Immunomodulatory and protective effect of probiotic Lactobacillus casei against Candida albicans infection in malnourished mice

The effect of Lactobacillus casei CRL 431 (Lc), when administered as a supplement to a repletion diet, on the resistance of malnourished mice to Candida albicans infection was studied. Weaned mice were malnourished by being given a protein-free diet (PFD) for 21 days. The malnourished mice were then fed a balanced conventional diet (BCD) for 7 days or BCD for 7 days with supplemental Lc on days 6 and 7 (BCD+Lc). Malnourished (MNC) and well-nourished (WNC) mice were used as controls. At the end of the treatments the mice were infected intraperitoneally with C. albicans. Animals that had received probiotics had improved survival and resistance against this infection compared to those in the BCD and MNC groups. The number and fungicidal activity of phagocytes, and the concentrations of tumor necrosis factor-α, interferon-γ and interleukin-6 (IL-6), increased in blood and infected tissues in all experimental groups, but MNC mice showed lower concentrations than those in the WNC group. BCD and BCD+Lc mice showed higher concentrations of these variables than those in the MNC group, but only the BCD+Lc group presented values similar to the WNC mice. Malnutrition also impaired the production of IL-17 and IL-10 in response to infection. Both repletion treatments normalized IL-17 concentrations, but IL-10 in the BCD+Lc group was significantly higher than in WNC mice. The addition of L. casei to the repletion diet normalized the immune response against C. albicans, allowing efficient recruitment and activation of phagocytes, as well as effective release of pro-inflammatory cytokines. In addition, probiotic treatment induced an increase in IL-10 concentrations, which would have helped to prevent damage caused by the inflammatory response.     

SCID小鼠肠道菌群失调对深部白色念珠菌感染的影响研究

目的建立重度联合免疫缺陷（SCID）小鼠白色念珠菌感染模型,探讨肠道菌群失调与深部白色念珠菌感染的联系。方法SCID小鼠随机口服万古霉素水溶液7d,饥饿24h后给予白色念珠菌灌胃,建立小鼠白色念珠菌感染模型,观察小鼠死亡情况。荧光定量PCR检测肠道细菌总量、基质辅助激光解析电离飞行时间质谱仪鉴定肠道菌群种类,并应用扫描电镜观察肠壁黏膜组织超微结构的改变。结果应用万古霉素可致肠道菌群失调,肠道黏膜完整性受损。在万古霉素致肠道菌群失调的基础上,外源性白色念珠菌攻击可加重肠道菌群失调和肠壁黏膜损伤程度,促进深部白色念珠菌感染的发生。结论肠道菌群失衡可以导致深部白色念珠菌感染的发生,肠壁黏膜的完整性可能参与了肠道白色念珠菌播散过程。     

腹部大手术术后深部真菌感染的临床与真菌学分析

目的探讨腹部大手术术后深部真菌感染的危险因素及诊治措施。方法回顾性分析2006年1月～2009年6月期间我科腹部大手术术后合并深部真菌感染48例患者的临床和真菌学资料。结果 48例真菌感染患者共分离出56株菌株,其中白念珠菌占41.1%,是最主要的致病菌株。患者基础疾病和术后长期、多种广谱抗生素联合使用是深部真菌感染的重要因素。结论深部真菌感染是腹部大手术术后的重要并发症,白念珠菌仍然是主要病原菌。对术后患者深部真菌感染应采取积极预防、及时发现和有效治疗。     

铜绿假单胞菌菌苗预防白色念珠菌感染的实验研究

目的利用铜绿假单胞菌菌苗探索阻断白色念珠菌感染的影响。方法家兔耳皮内注射白色念珠菌,进行活菌攻击,建立动物感染模型。观察用铜绿假单胞菌菌苗免疫的家兔状况;用试管凝集法测定家免的抗体效价;采用小白鼠体内吞噬法测定免疫组和空白对照组巨噬细胞的吞噬百分率和吞噬指数,以得出铜绿假单胞菌菌苗的免疫性。结果观察发现：与对照组比较,铜绿假单胞菌菌苗免疫组抗体效价,巨噬细胞的吞噬能力明显提高;注射区局部感染及全身症状明显减轻;肾脏感染灶减少,病理切片显示病情较轻。结论铜绿假单胞菌菌苗能够减轻注射区局部感染及全身症状,对肾脏的播散性白色念珠菌感染具有一定的阻断作用。     

Miconazole nitrate bearing ultraflexible liposomes for the treatment of fungal infection

Miconazole nitrate is a widely used antifungal agent, but its use in topical formulations is not efficacious because deep seated fungal infections are difficult to treat with conventional topical formulation. Miconazole nitrate loaded ultraflexible liposomes have been prepared and their topical performance has been compared with conventional liposomes containing miconazole nitrate. Various ultraflexible liposomal formulations were prepared and extensively characterized for vesicular shape, size, entrapment efficiency, degree of deformability and in-vitro skin permeation through rat skin. Higher rate of drug transfer across the skin with ultraflexible liposomal formulations of miconazole nitrate suggests that the drug in its lipo-solubilized state might have gained facilitated entry into the tough barrier consisting of subcutaneous. In-vivo study showed better antifungal activity as compared to traditional liposomes and plain drug solution. This was confirmed through fluoroscence microscopy. It is concluded that prepared ultraflexible liposomes can facilitate improved and localized drug action in the skin, thus providing a better option to deal with deep seated skin problems.     

Protective effect of fungal extracellular vesicles against murine candidiasis

Extracellular vesicles (EVs) are lipid bilayered compartments released by virtually all living cells, including fungi. Among the diverse molecules carried by fungal EVs, a number of immunogens, virulence factors and regulators have been characterised. Within EVs, these components could potentially impact disease outcomes by interacting with the host. From this perspective, we previously demonstrated that EVs from Candida albicans could be taken up by and activate macrophages and dendritic cells to produce cytokines and express costimulatory molecules. Moreover, pre‐treatment of Galleria mellonella larvae with fungal EVs protected the insects against a subsequent lethal infection with C. albicans yeasts. These data indicate that C. albicans EVs are multi‐antigenic compartments that activate the innate immune system and could be exploited as vaccine formulations. Here, we investigated whether immunisation with C. albicans EVs induces a protective effect against murine candidiasis in immunosuppressed mice. Total and fungal antigen‐specific serum IgG antibodies increased by 21 days after immunisation, confirming the efficacy of the protocol. Vaccination decreased fungal burden in the liver, spleen and kidney of mice challenged with C. albicans. Splenic levels of cytokines indicated a lower inflammatory response in mice immunised with EVs when compared with EVs + Freund's adjuvant (ADJ). Higher levels of IL‐12p70, TNFα and IFNγ were detected in mice vaccinated with EVs + ADJ, while IL‐12p70, TGFβ, IL‐4 and IL‐10 were increased when no adjuvants were added. Full protection of lethally challenged mice was observed when EVs were administered, regardless the presence of adjuvant. Physical properties of the EVs were also investigated and EVs produced by C. albicans were relatively stable after storage at 4, ?20 or ?80°C, keeping their ability to activate dendritic cells and to protect G. mellonella against a lethal candidiasis. Our data suggest that fungal EVs could be a safe source of antigens to be exploited in vaccine formulations.     

The effect of denture adhesives on Candida albicans growth in vitro

doi: 10.1111/j.1741‐2358.2011.00478.x
The effect of denture adhesives on Candida albicans growth in vitro Objective: Denture‐wearing favours the growth of Candida. In view of the fact that many denture wearers regularly use adhesives to enhance denture retention, stability and function, the aim of this work was to study the effect of denture adhesives on Candida albicans growth in vitro. Materials and methods: The denture adhesives tested were Corega^® cream, Kukident^® cream, Novafix^® cream, Polident^® cream, Protefix^® cream, Steradent^® cream, Aderyn^® powder, Corega^® ultra powder, Protefix^® powder and Corega^® strip. C. albicans growth curves were obtained in the presence or absence of a 1% solution of the denture adhesive diluted in Sabouraud broth. Macro‐ and microscopic morphological changes in C. albicans were analysed, as was microbial contamination of the denture adhesive. Results: Most of the denture adhesives studied induced morphological changes in C. albicans cells and colonies, but only two had any significant inhibitory effect on yeast growth. Kukident^® cream markedly inhibited C. albicans growth in a concentration‐dependent way, reducing the growth rate by 95%, whereas Corega^® cream also inhibited C. albicans growth but in a non‐concentration‐dependent way, reducing the growth rate by 37%. In addition, denture adhesives available as powders had detectable microbial contamination. Conclusion: Some commercially available denture adhesives showed microbial contamination and some had significant inhibitory effect on C. albicans growth.     

紫外线对白念珠菌的影响

紫外线作为重要的环境因子之一,能显著影响包括白念珠菌在内的多种生物的生长及生理过程.研究发现白念珠菌受紫外线照射后菌丝形成被抑制,孢子形成增多且没有向光性;脉冲紫外线辐射可通过多靶点程序使白念珠菌失活;rad 51缺陷株比rad 52缺陷株更易受紫外线损害,同时紫外线可导致白念珠菌杂合性丢失.研究还发现UVC治疗可明显减少烧伤后真菌微生物感染,核黄素/UVA治疗可明显抑制白念珠菌生长.因此紫外线对白念珠菌有一定的抑制作用.     

Early inflammatory responses to Candida albicans infection in inbred and complement-deficient mice

Abstract Inflammatory responses that developed in the footpad during the first 48 h after inoculation of Candida albicans were compared in six genetically defined inbred strains of mice. Tissue responses consisted predominantly of accumulations of polymorphonuclear leucocytes, the magnitude of which was significantly less in mice lacking the fifth component of complement (C5). Despite this, there was no difference between C5-sufficient and C5-deficient mice in the total infectious burden, nor did depletion of complement by treatment with cobra venom factor cause any detectable reduction in the numbers of inflammatory cells in the area of the lesion. Ablation of granulocytes had no significant effect on the fungal burden over the period of the experiment. Immunisation provided some protection against tissue damage, but did not reduce the number of yeasts at the site of infection.     

Lasioglossins LLIII affect the morphogenesis of Candida albicans and reduces the duration of experimental vaginal candidiasis in mice

Lasioglossins are a group of peptides with identified antimicrobial activity. The inhibitory effects of two synthetic lasioglossin derivatives, LLIII and D‐isomeric variant LLIII‐D, on morphological changes in Candida albicans in vitro and the effect of local administration of LLIII during experimental murine candidiasis were investigated. C. albicans blastoconidia were grown in the presence of lasioglossin LLIII or LLIII‐D at concentrations of 11.5 μM and 21 μM, respectively, for 1, 2 and 3 days and their viability determined by flow cytometry using eosin Y staining. Morphological changes were examined by light and fluorescent microscopy. The Candida‐inhibitory effect of daily intravaginal administration of 0.7 or 1.4 μg of LLIII was assessed in mice with experimentally‐induced vaginal candidiasis. LLIII and LLIII‐D lasioglossins exhibited candidacidal activity in vitro (>76% after 24 hr and >84% after 48 hr of incubation). After 72 hr incubation of Candida with low concentration of lasioglossins, an increase in viability was detected, probably due to a Candida antimicrobial peptides evasion strategy. Furthermore, lasioglossins inhibited temperature‐induced morphotype changes toward hyphae and pseudohyphae with sporadic occurrence of atypical cells with two or enlarged nuclei, suggesting interference with mitosis or cytokinesis. Local application of LLIII reduced the duration of experimental candidiasis with no evidence of adverse effects. Lasioglossin LLIII is a promising candidate for development as an antimicrobial drug for treating the vaginal candidiasis.

     

Antifungal drugs effect adherence of Candida albicans to acrylic surfaces by changing the zeta-potential of fungal cells

The effect of sub-inhibitory concentrations of antifungal drugs on the adherence of Candida albicans to acrylic surfaces was investigated. Among five antifungals tested, azalomycin F and aculeacin A significantly enhanced the adherence. The zeta-potential of fungal cells was affected by antifungal drugs, whereas no significant change in cell surface hydrophobicity was observed. The relationship obtained between the change in the adherence and that in zeta-potential suggests that the enhanced adherence was caused by decreased electric repulsive forces.     

三颗针对白假丝酵母的抑制作用

目的 研究三颗针对白假丝酵母的抑制作用。方法 采用溴化噻唑蓝四唑法（MTT）和琼脂平板法测定三颗针对白假丝酵母的最低抑菌浓度（MIC）和最低杀菌浓度（MFC）;利用倒置荧光显微镜观察三颗针对白假丝酵母菌丝形成的影响;采用MTT法测定三颗针对白假丝酵母菌丝萌发不同时期的抑制率。结果 三颗针对白假丝酵母具有较强的抑制作用。其中,采用琼脂平板法测得三颗针抑制该菌的MIC为2 mg/mL,MFC为4 mg/mL。三颗针能减缓该菌的生长速度或使其停止生长,且浓度越高作用效果越明显。其中4 mg/mL的三颗针作用白假丝酵母6 h后,能够完全抑制菌丝形成。对于24 h后已经萌发为菌丝态的白假丝酵母,三颗针可抑制菌丝的继续生长,与对照组相比,4 mg/mL的三颗针对于萌发2 h后的白假丝酵母的菌丝抑制率为76.7%（P<0.01）。结论 三颗针能抑制白假丝酵母菌丝的生长和萌发。     

大蒜素对白念珠菌形态转换的影响研究

目的探讨大蒜素对白念珠菌形态转换的影响及其作用机制。方法倒置显微镜观察白念珠菌菌丝形成的体外动力学过程;采用CLSI-M27-A3微量液基稀释法检测大蒜素对白念珠菌的最小抑菌浓度(minimum inhibitory concentration,MIC);倒置显微镜观察不同浓度大蒜素对白念珠菌在Spider液体培养基中菌丝形成的影响;qRT-PCR法检测在不同浓度大蒜素作用下白念珠菌菌丝相关基因HWP1、ALS1、EFG1、PDE2表达水平的变化。结果白念珠菌在Spider液体培养基中6 h时出现较长菌丝,24 h后镜下可见大量念珠菌菌丝包裹酵母细胞,紧密交错;大蒜素对白念珠菌的MIC值为25μg/mL;倒置显微镜观察(25~100)μg/mL浓度的大蒜素能明显抑制Spider液体培养基中白念珠菌菌丝的生长;qRT-PCR结果显示,在(25~100)μg/mL浓度的大蒜素作用下,白念珠菌菌丝相关基因表达下调。结论大蒜素能有效抑制白念珠菌的形态转换,其作用机制可能与调节菌丝形成相关基因的表达水平有关。     

不同消毒方式对硅橡胶类义齿软衬材料消毒效果的实验研究

目的 对比微波、紫外线和环氧乙烷对Silagum硅橡胶义齿软衬材料的消毒效果.方法 将高温消毒后的软衬硅橡胶试件染菌(白色念珠菌),分别进行不同时间的紫外线、微波、环氧乙烷消毒,消毒后的试件用无菌生理盐水冲洗,无菌玻璃平皿收集冲洗液,接种环将冲洗液接种于沙氏琼脂平板培养基,37℃恒温培养24～48 h观察,菌落计数,评价消毒效果.结果 环氧乙烷(EO浓度600 mg/L)肖毒60 min灭菌率为100％;紫外线(功率40 W、波长253.7 nm、距离1 m)照射40 min可杀灭99％的白色念珠菌、50 min可完全杀灭;微波(800 W V 100％)照射1 min灭菌率为93％,照射2 min灭菌率为100％.结论 三种消毒方法都具有消毒效果,且消毒时间越长消毒效果越好;环氧乙烷、紫外线、微波消毒口腔硅橡胶义齿软衬材料可与高温高压灭菌达到同样的效果,而且方便、有效,是良好的软衬材料的消毒方法.     

Modulation of neutrophil function in host defense against disseminated Candida albicans infection in mice

Neutrophils (PMNs) constitute the main mechanism of host defense against acute invasive and disseminated candidiasis. Recent studies have demonstrated that tumor necrosis factor-alpha (TNFalpha), interleukin-6 (IL-6) and granulocyte colony-stimulating factor (G-CSF) play an important role in the recruitment of PMNs at the site of invasive Candida infection. In the absence of either TNFalpha or IL-6, the course of experimental disseminated candidiasis is more severe, due to defective PMN recruitment. Treatment of mice with recombinant G-CSF (rG-CSF) leads to a significantly reduced mortality during disseminated candidiasis. The outgrowth of Candida albicans from the organs of rG-CSF-treated mice is significantly decreased. Treatment with the combination of rG-CSF and fluconazole has an additive effect on the reduction of fungal load in the organs. In subacute or chronic disseminated Candida infection, rG-CSF is less effective, indicating that neutrophil recruitment and activation are crucial in acute, life-threatening candidiasis, whereas other host defense mechanisms control the outcome of less overwhelming invasive Candida infection.     

Effect of suramin on the human pathogen Candida albicans: implications on the fungal development and virulence

Candida albicans is an opportunistic pathogen that is of growing medical importance because it causes superficial, mucosal and systemic infections in susceptible individuals. Here, the effect of suramin, a polysulfonated naphthylurea derivative, on C. albicans development and virulence was evaluated. Firstly, it was demonstrated that suramin (500 microM) arrested its growth, showing a fungicidal action dependent on cell number. Suramin treatment caused profound changes in the yeast ultrastructure as shown by transmission electron microscopy. The more important changes were the enlargement of the fungi cytoplasmic vacuoles, the appearance of yeasts with an empty cytoplasm resembling ghost cells and a reduction in cell wall thickness. Suramin also blocked the transformation of yeast cells to the germ-tube and the interaction between C. albicans and epithelial cells. In order to ascertain that the action of suramin on C. albicans growth is a general feature instead of being strain-specific, the effects of suramin on 14 oral clinical strains isolated from healthy children and HIV-positive infants were analyzed. Interestingly, the strains of C. albicans isolated from HIV-positive patients were more resistant to suramin than strains isolated from healthy patients. Altogether, the results produced here show that suramin interfered with essential fungal processes, such as growth, differentiation and interaction with host cells.