Allergic bronchopulmonary aspergillosis in children

Allergic bronchopulmonary aspergillosis (ABPA) is a disease of the lungs resulting from a hypersensitivity reaction to spores of Aspergillus fumigatus. Here we report the case of a 13‐year‐old girl with ABPA who presented with productive cough, bronchiectasis and decline in lung function, and review the clinical features and treatment for pediatric ABPA.     

Efficacy of oleylphosphocholine (OlPC) in vitro and in a mouse model of invasive aspergillosis

Invasive aspergillosis (IA) has become increasingly common and is characterised by high morbidity and mortality. Upcoming resistance threatens treatment with azoles and highlights the continuous need for novel therapeutics. This laboratory study investigated the in vitro and in vivo potential of the alkylphospholipid oleylphosphocholine (OlPC) against Aspergillus. In vitro activities of OlPC, miltefosine, posaconazole and voriconazole were determined for Aspergillus fumigatus, A. niger, A. terreus and A. flavus. In vivo efficacy of OlPC was evaluated in a systemic A. fumigatus mouse model, adopting a short‐term and long‐term oral or intraperitoneal dosing regimen. OlPC showed good in vitro activity against A. fumigatus (IC₅₀ = 1.04 μmol l^?1). Intraperitoneal administration of 50 mg kg^?1 day^?1 OlPC significantly reduced the fungal organ burdens at 4 days post‐infection (dpi). Although 5‐ and 10‐day OlPC treatment improved survival, organ burdens were not affected at 10 and 15 dpi. While this study showed excellent in vitro activity of OlPC against Aspergillus spp., its therapeutic efficacy in an acute mouse model for IA was less convincing. Given the limited therapeutic options in the current antifungal market for invasive infections, OlPC activity should be assessed in a less stringent in vivo model, potentially in combination treatment with other already marketed antifungal drugs.     

Nebulised liposomal amphotericin B for Aspergillus lung diseases: case series and literature review

Over the past 10 years the incidence of Aspergillus spp. has significantly increased, and it is now the most widespread air transmission fungal pathogen in developed countries. Whatever the clinical expression of the pulmonary disease and despite recent progress in antifungal drug therapy, morbidity and mortality related to aspergillosis lung disease still constitute a serious threat for immunosuppressed or mildly immunocompromised patients. Moreover, the treatments currently used have many limitations due to adverse effects and drug interactions. Finally, subjects exposed to azoles present an increased risk of Aspergillus‐resistant strain emergence. We have reported five cases with aspergillosis lung diseases that were either difficult to control or in which patients had a contra‐indication to triazole therapy, but which showed durable improvement following the administration of nebulised liposomal amphotericin B. Our alternative strategy could be of interest for patients with aspergillosis lung disease who otherwise cannot be conventionally treated by triazoles.     

Diagnostic accuracy of the Aspergillus‐specific bronchoalveolar lavage lateral‐flow assay in haematological malignancy patients

We evaluated the performance of the Aspergillus‐specific lateral‐flow device (LFD) test for diagnosing invasive pulmonary aspergillosis (IPA) in patients with underlying haematological malignancies. Participating centres were the two Austrian University Hospitals of Graz and Innsbruck. LFD performance was evaluated with 95 bronchoalveolar lavage fluid (BALF) samples from 72 patients collected prospectively in Graz, and with 24 BALF bio bank samples from 23 patients (21 samples with probable IPA) in Innsbruck. Invasive fungal infections were classified according to the revised European Organization of Research and Treatment of Cancer/Mycoses Study Group criteria. Overall, 27 patients (30 samples) had probable IPA, 32 (43 samples) possible and 36 (46 samples) did not fulfil IPA criteria. The vast majority of patients – in particular those with probable IPA – received mould‐active treatment before bronchoscopy. Sensitivity, specificity, positive predictive value and negative‐predictive‐value for probable IPA diagnosis using the BALF‐LFD test were 71%, 76%, 35% and 94% for the Graz cohort. Sensitivity of the BALF‐LFD test for probable IPA was 57% in Innsbruck bio bank samples. Our results indicate that the BALF‐LFD‐test provides fast results with moderate sensitivities in patients with underlying haematological malignancies. Similar to other diagnostic tests and biomarkers sensitivity of the test may be influenced by ongoing systemic mould‐active treatment.     

八种化学消毒剂对黑曲霉菌杀灭效果的研究

目的了解8种常用化学消毒剂对黑曲霉菌的杀灭效果。方法用悬液定量杀菌试验法进行了观察。结果75%乙醇作用1 m in、4000 mg/L长链季铵盐作用5 m in、4000 mg/L聚醇醚碘作用45 m in、250 mg/L二氧化氯作用2.5 m in、3000 mg/L有效氯的84消毒液作用10 m in,对黑曲霉菌的杀灭对数值均>4.00。用8000 mg/L醋酸氯己定作用20 m in、5000 mg/L对氯间二甲苯酚作用30 m in、4000 mg/L聚维酮碘作用60 m in,对黑曲霉菌的杀灭对数值依次分别为0.60、0.86、0.83。结论在常规使用浓度下,乙醇、长链季铵盐、聚醇醚碘、二氧化氯和84消毒液对黑曲霉菌杀灭效果较好;醋酸氯己定、对氯间二甲苯酚、聚维酮碘对黑曲霉菌杀灭效果较差。     

Heat shock protein 90 (Hsp90): A novel antifungal target against Aspergillus fumigatus

Abstract

Invasive aspergillosis is a life-threatening and difficult to treat infection in immunosuppressed patients. The efficacy of current anti-Aspergillus therapies, targeting the cell wall or membrane, is limited by toxicity (polyenes), fungistatic activity and some level of basal resistance (echinocandins), or the emergence of acquired resistance (triazoles). The heat shock protein 90 (Hsp90) is a conserved molecular chaperone involved in the rapid development of antifungal resistance in the yeast Candida albicans. Few studies have addressed its role in filamentous fungi such as Aspergillus fumigatus, in which mechanisms of resistance may differ substantially. Hsp90 is at the center of a complex network involving calcineurin, lysine deacetylases (KDAC) and other client proteins, which orchestrate compensatory repair mechanisms of the cell wall in response to the stress induced by antifungals. In A. fumigatus, Hsp90 is a trigger for resistance to high concentrations of caspofungin, known as the paradoxical effect. Disrupting Hsp90 circuitry by different means (Hsp90 inhibitors, KDAC inhibitors and anti-calcineurin drugs) potentiates the antifungal activity of caspofungin, thus representing a promising novel antifungal approach. This review will discuss the specific features of A. fumigatus Hsp90 and the potential for antifungal strategies of invasive aspergillosis targeting this essential chaperone.     

曲霉菌临床分离株的分子鉴定及体外抗真菌药物敏感性分析

目的 研究曲霉菌临床分离株的分子鉴定及体外抗真菌药物的敏感性。方法 采用Internal Transcribed Spacer（ITS）及β-tubulin基因部分测序对临床分离的53株经形态学鉴定为曲霉的菌株进行分子鉴定,并按照美国临床实验室标准化协会（CLSI）推荐的M38-A2微量液基稀释法测定唑类（伊曲康唑、伏立康唑、泊沙康唑）,两性霉素B和棘白菌素类（米卡芬净、阿尼芬净和卡泊芬净）共7种抗真菌药物对所有菌株的最低抑菌浓度（minimum inhibitory concentration, MIC）或最低有效浓度（minimum effective concentration, MEC）。结果 53株曲霉菌包含烟曲霉复合体22株（烟曲霉21株和仑图卢斯曲霉1株）,黑曲霉复合体23株（塔宾曲霉16株、黑曲霉5株和百岁兰曲霉2株）及土曲霉和黄曲霉各4株;米卡芬净、阿尼芬净、卡泊芬净、泊沙康唑、伏立康唑、伊曲康唑、两性霉素B对曲霉菌株的MIC90为0.031、0.031、0.25、0.5、0.5、1、2μg/mL。除1株烟曲霉对伊曲康唑（MIC≥16μg/mL）和伏立康唑（MIC为2μg/mL）耐药及1株仑图卢斯曲霉对两性霉素B耐药（MIC为8μg/mL）外,其余菌株对7种药物均敏感。4种曲霉复合体对7种抗真菌药物的敏感性差异有统计学意义（P＜0.05）。结论 棘白菌素类药物体外抗曲霉菌活性最好,两性霉素B体外抗曲霉菌活性相对较差,唑类药物介于以上两者之间。不同种曲霉菌对不同抗真菌药物的敏感性存在差别。     

健康人外周血烟曲霉硫氧还蛋白还原酶抗原特异性T 淋巴细胞检测与结果评价

目的：为评估烟曲霉硫氧还蛋白还原酶（ thioredoxin reductase , TR）抗原是否具有免疫保护作用,文中建立并优化检测健康人外周血单个核细胞（peripheral blood mononuclear cell , PBMC）中TR抗原特异性T淋巴细胞（TR／AST）分泌干扰素γ（interferon γ, IFN-γ）与白细胞介素-4（interleukin-4, IL-4）的酶联免疫斑点法（enzyme linked immunospot assay , ELISPOT）检测技术,以探讨TR抗原特异性T淋巴细胞在侵袭性曲霉病（ invasive aspergillosis , IA）中的作用。方法采用方阵滴定法优化ELISPOT反应条件。以烟曲霉TR为特异性刺激物,阳性细胞刺激剂为对照,用ELISPOT技术检测20名健康人外周血PBMC分泌IFN-γ、IL-4的阳性斑点形成细胞（spot forming cell, SFC）频数。结果方阵滴定法结果显示,10μg TR抗原、健康人PBMC终浓度为3×105／孔时ELISPOT检测结果最佳。20名健康人特异性分泌IFN-γ和IL-4的SFC频数分别为15（3．5,59．5）个和0（0,0）个,IFN-γ的SFC频数显著高于IL-4,差异有统计学意义（P＜0．001）。 TR可刺激所有20名健康人产生IFN-γ应答,其中9例产生强IFN-γ应答（SFC＞20个／3×105 PBMC）,占45％;而19名健康人在TR刺激后不产生IL-4应答,仅1例产生弱IL-4应答（SFC＝1）。结论烟曲霉TR抗原诱导的T细胞免疫以Th1型免疫应答占绝对优势,因此,该抗原具有成为保护性抗原的潜能。     

重症肺部感染后继发曲霉菌感染的临床研究

目的研究重症肺部感染后患曲霉菌感染的危险因素、临床特征、影像学特点,以做到早期诊断和治疗。方法回顾分析2005年1月—2011年12月在呼吸重症监护室（RICU）的重症肺部感染继发曲霉菌感染患者,随机抽取同一时期未并发真菌感染的重症肺炎为对照组。记录患者临床资料,包括一般资料、基础疾病、治疗相关因素进行统计分析,以及血液指标、细菌培养结果和影像学资料。结果监护病房住院天数、广谱抗生素、糖皮质激素、机械通气（MV）、感染性休克、肝功能不全、糖尿病、免疫性疾病以及慢性呼吸道疾病（CRD）在2组比较中差异有统计学意义（P〈0．05）;而年龄、留置静脉导管、肠外营养以及实体肿瘤之间差异无统计学意义（P〉0．05）。临床以发热、呼吸困难及肺部哮鸣音为主;外周血白细胞升高、CRP、IGE升高占较大比例;同时影像学具有不典型性,以肺纹理增重、片状渗出和实变等非特异性表现。抢救成功8例中7例为伏立康唑治疗。结论重症肺部感染后存在上述相关危险因素时需注意易患曲霉菌感染可能;由于其临床表现及影像学具有不典型性,因此临床医师应认识其好发因素、观察临床病情的变化、多次查痰培养,同时气管镜检查观察黏膜、PBS及活检获得病理不失为一个比较安全的方法,抢先治疗成为降低病死率的关键。