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【白色念珠菌myristoylCoA的选择性肽和拟肽抑制剂：蛋白N-肉豆蔻酰基转移酶：一种抗真菌治疗的新方法。】
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DOI：10.1002/(SICI)1097-0282(1997)43:1<43::AID-BIP5>3.0
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BACKGROUND & AIMS: MyristoylCoA:protein N-myristoyltransferase (NMT) catalyzes the cotranslational covalent attachment of a rare cellular fatty acid, myristate, to the N-terminal Gly residue of a variety of eukaryotic proteins. The myristoyl moiety is often essential for expression of the biological functions for these proteins.
Attachment of C14:0 alone provides barely enough hydrophobicity to allow stable association with membranes. The partitioning of N-myrisotylproteins is therefore often modulated by "switches" that function through additional covalent or noncovalent modifications. Candida albicans, the principal cause of systemic fungal infection in immunocompromised humans, contains a single NMT gene that is essential for its viability. The functional properties of the acylCoA binding site of human and C. albicans NMT are very similar. However, there are distinct differences in their peptide binding sites. An ADP ribosylation factor (Arf) is included among the few cellular protein substrates of the fungal enzyme. Alanine scanning mutagenesis of an octapeptide derived from an N-terminal Arf sequence (GLYASKLS-NH2) disclosed that Gly1, Ser5, and Lys6 play predominant roles in binding. ALYASKLS-NH2 is an inhibitor competitive for peptide [Ki(app) = 15.3 +/- 6.4 microM] and noncompetitive for myristoylCoA. Remarkably, replacement of the N-terminal tetrapeptide with an 11-aminoundecanoyl group results in a competitive inhibitor (11-aminoundecanoyl-SKLS-NH2) that is approximately 40-fold more potent [Ki(app) = 0.40 +/- 0.03 microM] than the starting octapeptide. Removal of Leu-Ser from the C-terminus generates a competitive dipeptide inhibitor (11-aminoundecanoyl-SK-NH2) with a Ki(app) of 11.7 +/- 0.4 microM, equivalent to that of the starting octapeptide. A derivative dipeptide inhibitor containing a C-terminal N-cyclohexylethyl lysinamide moiety has the advantage of being more potent (IC50 = 0.11 +/- 0.03 microM) and resistant to digestion by cellular carboxypeptidases. Rigidifying the flexible aminoundecanoyl chain results in very potent general NMT inhibitors (IC50 = 40-50 nM). Substituting a 2-methylimidazole for the N-terminal amine and adding a benzylic alpha-methyl group with R stereochemistry to the rigidifying element produces even more potent inhibitors (IC50 = 20-50 nM) that are up to 500-fold selective for the fungal compared to human enzyme. A related less potent member of this series of compounds is fungistatic. Its growth inhibitory effects are associated with a reduction in cellular protein N-myristoylation, judged using cellular Arf as a reporter. These studies establish that NMT is a new antifungal target.
背景与目标： MyristoylCoA ：蛋白N-肉豆蔻酰基转移酶（NMT）催化稀有细胞脂肪酸肉豆蔻酸酯与多种真核蛋白N-末端Gly残基的共翻译共价连接。肉豆蔻酰基部分通常对于表达这些蛋白质的生物学功能是必不可少的。
仅C14 ：0的附件仅提供了不足以使与膜稳定结合的疏水性。因此，N-肉豆蔻酰蛋白的分区通常由通过附加的共价或非共价修饰起作用的“开关”调节。白色念珠菌是免疫力低下的人体内全身真菌感染的主要原因，它包含一个对其生存能力至关重要的单个NMT基因。人和白色念珠菌NMT的酰基辅酶A结合位点的功能特性非常相似。但是，它们的肽结合位点存在明显差异。 ADP核糖基化因子（Arf）包括在真菌酶的几种细胞蛋白底物中。来自N末端Arf序列（GLYASKLS-NH2）的八肽的丙氨酸扫描诱变显示，Gly1，Ser5和Lys6在结合中起主要作用。 ALYASKLS-NH2是一种对肽[Ki（app）= 15.3 /-6.4 microM]具有竞争性的抑制剂，对肉豆蔻酰辅酶A不具有竞争性。值得注意的是，用11-氨基十一烷酰基取代N末端四肽会产生竞争性抑制剂（11-氨基十一烷酰基-SKLS-NH2），其效力比[Ki（app）= 0.40 /-0.03 microM]高40倍左右。起始八肽。从C末端去除Leu-Ser会产生竞争性的二肽抑制剂（11-氨基十一烷酰基-SK-NH2），Ki（app）为11.7 /-0.4 microM，与起始八肽相当。含有C-末端N-环己基乙基赖氨酰胺部分的衍生物二肽抑制剂具有更有效的优势（IC 50 ＝ 0.11 /0.03μM），并且对细胞羧肽酶的消化具有抵抗力。刚性化柔性氨基十一烷酰基链会产生非常有效的常规NMT抑制剂（IC50 = 40-50 nM）。用2-甲基咪唑代替N端胺，并在刚性元素中添加具有R立体化学的苄基α-甲基，可产生更强效的抑制剂（IC50 = 20-50 nM），对真菌的选择性高达500倍与人类酶相比。该系列化合物的一个相关的效力较弱的成员是抑真菌的。使用细胞Arf作为报告基因判断，其生长抑制作用与细胞蛋白N-肉豆蔻酰化的减少有关。这些研究确定NMT是一种新的抗真菌靶标。
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【念珠菌性脊椎炎和硬膜外脓肿】
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BACKGROUND & AIMS: :Candida spondylodiscitis associatd with epidural abscess is rarely seen. We present a patient with Hodgkin lymphoma who received chemotherapy and developed systemic Candida infection, which was complicated by Candida spondylodiscitis and epidural abscess.背景与目标： ：与硬膜外脓肿相关的念珠菌脊椎盘炎很少见。我们介绍了一名霍奇金淋巴瘤患者，该患者接受了化疗并发展为全身性念珠菌感染，并发念珠菌脊椎盘炎和硬膜外脓肿。
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3 Viable quantitative PCR for assessing the response of Candida albicans to antifungal treatment.
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【可行的定量PCR，用于评估白色念珠菌对抗真菌治疗的反应。】
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DOI：10.1007/s00253-012-4524-z
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BACKGROUND & AIMS: :Propidium monoazide (PMA) or ethidium bromide monoazide (EMA) treatment has been used before nucleic acid detection methods, such as PCR, to distinguish between live and dead cells using membrane integrity as viability criterion. The performance of these DNA intercalating dyes was compared in many studies utilizing different microorganisms. These studies demonstrated that EMA and PMA differ in their abilities to identify nonviable cells from mixed cell populations, depending on the microorganism and the nature of the sample. Due to this heterogeneity, both dyes were used in the present study to specifically distinguish dead from live Candida albicans cells using viable quantitative PCR (qPCR). The viable qPCR was optimized, and the best results were obtained when pre-treating the cells for 10 min in the dark with 25 μM EMA followed by continuous photoactivation for 15 min. The suitability of this technique to distinguish clotrimazole- and fluconazole-treated C. albicans cells from untreated cells was then assessed. Furthermore, the antifungal properties of two commercial essential oils (Thymus vulgaris and Matricaria chamomilla) were evaluated. The viable qPCR method was determined to be a feasible technique for assessing the viability of C. albicans after drug treatment and may help to provide a rapid diagnostic and susceptibility testing method for fungal infections, especially for patients treated with antifungal therapies.背景与目标： 在核酸检测方法（例如PCR）之前，已经使用单叠氮化丙锭（PMA）或溴化乙叠氮化单乙胺（EMA）处理，以膜完整性作为生存力标准来区分活细胞和死细胞。在许多利用不同微生物的研究中，对这些DNA嵌入染料的性能进行了比较。这些研究表明，EMA和PMA从混合细胞群体中鉴定出无活力细胞的能力有所不同，具体取决于微生物和样品的性质。由于这种异质性，在本研究中使用了两种染料，以使用可行的定量PCR（qPCR）特异性区分死者和活白色念珠菌细胞。优化了可行的qPCR，当在黑暗中用25μMEMA在黑暗中预处理细胞10分钟，然后连续光激活15分钟时，可获得最佳结果。然后评估了该技术从未处理的细胞中区分出克霉唑和氟康唑处理过的白色念珠菌细胞的适用性。此外，还评估了两种市售精油（百里香和母菊）的抗真菌性能。确定可行的qPCR方法是评估药物治疗后白色念珠菌生存力的可行技术，并且可能有助于为真菌感染（尤其是抗真菌治疗的患者）提供快速的诊断和药敏测试方法。 -
【副念珠菌中基因敲除系统的开发揭示了BCR1在生物膜形成中的保守作用。】
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DOI：10.1128/EC.00136-07
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BACKGROUND & AIMS: :Candida parapsilosis is an important cause of candidiasis, yet few molecular tools are available. We adapted a recyclable nourseothricin resistance marker gene (SAT1) originally developed for use with C. albicans in order to generate gene knockouts from C. parapsilosis. We first replaced the promoters driving expression of the FLP recombinase and the SAT1 genes with the equivalent sequences from C. parapsilosis. We then used the cassette to generate a homozygous knockout of C. parapsilosis URA3. The ura3 knockouts have altered colony morphologies. We also knocked out both alleles of an ortholog of BCR1, a gene encoding a transcription factor known to be required for biofilm development in C. albicans. We show that C. parapsilosis BCR1 is necessary for biofilm development in C. parapsilosis and for expression of the cell wall protein encoded by RBT1. Our results suggest that there are significant similarities in the regulation of biofilms between the two species, despite the fact that C. parapsilosis does not generate true hyphae and that BCR1 regulates the expression of many hypha-specific adhesins in C. albicans.背景与目标： 念珠菌：念珠菌是念珠菌病的重要原因，但是分子工具很少。我们调整了最初为白念珠菌而开发的可回收的神经丝菌素抗性标记基因（SAT1），以便从副念珠菌中产生基因敲除。我们首先用来自C. parapsilosis的等效序列替换了驱动FLP重组酶和SAT1基因表达的启动子。然后，我们使用盒式磁带产生了C. parapsilosis URA3的纯合敲除。 ura3基因敲除改变了菌落的形态。我们还敲除了BCR1直系同源基因的两个等位基因，BCR1是编码白色念珠菌生物膜发育所需的转录因子的基因。我们表明，C。parapsilosis BCR1是必要的生物膜发育在C. parapsilosis和由RBT1编码的细胞壁蛋白的表达。我们的结果表明，尽管事实上副寄生念珠菌不会产生真正的菌丝并且BCR1调节了白色念珠菌中许多菌丝特异性粘附素的表达，但两种物种在生物膜的调控上存在着显着的相似之处。
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【Galectin-3诱导表达特定β-1,2-连接的甘露聚糖的念珠菌死亡。】
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DOI：10.4049/jimmunol.177.7.4718
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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。
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【Psd1对白色念珠菌浮游细胞和生物膜的影响。】
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DOI：10.3389/fcimb.2017.00249
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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。
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【在抗逆转录病毒疗法时代，来自HIV感染患者的口腔念珠菌分离株的种类分布和抗真菌药敏性。】
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DOI：10.1007/s11046-006-0032-y
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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。
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【评价Mueller-Hinton-琼脂作为生产白色念珠菌和dubliniensis念珠菌的简单培养基。】
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DOI：10.1111/j.1439-0507.2007.01469.x
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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。
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9 New antifungal agents that inhibit the growth of Candida species: dichlorinated 8-quinolinols.
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【抑制念珠菌物种生长的新型抗真菌剂：二氯8-喹啉醇。】
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DOI：10.1023/a:1007120913871
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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。 -
【白色念珠菌在新培养基上形成胚芽管和衣原体。】
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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。
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【碳源调节了光滑念珠菌生物膜细胞对氟康唑的转录反应。】
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DOI：10.1038/s41522-020-0114-5
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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。
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【副念珠菌ATCC 7330不对称还原3-氧代丁酸烷基酯：深入了解生物催化反应的溶剂和底物。】
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DOI：10.1007/s12010-013-0379-8
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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。
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【银纳米颗粒对白色念珠菌的抗真菌活性和作用方式。】
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DOI：10.1007/s10534-008-9159-2
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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。
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【在社会经济上处于不利地位的美国孕妇的口腔健康和念珠菌运输。】
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DOI：10.1186/s12884-019-2618-7
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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。 -
【转载两种比较方法以鉴定白色念珠菌和酿酒酵母中单倍型和点突变的比较。】
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DOI：10.1016/j.mimet.2013.09.010
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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。
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