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We report 2 cases of pulmonary aspergillosis treated successfully by combining micafungin and traconazole. Case 1: A 51-year-old man with hemoptysis and dyspnea on effort treated for pulmonary tuberculosis and aspergillosis was found on chest CT on admission to have a fungus ball in the left upper lobe and increasing consolidation around the cavity of both lung fields. Bronchoscopy proved positive for aspergillus PCR in bronchial lavage. He was diagnosed with chronic necrotizing pulmonary aspergillosis, based on clinical and radiological findings and the positive reaction for aspergillus PCR. He was treated with micafungin alone at first, this proved ineffective, so itraconazole was added, resulting in improvement. Case 2: A 24-year-old woman with stabilized Hodgkin's disease (mixed). She had suffered from a cough and back pain, and chest CT showed increasing consolidation inside and around a giant bulla. She was diagnosed with chronic necrotizing pulmonary aspergillosis, based on isolation for Aspergillus sp. in sputum culture and a positive reaction for Aspergillus antigen in bronchial lavage and Aspergillus antibody in serum. She was treated with the combined micafungin and itraconazole, which rapidly improved symptoms and radiological findings. Pulmonary aspergillosis therapy is often difficult, because delivery of the drug to the infection site is limited and drug tolerance is poor. We found that combination micafungin and itraconazole therapy is tolerable and effective in these cases.

We here reported two Japanese cases of mixed infections of plasmodium species, whose DNAs were detected using the PCR test. One case was a 31 year-old male, who presented fever and fatigue, and had a travel history to Kenya, Cameroon and Indonesia. Smear test of his peripheral blood found the presence of Plasmodium vivax, while nested-PCR diagnosis detected the DNAs both P. vivax and Plasmodium malariae. The other was a 54 year-old female suffering from general fatigue. She had been treated with chloroquine for falciparum malaria in Indonesia two weeks before. Malaria antigen test showed positive although no Plasmodium organisms were found in the smear test. The nested PCR detected the DNA of Plasmodium ovale in addition to that of Plasmodium falciparum. In conclusion, the PCR test is helpful and useful for detection of mixed infections of Plasmodium species.

The patient was 39-year-old male who had been administrated 20 mg of prednisolone for control of chronic eosinophilic pneumonia. He consulted the hospital with fever, headache and gait disturbance. The laboratory data of peripheral blood revealed a smoldering adult T cell leukemia. Computed tomogram of the chest and MRI of the brain revealed a mass in the right middle lobe of the lung and a brain abscess in the left hemisphere respectively. Biopsied specimens from the lung and brain abscess showed an Aspergillus like fungus. In spite of placement of an Ommaya reservoir for administration of AMPH-B and control of intracranial pressure, he died. During the course, specific antigen and specific gene were not detected in the peripheral blood, and no viable organism was isolated from the specimens. Post mortem examination revealed multiple nodular lesions in the lung, parietal pleura, liver, heart and kidney. After autopsy, disseminated aspergillosis was confirmed through a tissue examination using nested PCR for Aspergillus DNA. In this case, we think that viable fungi could endure in the tissue while circulating Aspergillus markers remained undetectable.

We report an atypical case of cat scratch disease (CSD), accompanied with encephalopathy that is a rare complication of CSD. A 17-year old man consulted a doctor for his right axillary lymphadenopathy. The history of his contact with cats and the sign of lymphnode swelling and fever suggested a suspect of cat scratch disease. Administration of ampicillin improved his clinical symptoms, but a few days later he suddenly fell into coma after an episode of convulsion. The CT scan of the brain and laboratory tests showed no significant findings except the slightly elevated cell counts and concentration of protein in his cerebrospinal fluid. He was referred to our hospital on the next day for further examinations and treatments for his coma of unknown cause. The physical examination on admission revealed slight neck stiffening and hypertonicity of his right lower limb, but radiological and laboratory tests showed no significant findings. He gradually recovered from his coma without apparent sequelae in three weeks. Indirect fluorescence antibody titers for CSD in his serum showed a significant elevation to 1:160 of IgM and 1:512 of IgG, and his clinical features were compatible to these of CSD with complications of the central nervous system.

We described the clinical and bacteriological features of 12 cases of liver abscess caused by Streptococcus milleri group (SMG) during a 6-year period from 1993 to 1998. The gender was 11 males and 1 female with their ages ranging from 39 to 76 years old (mean: 53.4). The common symptoms were fever (100%), abdominal pain (67%), and appetite loss (58%). Nine cases had underlying diseases such as carcinomas and diabetes mellitus. Predominant causes of the liver abscess were cryptogenic (42%) and biliary tract disease (33%). Three patients died of an exacerbation of the carcinoma. Eight cases (67%) was single infection of SMG and no mixed infection with anaerobes. No strains isolated in this series showed resistance against penicillin G and ampicillin. SMG was highly isolated from the blood culture in eight of the 11 cases (73%). Liver abscess should be taken into consideration as one of the causes of SMG septicemia.

Loiasis is quite common in the endemic regions of Central and West Africa. But only three cases were reported in Japan. This is a report of a 28 year old male from Gabon infected with Loa loa with eye symptoms as the chief complaint. For the first time in Japan he was treated with Ivermectin (IVM) which is recently attracting attention as the drug for filariasis world wide. IVM therapy was effective, and decreased the counts of microfilarias in the patient's blood. No adverse effect was seen in this patient. This case suggested that IVM is an useful drug for loiasis, and further study is warranted.

We have evaluated urine specimens of presumptive cases of legionnaires' disease (110 cases, 173 sample), collected in the past eight years (April, 1990-August, 1998) with the Binax EIA kit which detects the soluble antigen of Legionella pneumophila serogroup (SG) 1, and the Biotest EIA kit which detects Legionella species. Seven cases (19 specimens) were positive for the Binax EIA kit, and nine cases (22 specimens) were positive for the Biotest EIA kit. The sensitivity for culture, PCR, IFA method were 100%, 100%, and 50%, the specificity for these method were 93%, 97.1%, and 90% respectively. Overall agreements for these method were 93.5%, 97.4%, 86.8%, these results suggested that the urinary antigen detection test had high sensitivity and specificity. Our study indicated that concentrated urine samples increase sensitivity. We also evaluated the capabilities of both EIAs to detect soluble antigens were extracted from bacterial suspension of 18 strains of 5 Legionella species by heating. Both assays detected L. pneumophila serogroups 1 to 14, L. bozemanii. The Binax EIA proved to be useful as the Biotest EIA for diagnosis of legionellosis caused by Legionella species and serogroups other than L. pneumophila serogroup 1. Some cases have been shown to excrete antigen for prolonged period of times despite recovery from infection, so that the patient's history should be sought. The urine antigen detection EIA methods proved to be rapid and easy to use, detect antigen in the early stage of the disease with high sensitivity and specificity. Its use for the definition of legionellosis should be considered in Japan.

We studied the polymerase chain reaction (PCR) method with 5 strains of Legionella pneumophila serogroup 4, 7 strains of L. pneumophila serogroup 5, 7 strains of L. pneumophila UT and 15 strains of Legionella like organisms (LLO). We used mip-primers reported by Mahbubani et al and LEG-primers reported by Starnbach et al. Positive PCR results were obtained by all strains of L. pneumophila serogroup 4, L. pneamophila serogroup 5, L. pneumophila UT and 11 strains of LLO using mip-primers. Using LEG-primers, positive results were obtained by 2, 6, 5 and 8 strains respectively. All strains that showed positive results using LEG-primer PCR, were contained by the strains that showed positive results using mip-primer PCR. Conversely, there were some strains that were positive in mip-primer PCR and negative in LEG-primer simultaneously. Therefore, mip-primers is recommended to detect L. pneumophila in clinical specimens by PCR method.

We had examined antibody titers against Legionella spp. of patients' sera which were mainly sent from other hospitals, performed with the indirect fluorescent antibody (IFA) method. The clinical status of the cases diagnosed as Legionella pneumonia serologically, were also studied. Out of 105 cases with clinically suspected Legionella pneumonia, 15 cases (14.3%) were seropositive. In 9 out of the 15 cases (60.0%) were caused by Legionella pneumophila serogroup 1. Clinical outline of these 15 cases did not contradict those reported in the literature, and erythromycin was effective in many cases. Significant rises (more than four times) of the titer were observed 3 to 4 weeks after onset in most of cases. We would like to emphasize that this should be performed serodiagnosis of Legionella pneumonia.

A 51-year-old male was admitted because of cough and high fever. His chest X-ray and CT revealed miliary shadows, he was administered some antibiotics. But on the 8th day, his chest X-ray revealed ground glass shadows and he was under severe hypoxemia. We suspected miliary tuberculosis with acute respiratory failure due to his clinical course. The intensive therapy with antituberculosis drugs, hydrocortisone (1 g/day) was started and intubation was performed. The smear of the clinical samples did not show acid-fast bacilli on admission. However, by the polymerase chain reaction amplification method specific DNA fragments of Mycobacterium tuberculosis complex were detected from BALF and bone marrow. He was extubated after the 13th day and he recovered. It is difficult to separate and identify tubercle bacilli rapidly, but PCR is available for early diagnosis.

We evaluated the sensitivity and specificity of polymerase chain reaction (PCR) method in the detection of Pneumocystis carinii, and compared its clinical usefulness with that of a previous method, toluidine blue-O (TBO) staining in diagnosis of P. carinii pneumonia (PCP) using clinical samples such as sputum and bronchoalveolar lavage fluid (BALF). Using P. carinii obtained from BALF of the infected rats, we showed that PCR showed 100 fold more sensitivity than TBO staining as indicated by the comparison of their detection limits (78 cysts/ml vs 7800 cysts/ml). In addition, the detection using PCR in this study was specific for P. carinii because all other pathogens examined were not positive. In the clinical study we analyzed 44 patients including 10 with definite PCP, 8 with portable PCP and 26 with other diagnosis. In the sputum, the sensitivity and specificity were 100% and 95%, respectively, in PCR, while 13% and 100% in TBO staining. These results revealed that PCR was much superior to TBO staining in sensitivity especially when the sputum was used as samples, and suggested that examination of the sputum by PCR was clinically very useful in early diagnosis of P. carinii pneumonia.

We investigated the prophylactic and therapeutic effects of biological response modifiers (rHG-CSF, M-CSF, rhIL-2) on pulmonary candidiasis in neutropenic mice. Cyclophosphamide treated mice were injected by the intratracheal route with 5 x 10(6) Candida yeast cells. Prophylactic treatment with rhG-CSF afforded significant protection against pulmonary candidiasis in neutropenic mice. Treatment with rhG-CSF also increased the number of peripheral blood neutrophils. The histopathological investigations in our experiments showed that the assembly of PMNs to the infected lung at 24 hrs after bacterial challenge was more remarkable in the rhG-CSF treated mice than that in the vehicle alone. Number of viable candida cells in the infected lung in the rhG-CSF treated mice were significantly decreased. The combination of rhG-CSF and fluconazole was more effective than those of each monotherapy. Prophylactic treatment with M-CSF or rhIL-2 had no influence on pulmonary candidiasis. These results show the possibility that rhG-CSF could be of help for treating human deep candidiasis not successfully treated with antimicrobial agents alone.

We studied the usefulness of two sets of primers in macrophage infectivity potentiator gene sequence of Legionella pneumophila. PCR by 1st step primers produced 649 bp DNA bands and 2nd step primers 489 bp DNA bands. Two step PCR by these primers produced 489 bp DNA bands specific for L. pneumophila. Two step PCR detected 10 fg of pure DNA extracted from L. pneumophila, Philadelphia-1 strain. PCR sensitivity by these primers was superior to former primers reported by us using intratracheal aspirates collected from the patient with L. pneumophila serogroup 2 pneumonia.

We evaluated the usefulness of a Nested PCR method for detecting Legionella pneumophila. This method resulted in L. pneumophila specific detection as far as we evaluated. The first and second step PCR achieved the sensitivity as small as 10 pg and 10 fg of the target DNA, respectively. In the detection from Legionella seeded sputa, the method could detect 0.1 cfu/ml of the bacteria, and it took about 12 hours to detect the target DNA. We demonstrated that the Nested PCR method was superior in sensitivity and rapidity for isolation of the bacteria to the conventional using low pH treatment and selective media for Legionella.

We have tried to characterize the blastogenic responses of murine spleen lymphoid cells from BALB/c mice immunized with Legionella pneumophila serogroup (SG) 1 (Philadelphia 1 strain) and non-treated mice. Lymphoid cells from immunized mice showed stronger blastogenic responses following stimulation with concanavalin A or formalin-treated L. pneumophila SG1 whole cell antigen than those showed by lymphoid cells from non-treated mice. These cells from immunized mice also responded strongly when stimulated in vitro with other SGs of L. pneumophila, while these responded weakly when stimulated with other species of Legionella. Serum antibody titers of immunized mice against each SG of L. pneumophila were examined and the cross reactions were also recognized. However, the relatedness of serum antibody titers and the blastogenic responses against each serogroup of L. pneumophila was small. The epitopes recognized by the cellular immunity might be different in part from those recognized by serum antibodies, and investigations should be made on what the cellular immunity recognizes and how it works.