In situ detection of Aspergillus 18S ribosomal RNA in invasive pulmonary aspergillosis.

OBJECTS: We attempted to evaluate the usefulness of in situ hybridization (ISH) in the specific diagnosis of Aspergillus pulmonary infection. METHODS: We used an ISH technique using a multiple digoxigenin-incorporating probe, which was constructed by means of the polymerase chain reaction (PCR) from the 18S ribosomal RNA of Aspergillus fumigatus. MATERIALS: We studied twelve formalin-fixed, paraffin-embedded lung tissue sections from autopsy-confirmed invasive pulmonary aspergillosis (IPA) (5 acute myelocytic leukemias, 2 acute lymphocytic leukemias, 2 chronic myelocytic leukemias, 1 adult T-cell leukemia, 1 non-Hodgkin's lymphoma and 1 chronic obstructive pulmonary disease.), and 18 sections from other pulmonary infections as control. RESULTS: ISH using the probe and a low-viscosity hybridization buffer solution (LV) positively stained hyphal elements in 12 of 12 autopsy lung tissue specimens from subjects with IPA, while ISH using the probe and a high viscosity hybridization buffer solution (HV) positively stained the hyphal elements in 6 of 12. Specifically, ISH (LV) demonstrates hyphal elements of Aspergillus spp. in the center of Aspergillus abscess. While, ISH (HV) can detect hyphal elements located in the periphery of a suppurative abscess as well as those in the blood vessel. Conversely, ISH did not show positive results for any of the autopsy tissue specimens from subjects with other fungal pneumonia infections (Candida n=5, Mucor n=2, Cryptococcus n=2, and Pseudallescheria n=1), Pneumocystis carinii pneumonia (n=5), and cytomegalovirus pneumonia (n=3). Dual staining by means of ISH and immunohistochemistry (IHC) using anti-neutrophil elastase (NE) and anti-CD68 monoclonal antibodies showed that NE positive cells were localized at the edge of the radial growth of the organism, but CD68 positive cells were located around the center of the abscess. The accumulation of NE positive cells was rarely seen in half of the cases (6/12). In contrast, CD68 positive cells were routinely present in the center of the abscess (12/12). CONCLUSION: ISH in conjunction with IHC is a useful tool for differentiating Aspergillus spp. from other fungal genera in tissue sections from patients with IPA and may have a certain role in the evaluation of the interactions between organisms and recruiting inflammatory cells.     

A case of bronchocentric granulomatosis associated with uveitis]

A 32-year-old female was admitted to our hospital because of abnormal pulmonary shadows and a decrease in visual acuity. Analysis of peripheral blood revealed eosinophilia, and chest roentgenogram demonstrated multiple infiltrates in the right upper lung field. Pathological examination of transbronchial lung biopsy specimens revealed necrotizing granulomatous lesions in the walls of bronchioles, and a definitive diagnosis of bronchocentric granulomatosis was made. The cause of bronchocentric granulomatosis in this patient was suggested to be an allergic reaction to Aspergillus because of positive response to skin test for Aspergillus. Although it is reported that extrapulmonary involvement is rare in bronchocentric granulomatosis, the present case was associated with uveitis, and to our knowledge is the first reported case.     

Tissue culture isolation of Toxoplasma from blood of a patient with AIDS

A man with acquired immunodeficiency syndrome and multiple opportunistic infections developed central nervous system signs, and results of computed tomography suggested brain abscess. Histologic study and culture of material obtained from the brain biopsy specimen revealed Aspergillus. The clot from a peripheral venous blood sample obtained at the time of biopsy was processed and cultured on a monolayer of L cells. Five days later, Toxoplasma tachyzoites were identified in the cell culture. We bring this case to the attention of the medical community to point out that inoculation of tissue cell cultures as for viral isolation may aid in the diagnosis of Toxoplasma infection in these and other immunocompromised patients.     

Cerebral aspergillosis due to Aspergillus fumigatus in AIDS patient: first culture-proven case reported in Brazil

Cerebral aspergillosis is a rare cause of brain expansive lesion in AIDS patients. We report the first culture-proven case of brain abscess due to Aspergillus fumigatus in a Brazilian AIDS patient. The patient, a 26 year-old male with human immunodeficiency virus (HIV) infection and history of pulmonary tuberculosis and cerebral toxoplasmosis, had fever, cough, dyspnea, and two episodes of seizures. The brain computerized tomography (CT) showed a bi-parietal and parasagittal hypodense lesion with peripheral enhancement, and significant mass effect. There was started anti-Toxoplasma treatment. Three weeks later, the patient presented mental confusion, and a new brain CT evidenced increase in the lesion. He underwent brain biopsy, draining 10 mL of purulent material. The direct mycological examination revealed septated and hyaline hyphae. There was started amphotericin B deoxycholate. The culture of the material demonstrated presence of the Aspergillus fumigatus. The following two months, the patient was submitted to three surgeries, with insertion of drainage catheter and administration of amphotericin B intralesional. Three months after hospital admission, his neurological condition suffered discrete changes. However, he died due to intrahospital pneumonia. Brain abscess caused by Aspergillus fumigatus must be considered in the differential diagnosis of the brain expansive lesions in AIDS patients in Brazil.     

Additional value of K-ras point mutations in bronchial wash fluids for diagnosis of peripheral lung tumours.

The purpose of this study was to examine the additional diagnostic value of K-ras point mutations in the clinical diagnosis of peripheral lung tumours. To this end, bronchial wash fluids obtained during bronchoscopy from patients suspected of having lung cancer were studied. Only those patients were investigated for whom the cytological diagnosis was not conclusive for malignancy. As a control group, patients without lung cancer were investigated. The method of "point mutation detection using the exonuclease amplification coupled capture technique" (Point-EXACCT) for analysis of K-ras codon 12 was performed in bronchial wash fluids and the corresponding tumour tissue, if available. K-ras point mutations were identified in 4 out of 19 (21%) bronchial wash fluids from patients without a decisive diagnosis of malignancy. The diagnosis of malignancy was further based on cytological examination of bronchial brush specimens, perthoracic needle aspiration, histological investigation of biopsy and resection specimens, needle aspiration of a lymph node in the neck and pleural fluid examination. Four of the patients who were K-ras-positive yielded positive malignant tissue via bronchoscopy even though the bronchial wash was negative for malignancy. The bronchial wash was positive for K-ras in two of the four patients whose tumour tissue demonstrated the K-ras mutations. Analysis of bronchial wash fluids from 11 patients without lung cancer revealed no K-ras codon 12 mutations. In conclusion, K-ras point mutations can be identified in bronchial wash fluids obtained during bronchoscopic procedures. K-ras can be used as a biomarker in the clinical diagnosis of lung cancer and may serve as an adjunct to cytology in lung cancer diagnosis.     

Aspergillus fumigatus epidural abscess in a renal transplant recipient.

An epidural abscess caused by Aspergillus fumigatus occurred in a recipient of a cadaveric, renal allograft. The patient had persistent back pain and a peripheral neuropathy that involved the lower extremities. Signs of spinal cord compression evolved. No definite portal of entry was found. Diagnosis was made by histologic examination and culture of a biopsy specimen. Therapy, consisting of aggressive surgical debridement, intravenous amphotericin B, and oral flucytosine was unsuccessful in eradicating the organism. At postmortem examination, Aspergillus was identified at the abscess site. To our knowledge, aspergillosis presenting as an epidural abscess in the immunosuppressed, renal transplant recipient has not previously been reported and should be considered in the differential diagnosis of back pain and peripheral neuropathy in such a patient.     

Case of tuberculous psoas abscess successfully treated with surgery during antituberculosis therapy for miliary tuberculosis

A case of tuberculous psoas abscess complicated during antituberculosis therapy for miliary tuberculosis and successfully treated with surgery was reported. A 20-year-old man visited our hospital because of fever lasting for 3 months. Chest radiography showed miliary nodules in both lungs and transbronchial lung biopsy revealed granuloma. Magnetic resonance imaging of the head showed small lesions in the brain. Computed tomography of the abdomen showed an enlarged paraaortic lymph node and a nodule in the spleen. Needle biopsy of the lymph node revealed necrotic tissue. Mycobacterium tuberculosis was not isolated; however, miliary tuberculosis was highly suspected based on clinical and radiographic findings. Once antituberculosis therapy was initiated with isoniazid, rifampicin, streptomycin, and pyrazinamide, the fever subsided. In spite of improvement of general radiographic findings, a new abscess was found in the right psoas major muscle after 8 months of therapy by computed tomography. A sample of the abscess showed a positive smear, negative culture, and positive PCR test for M. tuberculosis. Although antituberculosis therapy continued for another 6 months, the abscess enlarged to 7 cm and new retroperitoneal lymph nodes also appeared. Surgical drainage and curettage of the abscess was performed. Intra- and post-operative specimens were negative for bacteria, fungi, and M. tuberculosis. The patient was treated with isoniazid, rifampicin, and ethambutol for one year postoperatively. The disease disappeared without any evidence of relapse for 2.5 years after surgery.     

Aspergillus flavus mitral valve endocarditis after lung abscess

A 16-year-old male with bone marrow failure due to chemotherapy for recurrent acute lymphoblastic leukemia developed an abscess in the lower lobe of the left lung draining through a bronchogastric fistula, as well as mitral valve endocarditis with large vegetations. After a course of antifungal therapy, the left lobe was removed and the fistula closed. The mitral valve was then replaced, after a failed attempt at valve repair, by a mechanical, double-leaflet prosthesis. Microscopy of the lung and heart specimens disclosed hyphae. Cultures of both specimens on Sabouraud's medium recovered a fungus, which was identified by culturing on Czapek's medium as Aspergillus flavus. Despite further antifungal therapy, fatal embolism developed. The emboli contained the same A. flavus as the valve and lung specimens. This case confirms the grim prognosis of primary Aspergillus endocarditis in immunocompromised patients, and suggests that delayed surgical treatment and the presence of another focus of Aspergillus infection may increase the risk of death.     

A case of allergic bronchopulmonary aspergillosis with no history of bronchial asthma]

A 53-year-old male was admitted to our hospital because of an abnormal shadow in the left upper lung. Bronchofiberscopy revealed edematous mucosa and pus at the orifice of the left upper lobe bronchus. Pathological examination revealed bronchial inflammatory change with infiltration of eosinophils and also the existence of aspergillus in the pus. Mild eosinophilia and elevation of serum IgE level were observed in the peripheral blood, and serum precipitin against Aspergillus fumigatus was positive. Bronchogram showed central bronchiectasis, and the diagnosis of allergic bronchopulmonary aspergillosis (ABPA) was made. In cases of ABPA, bronchial asthma is usually present prior to presentation, but this patient had no history of asthma even though airway hyperresponsiveness to methacholine was confirmed.     

Successful treatment of invasive mould infection affecting lung and brain in an adult suffering from acute leukaemia

We describe in detail a 67-yr-old woman who was treated with a cytostatic combination chemotherapy for newly diagnosed common-acute lymphoblastic leukaemia. At the end of induction therapy, the patient acquired invasive mould infection affecting lung and brain. The patient entered complete remission of her leukaemia. Treatment with liposomal amphotericin B was initiated along with surgical excision of the fungal brain abscess. Intrathecal instillation of amphotericin B deoxycholate was started using an Ommaya reservoir because of an anatomical connection between the postoperative cavity and the ventricle. Full dose cytostatic chemotherapy was continued with little delay. A computerised tomography scan of the chest performed 2 months later revealed no fungal abscesses. Magnetic resonance imaging of the brain did not reveal any fungal manifestation. During maintenance therapy/week 69, the patient relapsed from leukaemia. High doses of intravenous liposomal amphotericin B were administered prophylactically. The patient's leukaemia proved refractory to reinduction chemotherapy and the patient died from pneumonia 8 wk later. Post mortem microbiological investigation and histopathological examination of lung and brain tissue did not reveal any macroscopical or microscopical fungal manifestations. This case underlines the feasibility and successful application of combined antileukaemic, antifungal and surgical therapy in a patient with acute leukaemia.     

Multiple Aspergillus brain abscesses complicated by bronchial asthma]

A 35-year-old man was hospitalized for the treatment of severe asthma attack. His condition improved with intensive steroid chemotherapy under artificial ventilation. On the 12th hospital day, he was taken off respirator support but lost consciousness afterward. Computed tomography of the brain disclosed multiple hypodense lesions with bleeding. T1-weighted magnetic resonance imaging disclosed low-intensity lesions containing high-intensity areas. T2-weighted images showed heterogeneous high-intensity lesions. The autopsy specimen demonstrated multiple brain abscesses. Histologic examination revealed branching fungal hyphae in abscess walls and also extending through arterial walls with emboli. These findings yielded a diagnosis of multiple Aspergillus infarct abscesses of the brain.     

Chronic myelomonocytic leukemia associated with translocation 1;7, marked platelet aggregation and cryofibrinogenemia: a case report]

A 64-year-old male was admitted in September 1989 with complaints of fever and muscular weakness in the extremities. A peripheral blood examination on admission revealed WBC 10,300/microliters (monocytes 32%), RBC 195 x 10(4)/microliters, Hb 7.9 g/dl, Plt 12.8 x 10(4)/microliters with trilineage dysplasia. Bone marrow biopsy was normoplastic marrow with 25.7% of monocytes including immature blasts. Cytochemical analysis of the monocytes showed positive for peroxidase and dual esterase staining. Chromosomal analysis of peripheral blood revealed 46, XY, -7, +der(1) t(1;7)(p11;p11). A diagnosis of chronic myelomonocytic leukemia was made. Hemostatic studies revealed cryofibrinogenemia, marked platelet aggregation on blood smear, hyperfibrinogenemia and a marked increase in maximal amplitude of thrombelastogram. Treatment with prednisolone and VP16, resulted in a reduction of peripheral monocytes and a disappearance of cryofibrinogen, marked platelet aggregation and a decrease in muscular weakness. Nine months after diagnosis he died of DIC, pneumonia, lung abscess and sepsis.     

An autopsy case of Aspergillus pneumonia after inhalation of a great deal of dust in a mushroom factory]

A man in his forties with alcoholic liver injury was referred to our hospital because of progressive dyspnea and diffuse infiltrative shadows in both lung fields. Two weeks before admission he had carried a lot of rotten Enoki mushrooms in his factory, and then inhaled a great deal of airborne dust. Serological examination showed extremely elevated beta-D-glucan and positive Aspergillus antigen. Culture of dust collected from the floor of the factory proved to be positive for Aspergillus fumigatus. Aspergillus pneumonia was diagnosed and treatment with anti-fungal drugs was started. In spite of intensive treatment he died of respiratory failure. The autopsy disclosed multiple abscess formation containing hyphae of Aspergillus fumigatus and granulomatous reaction with multinuclear giant cells. However, there was no Aspergillus invasion in other organs. This was a rare case of invasive pulmonary aspergillosis that developed after inhalation of organic dust in a non-immunocompromised host.     

Successful treatment of cerebral aspergillosis with a novel triazole (voriconazole) in a patient with acute leukaemia

Invasive aspergillosis is an increasing problem in patients with acute leukaemia, bone marrow transplantation, immunosuppression after solid organ transplantation, or acquired immunodeficiency syndrome. Despite available antifungal treatment, the mortality approaches 100% in patients with dissemination of the infection into the central nervous system (CNS). Using a novel triazole, voriconazole, we successfully treated an Aspergillus brain abscess in a patient with acute leukaemia. Drug levels above the minimal fungicidal concentration for Aspergillus species were detected in cerebrospinal fluid (CSF) specimens, and the treatment achieved an objective response.     

A case of non-invasive pulmonary aspergillosis that rapidly deteriorated]

The pulmonary diseases caused by the Aspergillus species include invasive forms, for example, invasive pulmonary aspergillosis, chronic necrotizing pulmonary aspergillosis, and non-invasive pulmonary aspergillosis. Though these forms are defined pathologically by the presence of the Aspergillus species that invades the lung tissue, they are used as clinical entities. We report a case of non-invasive pulmonary aspergillosis which, from the clinical data, appeared likely to be misdiagnosed as the chronic invasive form. A 45 year-old man received chemoradiotherapy for lung cancer as well as undergoing an left upper lobectomy. Two weeks after the surgery the patient developed a cough, high fever and chest pain. Chest radiography and chest computed tomography showed a rapidly enlarging cavity with an internal mass and infiltration in the left lower lung field. A transbronchial biopsy specimen of the cavity wall showed fungal hyphae. Bronchial washing culture grew Aspergillus fumigatus. Itraconazole and amphotericin B were administered, but the patient's condition did not improve. A left lower lobectomy was performed. The histologic findings showed that the fungal hyphae were only on the surface of the cavity wall, and were surrounded by necrosis and widespread inflammatory cell infiltration. No fungal invasion of the viable lung tissue was seen. The area of infiltration revealed an organizing pneumonia without Aspergillus or other organisms. Our final diagnosis was non-invasive pulmonary aspergillosis. There has been no recurrence of the lung cancer or of the pulmonary aspergillosis in the three years since surgery. It is reported that non-invasive pulmonary aspergillosis passes through a period so active that it seems to be the invasive form for its entire clinical course. To avoid confusion in diagnosis, establishment of a comprehensive clinical classification of pulmonary aspergillosis will be needed.     

A case of pulmonary actinomycosis with a unique finding in the chest MR image]

A 57-year old man, who was complaining of a productive cough and right shoulder pain, was admitted to our hospital because of an irregularly shaped mass located at rt. S1 on a chest radiograph. Bronchoscopy revealed no evidence suggesting lung cancer or any specific infection, either pathologically or microbiologically. CT-guided biopsy revealed changes resembling lymphocytic or plasmocytic interstitial pneumonitis with thickening of the alveolar septum and with accumulations of mononuclear cells and plasma cells, indicating the proliferation of bronchus-associated lymphoid tissue (BALT system). Since no definitive diagnosis was considered possible, a right upper lobectomy was performed. Histopathologic examination of tissue from the right upper lobe revealed sulfur granules and branching Gram-positive filamentous bacteria, and the condition was pathologically diagnosed as pulmonary actinomycosis. In the center of the mass lesion, the patient's chest MRI showed a very small area with a low signal intensity in T1- and a high signal in T2-weighted images, which suggested an accumulation of fluid in the actinomycotic abscess. As detailed MR findings in this condition have not been well described in the literature, the MRI evidence seen in this case may be useful for the diagnosis of actinomycosis.     

A case of pulmonary tuberculosis complicated with subcutaneous phaeohyphomycosis

A 78-year-old male was admitted to our hospital because of fever, sputum and cough. Chest X-ray showed infiltrative shadows in the right lung field. Smears of his sputum were positive for acid-fast bacilli. We found multiple subcutaneous abscesses on the right distal forearm. Microscopic examination of skin biopsy specimens revealed granulation tissues with the proliferation of epitheloid cells with the scattering infiltrations of neurophils, giant cells and histiocytic cells. The examination of the PAS stained specimen revealed fungal elements and a black fungus, Exophiala jeanselmei, was isolated by the cultures of pus from the abscess. He was diagnosed as pulmonary tuberculosis complicated with subcutaneous phaeohyphomycosis caused by Exophiala jeanselmei and was successfully treated with anti-tuberculosis drugs and anti-fungal agent, 5-fluorocytosine.     

侵袭性深部酵母菌感染致溶骨性破坏

报告我科近期收治1例侵袭性深部酵母菌感染引起溶骨性破坏的病例。初病时曾怀疑肺结核、肺癌骨转移,后骨髓、血及皮下脓肿穿刺液均培养出同一种酵母菌,同时发现全身多处骨质破坏。结合复习文献探讨了真菌感染致溶骨性损害以马尔尼菲青霉菌感染引起多见,由于临床无特征性,常易误诊。确诊需作真菌培养。此真菌特征为温度双相性,可产生红色色素。两性霉素B脂质体及伊曲康唑治疗有效。     

电视胸腔镜治疗28例慢性肺脓疡的体会

目的探讨经内科常规治疗无效的慢性肺脓肿患者应用电视胸腔镜技术治疗的疗效。方法1997年10月～2007年10月期间，28例慢性肺脓肿患者应用电视胸腔镜技术实施脓肿切开、持续胸腔冲洗引流术的疗效进行回顾性分析。结果24例患者治愈，4例显效，无手术死亡及术中并发症发生。胸部失血量为60～190ml，平均（136±84）ml。术后第1d引流量为60-450ml，平均（278±112）ml，术后胸腔引流时间3—13d，平均（4．35±2．14）d，有3例出现持续漏气〉7d；术后静脉应用抗生素时间4～10d，平均（4．43±1．82）d；术后住院日为7—16d，平均（10．7±2．33）d。出院前复查胸片示脓腔消失，术后随访3～18个月，未见复发病例。结论：电视胸腔镜下切开慢性肺脓肿持续胸腔冲洗引流术具有显露清楚、创伤小、并发症少、恢复快、住院时间短的特点，而且完全可以达到常规开胸手术效果。     

A case of pulmonary aspergillosis accompanied with eosinophilic pneumonia]

An infiltrative shadow was observed in the upper lobe of the left of a 60-year-old man lung during the course of paroxysmal tachycardia. Fever and coughing occurred one month later, and ground-glass opacities (GGOs) were found extensively in the lower left lung field. Although his condition strongly suggested allergic bronchopulmonary aspergillosis (ABPA) due to significantly increased eosinophil count in the peripheral blood and bronchoalveolar lavage fluid of the lower lobe of the left lung, anti-aspergillus antibody precipitation, and positive immediate intradermal test, the Rosenberg diagnostic criteria for ABPA were not satisfied. He was given a diagnosis of eosinophilic pneumonia, and GGO disappeared when itraconazole and prednisolone were administered. However, since the shadow in the upper lobe of the left lung later increased and percutaneous thoracocentesis revealed the presence of Aspergillus fumigatus, prednisolone treatment was discontinued and micafungin was administered. Nonetheless, the shadow spread into both lungs and regardless of treatment with amphotericin-B, the patient died. Although the present case is different from a typical ABPA, it indicates that allergic reaction can be induced by Aspergillus infection in the lung field, apart from the central airway. In addition, the development of an appropriate therapeutic strategy against both infection and allergy in such cases is essential.