infected patients oral cavities and we accessed in vitro susceptibility pattern of the Candida isolates to four antifungal agents. Out of 99 patients investigated, 62 (62.6%) were colonized with yeasts. C. albicans was the prevailing species (50%). C. dubliniensis isolates were not recovered in our study. We verified that 8.1% of the yeasts isolated were resistant to fluconazole, 8.1% to itraconazole and 3.2% to voriconazole. The isolates demonstrated very low voriconazole MICs, in which 79% (49/62) presented values of 0.015 mug/ml. All Candida isolates were susceptible to amphotericin B. The results reported here showed that although C. albicans continues to be present in one-half of oral Candida carriage of HIV-infected patients, Candida non-albicans species are increasing among these patients. Besides, the findings of resistant isolates endorse the role of antifungal susceptibility testing whenever antifungal treatment with azoles is planned.

背景与目标： ：在这项研究中，我们调查了念珠菌属（包括C. dubliniensis）的酵母菌定殖，分离出了HIV感染患者的口腔，并研究了念珠菌分离株对四种抗真菌剂的体外敏感性模式。在调查的99位患者中，有62位（62.6％）被酵母菌定植。白色念珠菌是主要的物种（50％）。 C. dubliniensis分离株未在我们的研究中回收。我们证实，分离出的酵母中有8.1％对氟康唑有抵抗力，对伊曲康唑有8.1％的抗性，对伏立康唑有3.2％的抗性。分离物显示伏立康唑的MIC非常低，其中79％（49/62）的值为0.015杯/毫升。所有念珠菌分离株均对两性霉素B敏感。此处报告的结果表明，尽管白色念珠菌仍存在于感染了HIV的患者的一半念珠菌口服口服药物中，但这些患者中念珠菌中非白色念珠菌的种类有所增加。此外，每当计划使用吡咯类药物进行抗真菌治疗时，耐药菌的发现都支持抗真菌药敏试验的作用。

BACKGROUND & AIMS: :Candida albicans is the most frequently isolated yeast species from clinical specimens. A classical rapid presumptive differentiation from non-albicans species is based on its ability to produce germ tubes after incubation in human serum. The only non-albicans Candida species producing germ tubes is Candida dubliniensis. In this study, we evaluated Mueller-Hinton-agar (MH-agar) as a medium for germ tube formation of C. albicans and C. dubliniensis. A total of 859 yeast isolates from stool samples, including 632 strains of C. albicans, 10 C. dubliniensis and 217 other yeast strains from 20 different species, were grown on Sabouraud glucose (2%) agar at 37 degrees C for 24-72 h. Species were identified by standard methods. For the germ tube test (GTT), an inoculum from a single colony was streaked onto a MH-agar plate and covered by a sterile coverslip. After incubation at 37 degrees C for 2 h, the MH plates were examined using a light microscope at x200. The GTT was positive in 578 of 632 C. albicans strains (sensitivity 91.5%), in six of 10 C. dubliniensis strains (sensitivity 60.0%), and in none of the other yeast strains. MH-agar is a suitable medium for the GTT and the presumptive identification of C. albicans. It is safer to use than human serum and is widely available in microbiology laboratories.

背景与目标： ：白色念珠菌是从临床标本中最常分离的酵母菌种。与非白色物种之间的经典快速推定区分是基于其在人血清中温育后产生胚芽管的能力。唯一能产生胚芽管的非白色念珠菌种是假丝酵母。在这项研究中，我们评估了Mueller-Hinton-琼脂（MH-琼脂）作为白色念珠菌和dubliniensis胚芽管形成的培养基。从粪便样品中分离出总共859种酵母菌，其中包括632株白色念珠菌，10株杜氏藻和217种来自20个不同物种的酵母菌株，在37℃的Sabouraud琼脂上（2％）琼脂生长24-24-72 H。通过标准方法鉴定物种。对于生殖管测试（GTT），将来自单个菌落的接种物划线接种到MH-琼脂平板上，并盖上无菌盖玻片。在37℃下孵育2小时后，使用光学显微镜在x200下检查MH板。在632株白色念珠菌菌株中的578株（敏感性为91.5％），10株杜布利尼酵母菌株中的6株（敏感性为60.0％）中，其他酵母菌株中，GTT均为阳性。 MH-琼脂是GTT和白色念珠菌的推定鉴定的合适培养基。它比人血清更安全使用，并且在微生物学实验室中广泛使用。

BACKGROUND & AIMS: :Five dichlorinated 8-quinolinols (2,5- 5,6-, 3,5-, 3,7-, and 4,5-dichloro-8-quinolinol) were tested against Candida albicans and C. tropicalis in Sabouraud dextrose broth with and without bovine serum. The 5,6-, 3,5-, and 3,7-dichloro-8-quinolinols proved to be more effective than the control, 5-fluorocytosine. In cytotoxicity tests employing baby hamster kidney (BHK) cells, all test agents proved to be more cytotoxic than the control. However, the minimum inhibitory concentration (MIC) of 3,5-dichloro-8-quinolinol to both fungi was only one tenth the cytotoxic dose, suggesting that the compound may be useful as a topical or systemic antifungal agent.

背景与目标： ：在Sabouraud葡萄糖肉汤中测试了五种二氯8-喹啉醇（2,5- 5,6-，3,5-，3,7-和4,5-二氯8-喹啉醇）对白色念珠菌和热带念珠菌的抵抗力有和没有牛血清。 5,6-，3,5-和3,7-二氯-8-喹啉醇被证明比对照的5-氟胞嘧啶更有效。在使用小仓鼠肾脏（BHK）细胞的细胞毒性测试中，所有测试药物均被证明比对照更具细胞毒性。但是，3,5-二氯-8-喹啉醇对两种真菌的最小抑制浓度（MIC）仅是细胞毒性剂量的十分之一，表明该化合物可用作局部或全身性抗真菌剂。

BACKGROUND & AIMS: :A new medium composed of "cream of rice" infusion, oxgall, Tween 80, and agar is described for the sequential development of germ tubes and chlamydospores by Candida albicans. The procedure used (Dalmau's technique) is an improvement over the fluid substrate procedures previously advocated for germ tube formation. That the same preparation is then used for chlamydospore production is of practical importance for the clinical mycology laboratory.

背景与目标： ：描述了一种由“米糊”输液，oxgall，Tween 80和琼脂组成的新培养基，用于白色念珠菌对胚芽管和衣原体的连续发育。所使用的程序（Dalmau的技术）是对先前提倡用于胚芽管形成的流体基质程序的一种改进。然后将相同的制剂用于衣原体生产对于临床真菌学实验室来说具有实际重要性。

BACKGROUND & AIMS: :Candida glabrata is an important human fungal pathogen known to trigger serious infections in immune-compromised individuals. Its ability to form biofilms, which exhibit high tolerance to antifungal treatments, has been considered as an important virulence factor. However, the mechanisms involving antifungal resistance in biofilms and the impact of host niche environments on these processes are still poorly defined. In this study, we performed a whole-transcriptome analysis of C. glabrata biofilm cells exposed to different environmental conditions and constraints in order to identify the molecular pathways involved in fluconazole resistance and understand how acidic pH niches, associated with the presence of acetic acid, are able to modulate these responses. We show that fluconazole treatment induces gene expression reprogramming in a carbon source and pH-dependent manner. This is particularly relevant for a set of genes involved in DNA replication, ergosterol, and ubiquinone biosynthesis. We also provide additional evidence that the loss of mitochondrial function is associated with fluconazole resistance, independently of the growth condition. Lastly, we propose that C. glabrata Mge1, a cochaperone involved in iron metabolism and protein import into the mitochondria, is a key regulator of fluconazole susceptibility during carbon and pH adaptation by reducing the metabolic flux towards toxic sterol formation. These new findings suggest that different host microenvironments influence directly the physiology of C. glabrata, with implications on how this pathogen responds to antifungal treatment. Our analyses identify several pathways that can be targeted and will potentially prove to be useful for developing new antifungals to treat biofilm-based infections.

背景与目标： ：Candida glabrata是一种重要的人类真菌病原体，已知会在免疫受损的个体中引发严重感染。它形成生物膜的能力被证明是重要的毒力因子，该生物膜对抗真菌治疗表现出很高的耐受性。但是，涉及生物膜抗真菌性的机制以及宿主生态位环境对这些过程的影响仍知之甚少。在这项研究中，我们对暴露于不同环境条件和限制条件下的光滑小球藻生物膜细胞进行了全转录组分析，以鉴定参与氟康唑耐药性的分子途径，并了解酸性pH壁ni与乙酸的存在如何相关，能够调节这些响应。我们显示氟康唑治疗诱导碳表达和pH依赖方式的基因表达重编程。这与涉及DNA复制，麦角固醇和泛醌生物合成的一组基因特别相关。我们还提供了其他证据，表明线粒体功能的丧失与氟康唑耐药性有关，而与生长条件无关。最后，我们提出C. glabrata Mge1，参与铁代谢和线粒体蛋白质进口的陪伴酮，是通过降低代谢通向有毒甾醇的通量来调节氟康唑对碳和pH的敏感性的关键调节剂。这些新发现表明，不同的宿主微环境直接影响了光滑念珠菌的生理，并对该病原体对抗真菌治疗的反应产生了影响。我们的分析确定了几种可靶向的途径，并可能被证明对开发新的抗真菌药以治疗基于生物膜的感染很有用。

BACKGROUND & AIMS: :Asymmetric reduction of alkyl-3-oxobutanoates mediated by Candida parapsilosis ATCC 7330 resulted in optically pure alkyl-3-hydroxybutanoates in good yields (up to 72%) and excellent enantiomeric excess (up to >99 %). A detailed and systematic optimisation study was necessary and was carried out to avoid the undesired transesterification reaction during the course of asymmetric reduction. Under optimised conditions, the (S)-alkyl hydroxyesters were produced predominantly except for the methyl ester which formed the (R)-enantiomer. To the best of our knowledge, the biocatalytic asymmetric reduction of isoamyl-3-oxobutanoate to (S)-isoamyl-3-hydroxybutanoate is reported here for the first time.

背景与目标： ：由副念珠菌ATCC 7330介导的3-氧代丁酸烷基酯的不对称还原导致光学纯的-3-羟丁酸烷基酯具有良好的收率（高达72％）和出色的对映体过量（高达> 99％）。有必要进行详细而系统的优化研究，以避免在不对称还原过程中发生不希望的酯交换反应。在最佳条件下，主要生成（S）-烷基羟基酯，除了形成（R）-对映异构体的甲酯以外。据我们所知，这是首次报道将3-氧代丁酸异戊酯生物催化不对称还原为（S）-3-羟丁酸异戊酯。

BACKGROUND & AIMS: :In this study, the antifungal effects of silver nano-particles (nano-Ag) and their mode of action were investigated. Nano-Ag showed antifungal effects on fungi tested with low hemolytic effects against human erythrocytes. To elucidate the antifungal mode of action of nano-Ag, flow cytometry analysis, a glucose-release test, transmission electron microscopy (TEM) and the change in membrane dynamics using 1,6-diphenyl-1,3,5-hexatriene (DPH), as a plasma membrane probe, were performed with Candida albicans. The results suggest nano-Ag may exert an antifungal activity by disrupting the structure of the cell membrane and inhibiting the normal budding process due to the destruction of the membrane integrity. The present study indicates nano-Ag has considerable antifungal activity, deserving further investigation for clinical applications.

背景与目标： ：在这项研究中，研究了银纳米颗粒（nano-Ag）的抗真菌作用及其作用方式。纳米银对真菌显示出抗真菌作用，对人红细胞的溶血作用低。为了阐明纳米银的抗真菌作用模式，流式细胞仪分析，葡萄糖释放测试，透射电子显微镜（TEM）以及使用1,6-二苯基-1,3,5-己三烯（DPH）的膜动力学变化），作为白色质假丝酵母进行质膜探针。结果表明，纳米银可能会破坏细胞膜的结构，并由于破坏膜的完整性而抑制正常的出芽过程，从而发挥抗真菌活性。本研究表明纳米银具有相当大的抗真菌活性，值得临床应用进一步研究。

BACKGROUND & AIMS: BACKGROUND:Despite the well-documented associations between poor maternal oral health and increased risk for adverse birth outcomes and dental caries in children after birth, prenatal oral health care is under-utilized, especially among the underserved population. In addition, oral Candida has recently been suggested as a potential culprit for children's dental caries, with evident maternal contributions. Therefore, this study aimed to obtain epidemiological data on the oral health and oral Candida carriage in a cohort of underserved US pregnant women, and reveal factors associated with their oral Candida carriage. METHODS:Demographic-medical-oral hygiene practice data were collected. Comprehensive oral examination was conducted. Caries status and plaque index were recorded. Oral samples (saliva, plaque and swab) were processed to identify Candida species and Streptococcus mutans by culturing-dependent and -independent methods. Multiple logistic regression analyses were used to identify factors associated with oral Candida carriage and caries severity. RESULTS:Eighty-two socioeconomically disadvantaged women (48 pregnant and 34 non-pregnant) were enrolled. More pregnant women (79.1%) had > = 1 untreated decayed tooth when compared to their non-pregnant counterparts (47.1%) (p = 0.01). The average number of decayed teeth in pregnant and non-pregnant women was 3.9 and 3.1 (p > 0.05). Caries severity was positively associated with race (African American vs. white), plaque index and salivary Candida albicans level. C. albicans was the most predominant/abundant Candida strain, with cheek and tonsil as the most common colonized sites. The detection of C. albicans was 56%/56% in saliva and 40%/47% in plaque of the pregnant and non-pregnant groups, respectively. Study women's oral Candida carriage is positively associated with hypertension [p = 0.03, odds ratio = 14.47(1.28, 163.51)], decayed teeth number [p = 0.04, odds ratio = 1.31 (1.01,1.69)] and salivary S. mutans level [p = 0.03, odds ratio = 4.80 (1.18-19.43)]. CONCLUSIONS:Socioeconomically disadvantaged US women are in need of improved prenatal oral health, a large proportion of them have untreated decayed teeth and high carriage of oral Candida. Due to the observed significant association between the decayed teeth number and oral Candida carriage, providing oral health care during pregnancy (including limiting decayed teeth) will not only improve women's oral health, but also present as a promising approach to reduce oral Candida carriage in women.

背景与目标： 背景：尽管有据可查的孕产妇口腔健康状况与出生后儿童不良出生结局和龋齿的风险增加之间存在相关性的文献记载，但产前口腔保健服务的利用不足，尤其是在服务不足的人群中。此外，最近有人提出口服念珠菌可能是儿童龋齿的潜在罪魁祸首，并具有明显的产妇贡献。因此，本研究旨在获得服务不足的美国孕妇队列中口腔健康和口腔念珠菌携带的流行病学数据，并揭示与其口腔念珠菌携带有关的因素。
方法：收集人口统计学，医学和口腔卫生实践数据。进行了全面的口腔检查。记录龋病状态和斑块指数。通过培养依赖性和非依赖性方法，对口腔样品（唾液，菌斑和拭子）进行处理，以鉴定念珠菌和变形链球菌。使用多重逻辑回归分析来确定与口腔念珠菌携带和龋齿严重程度相关的因素。
结果：招募了82名在社会经济方面处于不利地位的妇女（48名孕妇和34名非孕妇）。与未怀孕的孕妇（47.1％）相比，更多的孕妇（79.1％）的未治疗蛀牙> = 1（p = 0.01）。孕妇和非孕妇的平均蛀牙数量分别为3.9和3.1（p> 0.05）。龋齿严重程度与种族（非裔美国人与白人），斑块指数和唾液念珠菌水平呈正相关。白色念珠菌是最主要/最丰富的念珠菌菌株，脸颊和扁桃体是最常见的定植部位。在孕妇和非孕妇组的唾液中检出白色念珠菌的比例分别为56％/ 56％和在斑块中的检出比例为40％/ 47％。研究妇女的口腔念珠菌携带与高血压[p = 0.03，比值比= 14.47（1.28，163.51）]，蛀牙数[p = 0.04，比值= 1.31（1.01,1.69）]和唾液变形链球菌含量呈正相关。 [p = 0.03，优势比= 4.80（1.18-19.43）]。
结论：在社会经济上处于不利地位的美国妇女需要改善产前口腔健康，其中很大一部分妇女未经治疗的蛀牙和口腔念珠菌的大量携带。由于观察到龋齿数目与口腔念珠菌运输之间存在显着关联，因此在怀孕期间提供口腔保健（包括限制蛀牙）不仅可以改善女性的口腔健康，而且是减少女性口腔念珠菌运输的一种有前途的方法。

BACKGROUND & AIMS: :The human fungal pathogen Candida albicans displays a very high degree of plasticity, including the types of genomic changes frequently observed with cancer cells, such as gross chromosomal rearrangements, aneuploidy, and loss of heterozygosity. Despite its relevance to every aspect of genetics and evolution of this pathogen, our understanding of the mutation process and its bearing on organismal fitness remains quite limited. Here, we have evaluated and compared two approaches to estimate the mutation frequency at three ORFs/regions (HIS4, CEN4 and EST2) of the C. albicans genome. Sequencing of individual DNA molecules (clone-by-clone sequencing) identified de novo mutations at these DNA regions, whose frequency was similar to that observed for S. cerevisiae at homolog sites following the same approach. However, mutations were not detected when the same regions were directly sequenced from the pooled DNA. In addition, in the absence of the homologous recombination protein Rad52, mutation frequency within these sites remained unaltered. The use of an alternative polymerase also found mutations. These results suggest that at least some mutations are artifacts caused by the polymerase used, advising that post-PCR procedures might generate mutations which may become undistinguishable from the genuine mutations and thus may interfere with mutational analysis. Furthermore, we recommend that new mutations found in the sequences of cloned alleles used for the determination of haplotypes should be contrasted with the sequence yielded by the pooled DNA.

背景与目标： ：人类真菌病原体白色念珠菌具有很高的可塑性，包括癌细胞经常观察到的基因组变化类型，例如总体染色体重排，非整倍性和杂合性丧失。尽管它与这种病原体的遗传学和进化的各个方面都相关，但我们对突变过程及其对机体适应性的了解仍然十分有限。在这里，我们评估和比较了两种方法来估计白色念珠菌基因组的三个ORF /区域（HIS4，CEN4和EST2）的突变频率。单个DNA分子的测序（逐个克隆测序）在这些DNA区域识别出从头突变，其频率与酿酒酵母在同源位点处观察到的频率相似。但是，当从合并的DNA直接对相同区域进行测序时，未检测到突变。另外，在缺少同源重组蛋白Rad52的情况下，这些位点内的突变频率保持不变。使用替代的聚合酶也发现了突变。这些结果表明，至少某些突变是由使用的聚合酶引起的伪像，建议PCR后操作可能会产生突变，这些突变可能与真正的突变无法区分开，因此可能会干扰突变分析。此外，我们建议将用于确定单倍型的克隆等位基因序列中发现的新突变与汇集的DNA产生的序列进行对比。

BACKGROUND & AIMS: :Two isolates of Candida glabrata from the same stool sample from a bone marrow transplant recipient treated with fluconazole, and designated 1084-L for large colonies on yeast extract-peptone-dextrose-agar and 1084-S for small colonies, were analysed. In-vitro susceptibility tests with a commercially available disk diffusion procedure showed that isolate 1084-L had a susceptibility pattern typical of wild-type strains of C. glabrata with sensitivity to polyenes and the presence of resistant colonies randomly distributed within the inhibition zones for all azole compounds except tioconazole. In contrast, isolate 1084-S, which was found by pulsed-field gel electrophoresis and random amplification of polymorphic DNA to be genetically closely related to isolate 1084-L, exhibited cross-resistance to the azole compounds except tioconazole. Determination of MICs by the E-test method confirmed these results, showing that isolate 1084-S had greater sensitivity to amphotericin B and complete resistance to ketoconazole and fluconazole. Growth on agar plates containing glucose or glycerol as the sole carbon source suggested that the resistant isolate had a respiratory deficiency, which was further demonstrated by flow cytometric analysis of the fluorescence of rhodamine 123-stained blastoconidia. Restriction endonuclease analysis of mitochondrial DNA (mtDNA) established the mitochondrial origin of the respiratory deficiency. However, PCR amplification of the mtDNA with primers ML1 and ML6, as well as transmission electron microscopy, suggested a partial deletion of the mtDNA analogous to that described for rho- petite mutants of Saccharomyces cerevisiae. Together, these results provided evidence that the selection of azole-resistant petite mutants of C. glabrata may occur in vivo after fluconazole administration, which might explain, therefore, clinical failure of antifungal therapy.

背景与目标： ：分析了来自两个接受氟康唑处理的骨髓移植受者相同粪便样品的光滑念珠菌分离物，分别将其在酵母提取物-pe-葡萄糖-琼脂上的大菌落命名为1084-L，在小菌落上命名为1084-S。使用市售的磁盘扩散程序进行的体外药敏试验表明，分离株1084-L具有典型的光滑毛孢菌野生型药敏模式，对多烯敏感，并且所有菌落均在抑制区内随机分布有抗性菌落。噻康唑以外的其他唑化合物。相反，通过脉冲场凝胶电泳和多态性DNA的随机扩增发现分离株1084-S在遗传上与分离株1084-L密切相关，除噻康唑外，对唑类化合物表现出交叉耐药性。通过E-test法测定MIC证实了这些结果，表明分离物1084-S对两性霉素B的敏感性更高，对酮康唑和氟康唑的完全耐药性。在含有葡萄糖或甘油作为唯一碳源的琼脂平板上生长表明该抗性分离株具有呼吸道缺陷，这通过若丹明123染色的球孢子菌荧光的流式细胞术分析进一步证实。线粒体DNA（mtDNA）的限制性核酸内切酶分析确定了呼吸不足的线粒体起源。但是，用引物ML1和ML6进行mtDNA的PCR扩增以及透射电镜观察，表明部分mtDNA缺失，类似于酿酒酵母变节菌突变体中所述。在一起，这些结果提供了证据，在给予氟康唑后，可能在体内发生了光滑毛孢梭菌的耐唑性娇小突变体的选择，因此，这可能解释了抗真菌治疗的临床失败。

BACKGROUND & AIMS: :Candida albicans is considered the main pathogenic yeast responsible for a multitude of infective disorders. However, other yeasts, such as Candida famata, are being recognized as potential emerging pathogens that cause several types of infections in humans and animals. Consequently, we have investigated the adhesion and internalization of Candida famata into monocytes and epithelial cells. The interaction of the yeast with the cells is very rapid and takes place during the first 15 min of injection. However, the affinity of the yeast for the cells varies, THP-1 (human monocytes) being the highest and followed in decreasing order by HeLa (human carcinoma), HaCaT, and Pam-212 (human and mouse keratinocytes, respectively). Heat inactivation or treatment with nystatin, significantly decreases yeast adhesion to cells. Immunofluorescence, as well as scanning and transmission electron microscopy, reveals that cell lines are able to internalize C. famata. At 48 h after infection, most of the yeasts located inside cells appear degraded, but some yeasts recovered from lysed cells, were still viable. Adhesion and internalization of C. famata into HeLa cells were found to be lower than those of C. albicans and C. glabrata, but higher than those of S. cerevisiae. In addition, infection with C. famata results in actin microfilaments rearrangement. This article presents novel data in the interaction of this pathogenic yeast with mammalian cells.

背景与目标： ：白色念珠菌被认为是导致多种感染性疾病的主要致病酵母。但是，其他酵母，例如念珠菌，也被认为是潜在的新兴病原体，可引起人类和动物的几种感染。因此，我们调查了念珠菌进入单核细胞和上皮细胞的黏附和内在化。酵母与细胞的相互作用非常迅速，并在注射的前15分钟内发生。但是，酵母对细胞的亲和力各不相同，THP-1（人类单核细胞）最高，依次为HeLa（人类癌），HaCaT和Pam-212（分别为人类和小鼠角质形成细胞）。热灭活或用制霉菌素处理，会显着降低酵母菌对细胞的粘附力。免疫荧光以及扫描和透射电子显微镜显示，细胞系能够内化法氏梭菌。感染后48小时，位于细胞内的大多数酵母菌似乎已降解，但从裂解细胞中回收的一些酵母菌仍能存活。发现C. famata在HeLa细胞中的黏附和内在化低于白色念珠菌和光滑念珠菌，但高于酿酒酵母。另外，用法氏梭状芽胞杆菌感染导致肌动蛋白微丝重排。本文介绍了这种病原酵母与哺乳动物细胞相互作用的新数据。

BACKGROUND & AIMS: :Oropharyngeal candidiasis (OPC) caused by Candida albicans is a significant problem in human immunodeficiency virus (HIV)-infected persons. Recognizing the paucity of information on innate and/or adaptive mucosal host defenses against C. albicans, we recently reported that human and nonhuman primate and mouse vaginal epithelial cells inhibit the growth of C. albicans in vitro. In the present study, oral epithelial cells collected from saliva of healthy volunteers and a purified oral epithelial cell line were found to inhibit blastoconidia and/or hyphal growth of several Candida species. Cell contact was a strict requirement for the epithelial cell anti-Candida activity; neither saliva nor culture supernatants alone inhibited Candida growth, and addition of saliva to the coculture did not modulate the epithelial cell activity. Finally, epithelial cell anti-Candida activity was significantly lower in HIV-infected persons with OPC. Together, these results suggest that oral epithelial cells may play a role in innate resistance against OPC.

背景与目标： ：白色念珠菌引起的口咽念珠菌病（OPC）在人类免疫缺陷病毒（HIV）感染者中是一个重大问题。认识到关于白念珠菌的固有和/或适应性粘膜宿主防御的信息很少，我们最近报道了人和非人的灵长类和小鼠阴道上皮细胞在体外抑制白念珠菌的生长。在本研究中，发现从健康志愿者的唾液中收集的口腔上皮细胞和纯化的口腔上皮细胞系可抑制几种念珠菌的胚泡和/或菌丝生长。细胞接触是上皮细胞抗念珠菌活性的严格要求。唾液和培养物上清液都不能单独抑制念珠菌的生长，向共培养物中添加唾液并不能调节上皮细胞的活性。最后，在HIV感染的OPC患者中，上皮细胞的抗念珠菌活性显着降低。总之，这些结果表明口腔上皮细胞可能在对OPC的先天性​​抵抗中发挥作用。

BACKGROUND & AIMS: :Candida albicans cell wall constitutes a sensitive boundary that undergoes molecular changes upon environmental injuries. Antimycotics exert an intense action on cell wall eliciting both qualitative and quantitative changes of resident proteins. The emergence of drug resistance is marked by a modulation of cell wall proteomic profile. In this study, we monitored, at the proteome level through a two-dimensional gel electrophoresis-based approach, differences of cell wall proteins in sensitive and resistant strains of C. albicans, and variations occurring upon treatment of these strains with antifungal drugs. We identified Rhd3/Pga29, a glycophosphatidylinositol (GPI)-anchored protein, as the main over-expressed protein in micafungin resistant strain with respect to the sensitive control cells. A further increase of Rhd3/Pga29 took place when these resistant strains were treated with sub-lethal dose of micafungin. These results were also confirmed in other two clinical isolates resistant to caspofungin. Results were validated by Western blot analyses and RT-PCR and immunoelectron microscopy images confirmed the increase of the Rhd3/Pga29 on the cell wall as well as in the cytosolic compartment of the micafungin-treated resistant cells. Rhd3/Pga29 over-expression upon echinocandin treatment could represent a strategy of C. albicans to counteract the toxic action of this drug. A role of this protein has also been claimed in the virulence of the fungus, suggesting an involvement of Rhd3/Pga29 in the relationship between C. albicans and the host.

背景与目标： ：白色念珠菌细胞壁构成敏感边界，在环境受伤时会发生分子变化。抗真菌药对细胞壁产生强烈作用，引起驻留蛋白的质和量变化。耐药性的出现以细胞壁蛋白质组学特征的调节为标志。在这项研究中，我们通过基于二维凝胶电泳的方法在蛋白质组水平上监测了白色念珠菌敏感菌株和耐药菌株中细胞壁蛋白的差异，以及用抗真菌药物治疗这些菌株时发生的变化。我们确定Rhd3 / Pga29，糖磷脂酰肌醇（GPI）锚定的蛋白质，相对于敏感的对照细胞，在米卡芬净抗性菌株中主要过表达的蛋白质。当用亚致死剂量的米卡芬净治疗这些耐药菌株时，Rhd3 / Pga29进一步升高。这些结果在其他两种对卡泊芬净有抗药性的临床分离株中也得到了证实。结果通过Western印迹分析和RT-PCR验证，免疫电子显微镜图像证实在米卡芬净处理过的耐药细胞的细胞壁上以及胞质区室中Rhd3 / Pga29的增加。棘皮菌素治疗后Rhd3 / Pga29的过表达可能代表白色念珠菌抵消该药物毒性作用的策略。该蛋白在真菌的毒性中也被要求发挥作用，表明Rhd3 / Pga29参与了白色念珠菌与宿主之间的关系。