Elevated levels of soluble adhesion molecules in sera and BAL fluid of individuals infected with human T-cell lymphotropic virus type 1.

STUDY OBJECTIVE: T-lymphocytic alveolitis and increased levels of interleukin-2 receptor-alpha (CD25)-bearing T cells in the BAL fluid (BALF) of human T-cell lymphotropic virus type 1 (HTLV-1) carriers have been reported. Several chemokines and adhesion molecules may contribute to the accumulation of T lymphocytes in the lungs of HTLV-1 carriers. To clarify the correlation between adhesion molecules and HTLV-1-associated pulmonary disorders, we compared the distribution of T-lymphocyte subsets and soluble adhesion molecules, including soluble intercellular adhesion molecule (sICAM)-1, soluble vascular cell adhesion molecule-1 (sVCAM-1), soluble L-selectin (sL-selectin), soluble E-selectin (sE-selectin), and soluble P-selectin (sP-selectin), in BALF and peripheral blood, between HTLV-1 carriers and noninfected healthy subjects. DESIGN: Flow cytometric analysis with monoclonal antibodies to cell-surface antigens was used to identify T-lymphocyte subsets in BALF samples from HTLV-1 carriers (n = 13) and noninfected healthy control subjects (n = 10). The levels of various soluble adhesion molecules in serum and in BALF were estimated by enzyme-linked immunosorbent assay. RESULTS: Higher percentages of CD3+ cells, CD3-expressing human leukocyte antigen-DR antigen, and CD3+CD25+ cells were detected in the BALF of HTLV-1 carriers than in that of noninfected control subjects. The concentrations of sICAM-1, sVCAM-1, sL-selectin, SE- selectin, and sP-selectin in the sera of patients were significantly higher than those in noninfected healthy control subjects. The concentration of sICAM-1 in the BALF of patients was significantly higher than that in noninfected healthy control subjects, and the concentration of sICAM-1 correlated well with the percentage of CD3+CD25+ cells. CONCLUSION: The concentrations of adhesion molecules in the sera of and sICAM-1 in the BALF of HTLV-1 carriers were significantly higher than those in noninfected individuals, and the concentration of sICAM-1 correlated well with the percentage of CD3+CD25+ cells in BALF. Our results suggest a possible interaction between activated T cells bearing CD25 and soluble adhesion molecules, especially sICAM-1, which may contribute to the pulmonary involvement in HTLV-1 carriers.

Regulation of gamma-glutamylcysteine synthetase expression in response to oxidative stress.

Glutathione (GSH) is synthesized by the activity of two ATP-requiring GSH synthesizing enzymes. Gamma-glutamylcysteine synthetase (gamma-GCS) is the rate limiting enzyme for the GSH synthesis. Gamma-GCS is a heterodimer of heavy, catalytic subunit and light, regulatory subunit and responsive to many stresses, such as heat shock, oxidative stress or cytokines. To know the regulation of the expression of gamma-GCS gene, in the present study, we show evidences that gamma-GCS heavy subunit is upregulated by oxidative stress by ionizing radiation and TNF-alpha mediated by nuclear factor-kappaB (NF-kappaB), and impairment of the expression of gamma-GCS by TNF-alpha in diabetic condition. Furthermore we describe the importance of GSH in the regulation of NF-kappaB subunits.

An outbreak of Legionnaires' pneumonia in a nursing home.

An outbreak of Legionnaires' pneumonia occurred at a nursing home in December 1990. A 79-year-old female and a 73-year-old male clerk who were staying at the nursing home developed pneumonia with only a 5-day interval. Legionella pneumophila serogroup I was isolated from transtracheal aspirate of the former and sputum of the latter. After treatment with a combination of erythromycin and rifampicin both patients improved. Serological surveillance of inpatients and staff of the nursing home was performed in February 1991. Seven out of 51 samples (14.0%) showed a titer higher than 1:128 of anti-Legionella pneumophila serogroup I antibody determined by indirect immunofluorescence; two of these seven complained of respiratory symptoms. Molecular epidemiology analyzed by restriction endonuclease digestion of isolated L. pneumophila showed an identical pattern which suggested a common origin.

Fatal pneumonia caused by Corynebacterium group JK after treatment of Staphylococcus aureus pneumonia.

A 76-year-old man who was admitted to the hospital because of chronic renal insufficiency and chronic hepatitis died of Corynebacterium group JK pneumonia, after showing a slight improvement by treatment of Staphylococcus aureus with sulbactam/cefoperazone and minocycline. Transtracheal aspiration (TTA) just before his death revealed numerous gram-positive bacilli phagocytized by many neutrophils and more than 10(8) colony forming units (CFU)/ml of Corynebacterium group JK. A drug susceptibility test showed Corynebacterium group JK was resistant to many antibiotics, with the exception of vancomycin and amikacin.

Antibody response to Mycoplasma bovis of calves introduced to a farm contaminated with the organism.

Antibody against Mycoplasma bovis in sera of 48 calves introduced to a farm, in which calf pneumonia associated with M. bovis had been occurring in the last 3 years, was detected by an indirect hemagglutination test. Significant rises of antibody titers in sera of calves belonging to the groups A (16 calves) and B (14 calves) were recorded by day 60 post-introduction. On the other hand, a significant increase of antibody titers of 18 calves in group C, which had been administered antibiotics as a preventive therapy, was demonstrated at day 248 after arrival. These results indicated that the spread of M. bovis infection occurred easily on the contaminated farm, and a preventive therapy could delay the outbreak of calf pneumonia associated with M. bovis.

[The future of antifungal agents. Non azole antifungal agents].

We investigated the efficacy of non-azole antifungal agents. Long circulating immunoliposomal amphotericin B was potent in murine invasive pulmonary aspergillosis. The concentration of AMPH-B was still high in the lung after 6 hours of 34A-PEG-liposomal AMPH-B. Lipid nanosphere amphotericin B (NS-718) showed efficacy against pulmonary aspergillosis in rats and pulmonary cryptococcosis in mice. The renal toxicity of NS-718 was estimated to be lower than that of AMPH-B from the results of the toxicity study in the rat infusion model. FK 463, a novel (1,3)-beta-D-glucan synthase inhibitor, showed efficacy against azole-resistant Candida albicans in murine experimental disseminated candidiasis. FK463 could be a promising drug and the therapy of choice for azole resistant C. albicans infection.

[Detection of mecA gene of methicillin resistant staphylococci from oral swabs by polymerase chain reaction].

We evaluated the polymerase chain reaction method (PCR) for the rapid detection of methicillin resistant Staphylococcus aureus (MRSA), and compared it with the conventional culture method. Primers for amplification of mecA gene were synthesized by DNA synthesizer according to the published sequences of mecA gene. The results were as follows: Specificity of PCR was excellent, as there was no cross-reaction with any organism other than MRSA. Fifty colony forming units of MRSA were detected, indicating good sensitivity. We examined oral swabs from 13 bed-ridden patients (age range: 65-82 yrs) by PCR and the conventional culture method. The PCR elicited positive results in 6 out of 13 cases. MRSA or coagulase negative Staphylococci (CNS) in Mueller-Hinton agar containing 12.5 micrograms/ml methicillin were obtained from all of these PCR-positive specimens. Two of 7 PCR-negative specimens raised MRSA or CNS colonies, but the number of colonies from these specimens was below the sensitivity threshold of the PCR. The good specificity and sensitivity of the PCR method for the detection of mecA gene obtained in this study suggest the possibility of application of this method for detection of MRSA in clinical specimens.

[Case report: lung abscess caused by Streptococcus agalactiae].

Streptococcus agalactiae is a well-recognized cause of neonatal sepsis and meningitis. In adults, infections by S. agalactiae are rare. We report an adult case of lung abscess and pyogenic spondylitis caused by S. agalactiae. A 51-year-old male was admitted to our hospital because of an abnormal shadow in the chest and lumbago on May 25, 1995. He was diagnosed as lung abscess from the chest roentgenogram and CT scan and the subcutaneous pus was aspirated. The pus culture was only positive for S. agalactiae. He was treated with IPM/CS 1 g/day and CLDM 1.2 g/day and the abscess was drained. MRI showed his lumbago was caused by pyogenic spondylitis. The underlying disease of this case was diabetes mellitus. He recovered from the infections with in about 10 weeks of antibiotic treatment.

[Antigen detection in the murine models of systemic candidiasis].

We compared two latex agglutination tests for serum candidal antigen in murine models of systemic infection and gastrointestinal colonization. In the intravascular-infection model, mannan detection test was positive at 2 hours after inoculation, but Cand-Tec was less sensitive than mannan detection test in this model. In the gastrointestinal colonization model followed by systemic infection in neutropenic mice, the mannan antigen test became positive after 3 weeks of colonization with increased number of Candida of the stool, and Cand-Tec became positive after 5 weeks in neutropenic mice.

[Bacteriological studies on pharyngeal and tracheal colonization in patients of operations and patients in ICU].

Bacterial colonization of the pharynx before and one week after operation, and contamination of the trachea during intubation in 7 patients who underwent operations were investigated. And bacterial colonization of pharynx, tracheas, and mechanical-ventilators in 7 ICU patients was investigated. Flora of palms of 10 ICU personnel was also examined: 1) Oral bacteria began to be found in the trachea several hours after intubation. 2) In oropharyngeal flora, one week after operation, Neisseria and anaerobes which belong to normal flora decreased because of antibiotics, but no new resistant bacteria appeared. 3) Although isolates from pharynx of preoperative patients were normal flora, those of ICU patients were Staphylococcus epidermidis, Enterococcus spp., gram-negative rods such as Pseudomonas aeruginosa, and yeast liked organisms etc, which were resistant to antibiotics. 4) From the trachea of intubated patients in ICU, oral bacteria were isolated. 5) Pseudomonas pickettii was isolated from the expiratory side of ventilator circuits in one patient, but it was not isolated from the patient. 6) From hands of ICU personnel, gram-positive bacteria such as coagulase negative staphylococci, Coryne bacterium spp. and Bacillus spp. were isolated. 7) One patient in ICU, who developed pneumonia due to resistant Bacteroides fragilis following oropharyngeal colonization, was reported. These studies suggested the importance of environmental control, careful selection of antibiotics, and attention to change of flora.

[Four cases of respiratory tract infections caused by Corynebacterium pseudodiphtheriticum].

Four cases of respiratory tract infections caused by Corynebacterium pseudodiphtheriticum were reported. The first two patients developed pneumonia with Corynebacterium pseudodiphtheriticum during steroid therapy used against their underlying diseases. The other two patients had acute exacerbation of chronic pulmonary diseases caused by C. pseudodiphtheriticum. These four patients improved by antibiotic therapy. Though nondiphtheria corynebacteria are regarded as "normal flora" when they are isolated from sputum, they should be recognized as potential pathogens.

[A case of bacterial meningitis induced by strongyloidiasis].

A 75-year-old female with diabetes mellitus, who was born and lived in West north Kyusyu, was admitted to our hospital, because of unconsciousness and loss of appetite. The physical examination showed neck stiffness and a high fever. The laboratory data showed accentuation of inflammatory reaction and azotemia and positive HTLV-1 antibody. The spinal fluid showed increase of cell count and amount of protein. A stool and sputum smear revealed rhabditis form larvae of the nematode. Antibiotics and ivermectin were administered for the bacterial meningitis and hyperinfection of the strongyloides, respectively. Consequently, meningitis and strongyloidiasis improved. It was considered that the patient was infected with strongyloides from her husband who serve in the army during World-War II, and hyperinfection of strongyloides resulted from the immunosuppressive state of diabetes mellitus. Ivermectin, and anti-strongyloides agent, was effective, and no side effects were seen. However, the therapeutic resistance in this case was associated with the positive HTLV-1 antibody.

[The combination therapy of clarithromycin and sparfloxacin for pulmonary Mycobacterium gordonae infection].

Seventy years old woman had fever and hemosputum at May 1997. She was diagnosed as mycobacteriosis because of the positive acid fast bacilli smear from sputum. Mycobacterium gordonae was isolated from sputum, gastric juice, and bronchial aspirate. The combination therapy of isoniazid, rifampicin, ethambutol, and clarithromycin was administrated; however, M. gordonae was not eradicated from sputum. Sparfloxacin was administered instead of isoniazid based on the result of drug susceptibility test. The smear became negative and M. gordonae was eradicated from sputum one month after the initiation of treatment with the combination of clarithromycin and sparfloxacin.

[The comparison of drug susceptibility of antituberculous agents against colonizing M. intracellulare and infectious M. intracellulare].

The susceptibility of antituberculous agents against colonized M. intracellulare which were isolated from patients with pulmonary disease and infectious M. intracellulare (pathogens) isolated from atypical mycobacterial culture was investigated. Aminoglycosides were more potent against colonized organisms than against pathogens. Isoniazid, ethambutol and rifampicin showed less potent antimicrobial activity against both colonized organism and pathogens as compared to aminoglycosides. On the contrary susceptibility of cycloserine against colonized organisms was as potent as against pathogens.

[Evaluation for rapid detection of rifampicin-resistant mycobacterium tuberculosis by polymerase chain reaction-single strand conformation polymorphism].

We evaluated usefulness of the rapid diagnostic method for detection of rifampicin (RFP)-resistant Mycobacterium tuberculosis, which was based on polymerase chain reaction. The MICs of RFP were measured for 38 clinical isolates of Mycobacterium tuberculosis which were suspected to be RFP-resistant organisms, and 12 strains were found to be resistant to RFP. The PCR primers used were the same as those reported by Telenti et al, which were targeting the RNA polymerase beta subunit gene (rpoB). We confirmed that this gene was possessed by all the strains tested. Eight strains out of the 12 strains with RFP-resistant phenotype were demonstrated to have a point mutation or some alterationin the rpoB gene on the basis of PCR-single strand conformation polymorphism (SSCP). Thus, the sensitivity of our method was calculated as 67%. In addition, we could not detect any alterations in the rpoB gene by all RFP-susceptible strains. These results indicated that rapid detection of the RFP-resistant Mycobacterium tuberculosis was possible directly from clinical specimens by using PCR-SSCP technique.

[Rapid identification of mycobacteria by combined method of polymerase chain reaction and the gen-probe DNA hybridization system].

We developed the rapid detection and identification method of mycobacteria, involving amplification of mycobacterial 16S rRNA gene by nested PCR and identification of M. tuberculosis complex or M. avium-intracellulare complex (MAC) by hybridization protection assay (HPA) using the acridinium-ester (AE) labeled DNA probe. The specificity of the nested PCR combined with DNA probe test was excellent in terms of detection of mycobacterial organisms and identification of M. tuberculosis or MAC. The detection limits of the present method were 10 fg DNA for M. tuberculosis, and 100 fg DNA for MAC, respectively. We further investigated on the optimum temperature for hybridization in HPA with AE labeled DNA probe because there was the difference in the mode of DNA-RNA hybridization from that of DNA-DNA hybridization. In our method, the optimum temperature of hybridization was estimated as 55 +/- 1 degrees C. In preliminary experiments on two clinical cases, we practically detected and identified M. tuberculosis and MAC in clinical specimens, such as sputa, by using this newly devised method. We concluded that this method is useful for rapid detection and identification of M. tuberculosis and MAC in clinical specimens.

[A rare case of lung adenocarcinoma in cavity wall of pulmonary aspergilloma].

A 67-year-old man was admitted with the complaint of hemosputum. Chest X-ray films resulted in a diagnosis of pulmonary aspergilloma, and treatment with intravenous amphotericin B was initiated. However, therapy was discontinued due to renal insufficiency, an adverse effect of amphotericin B. The size of the fungus ball and cavity increased despite treatment with oral itraconazole (200 mg/day). Cavernostomy was performed and the fungus ball was removed from the upper lobe of the left lung. Computed tomographic scans disclosed thickening of the remaining wall of the cavity, with destruction of the ribs. The patient experienced worsening respiratory distress and died. Necropsy revealed adenocarcinoma of the left lung.

[Correlations between drug plasma concentration and adverse effects in patients treated with itraconazole for pulmonary aspergilloma].

We measured the plasma concentration of itraconazole (ITCZ) in 18 patients who received ITCZ for the treatment of pulmonary aspergilloma. Abnormal laboratory values were observed in 4 out of 10 patients who received 200 mg/day, 1 out of 3 patients who received 300 mg/day, and 2 out of 5 patients who received 400 mg/day. Four patients discontinued ITCZ therapy because of adverse effects following the administration of 200 mg/day or 400 mg/day. The mean plasma ITCZ concentration was 622 ng/ml in patients treated with less than 4 mg/kg, and 1,352 ng/ml in patients treated with more than 4 mg/kg of ITCZ. The sensitivity of Aspergillus species to ITCZ was measured with the NCCLS microdilution method, using alamar blue indicator. The MIC50 of ITCZ was 0.5 microg/ ml for 25 strains of A. fumigatus, 4 microg/ml for 15 strains of A. niger, and 0.25 microg/ml for 10 strains of A. flavus. In conclusion, this study underscored the necessity of monitoring the plasma concentration of ITCZ for effective treatment of patients with pulmonary aspergilloma.

Effects of macrolides on pneumolysin of macrolide-resistant Streptococcus pneumoniae.

To clarify the discrepancy between increasing resistance and conservative clinical effects of macrolides on macrolide-resistant Streptococcus pneumoniae, the authors evaluated the effects of sub-minimum inhibitory concentrations of macrolides on pneumolysin. In vitro, S. pneumoniae was incubated with 1, 2 and 4 microg.mL(-1) of clarithromycin (CLR) and azithromycin (AZM) for 8 h. Western blot analysis and haemolytic assay were performed to examine the production and activities of pneumolysin. In vivo, mice were infected with S. pneumoniae intra-nasally and treated with CLR (40 or 200 mg.kg(-1) twice daily) or AZM (40 or 200 mg.kg(-1) once daily) orally for 7 days. After 72 h post-infection, western blot analysis was performed to examine pneumolysin production in lungs. Survival rates were observed for 10 days. In vitro, every concentration of macrolide inhibited pneumolysin production more than the control. CLR (2 and 4 microg.mL(-1)) and AZM (4 microg.mL(-1)) reduced the pneumolysin activities more than the control. In vivo, macrolides (200 mg.kg(-1)) reduced pneumolysin in murine lungs more than the control. CLR (40 and 200 mg.kg(-1)) and AZM (200 mg.kg(-1)) improved the survival rates more than the control. The study results show that sub-minimum inhibitory concentrations of macrolides reduced pneumolysin. This might be related to the effectiveness of macrolides against pneumonia caused by high-level macrolide-resistant Streptococcus pneumoniae. Further investigations are necessary to evaluate the effects of macrolides on macrolide-resistant Streptococcus pneumoniae.