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Biomedical subjects

Keizo Yamaguchi

Publications and source records attributed to Keizo Yamaguchi.

At least 73 records · Page 4Linked to original sources
PubMed2004

Regulatory effects of macrolides on bacterial virulence: potential role as quorum-sensing inhibitors.

Pseudomonas aeruginosa is an opportunistic pathogen, and this organism is a major cause of pulmonary damage and mortality in patients with cystic fibrosis (CF), diffuse panbronchiolitis (DPB) and other forms of bronchiectasis. A break-through in the treatment of DPB and associated chronic P. aeruginosa pulmonary infection was realized when a patient with DPB improved dramatically after treatment with erythromycin for years. Now, long-term macrolide therapy has become a first line of treatment in DPB patients, and the immunomodulatory properties have now been extended to other clinical settings, including CF. An important factor in the pathogenesis of chronic P. aeruginosa infection is a bacterial cell-to-cell signaling mechanism, referred to as "quorum sensing", which enables bacteria to coordinately turn on and off specific virulence genes through the production of autoinducer molecules. Interference or blocking of quorum-sensing systems has been considered an attractive therapeutic strategy. Clinical and basic science data suggests the potential of macrolides as relevant inhibitors of the Pseudomonas quorum-sensing system. In fact, certain macrolides strongly suppressed quorum-sensing associated genes and autoinducer production, in addition to inhibition of a variety of virulence factors. In this review, clinical efficacy of macrolides on DPB and CF patients will be briefly summarized. Additionally, the mechanisms of action of macrolides will be discussed from the standpoint of sub-MIC macrolide effects on P. aeruginosa, particularly the ability of this antibiotic to suppress quorum-sensing systems, which may be crucial in the pathogenesis of chronic P. aeruginosa infection.

Anti-Bacterial Agents↗
PubMed2004

[Nationwide surveillance of parenteral antibiotics containing meropenem activities against clinically isolated strains in 2002].

The antibacterial activity of meropenem (MEPM) and other parenteral antibiotics against clinical isolates of 899 strains of Gram-positive bacteria, 1500 strains of Gram-negative bacteria, and 158 strains of anaerobic bacteria obtained from 28 medical institutions during 2002 was measured. The results were as follows; 1. MEPM was more active than other carbapenem antibiotics against Gram-negative bacteria, especially against enterobacteriaceae and Haemophilus influenzae. MIC90 of MEPM against Pseudomonas aeruginosa was the lowest of the drugs tested. MEPM showed low cross-resistant rate against both imipenem-resistant P. aeruginosa and ciprofloxacin-resistant P. aeruginosa. MEPM was active against most of the species tested in Gram-positive and anaerobic bacteria, except for multi-drug resistant strains including methicillin-resistant Staphylococcus aureus (MRSA), methicillin-resistant Staphylococcus epidermidis (MRSE). 2. The proportion of extended-spectrum beta-lactamase (ESBL) strains was 3.1% (4 strains) in Escherichia coli and 1.9% (2 strains) in Klebsiella pneumoniae. Carbapenems including MEPM were active against these ESBL strains. In conclusion, the results from this surveillance study suggest that MEPM retains its potent and broad antibacterial activity and therefore is a clinically useful carbapenem; at present, 7 years after available for commercial use.

Bacteria↗
PubMed2004

[Comparative studies on activities of antimicrobial agents against causative organisms isolated from patients with urinary tract infections (2002). I. Susceptibility distribution].

The bacterial strains isolated from 491 patients diagnosed as having urinary tract infections (UTIs) in 13 institutions in Japan were supplied between August 2002 and July 2003. The susceptibilities of them to many kinds of antimicrobial agents were investigated. Of them, 578 strains were estimated as causative bacteria and used for the investigation. The number of them was 578 strains consisting of 177 gram-positive bacterial strains (30.6%) and 401 gram-negative bacterial strains (69.4%). Against Staphylococcus aureus, vancomycin (VCM) showed a strong activity and prevented the growth of all strains with 1 microg/mL. The susceptibility of Staphylococcus epidermidis to cephems including cefotiam (CTM) was relatively good. Against Enterococcus faecalis, ampicillin (ABPC), imipenem (IPM), and VCM showed the strongest antibacterial activity (MIC90: 2-4 microg/mL). In addition, the low sensitive strains (MIC: > or = 256 microg/mL) to clarithromycin (CAM) were detected at 48.3% but none to cefozopran (CZOP). The antibacterial activity of cephems to Escherichia coli was generally good, and especially CZOP and cefpirome (CPR) showed the highest activity (MIC90: < or = 0.125 microg/mL). Quinolone resistant E. coli was detected at frequency of 13.5%, which was higher than that in the last year. The antibacterial activity of cephems to Citrobacter freundii was generally low but CZOP and CPR had a strong acitivity (MIC90: 0.25 and 0.5 microg/mL, respectively). The antibacterial activity of cephems to Klebsiella pneumoniae was good and especially cefmenoxime (CMX), cefixime (CFIX), flomoxef (FMOX), CPR, and CZOP showed stronger activity (MIC90: < or = 0.125 microg/mL). Against Serratia marcescens, meropenem (MEPM) had the highest antibacterial activity followed by CPR and CZOP. Against Proteus mirabilis, CMX, ceftazidime (CAZ), CPR, MEPM, carumonam (CRMN), and levofloxacin (LVFX) showed the strongest activity (MC90: < or = 0.125 microg/mL). Among other cephems, CZOP and CFIX were also strong (MIC90: 0.25 microg/mL). The antibacterial activity of the drugs to Pseudomonas aeruginosa was generally low, and MIC90 of all the drugs were ranged from 64 to > or = 256 microg/mL except IPM and amikacin (AMK) having 16 microg/mL. The antibacterial activity of CZOP was relatively good (MIC50: 8 microg/mL).

Anti-Bacterial Agents↗
PubMed2004

[Comparative studies on activities of antimicrobial agents against causative organisms isolated from patients with urinary tract infections (2002). II. Background of patients].

A total of 615 bacterial strains isolated from 491 patients diagnosed as having urinary tract infections (UTIs) in 13 institutions in Japan were supplied between August 2002 and July 2003. Then, the clinical background of patients were investigated such as sex, age, and type of infections and kind of bacteria, frequency of bacteria isolation by age and infections, bacteria and infections by timing of antibiotics administration, and bacteria and infections by surgical procedures. The bacterial strains were divided by the age and sex of the patients and the types of infections. In males, the number of patients aged less than 50 years was few and the complicated UTIs without indwelling catheter was observed most frequently. In the present time, the bacteria most frequently isolated were Escherichia coli. Pseudomonas aeruginosa and Enterococcus faecalis were also relatively frequently isolated. E. coli decreased and P. aeruginosa increased with the complication of the infections. E. faecalis tended to be isolated more frequently in the complicated UTIs than in the uncomplicated UTIs. With respect to the relation of these results to the age of the patients, in the uncomplicated UTIs, the isolation frequency of E. coli was the highest in all age groups, accounting for 70.8% in 20-49 years old. In the complicated UTIs without indwelling catheter, the isolation frequency of E. coli decreased with aging of the patients and was lower than that of E. faecalis in the patients aged 70 or more (18.3 vs 20.6%). In the complicated UTIs with indwelling catheter, the isolation frequency of E. coli was lower than in the uncomplicated UTIs in all age groups and P. aeruginosa and E. faecalis were more frequently isolated. In comparison of causative bacteria in UTIs between before and after the administration of antibiotics, E. coli decreased and P. aeruginosa increased after the administration in any UTIs. In comparison of causative bacteria in UTIs with or without surgery, E. coli was more frequently isolated in the patients without surgery, while P. aeruginosa was more in the patients with surgery in any UTIs. E. faecalis was more frequently isolated in the patients with surgery in uncomplicated and complicated UTIs without indwelling catheter and in the patients without surgery in complicated UTIs with indwelling catheter.

Age Factors↗
PubMed2004

[Comparative studies on activities of antimicrobial agents against causative organisms isolated from patients with urinary tract infections (2002). III. Secular changes in susceptibility].

The bacteria (Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Klebsiella spp. and Pseudomonas aeruginosa) isolated from patients diagnosed as urinary tract infections (UTIs) in 13 institutions in Japan were supplied between August 2002 and July 2003. The susceptibilities of these bacteria to various antimicrobial agents were examined. The bacteria were divided into 2 groups consisting of uncomplicated UTIs and complicated UTIs (with and without indwelling catheter) based on their isolation origins. The results were compared with those obtained between 1993 and 2001. The drug sensitivity of S. aureus in this year was similar to those in up to the previous year and S. aureus showed the best susceptibility to vancomycin. The drug sensitivity of E. faecalis in this year also was similar to those in up to the previous year. The drug sensitivity of E. coli in this year was generally good except penicillins and was similar to those in up to the previous year. Among cephems, cefozopran (CZOP) and cefpirome (CPR) showed the highest potency (MIC90: < or = 0.125 microg/mL). An antibacterial activity of cefotiam (CTM) was similar to it in 10 years ago and was fine (MIC90: < or = 1 microg/mL). The sensitivity of E. coli to carbapenems and carumonam (CRMN) also was good like to CZOP. However, the sensitivity of the complicated UTIs group to quinolones decreased after 2000 and was suggested to develop the resistance to the drug. The drug sensitivity of Klebsiella spp. in this year also was similar to those in up to the previous year. The bacteria showed good susceptibility (MIC: < or = 0.125 microg/mL) to cefmenoxime (CMX), CPR, cefixime (CFIX), flomoxef (FMOX), and CZOP among cephems. The drug sensitivity of P. aeruginosa was generally low. Most of the bacteria were little sensitive to cephems except CZOP and ceftazidime (CAZ). The sensitive bacteria to CZOP and ceftazidime (CAZ) were observed to be 26.8% (15/56 strains) and 39.3% (22/56 strains) in complicated UTIs group, respectively. The sensitivity profile of P. aeruginosa to the other tested drugs was not much different from that in up to the previous year. However, the sensitivity of the bacteria to carbapenems tended to decrease after 2000, and the low sensitive strains (MIC: > or = 256 microg/mL) were detected at 22.2% (2/9 strains) in the uncomplicated UTIs group and 3.6% (2/56 strains) in the complicated UTIs group.

Anti-Bacterial Agents↗
PubMed2003

Crystal structure of extended-spectrum beta-lactamase Toho-1: insights into the molecular mechanism for catalytic reaction and substrate specificity expansion.

The crystallographic structure of the class A beta-lactamase Toho-1, an extended-spectrum beta-lactamase with potent activity against expanded-spectrum cephems, has been determined at 1.65 A resolution. The result reveals that the Lys73 side chain can adopt two alternative conformations. The predominant conformation of Lys73 is different from that observed in the E166A mutant, indicating that removal of the Glu166 side chain changes the conformation of the Lys73 side chain and thus the interaction between Lys73 and Glu166. The Lys73 side chain would play an important role in proton relay, switching its conformation from one to the other depending on the circumstances. The electron density map also implies possible rotation of Ser237. Comparison of the Toho-1 structure with the structure of other class A beta-lactamases shows that the hydroxyl group of Ser237 is likely to rotate through interaction with the carboxyl group of the substrate. Another peculiarity is the existence of three sulfate ions positioned in or near the substrate-binding cavity. One of these sulfate ions is tightly bound to the active center, while the other two are held by a region of positive charge formed by two arginine residues, Arg274 and Arg276. This positively charged region is speculated to represent a pseudo-binding site of the beta-lactam antibiotics, presumably catching the methoxyimino group of the third-generation cephems prior to proper binding in the substrate-binding cleft for hydrolysis. This high-resolution structure, together with detailed kinetic analysis of Toho-1, provides a new hypothesis for the catalytic mechanism and substrate specificity of Toho-1.

Amino Acid Substitution↗
PubMed2003

A detailed kinetic study of Mox-1, a plasmid-encoded class C beta-lactamase.

Surveys of beta-lactamases in different parts of the world show an important increase in class C beta-lactamases, thus the study of these enzymes is becoming an important issue. We created an overproduction system for Mox-1, a plasmid class C beta-lactamase, by cloning the gene encoding this enzyme, and placing it under the control of a T7 promoter, using vector pET 28a. The enzyme, purified by ion exchange chromatography, was used to obtain the molecular mass (38246), the N-terminal sequence (GEASPVDPLRPVV), and pI (8.9), and to perform a detailed kinetic study. Cephalotin was used as reporter substrate in the case of poor substrates. The kinetic study showed that benzylpenicillin, cephalotin, cefcapene and moxalactam were good substrates for Mox-1 (k(cat)/K(m) values >2.5 x 10(6) M(-1) s(-1)). On the other hand, ceftazidime and cefepime were poor substrates for this enzyme (K(m) values >200 microM). Clavulanic acid had no inhibitory effect on Mox-1 (K(m)=30.2 mM), however aztreonam behaved as an inhibitor of Mox-1 (K(i)=2.85 microM).

Amino Acid Sequence↗
PubMed2003

Hyperoxia mediates acute lung injury and increased lethality in murine Legionella pneumonia: the role of apoptosis.

Legionella pneumophila is a major cause of life-threatening pneumonia, which is characterized by a high incidence of acute lung injury and resultant severe hypoxemia. Mechanical ventilation using high oxygen concentrations is often required in the treatment of patients with L. pneumophila pneumonia. Unfortunately, oxygen itself may propagate various forms of tissue damage, including acute lung injury. The effect of hyperoxia as a cofactor in the course of L. pneumophila pneumonia is poorly understood. In this study, we show that exposure to hyperoxic conditions during the evolution of pneumonia results in a marked increase in lethality in mice with Legionella pneumonia. The enhanced lethality was associated with an increase in lung permeability, but not changes in either lung bacterial burden or leukocyte accumulation. Interestingly, accelerated apoptosis as evidenced by assessment of histone-DNA fragments and caspase-3 activity were noted in the infected lungs of mice exposed to hyperoxia. TUNEL staining of infected lung sections demonstrated increased apoptosis in hyperoxic mice, predominantly in macrophages and alveolar epithelial cells. In vitro exposure of primary murine alveolar epithelial cells to Legionella in conjunction with hyperoxia accelerated apoptosis and loss of barrier function. Fas-deficient mice demonstrated partial resistance to the lethal effects of Legionella infection induced by hyperoxia, which was associated with attenuated apoptosis in the lung. These results demonstrate that hyperoxia serves as an important cofactor for the development of acute lung injury and lethality in L. pneumophila pneumonia. Exaggerated apoptosis, in part through Fas-mediated signaling, may accelerate hyperoxia-induced acute lung injury in Legionella pneumonia.

Acute Disease↗
PubMed2003

Evaluation of antimicrobial agents using an experimental pulmonary superinfection model with Aspergillus fumigatus and Pseudomonas aeruginosain leukopenic mice.

The therapeutic efficacy of amphotericin B (AmB), imipenem/cilastatin (IPM/CS), pazufloxacin (PZFX) mesilate, and combinations of these, was evaluated using an experimental pulmonary superinfection model in mice caused by Pseudomonas aeruginosa and Aspergillus fumigatus. The superinfected mice died within 3 days. Although the viable cell count of P. aeruginosa increased markedly from 10(3) to 10(8) CFU/lung on day 2 after infection, that of A. fumigatus decreased from 10(3) to 10(2) CFU/lung on that day, showing that P. aeruginosa facilitated the mortality in the superinfection. Extensive necrosis in the lung parenchyma and moderate hyphae proliferation of A. fumigatus were observed on day 2 after infection. Mice treated with PZFX mesilate (50 mg/kg per day) and the combination of PZFX mesilate (50 mg/kg per day) - AmB (2.5 mg/kg per day) showed prolonged survival in comparison to untreated control mice ( P < 0.05). In the PZFX mesilate-treated group, no significant necrosis was observed, but necrosis due to the hyphae proliferation of A. fumigatus was still observed in the lung parenchyma on day 6 after infection. However, neither significant necrosis nor hyphae proliferation of A. fumigatus was observed in mice treated with the combination of PZFX mesilate - AmB. On the other hand, the survival rates of mice treated with AmB (2.5 mg/kg per day), IPM/CS (50 mg/kg per day), and the IPM/CS-AmB combination were all less than 10%. The viable cell count of P. aeruginosa decreased in PZFX mesilate-alone group and in the combination of PZFX mesilate - AmB group, but no significant decrease in this count was observed in the IPM/CS and combination of IPM/CS-AmB group. The viable count of A. fumigatus was increased in the IPM/CS, PZFX mesilate-alone, and combination of IPM/CS-AmB groups, but the count was suppressed in the AmB-alone and the combination of PZFX mesilate - AmB group. In conclusion, this superinfection model would be useful to evaluate the therapeutic potential of combinations of antibacterial and antifungal agents, and the scheduling of drug administration in terminal infections caused by P. aeruginosa and A. fumigatus.

Animals↗
PubMed2003

Isolation of Helicobacter cinaedi from blood of an immunocompromised patient in Japan.

We report the isolation of Helicobacter cinaedi (previously called " Campylobacter-like organism") from the blood of an immunosuppressed Japanese patient receiving immunosuppressive therapy after renal transplantation. The identification of H. cinaedi was based on the findings of microscopic examinations, biochemical properties, and 16S rRNA gene nucleotide sequences. H. cinaedi bacteremia should be considered as one of the critical infectious diseases in immunocompromised patients, and the sequencing of 16S rRNA may be a useful method to confirm the identification of this organism.

Adult↗
PubMed2003

A case of Legionella pneumophila pneumonia complicated by miliary tuberculosis.

A 47-year-old woman was admitted to hospital with severe Legionella pneumonia. The respiratory symptoms improved dramatically and the X-rays revealed a decrease in the diffuse chest infiltrates after treatment with erythromycin and rifampicin. However, chest CT scans showed that the reticulonodular opacities persisted for several weeks after the onset of pneumonia. Two months after admission, the chest X-rays showed the progression of small nodules in both lungs and there was increasing respiratory distress. A diagnosis of miliary tuberculosis was confirmed. The present case should alert physicians to this potentially confusing combination of respiratory pathogens.

Diagnosis, Differential↗
PubMed2003

Factors influencing survival of Legionella pneumophila serotype 1 in hot spring water and tap water.

The factors involved in the survival of Legionella pneumophila in the microcosms of both hot spring water and tap water were studied by examining cultivability and metabolic activity. L. pneumophila could survive by maintaining metabolic activity but was noncultivable in all microcosms at 42 degrees C, except for one microcosm with a pH of <2.0. Lower temperatures supported survival without loss of cultivability. The cultivability declined with increasing temperature, although metabolic activity was observed at temperatures of up to 45 degrees C. The optimal range of pH for survival was between 6.0 and 8. The metabolic activity could be maintained for long periods even in microcosms with high concentrations of salt. The cultivability of organisms in the post-exponential phase in a tap water microcosm with a low inoculum size was more rapidly reduced than that of organisms in the exponential phase. In contrast, the loss of cultivability in microcosms of a high inoculum size was significant in the exponential phase. Random(ly) amplified polymorphic DNA analysis of microcosms where cultivability was lost but metabolic activity was retained showed no change compared to cells grown freshly, although an effect on the amplified DNA band pattern by production of stress proteins was expected. Resuscitation by the addition of Acanthamoeba castellanii to the microcosm in which cultivability was completely lost but metabolic activity was maintained was observed only in part of the cell population. Our results suggest that L. pneumophila cell populations can potentially survive as free organisms for long periods by maintaining metabolic activity but temporarily losing cultivability under strict environments and requiring resuscitation by ingestion by amoebas.

Acanthamoeba↗
PubMed2003

The Pseudomonas aeruginosa autoinducer N-3-oxododecanoyl homoserine lactone accelerates apoptosis in macrophages and neutrophils.

Quorum-sensing systems are critical regulators of the expression of virulence factors of various organisms, including Pseudomonas aeruginosa. Las and Rhl are two major quorum-sensing components, and they are regulated by their corresponding autoinducers, N-3-oxododecanoyl homoserine lactone (3-oxo-C(12)-HSL) and N-butyryl-L-homoserine lactone (C(4)-HSL). Recent progress has demonstrated the potential of quorum-sensing molecules, especially 3-oxo-C(12)-HSL, for modulation of the host immune system. Here we show the specific ability of 3-oxo-C(12)-HSL to induce apoptosis in certain types of cells. When bone marrow-derived macrophages were incubated with synthetic 3-oxo-C(12)-HSL, but when they were incubated not C(4)-HSL, significant loss of viability was observed in a concentration (12 to 50 micro M)- and incubation time (1 to 24 h)-dependent manner. The cytotoxic activity of 3-oxo-C(12)-HSL was also observed in neutrophils and monocytic cell lines U-937 and P388D1 but not in epithelial cell lines CCL-185 and HEp-2. Cells treated with 3-oxo-C(12)-HSL revealed morphological alterations indicative of apoptosis. Acceleration of apoptosis in 3-oxo-C(12)-HSL-treated cells was confirmed by multiple criteria (caspases 3 and 8, histone-associated DNA fragments, phosphatidylserine expression). Structure-activity correlation experiments demonstrated that the fine structure of 3-oxo-C(12)-HSL, the HSL backbone, and side chain length are required for maximal activity. These data suggest that Pseudomonas 3-oxo-C(12)-HSL specifically promotes induction of apoptosis, which may be associated with 3-oxo-C(12)-HSL-induced cytotoxicity in macrophages and neutrophils. Our data suggest that the quorum-sensing molecule 3-oxo-C(12)-HSL has critical roles in the pathogenesis of P. aeruginosa infection, not only in the induction of bacterial virulence factors but also in the modulation of host responses.

4-Butyrolactone↗
PubMed2003

A large outbreak of Legionnaires' disease due to an inadequate circulating and filtration system for bath water--epidemiologic manifestations.

OBJECTIVE: To study the epidemiologic manifestations of a large outbreak of Legionnaires' disease due to an inadequate circulating and filtration system for bath water. PATIENTS: In June 2000 at Ishioka City, Ibaraki Prefecture, a large outbreak of Legionnaire's disease occurred, as a result of an inadequate circulating and filtration system for communal bath water. This outbreak was the worst ever experienced in Japan, involving a total of 34 patients (20 confirmed, 14 probable), 3 of whom died. MEASUREMENTS AND RESULTS: Legionella pneumophila serogroup 1 was isolated from sputum culture in two patients. Bacteriological culture of the public bath water subsequently yielded large numbers of Legionella species. Cleavage of genomic DNA showed that restriction fragment patterns coming from clinical and environmental isolates of L. pneumophila serogroup 1 were closely related, focusing the inquiry to a public bathhouse where a circulating filtration system was suspected as the source of infection. CONCLUSIONS: It was later concluded that the circulating filtration system adopted for bath water was marred by a serious design flaw that subsequently caused the mass outbreak. Specifically, a line of the bath water was being returned to the bath without undergoing heat exchange or sterilization by chlorine; and the Legionella species that had proliferated in the filter and the bright stone filtration unit were allowed to return to the bath, eventually culminating in a mass infection.

Adult↗
PubMed2003

[A major outbreak of Legionnaire's disease due to a public bathhouse: clinical examination].

The largest epidemic outbreak of legionnaires' disease occurred in Japan, where there were 45 cases, including three deaths. There was a high degree of homology at the genetic level between Legionella pneumophila sero-group 1 isolated from two patients and water samples collected from a public bath, and this highlighted the public bath visited by the patients as the source of the infection. We report our clinical observations of 34 cases with a pneumonia type (20 definite and 14 probable) of the 42 cases for whom a diagnosis was made and follow-up care was subsequently provided at our hospital or hospital-affiliated facilities, out of the initial total of 45 cases. Twenty-one cases had underlying diseases, but the disease was mild in all patients. Three cases showed different neuropsychiatric symptoms such as reduced capacity for memorization, somnolence or delirium. Diarrhea was found in three cases, and relative bradycardia in ten. Clinical laboratory test results indicated that a severe inflammatory reaction, renal or hepatic dysfunction, an electrolyte abnormality and abnormal urinalysis were common in the 34 cases. Chest radiography showed a shadow suggestive of infection of the pulmonary parenchyma with multilobar involvement in most cases and pleural effusion in nine cases.

Baths↗