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Purpose Short bowel syndrome (SBS) with intestinal failure (SBS-IF) requires long-term parenteral nutrition (PN). This study investigated the real-world etiologies of SBS, treatment patterns, and PN-related outcomes among adult patients with SBS-IF in Japan. Methods This retrospective, observational cohort study was based on data from April, 2008 to January, 2020 from one of the largest hospital-based claim databases in Japan. Analyzed patients were aged ≥ 16 years, had received continuous PN for ≥ 6 months, and had SBS or undergone SBS-related surgery with a diagnosis of a causative disease. The primary endpoint was PN weaning. Results We analyzed data for 393 patients. The most frequent causes of SBS-IF were ileus (31.8%), Crohn’s disease (20.1%), and mesenteric ischemia (16.0%). Of 144/393 (36.6%) patients who were weaned off their PN, 48 (33.3%) were subsequently restarted on PN. Of 276/393 (70.2%) patients whose PN was initiated in hospital, 156 (56.5%) transitioned to home management. The mean duration of initial PN was 450.4 and 675.5 days for patients who were able or unable to be weaned off PN, respectively. Sepsis (67.4%), catheter-related bloodstream infections (49.1%), and liver disorders (45.0%) were the most reported PN-related complications. Conclusions Most patients with SBS-IF in Japan could not be weaned off PN and suffered life-threatening complications.

A 66-year-old Japanese woman who was diagnosed with synovitis-acne-pustulosis-hyperostosis-osteitis (SAPHO) syndrome presented with bilateral blurred vision 4 months prior to visiting our hospital. She had visited a local ophthalmology clinic first. She was diagnosed with conjunctivitis and was prescribed antibacterial eye drops. The symptoms persisted in spite of treatment. She was then referred to our hospital. At her initial visit, the visual acuities were 0.6 in both eyes. A slit-lamp examination revealed bilateral shallow anterior chamber, and intraocular pressures of 18 mm Hg in the right eye and 16 mm Hg in the left eye. There were no cells in the anterior chamber. Fundus examination revealed bilateral annular choroidal detachment and serous retinal detachment. Fluorescein angiography showed leakage of dye from the retinal pigment epithelium (RPE) and indocyanine green angiography showed focal choroidal hypoperfusion. Optical coherence tomography showed wavy RPE line and blurry thick choroid. Systemic investigation by the physician demonstrated bilateral pleural effusions of unknown origin. The patient had a past history of breast cancer; however, no metastasis was identified via malignant cells through cytology, laboratory findings, radiographs, CT, and MRI. After the diagnosis of Vogt-Koyanagi-Harada (VKH) disease was made, the patient was treated with local and systemic steroid including high-dose intravenous corticosteroids, and 150 mg of cyclosporine per day. Seventy days after the second high-dose of intravenous corticosteroids, these medications brought a complete resolution of both choroidal and retinal detachment. VKH disease associated with SAPHO syndrome is rare. The combination of immunosuppressive drug and steroid might be helpful for severe cases of VKH disease.

Objective The present study aimed to estimate cumulative incidence of overweight and obesity and describe 5‐year longitudinal changes in body mass index (BMI) in a large occupational cohort in Japan. Methods Participants were 55 229 Japanese employees, who were aged 20‐59 years and attended at all subsequent annual health check‐ups between 2009 and 2014. Mixed model analysis was performed to examine the effects of age and cohort by gender on BMI change, with age as a random variable. Cumulative incidence of overweight (23.0≤ BMI

Metabolic syndrome (MetS) is an insulin-resistance syndrome deeply associated with obesity. The association between MetS and several clinical factors, including serum levels of adiponectin, C-reactive protein and insulin, was examined.The study was performed in 219 men aged 43.2 ± 6.6 years, who were employees of a company. The study measured the serum high sensitivity C-reactive protein (CRP) as an inflammatory marker, serum insulin as a marker of insulin resistance and serum high-molecular-weight adiponectin (HMW-adipo), in addition to the serum levels of liver enzymes and uric acid. MetS was diagnosed according to the National Cholesterol Education Programme ATPIII criteria.Log-transformed serum ALT, HMW-adipo, insulin and uric acid were found to be significantly related to the presence/absence of Mets by logistic regression analysis, with odds ratios (95% confidence intervals) of 25.9 (3.3-204.4), 0.10 (0.02-0.56), 7.5 (1.6-34.1) and 1.5 (1.04-2.1), respectively.Serum high HMW-adipo level was significantly negatively related to the presence of MetS, independently of the serum levels of insulin, liver enzymes and uric acid. Among the markers examined, serum HMW-adipo and ALT were considered as markers for MetS.

The antimicrobial susceptibility of 93 Acinetobacter baumannii complex isolates from clinical specimens collected nationwide between May and October 2009 were measured by microdilution antimicrobial susceptibility testing based on CLSI M100-S20. Beta-lactamase genes, including classes B and D and ISAbal in meropenem nonsusceptible, including intermediate or resistant isolates, were detected using PCR. Rates of isolates nonsusceptible to meropenem were 18%, to ciprofloxacin 41% and to amikacin 14%. L7-L8: The rate of multidrug-resistant Acinetobacter (MDRA) isolates which were resistant to all 3 antimicrobial agents was 4.3%. MDRA isolates were classified into ST92 by multilocus sequence typing. No metallo-beta-lactamase producer was seen among the 17 meropenem nonsusceptible isolates. The blaoxa-51-like carbapenemase gene and ISAbal were detected in all 17 isolates. ISAba1 upstream presence of the blaOXA-51-like gene was observed in 7 of 17 isolates and the blaOXA-23 like gene in 5 of 17. Consistent with overseas reports, our results confirm the existence of MDRA isolates and isolates harboring OXA carbapenemase genes in Japan. While resistance rates were lower than reports elsewhere, it is clear that resistance trends must be carefully monitored.

β-Lactamase-negative ampicillin-resistant (BLNAR) isolates of Haemophilus influenza e have been emerging in some countries, including Japan. The Clinical and Laboratory Standards Institute has only a susceptible MIC breakpoint (≤1 μg/ml) for piperacillin-tazobactam and a disclaimer comment that BLNAR H. influenzae should be considered resistant, which was adapted without presentation of data. In addition, fluoroquinolone-resistant H. influenzae isolates have recently been occasionally reported worldwide. To address these problems, we examined susceptibilities to β-lactams, including piperacillin-tazobactam, and ciprofloxacin by microdilution and disk diffusion (only for piperacillin-tazobactam) methods, against a total of 400 recent H. influenzae clinical isolates, including 100 β-lactamase-negative ampicillin-susceptible, β-lactamase-positive ampicillin-resistant, BLNAR, and β-lactamase-positive amoxicillin-clavulanate-resistant (BLPACR) isolates each. BLNAR and BLPACR isolates were tested by PCR using primers that amplify specific regions of the ftsI gene. We also detected mutations in quinolone resistance-determining regions (QRDRs) by direct sequencing of the PCR products of DNA fragments. Among β-lactams, piperacillin-tazobactam exhibited potent activity against all isolates of H. influenzae , with all MICs at ≤0.5 μg/ml (susceptible). A disk diffusion breakpoint for piperacillin-tazobactam of ≥21 mm is proposed. We confirmed that all BLNAR and BLPACR isolates had amino acid substitutions in the ftsI gene and that the major pattern was group III-like (87.5%). One ciprofloxacin-resistant isolate (MIC, 16 μg/ml) and 31 ciprofloxacin-susceptible isolates (MICs, 0.06 to 0.5 μg/ml) had amino acid changes in their QRDRs. Piperacillin-tazobactam was the most potent β-lactam tested against all classes of H. influenzae isolates. It is possible that fluoroquinolone-resistant H. influenzae will emerge since several clinical isolates carried mutations in their QRDRs.

As the most common cause of neonatal sepsis, Lancefield Group B Streptococcus (GBS) must be diagnosed as early as possible in pregnant women is prevent neonatal infection. A selective enrichment broth medium has been widely recommended to optimally recover GBS from genital and anorectal samples. To establish a culture suitable for screening vaginal swab specimens, we compared subcultures of three selective enrichment media to direct culture on agar medium. Vaginal swab samples were inoculated directly onto 5% sheep blood agar and into New Granada medium (Eiken), Lim broth (Becton, Dickinson, and Company), and Todd Hewitt broth with gentamicin and nalidixic acid (Becton, Dickinson, and Company, Todd). Of the 288 specimens tested, GBS was recovered from 43 samples (14.9%) on direct agar media, with 82 (28.5%), positive on New Granada medium subculture, 67 (23.3%) on Lim broth subculture, and 61 (21.2%) on Todd, subculture. These results demonstrates that selective enrichment broth media provides more superior sensitivity than direct agar media for detection of GBS colonization in vaginal specimens, underscoring the usefulness of selective enrichment broth media in GBS screening for vaginal swabs in pregnant woman.

The minimum inhibitory concentrations (MICs) of six antibiotics against 33 strains of Burkholderia cepacia isolated from 33 patients suspected of having sepsis or bacteremia were determined. All the B. cepacia isolates were resistant to gentamicin (MIC(90), >128 microg/ml), and 21.2% were resistant to imipenem (MIC(90), 16 microg/ml). They were susceptible to ciprofloxacin, ceftazidime, and minocycline (MIC(90), 2-8 microg/ml). The 33 blood isolates were classified into 12 types (A-L) by the pulsed-field gel electrophoresis pattern. All 11 of A type, 2/4 of B type, 3/4 of C type, 4/5 of E type, and both G type B. cepacia were isolated from different patients in the same hospital. These results suggest that B. cepacia can spread among patients with sepsis or bacteremia.

We conducted a study for the growth of and selectivity for the desired microorganisms using a newly developed selective culture medium for Helicobacter pylori, Columbia horse blood agar HP (CHBHP), at three different Japanese clinical laboratories, Hokkaido, Kanto and Kyusyu. When standard strains and clinical isolates of H. pylori were examined, the recovery of the organism on the CHBHP media was comparable to that of conventional selective and nonselective media. However, colonies were obviously larger on the CHBHP media. These media yielded the highest H. pylori positive rate for clinical specimens at all the three laboratories. The detection rate of the CHBHP media in H. pylori-positive specimens was higher than that of media commonly used at the three laboratories (98.1% to 100% vs. 88.0% to 96.2%). The CHBHP media also achieved a higher detection rate for specimens from H. pylori-infected animals. CHBHP media have an excellent growth supporting ability and selectivity originating from Columbia agar base and do not require the combined use of non-selective media for the growth and isolation of the organism, resulting in lower cost. Thus, they are useful media for the selective culture and isolation of H. pylori from clinical and animal specimens.

Sitafloxacin showed MICs of less than or equal to 0.5 μg/ml against 105 isolates of Helicobacter pylori , including 44 isolates with mutations in the gyrA gene. The highest MICs for garenoxacin and levofloxacin were 8 and 64 times, respectively, higher than the highest MICs observed for sitafloxacin.

The clinical isolates of Pseudomonas aeruginosa can be roughly classified into long- and short-LPS strains and LPS-deficient strains. Ionic binding of 3H-gentamicin was the highest in the long-LPS strains followed in descending order by short-LPS and LPS-deficient strains. However, PAC605 strain, completely lacking in the repeated units of O-polysaccharide and also lacking in some of the neutral sugar residues of the core oligosaccharide region, highly bound to 3H-gentamicin and it is suggested that the negatively charged sites on the deep-core oligosaccharide region and/or on lipid A participated in the binding to 3H-gentamicin. This manner of binding may be also applied to P. aeruginosa No. 45 (LPS-deficient), a clinical isolate.

We compared the Mycobacteria Growth Indicator Tube 960 (MGIT 960) and Ogawa medium (OM) for the detection of mycobacteria (acid fast bacteria: AFB) using 882 sputum specimens. Overall, 120 strains of AFB were isolated by the MGIT 960 system and 99 strains of AFB were isolated by using OM. As far as Mycobacterium tuberculosis is concerned, 88 and 71 isolates were achieved by the MGIT 960 and OM respectively. A total of 28 isolates (18 isolates of M. tuberculosis and 10 isolates of nontuberculous mycobacteria: NTM) were detected by the MGIT 960 only whereas only 2 isolates (1 M. tuberculosis and 1 NTM) were detected by OM only. Of these sputum specimens, 72 were smear positive for AFB. The rates of smear negative but culture positive specimens were 8.0% (65 out of 809) for the MGIT 960 system and 6.2% (50 out of 809) for OM. The contamination rate for MGIT 960 was only 1.2%. The average time required for detection of M. tuberculosis was 14.1 days by the MGIT 960 system and 24.6 days by OM. For the NTM, the average detection time were 8.3 days for the MGIT 960 system and 22.8 days for OM. These results indicate that the MGIT 960 system allows detection of mycobacteria significantly faster than OM.

In vitro activity of sitafloxacin (STFX) and various oral antimicrobial agents against bacterial isolates recovered from clinical specimens between January and December 2012, at different healthcare facilities in Japan was evaluated. A total of 1,620 isolates including aerobic and anaerobic organisms were available for the susceptibility testing using the microbroth dilution methods recommended by Clinical and Laboratory Standards Institute. The minimum inhibitory concentration of STFX at which 90% of isolates (MIC90) was 0.5 microg/mL for methicillin-susceptible Staphylococcus aureus and was 2 times lower than that of garenoxacin (GRNX), 4 times lower than that of moxifloxacin (MFLX), and 16 times lower than that of levofloxacin (LVFX). STFX inhibited the growth of all the isolates of Streptococcus pneumoniae at 0.06 microg/mL or less. The MIC90 of STFX was 0.03 microg/mL and was 2 times lower than that of GRNX, 4 times lower than that of MFLX, and 32 times lower than that of LVFX. Against Streptococcus pyogenes, the MIC90 of STFX was 0.06 microg/mL and was 2 times lower than that of GRNX, 8 times lower than that of MFLX, and 32 times lower than that of LVFX. The MIC90 of STFX was 2 microg/mL for Enterococcus faecalis, and was 4 times lower than that of GRNX, 8 times lower than that of MFLX, and 32 times lower than that of LVFX. The MIC90 of STFX for Escherichia coli was 2 microg/mL, and the MIC90(s) of other 10 species of Enterobacteriaceae which were the lowest values of the quinolones tested ranged from 0.03 to 1 microg/mL. The MIC90 of STFX for Pseudomonas aeruginosa isolates recovered from urinary infections was 4 microg/mL and was 32 times lower than those of GRNX, MFLX and LVFX. The MIC90 of STFX for P. aeruginosa isolates recovered from respiratory infections was 4 microg/mL and was 8 to 16 times lower than those of GRNX, MFLX, and LVFX. STFX inhibited the growth of all the isolates of Haemophilus influenzae at 0.004 microg/mL or less, and was 4 times lower than that of GRNX, 16 times lower than that of MFLX, and 8 times lower than that of LVFX. The MIC90 of STFX was 0.015 microg/mL for Moraxella catarrhalis, and was equal to that of GRNX, 4 times lower than those of MFLX and LVFX. The MIC90(s) of STFX ranged from 0.03 to 0.25 microg/mL for all the species of anaerobic bacteria and were the lowest values of all the antimicrobial agents tested. In conclusion, the activity of STFX against Gram-positive cocci was comparable or superior to those of GRNX, MFLX and LVFX. STFX showed the most potent activity against Gram-negative bacteria and anaerobic bacteria of all the antimicrobial agents tested in this study.

Fifty-nine strains of Pseudomonas aeruginosa, isolated from blood, urine, pus, sputum and feces, were divided into three groups on the basis of differences in their LPS composition, in long- and short-LPS strains and in LPS-deficient strains. The long-LPS strains were far more frequently isolated among blood, urine and fecal specimens than in the remainder, and the frequency of LPS-deficient strains was higher in sputum specimens than in the remainder. The drug resistance patterns of the three LPS-type strains to six different kinds of anti-pseudomonal drugs, including gentamicin, differed significantly.

A total of 21,609 faecal specimens obtained from patients with diarrhea mainly in Kanto area between June and September 1996 were investigated to identify the causative pathogens for diarrhea. One-hundred fifty-seven strains of Escherichia coli of 29 different O-serotypes were isolated as the causative pathogens, which were previously recognized to induce severe abdominal cramps and diarrhea. Of these, 114 strains, in which the possibility of enterohemorrhagic E. coli due to their O-serotypes was predicted, were examined for the producibility of Vero toxins. Twenty-six (76.5%) of the 34 strains of E. coli O157 produced the Vero toxins, and other 8 strains were the non-producers. Twenty of the 26 producers produced both VT1 and VT2, whereas the other 6 strains produced VT2 only. Furthermore, 4 strains of E. coli O26, and 1 strain each of E. coli O125 and O126 produced Vero toxins. Thirty-two of the 114 strains, isolated from the patients with diarrhea and selected as the enterohemorrhagic E. coli according to the specific O-serotypes, were actually confirmed produce Vero toxins. Thirty-four strains of E. coli O157 tested were susceptible to all antibiotics such as ampicillin, doxycylin, levofloxacin, fosfomycin, chloramphenicol and polymyxin B, and no strains resistant to levofloxacin, polymyxin B and fosfomycin were found.

It is well known that Helicobacter pylori produced a large amount of urease which plays an important role in the maintenance of infection and adhesion. Two thousand three hundred and thirty-one specimens of the gastrointestinal mucosa were obtained from patients with peptic ulcer to isolate H. pylori strains. Of these specimens, 1602 strains of H. pylori were isolated and 7 urease-negative H. pylori strains were found. Biological characteristics (except urease production) of urease-negative H. pylori were in accord with urease-positive reference strains (ATCC strains). Furthermore, these urease-negative strains were confirmed to be H. pylori strains by PCR. These results suggest that the emergence of urease-negative strains pose a new problem for H. pylori infection in patients with peptic ulcer.

We investigated annual changes in clarithromycin resistance and resistance genes in 579 strains of Streptococcus pyogenes isolated from patients with symptomatic respiratory tract infections who visited primary medical institutions during the 5-year period from 1998 to 2002. The minimum inhibitory concentrations (MICs) of clarithromycin for S. pyogenes were measured using the standard broth microdilution method according to the National Committee for Clinical Laboratory Standards (NCCLS) guidelines, and strains showing MICs of 1 microg/ml or greater were regarded as being resistant to clarithromycin, according to the resistance standard specified by the NCCLS. The rates of S. pyogenes resistance to clarithromycin were 7.3% overall, 5.8% in 1998, 4.9% in 1999, 7.7% in 2000, 6.4% in 2001, and 11.1% in 2002. While the annual rates fluctuated slightly each year, an overall tendency to increase was observed during the 5-year period. Regarding the macrolide-resistance genes in the macrolide-resistant strains, mefA/E (+)/ ermB(-) was the most common genotype detected in these strains, while the ermB (+)/ mefA/E (-) and mef A/E (-)/ ermB (-) genotypes were detected at about the same rate. The MICs of clarithromycin for the ermB (+) strains tended to be higher than those of the mefA/E (+) strains, but some mefA/E (-) / ermB (-) strains also exhibited high MICs of clarithromycin, similar to those of the ermB (+) strains. The above results indicate that the number of clarithromycin-resistant strains of S. pyogenes is gradually increasing and that the resistance is becoming stronger; thus, special attention must be paid to the appearance of macrolide-resistant strains of S. pyogenes.

We conducted a study assess the bactericidal activity of sitafloxacin (STFX) against Streptococcus pneumoniae isolates recovered from respiratory infections including penicillin-resistant (PRSP) isolates, macrolide resistant isolates possessing mefA and ermB resistance genes and quinolone resistance isolates with mutations in gyrA or gyrA and parC. Each isolate tested was grown in hemosupplemented Mueller-Hinton broth and adjusted to approximately 10(5) CFU/ mL. Isolates were than exposed to a Cmax antimicrobial blood level that would be attained with routine antimicrobial administration and an antimicrobial level that would be expected 4 hours post-Cmax (Cmax 4hr). Bactericidal activity was measured for up to 8 hours. Excluding a subset of S. pneumoniae isolates with mutations in the quinolone resistance determining region (QRDR), all quinolones showed bactericidal activity at Cmax and Cmax 4 hr antimicrobial concentrations for up to 8 hours. Against S. pneumoniae isolates with either gyrA or gyrA and parC mutations, bactericidal activity of STFX was shown for up to 4 to 8 hours following Cmax based on a limit of detection of1.3 log CFU/mL. Garenoxacin (GRNX) did not showed bactericidal activity below the limit of detection for up to 8 hours with exposure to Cmax and no bactericidal activity was seen with levofloxacin. When all quinolones tested where adjusted to concentrations corresponding to their MICs, STFX showed the most rapid bactericidal activity against PRSP. This rapid bactericidal activity in PRSP is a key to the effectiveness of STFX. Our findings show that beyond inhibition of bacterial replication by blocking their DNA replication pathway and synthesis of proteins, STFX demonstrated characteristics contributing to greater bactericidal activity compared to GRNX. In conclusion, of the newer quinolones, STFX showed the strongest bactericidal activity against S. pneumoniae isolates with mutations in the QRDR which indicates that it may show the most effective clinical utility among the quinolones in respiratory infections.

One hundred isolates of methicillin-resistant Staphylococcus aureus (MRSA) collected from hospitals located in different geographical areas of Japan were used in the present study. The susceptibilities of the strains to gentamicin (GM), erythromycin (EM), tetracycline (TC) and ofloxacin (OFLX) were determined and classified into twelve groups according to their differences in the patterns of the resistance to the four drugs. Of the 100 strains tested, 75 belonging to the main four groups were investigated for the relationships between their patterns of the drug-resistance and biological properties such as coagulase, enterotoxin and phage types, TSST-1 and beta-lactamase producibilities, and percentages of the strains carrying plasmids of large size (> or = 20,000 bp). The main four patterns of drug resistance of the 75 strains were as follows: the first group (33 strains resistant to GM, EM, TC and OFLX), the second group (15 strains resistant to EM, TC and OFLX), the third group (11 strains resistant to GM, EM and OFLX) and the fourth group (16 strains resistant to EM and OFLX) and the remaining 25 strains were divided into further groups. A considerable number of the strains in the third group differed markedly in some biological properties from those in the other groups; coagulase typing (III type-50%: the other groups-II type), enterotoxin typing (64%-non-production: the other groups-C type), TSST-1 producibility (36%-production: the other groups-76 to 88%-production) and phage typing (55%-non-typable: the other groups-80 to 97%-non-typable). In beta-lactamase and plasmids of large size, the first group consisted of the strains with beta-lactamase producibility at the ratio of 50:50, but the other groups consisted of most of the strains with the beta-lactamase producibility. Most of the strains belonging to the second and fourth groups carried large size of plasmids, but 36 and 46% of those belonging to the first and third groups did not carry them. On the other hand, our results of the susceptibility test for the 100 strains showed that a considerable number of strains were susceptible to GM (35%) and TC (42%) at therapeutically significant concentrations.

Pseudomonas aeruginosa strains isolated from patients with different types of the infections each consisted of LPSs different in chain sizes. The drug-susceptibility patterns of these strains of P. aeruginosa were investigated to clarify the relationship between the LPS-compositions and susceptibility to some kinds of anti-pseudomonal drugs. The susceptibilities of nineteen strains (seven long-LPS strains, four short-LPS strains and eight LPS-deficient strains) to piperacillin, ceftazidime, gentamicin, norfloxacin and polymyxin-B were determined and these strains were classified into six types (Types I-VI) according to their drug-resistance patterns. Six of the eight LPS-deficient strains were found to be highly resistant to gentamicin alone (Type IV). Four of the seven strains with the long-LPS and one strain with the short-LPS were resistant to three drugs such as piperacillin, ceftazidime and norfloxacin, and classified into Type I. These results indicated that the major part of the LPS-deficient strains and the considerable part of the long-chain LPS strains of P. aeruginosa tested had each characteristic profile in the drug resistance. The outer membrane proteins of thirteen strains, consisting of different types of LPS compositions, were analyzed by SDS-PAGE. The strains belonging to the same types of the drug-resistance patterns were found to have similar OMP-profiles, although a few exceptions were found. beta-lactamase and gentamicin-inactivating activities were determined for piperacillin-resistant and gentamicin-resistant strains, respectively. Of the piperacillin-resistant strains tested, the activity of beta-lactamase was high in one (No. 8) only, low in four and not found in four. The results showed that degrees of resistance of P. aeruginosa strains tested to piperacillin did not correlate to their producibility of beta-lactamase except one strain. Of the nine gentamicin-resistant strains tested, the gentamicin inactivating activity was high in one (No. 30) only, moderate in six and low in two. These results suggested that the significant levels of piperacillin- or gentamicin-resistance of P. aeruginosa isolated tested might be expressed each due to their decreased abilities for drug-permeabilities in addition to drug-inactivating activities such as beta-lactamase or gentamicin-modifying enzyme. In the case of some resistant strains, the resistance to piperacillin or gentamicin was not explained by the results of the present study. Therefore, we must investigate the possibility that other mechanisms participate in the resistance of these strains.

Lipopolysaccharide (LPS) compositions of P. aeruginosa isolated from from clinical sources such as blood, urine, pus, sputum, and feces were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and aminoglycoside-susceptibility and serotype of these isolates were investigated in this study. Fifty-nine isolates tested were divided into three groups according to the difference in their LPS compositions; 35 strains with the long-LPS chain (B-band LPS), 14 strains with the short chain (A-band LPS) and 10 LPS-deficient strains. The relationship between the LPS compositions and their sources of 59 strains were investigated. The majority of clinical isolates (12 of the 13 strains) from the blood samples possessed the long-LPS chain (B-band LPS) and the remaining possessed the short-LPS chain (A-band LPS). About 67% each of the isolates from urine and feces possessed the long-LPS chain, and the minor part of both groups possessed the short-LPS chain. In isolates from both sputum and pus samples, the long- and short-LPS chains were found at almost the same rate, and the LPS-deficient isolates were found in the sputum samples at a considerably high rate of 42%. The 19 of the 35 isolates with the long-LPS chain were susceptible to gentamicin (54%) and 12 isolates were resistant (34%). On the other hand, the 14 isolates with the short chain were divided roughly into three groups, gentamicin-resistant, and -susceptible groups and intermediate groups. It was also notable that 7 of the 10 LPS-deficient isolates were resistant to gentamicin.(ABSTRACT TRUNCATED AT 250 WORDS)

Bioluminescence resonance energy transfer coupled near-infrared quantum dots using glutathione-s-transferase (GST) tagged luciferase were synthesized as luminescent probes for in vivo imaging.

Multilayered, core/shell nanoprobes (MQQ-probe) based on magnetic nanoparticles (MNPs) and quantum dots (QDs) have been successfully developed for multimodality tumor imaging. This MQQ-probe contains Fe(3)O(4) MNPs, visible-fluorescent QDs (600 nm emission) and near infrared-fluorescent QDs (780 nm emission) in multiple silica layers. The fabrication of the MQQ-probe involves the synthesis of a primer Fe(3)O(4) MNPs/SiO(2) core by a reverse microemulsion method. The MQQ-probe can be used both as a fluorescent probe and a contrast reagent of magnetic resonance imaging. For breast cancer tumor imaging, anti-HER2 (human epidermal growth factor receptor 2) antibody was conjugated to the surface of the MQQ-probe. The specific binding of the antibody conjugated MQQ-probe to the surface of human breast cancer cells (KPL-4) was confirmed by fluorescence microscopy and fluorescence-activated cell sorting analysis in vitro. Due to the high tissue permeability of near-infrared (NIR) light, NIR fluorescence imaging of the tumor mice (KPL-4 cells transplanted) was conducted by using the anti-HER2 antibody conjugated MQQ-probe. In vivo multimodality images of breast tumors were successfully taken by NIR fluorescence and T(2)-weighted magnetic resonance. Antibody conjugated MQQ-probes have great potential to use for multimodality imaging of cancer tumors in vitro and in vivo.

The present study was designed to clarify whether anti-pseudomonal drugs affected in vitro and in vivo changes in serotypes of Pseudomonas aeruginosa isolates. Forty-two isolates of P. aeruginosa belonging to different serotype groups were each incubated in Mueller-Hinton broth including piperacillin, cefsulodin, ceftazidime, imipenem, gentamicin or norfloxacin at 1 approximately 4 MICs at 35 degrees C for 20 hours. The bacterial cells in the media were serotyped after the incubation. In each experiment with these six drugs, serotypes of 4 (9.5%) to 8 (19.0%) of the 42 isolates changed to other different groups. There was no relationship between the serotypes of the variants formed with the anti-pseudomonal drugs and kinds of the drugs. In the case of P. aeruginosa TA-2 isolate, the different concentrations (1/2 MIC and 2 x MIC) of cefsulodin induced the distinct changes in serotypes such as the poly-agglutinable (M, G) and non-typable groups, respectively. The time course for the formation of the variant cells was investigated during the incubation of P. aeruginosa TA-2 isolate in the presence of cefsulodin, respectively. In this case, the variant cells appeared 6 hours after incubation and continued to grow together with the intact cells. On the other hand, the variant cells with anti-pseudomonal drugs (cefsulodin, imipenem and gentamicin) were also formed in the model infections in mice. The present results indicated that the co-existence of colonies with different serotypes in some isolates of P. aeruginosa was partially due to the alternation of serotypes with anti-pseudomonal drugs given to patients with infection.

中山らが紹介したDYBOCのうち，迷路誘導型バランスボードを自作し所要時間と重心軌跡から標準値と学習効果について検討した。対象は健常青年者8名であった。結果;(1)初回測定値とパラメーターとの関連では重心動揺距離，所要時間とも身長，体重と相関を認めず，両測定値間に有意ではないが負の相関を認めた。(2)連続10回の各測定値の変動では重心動揺距離(F=3.52，p

The antimicrobial susceptibility of 93 Acinetobacter baumannii complex isolates from clinical specimens collected nationwide between May and October 2009 were measured by microdilution antimicrobial susceptibility testing based on CLSI M100-S20. Beta-lactamase genes, including classes B and D and ISAbal in meropenem nonsusceptible, including intermediate or resistant isolates, were detected using PCR. Rates of isolates nonsusceptible to meropenem were 18%, to ciprofloxacin 41% and to amikacin 14%. L7-L8: The rate of multidrug-resistant Acinetobacter (MDRA) isolates which were resistant to all 3 antimicrobial agents was 4.3%. MDRA isolates were classified into ST92 by multilocus sequence typing. No metallo-beta-lactamase producer was seen among the 17 meropenem nonsusceptible isolates. The blaoxa-51-like carbapenemase gene and ISAbal were detected in all 17 isolates. ISAba1 upstream presence of the blaOXA-51-like gene was observed in 7 of 17 isolates and the blaOXA-23 like gene in 5 of 17. Consistent with overseas reports, our results confirm the existence of MDRA isolates and isolates harboring OXA carbapenemase genes in Japan. While resistance rates were lower than reports elsewhere, it is clear that resistance trends must be carefully monitored.

In vitro activity of sitafloxacin (STFX) and various oral antimicrobial agents against bacterial isolates recovered from clinical specimens between January and December 2009, at different healthcare facilities in Japan was evaluated. A total of 1,620 isolates including aerobic and anaerobic organisms was available for the susceptibility testing using the microbroth dilution methods recommended by Clinical Laboratory Standard Institute. The minimum inhibitory concentration of STFX at which 90% of isolates (MIC90) was 0.06 microg/mL for methicillin-susceptible Staphylococcus aureus and was equal to that of garenoxacin (GRNX), 2 times lower than that of moxifloxacin (MFLX), and 8 times lower than that of levofloxacin (LVFX). STFX inhibited the growth of all the isolates of Streptococcus pneumoniae at 0.06 microg/mL or less. The MIC90s of STFX ranged from 0.03 to 0.06 microg/mL and were 1 to 2 times lower than those of GRNX, 2 to 4 times lower than those of MFLX, and 16 to 32 times lower than those of LVFX. Against Streptococcus pyogenes, the MIC90 of STFX was 0.06 microg/mL and was 2 times lower than that of GRNX, 4 times lower than that of MFLX, and 32 times lower than that of LVFX. The MIC90 of STFX was 0.25 microg/mL for Enterococcus faecalis, and was 2 times lower than those of GRNX and MFLX, and 8 times lower than that of LVFX. The MIC90 of STFX for E. coli was 2 microg/mL, and the MIC90s of other 10 species of Enterobacteriaceae which were the lowest values of the quinolones tested ranged from 0.03 to 1 microg/mL. The MIC90 of STFX for Pseudomonas aeruginosa isolates recovered from urinary infections was 8 microg/mL and was 16 times lower than those of GRNX, MFLX and LVFX. The MIC90 of STFX for P aeruginosa isolates recovered from respiratory infections was 2 microg/mL and was 32 times lower than those of GRNX and MFLX, and 16 times lower than that of LVFX. STFX inhibited the growth of all the isolates of Haemophilus influenzae at 0.004 microg/mL or less, and was 2 to 4 times lower than those of GRNX, 8 times lower than those of MFLX, and 4 times lower than those of LVFX. The MIC90 of STFX was 0.008 microg/mL for Moraxella catarrhalis, and was 2 times lower than that of GRNX, 8 times lower than those of MFLX and LVFX. The MIC90s of STFX ranged from 0.015 to 0.12 microg/mL for all the species of anaerobic bacteria and were the lowest values of all the antimicrobial agents tested. In conclusion, the activity of STFX against Gram-positive cocci was comparable or superior to those of GRNX, MFLX and LVFX. STFX showed the most potent activity against Gram-negative bacteria and anaerobic bacteria of all the antimicrobial agents tested in this study.

Pseudomonas aeruginosa Nos. 1 and 5, each co-existing growth-inhibitor-producing and -nonproducing cells, were used in this study. An equal number of both cells (each 10(8) CFU/mouse) was challenged intraperitoneally to mice, and these cells in the heart blood and kidneys of mice were determined. Furthermore, the effect of piperacillin, ceftazidime and sisomicin on the cell distribution in mice was studied in the model infection due to P. aeruginosa Nos. 1 and 5. As a control experiment both cells of P. aeruginosa No. 1 were each challenged intraperitoneally at a dose of 10(8) CFU/mouse to mice of two groups, but there were no marked differences between the two types in cell counts of the heart blood or kidneys 9 hours after challenge. When a concomitant challenge of both cells (each 10(8) CFU/mouse) was performed in mice, the number of growth-inhibitor-producing cells of the heart blood and kidneys was about 100 times greater than that of the non-producing cells. These in vivo results were well comparable to the previous in vitro results and indicated that the inhibitor affected the invasion of the non-producing bacteria in the body in the model infection due to P. aeruginosa isolates consisting of the two types of cells. Similar results were obtained in mice with the model infection due to P. aeruginosa No. 5. Anti-pseudomonal drugs such as piperacillin (50 mg/mouse) and ceftazidime (50 mg/mouse) and sisomicin (1 mg/mouse) were given intramuscularly to mice infected concomitantly with both cells of P. aeruginosa No. 1.(ABSTRACT TRUNCATED AT 250 WORDS)

The biological characteristics of individual colonies of Pseudomonas aeruginosa from 138 specimens were investigated. Of these isolates, 90 (65.2%) formed colonies of similar appearance and morphology, and 48 (34.8%) formed colonies which differed either in appearance or morphology. The individual colonies of 138 isolates were tested for serotype. The former 90 isolates formed only the colonies with one kind of serotype, whereas 17 of the latter 48 isolates formed the colonies with more than one kind of serotype. All the 9 isolates tested also differed in other biochemical characteristics: acid productions from xylose, mannitol and maltose, urease production and gelatin liquefaction. beta-Lactamase activity was investigated in 7 isolates forming colonies with more than one serotype. There were no marked differences in beta-lactamase activity among the different colonies in 5 isolates but marked differences among those in the other 2 isolates.

Forty-two isolates of P. aeruginosa from various infections were each incubated in Mueller-Hinton broth including piperacillin, cefsulodin, ceftazidime, imipenem, gentamicin or norfloxacin (1MIC-4MIC) at 35 degrees C for 18 hours, and serotyped using monoclonal antibodies. Serotypes of 4 (9.5%)-8 (19.0%) of the 42 isolates each changed to different groups after incubation. No relationship was found between serotypes of the formed variants and anti-pseudomonal drugs used. When P. aeruginosa TA-2 was exposed to cefsulodin at different concentrations (1MIC and 2MIC) under the above conditions, the distinct variants different in serotypes were formed according to the drug concentrations. Furthermore, P. aeruginosa TA-2 and TA-13 were incubated in Mueller-Hinton broth including cefsulodin (1/2 or 2MIC) and gentamicin (1/2MIC), respectively, and the growth curves of parent and variant cells were determined. In the experiments, the variants appeared 6 hours after onset of the incubation and grew with the parents. The present results may explain our findings previously reported; coexistence of colonies different in serotype of P. aeruginosa isolated from some individual patients. The results indicate the possibility that alteration in bacterial surfaces in association with changes in serotypes might occur in vivo in these patients infected with P. aeruginosa and treated with anti-pseudomonal drugs.

Beta-lactamase-negative ampicillin-resistant (BLNAR) isolates of Haemophilus influenzae have been emerging in some countries, including Japan. The Clinical and Laboratory Standards Institute has only a susceptible MIC breakpoint (or = 1 microg/ml) for piperacillin-tazobactam and a disclaimer comment that BLNAR H. influenzae should be considered resistant, which was adapted without presentation of data. In addition, fluoroquinolone-resistant H. influenzae isolates have recently been occasionally reported worldwide. To address these problems, we examined susceptibilities to beta-lactams, including piperacillin-tazobactam, and ciprofloxacin by microdilution and disk diffusion (only for piperacillin-tazobactam) methods, against a total of 400 recent H. influenzae clinical isolates, including 100 beta-lactamase-negative ampicillin-susceptible, beta-lactamase-positive ampicillin-resistant, BLNAR, and beta-lactamase-positive amoxicillin-clavulanate-resistant (BLPACR) isolates each. BLNAR and BLPACR isolates were tested by PCR using primers that amplify specific regions of the ftsI gene. We also detected mutations in quinolone resistance-determining regions (QRDRs) by direct sequencing of the PCR products of DNA fragments. Among beta-lactams, piperacillin-tazobactam exhibited potent activity against all isolates of H. influenzae, with all MICs ator = 0.5 microg/ml (susceptible). A disk diffusion breakpoint for piperacillin-tazobactam ofor = 21 mm is proposed. We confirmed that all BLNAR and BLPACR isolates had amino acid substitutions in the ftsI gene and that the major pattern was group III-like (87.5%). One ciprofloxacin-resistant isolate (MIC, 16 microg/ml) and 31 ciprofloxacin-susceptible isolates (MICs, 0.06 to 0.5 microg/ml) had amino acid changes in their QRDRs. Piperacillin-tazobactam was the most potent beta-lactam tested against all classes of H. influenzae isolates. It is possible that fluoroquinolone-resistant H. influenzae will emerge since several clinical isolates carried mutations in their QRDRs.

As the most common cause of neonatal sepsis, Lancefield Group B Streptococcus (GBS) must be diagnosed as early as possible in pregnant women is prevent neonatal infection. A selective enrichment broth medium has been widely recommended to optimally recover GBS from genital and anorectal samples. To establish a culture suitable for screening vaginal swab specimens, we compared subcultures of three selective enrichment media to direct culture on agar medium. Vaginal swab samples were inoculated directly onto 5% sheep blood agar and into New Granada medium (Eiken), Lim broth (Becton, Dickinson, and Company), and Todd Hewitt broth with gentamicin and nalidixic acid (Becton, Dickinson, and Company, Todd). Of the 288 specimens tested, GBS was recovered from 43 samples (14.9%) on direct agar media, with 82 (28.5%), positive on New Granada medium subculture, 67 (23.3%) on Lim broth subculture, and 61 (21.2%) on Todd, subculture. These results demonstrates that selective enrichment broth media provides more superior sensitivity than direct agar media for detection of GBS colonization in vaginal specimens, underscoring the usefulness of selective enrichment broth media in GBS screening for vaginal swabs in pregnant woman.

The in vitro antifungal activities of micafungin in comparison to caspofungin, fluconazole, itraconazole, voriconazole, and amphotericin B were evaluated against 93 Candida and 23 Aspergillus isolates recovered from pediatric patients with fungal infections. MICs were determined by the CLSI M27-A2 and M38-A for Candida and Aspergillus species, respectively. Micafungin showed potent activity against Candida albicans, Candida tropicalis, and Candida glabrata with a MIC range of

The antimicrobial activity of various antibiotics against clinical bacterial isolates recovered from patients with infectious diseases at the medical facilities in the Kanto region between March and September 2006 was evaluated. A total of 1030 clinical isolates were available for susceptibility tests: 420 aerobic Gram-positive organisms, 520 aerobic Gram-negative organisms, 30 anaerobic Gram-positive organisms and 60 anaerobic Gram-negative pathogens. Antimicrobial susceptibility data for Streptococcus pneumoniae and Haemophilus influenzae isolates from pediatric and adult patients were analyzed separately. Panipenem (PAPM), imipenem (IPM), meropenem (MEPM), biapenem (BIPM), doripenem (DRPM), cefozopran (CZOP), cefepime (CFPM), and sulbactam/cefoperazone (SBT/CPZ) were used as test antibiotics. PAPM, IPM and DRPM exhibited excellent in vitro antibacterial activities against methicillin-susceptible Staphylococcus, with all isolates exhibiting a MIC ofor =0.06 microg/mL. Against Streptococcus including penicillin-resistant S. pneumoniae, PAPM demonstrated the strongest antibacterial activity among the carbapenems with a MIC range ofor =0.06 to 0.12 microg/mL. Against Enterobacteriaceae, MEPM showed the strongest antibacterial activity, and PAPM had comparable activity to IPM. Against the extended-spectrum beta-lactamase producing Escherichia coli, Klebsiella species and Proteus species, the MICs for the cephems were high, however, those for the carbepenems were low. Against H. influenzae, PAPM had comparable activity to IPM. With respect to anaerobes, each of the carbapenems tested demonstrated almost the same strong antibacterial activity. In conclusion, 13 years has passed since PAPM was launched in 1993, PAPM still maintains potent antibacterial activity and is considered an effective antimicrobial agent for various types of infectious diseases.

We determined the susceptibility of bacteria which were isolated from the patients with respiratory infections between January and October 2005, to tosufloxacin and other fluoroquinolones. A total of 900 isolate including 300 Streptococcus pneumoniae, 100 Streptococcus pyogenes, 100 Moraxella catarrhalis, 200 Haemophilus influenzae, 100 Klebsiella pneumoniae and 100 Pseudomonas aeruginosa were tested. Tosufloxacin, gatifloxacin, levofloxacin, moxifloxacin, ciprofloxacin and prulifloxacin were used as the test antimicrobials. Tosufloxacin, gatifloxacin and moxifloxacin were potent antibiotics tested for the antibacterial activity against Streptococcus including penicillin-resistant S. pneumoniae; the MIC90 were 0.12-0.5/ micromL. Fluoroquinolones exerted the potent antibacterial activity against M. catarrhalis and H. influenzae; the MIC90 of fluoroquinolones tested wereor =0.06 microg/mL. Tosufloxacin, ciprofloxacin and prulifloxacin showed to be more active against K. pneumoniae and P. aeruginosa, but parts of some strains were resistant. These results indicate that tosufloxacin has the potent antibacterial activity against major organisms detected from patients with respiratory infections. Since it was approved in 1990, tosufloxacin was considered to be useful as a therapeutic antimicrobial for the treatment of respiratory infections.

Nadifloxacin (NDFX) is a fluoroquinolone antibiotic developed by Otsuka Pharmaceutical Co., Ltd. and is used as a topical drug for the treatment of infections in the field of dermatology. We investigated the susceptibility of a total of 575 strains (two kinds of Staphylococcus species and Propionibacterium species which were isolated from patients with dermatological infections for 3 periods, i.e., 1996, 2000 and 2005) to NDFX and other reference antibiotics. The minimum inhibitory concentration of the four antibiotics, NDFX, levofloxacin (LVFX), clindamycin (CLDM) and gentamicin (GM), against the test strains were determined by the agar dilution methods, in according with the Japan Society of Chemotherapy. The antibacterial activity of NDFX against methicillin-susceptible Staphylococcus aureus (MSSA), methicillin-resistant S. aureus (MRSA), Staphylococcus epidermidis and P. acnes was the most potent of all the antibiotics tested, and there were no test organisms which became resistant to NDFX with period. The MIC90 values of NDFX for the four test organisms isolated in 2005 were 0.05 microg/mL; MSSA, 1.56 microg/mL; MRSA, 0.78 microg/mL; S. epidermidis and 0.20 microg/mL; P. acnes, respectively. On the other hand, there were LVFX-, CLDM- and GM-resistant MRSA. The MIC50 values of CLDM and GM for MRSA were100 and 25 microg/mL, respectively. The MIC50 value of GM for P acnes was 12.5 microg/mL, but NDFX was potently active against these organisms as compared with these two antibiotics and the MIC50 values of NDFX were 0.05 microg/mL for MRSA and 0.20 microg/mL for P. acnes. These results suggest that NDFX is even at present useful as an antibiotic for the treatment of infections in the field of dermatology though it is more than 12 years since the approval to manufacture and sell the drug was obtained in 1993.

The minimum inhibitory concentrations (MICs) of tosufloxacin and other fluoroquinolone antimicrobials for Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella (Branhamella) catarrhalis, isolated, between January 2003 and July 2004, from patients suspected of having respiratory or otorhinological infections were determined. The results were compared with those for these organisms isolated in 1994, plus some H. influenzae strains isolated in 1998. Tosufloxacin was the most potent of all the antibiotics tested for antibacterial activity against S. pneumoniae (including penicillin-intermediate S. pneumoniae and penicillin-resistant S. pneumoniae). The MIC50 and MIC90 values did not differ from those obtained for the strains isolated in 1994. Fluoroquinolones exerted the most potent antibacterial activity against M. (B.) catarrhalis; the MICs for most of the strains were < or = 0.06 microg/ml; fluoroquinolones inhibited the growth of all the strains at 0.25 microg/ml or less. Fluoroquinolones showed the most potent antibacterial activity against H. influenzae strains isolated between 2003 and 2004, and in 1994, but, for one H. influenzae strain isolated, between 2003 and 2004, the MICs of fluoroquinolones were high. Some strains of S. pneumoniae and H. influenzae were resistant to fluoroquinolones. Genetic analysis showed that all of these strains had mutations in the quinolone resistance-determining region, but there were no differences according to the years of isolation. These results indicate that tosufloxacin has potent antibacterial activity against major organisms isolated from patients with respiratory or otorhinological infections; further, the results of the present study did not differ from those obtained about 10 years ago.

The susceptibility of clinical strains isolated from patients with infection to ceftriaxone (CTRX) and injectable beta-lactam antibiotics was determined in Japanese medical institutions in 2004. The in vitro antibacterial activity of the above antibiotics against clinical strains isolated in 2004 was compared with that against part of the clinical strains isolated about ten years ago (1994 - 1996). The antibacterial activity of CTRX against Streptococcus including penicillin-intermediate and penicillin-resistant Streptocuccus pneumoniae was potent and there were no large differences between clinical strains isolated in 1994 - 1996 and those in 2004 in the antibacterial activity of CTRX. CTRX inhibited the growth of all the strains of Haemophilus influenzae including beta-lactamase-negative, ampicillin-resistant (BLNAR) strains at 0.5 microg/mL or less and showed the potent antibacterial activity against these strains. In addition, since there are no large differences between strains isolated in the past and those isolated in 2004 in the antibacterial activity, CTRX is considered to be a useful antibiotic in the treatment of BLNAR infections which are increased by the causative organism of infections especially in the field of pediatrics in recent years. CTRX showed the excellent antibacterial activity against Escherichia coli and Klebsiella pneumoniae but one of the 50 strains of E. coli tested was highly resistant. The antibacterial activity of CTRX against Neisseria gonorrhoeae was the most potent in all the antibiotics tested. These results indicate that CTRX shows excellent antimicrobial activity against fresh strains isolated from patients with infections. CTRX is thus useful as a therapeutic antimicrobial for the treating a variety of infections.

The antimicrobial susceptibility of 240 isolates from the ophthalmological infections during July 2003 to March 2004 was determined to gatifloxacin (GFLX), levofloxacin, lomefloxacin and cefmenoxime applicable for ophthalmological infections. The in vitro activities of these drugs against the fresh isolates were compared. The quinolones including GFLX were potently active against Gram-positive bacteria, except for MRSA, a major causative pathogens for ophthalmological infection. When MIC ranges, MIC50 and MIC90 of three quinolones were compared, it was considered that the activity of GFLX was the most active of them. GFLX showed to be more active against opportunistic pathogens including Pseudomonas aeruginosa than other antimicrobial agents, and GFLX was especially potent against Streptococcus pneumoniae and Enterococcus faecalis. In conclusion, GFLX exhibits a potently active against fresh isolates from ophthalmological infections, and has an effective potential in the treatment of ophthalmological infections with the drug to administer eye drops.

The antimicrobial susceptibility of Streptococcus pneumoniae and Haemophilus influenzae isolates during January 2003 to July 2004 was determined to seven various antimicrobial drugs including cefteram (CFTM). The in vitro activities of these drugs against the fresh isolates were compared. The oral cephalosporins including CFTM were potently active against penicillin susceptible S. pneumoniae. The activity of CFTM and cefditoren was the most active among four oral cephalosporins. The susceptibilities of penicillin intermediate S. pneumoniae and penicillin resistant S. pneumoniae to antimicrobial agents were decreased. The MIC of CFTM was not beyond 4 microg/mL for any isolate of S. pneumoniae. The activity of CFTM was very high to beta-lactamase-negative and ampicillin-susceptible H. influenzae isolates. These MIC against all isolates were 0.03 microg/mL or less. The MIC of CFTM was not beyond 1 microg/mL for any isolate of beta-lactamase-positive H. influenzae or beta-lactamase-negative-ampicillin resistant H. influenzae. In conclusion, CFTM exhibits a potent activity against fresh isolates of S. pneumoniae and H. influenzae, and has a potential of effectiveness in the infections.

The effect of entry delayed blood culture bottles until the start of incubation for mechanical detection of organism were compared using 2 major blood culture systems; BACTEC 9240 system and BacT/ALERT 3D system. Total of 13 bacterial strains; 5 gram-positive cocci, 7 gram-negative bacilli and Candida parapsilosis which were isolated mainly from blood cultures were used as the test strains. BACTEC 92F, 93F and BacT/ALERT FA, FN bottles were used as the blood culture bottles. All the bottles inoculated with the test strains were incubated and evaluated immediately after standing at room temperature for 24, 42, 48, 54 or 72 hours, using the respective automated blood culture systems. All the bottles were subcultured. The effect of entry delay the blood culture bottles for the mechanical detection was observed in many gram-negative organisms in BACTEC 9240 system. The blood cultures were evaluated not to be positive in 4 of the 10 samples on delaying for 24 hours or in any of the samples on delaying for 42 hours in the BACTEC 92F bottles inoculated with Escherichia coli. In Serratia marcescens, the blood cultures were evaluated not to be positive in 5 of the 10 samples on delaying for 24 hours or in any of the samples on delaying for 42 hours in the BACTEC 92F bottles. In Klebsiella pneumoniae, the blood cultures were evaluated not to be positive in 9 of the 10 samples on delaying for 42 hours. In Enterococcus faecalis, Pseudomonas aeruginosa and Proteus mirabilis, the blood cultures were evaluated not to be positive in 5-6 of the 10 samples on delaying for 42 hours. On the other hand, the blood cultures were evaluated to be positive in most of the samples of Acinetobacter calcoaceticus (except 3 of the 10 samples which were evaluated not to be positive) on delaying for 42 hours in BacT/ALERT 3 D system. The samples except part of Streptococcus spp. were detected by subculture in both the bottles. These results indicate that the delayed time of blood culture bottles before inoculation with the test bacterial samples affects the positive detection of blood cultures markedly in the blood culture system. Therefore, the immediate incubation was considered to be necessary.

We evaluated the automated VITEK 2 system (bioMérieux) for antimicrobial susceptibility testing of vancomycin-resistant Enterococcus (VRE). The results obtained with the VITEK 2 system were compared to those obtained using two NCCLS reference methods. The VITEK 2 system produced MICs for penicillin G, erythromycin and vancomycin that were very similar to those of the reference agar-dilution test with all results being within a twofold dilution. When MICs of teicoplanin for these isolates were measured by the agar-dilution method and VITEK 2 system, there was one ‘very major’ error and seven ‘minor’ errors. There were no ‘major’ errors for any of the antibiotics tested. When the results obtained by the micro broth-dilution method were compared with those obtained by the VITEK 2 system, there was one ‘very major’ error for teicoplanin by the VITEK 2 system, as was the case with the agar-dilution method. There were two ‘minor’ errors for erythromycin and seven ‘minor’ errors for teicoplanin. There were no ‘major’ errors for any of the antibiotics tested. The 35 VRE strains identified phenotypically by the VITEK 2 Advanced Expert System included nine of Enterococcus faecalis and 23 of Enterococcus faecium. Neither Enterococcus avium nor Enterococcus hirae were identified. A total of 32 phenotypes were classified into 22 VanA and 10 VanB strains. PCR genotyping demonstrated 23 vanA + and nine vanB + strains. There were differences between the VITEK 2 system results and those of PCR. Overall, 54.3 % of the test results were obtained within 7 h. All MIC values for the 35 VRE isolates were determined within 13 h of completing incubation. The VITEK 2 system is a simple method for accurately detecting vancomycin-resistant strains of Enterococcus and can be used to rapidly determine MICs.

A total of 2865 strains of the causative organisms isolated from the patients with acute pharyngitis and tonsillitis at the primary medical institutions were used in this study. The MICs of levofloxacin (LVFX) and other oral antimicrobial drugs were determined and evaluated by the NCCLS guideline. LVFX, cefditoren (CDTR) and cefcapene (CFPN) were potently active against 773 isolates of Hemophilus influenzae, the MIC50S of LVFX beingor = 0.06 microgram/mL and also the same as the MIC90S of LVFX. LVFX was the most active against 496 isolates of Enterobacteriaceae. The MIC50S of LVFX wereor = 0.06 microgram/mL and were lower than those of CDTR, cefdinir (CFDN) and cefpodoxime (CPDX) (MIC50S: 0.5 microgram/mL). The MIC90S of these cephems were markedly higher than the respective MIC50S, whereas MIC50 of LVFX was 0.12 microgram/mL, only twice the MIC50. Against the majority of Streptococcus pyogenes (555 isolates) and Streptococcus spp. (495 isolates), CDTR, CFDN, CPDX and CFPN were highly active (MICs:or = 0.06 microgram/mL), and clarithromycin (CAM) and azithromycin (AZM) were also active against these organisms (MICs: 0.12 to 0.25 microgram/mL). Against S. pneumoniae (92 isolates), CDTR and CFDN were active (MIC50S: 0.12 and 0.25 microgram/mL, respectively). However, the MIC90S of these drugs were 4-8 times the MIC50S. Against Moraxella (Branhamella) catarrhalis (454 isolates), LVFX was potently active, the MIC90 of LVFX beingor = 0.06 microgram/mL and MIC90S of the other cephems being 0.5 microgram/mL or more. When the susceptibility of these strains to LVFX was evaluated by the NCCLS guideline, about 3% of other Streptococcus spp. were resistant to the drug but no test strains resistant to LVFX were detected in H. influenzae, S. pyogenes or Enterobacteriaceae. On the other hand, the percentages of strains susceptible to the cephems tested were 60-90%, which were quite different according to kinds of drugs and species used. Furthermore, the strains of S. pneumoniae resistant to CFDN and CPDX, and those to CAM and AZM were 21-25% and 50% or more, respectively, whereas no LVFX-resistant strains were detected. The major pathogens isolated from patients with pharyngitis and tonsillitis in the primary institutions were highly susceptible to LVFX. These results suggest that LVFX is a useful drug which is potently active against the strains resistant to oral cephem and macrolide antibiotics.

Capilia TB, a lateral flow immunochromatographic slide test kit for directly identifying Mycobacterium tuberculosis complex (MTC), was evaluated by using culture-positive specimens from Mycobacteria Growth Indicator Tubes (MGIT). Sputum specimens from patients suspected of having tuberculosis were treated with NALC-NaOH and cultivated in MGIT960. Liquid specimens were collected from the positive tubes and directly inoculated with Capilia TB. Liquid specimens were also directly tested with AccuProbe. Of the organisms isolated from the 100 MGIT positive tubes, M. tuberculosis complex was identified in 49 (49%) tubes with Capilia TB and not identified in 51 (51%) with Capilia TB. Mycobacterium avium-intracellulare complex (MAC) was identified in 46 (46%) with AccuProbe MAC and other acid-fast bacteria were identified in 5 (5%) by DNA-DNA hybridization method. There were one tube in which M. tuberculosis complex was detected with Capilia TB and M. tuberculosis complex was not detected with AccuProbe MTC, but no tubes in which M. tuberculosis complex was detected with AccuProbe MTC and M. tuberculosis complex was not detected with Capilia TB. Capilia TB is excellent in sensitivity and specificity and very suitable for rapid diagnosis of tuberculosis and is considered to contribute to public health intervention measures taken for the tuberculosis control in Japan.

Neisseria gonorrhoeae were isolated from pharyngeal specimens of male and female patients and also from urethral and cervical discharges of male and female patients, respectively, suspected of having gonococcal infections in a urologic clinic in Kawasaki City. Microbiological and epidemiological studies were performed in 127 male and 41 female patients. The specimens were streaked onto the modified Thayer-Martin Selective Agar and the plates were incubated at 35 degrees C for 48 h under an atmosphere of 10% CO2. In 127 male patients, N. gonorrhoeae were detected in 117 (92.1%) of the urethral specimens. In these patients, N. gonorrhoeae were detected in pharyngeal specimens from 14 (11.0%) patients, but the pathogen was also detected in urethral specimens from these patients without exception. In 41 female patients. N. gonorrhoeae were detected in 20 (48.8%) of the 41 cervical discharges. When the pharyngeal specimens were tested, N. gonorrhoeae were detected in 14 (34.1%) of the 41 specimens. N. gonorrhoeae was simultaneously detected only in pharyngeal and cervical specimens from 11 of the 41 female patients and the pathogen was detected only in pharyngeal specimens from other 3 patients. There were no marked differences in antimicrobial susceptibilities between N. gonorrhoeae isolates from pharyngeal specimens and those from urethral or cervical discharges in all the patients tested. The PFGE patterns of 50 gonococcal isolates (25 pairs) from 25 patients (14 males and 11 females) in whom N. gonorrhoeae were simultaneously detected from pharyngeal and urethral or cervical specimens were analyzed. In 24 of 25 patients. N. gonorrhoeae isolated from the pharyngeal and urethral or cervical specimens in the same patients showed the same PFGE patterns. However, the 25 pairs showed the different PFGE patterns. From these results it is clarified that N. gonorrhoeae are detected in the pharyngeal specimens from considerable numbers of patients with gonorrhea, and there is a possibility that the pathogens prevailing among the patients differ in genetic sources.

Antibacterial activities of gatifloxacin (GFLX) and other antibacterial drugs against various fresh clinical strains (800 isolates) isolated from specimens of patients in 2002 were compared. GFLX was more active than levofloxacin and ciprofloxacin against Gram-positive bacteria such as methicillin susceptible Staphylococcus aureus and Streptococcus pneumoniae. For these isolates, clarithromycin and azithromycin were less active (MIC90;16-64 micrograms/mL), GFLX was more active than cefdinir. For Escherichia coli, Klebsiella pneumoniae, Acinetobacter species, Haemophilus influenzae and Moraxella (Branhamella) catarrhalis, three quinolones including GFLX were potently active (MIC90;or = 0.06-0.5 microgram/mL). Pseudomonas aeruginosa isolated from urinary tract infections were resistant to three quinolones including GFLX (MIC90; 32-64 micrograms/mL), however P. aeruginosa isolated from respiratory and otolaryngological infections were more susceptible (MIC90; 0.5-2 micrograms/mL). Quinolones were less active against Neisseria gonorrhoeae as compared with the cephem antibiotics tested, but GFLX was the most active against N. gonorrhoeae among the quinolones tested. In this study, we investigated activity of GFLX against fresh clinical strains isolated early in 2002, GFLX is widely and potently active against S. aureus, S. pneumoniae and various Gram-negative bacteria.

The antibacterial activity of BAS-118, a new benzamide derivative, against Helicobacter pylori and other species of bacteria was investigated, as was the in vitro ability of the compound to induce drug resistance in H. pylori. The MICs of BAS-118 for 155 isolates of H. pylori, including 30 clarithromycin (CAM)-resistant isolates (MICor= 1.56 mg/L) and 25 metronidazole (MNDZ)-resistant isolates (MICor = 6.25 mg/L), and 29 reference strains of other genera were determined by an agar dilution method. The MIC(50), MIC(90) and MIC range of BAS-118 for 100 randomly selected isolates of H. pylori wereor =0.003, 0.013 andor =0.003-0.025 mg/L, respectively, with similar values obtained for CAM- and MNDZ-resistant isolates. Furthermore, MICs of BAS-118 for five H. pylori strains increased no more than two-fold after 10 serial passages in the presence of subinhibitory concentrations. BAS-118 exhibited a low antibacterial activity against the 29 non-H. pylori strains, with MICs ofor =8 mg/L. In summary, BAS-118 is a novel anti-H. pylori agent with a potent and selective antibacterial activity, which includes CAM- and MNDZ-resistant isolates. Furthermore, BAS-118 does not appear to induce drug resistance readily in vitro.

Helicobacter pylori (H.p.) bacteria are the major causes of gastro-duodenal disease, and some association with stomach cancer has been suggested. Recently, H.p. eradicating treatment has been practiced using antibiotics and proton-pump inhibitors. However, at the same time, some reports have been made on the side-effects of this treatment; allergic reactions and uneffective resistant bacteria. Under these circumstances, there is a strong need for medicines, which are less harmful to the body, can be administered repeatedly, are less expensive, and yet as effective as antibiotics in inhibiting the bacteria, and in fact, some studies have been undertaken in this regard. We placed our focus on Bainiku-ekisu (Concentrate of Japanese apricot juice) which, as a Japanese folk remedy, has been used for the treatment of gastritis and enteritis since ancient times, and studied its bacteriosterile affects. The major ingredients of Bainiku-ekisu are citric acid (32%) and malic acid (11%), while its pH represents strong acid. We measured the bacteriosterile effects of Bainiku-ekisu by culturing ten H.p. strains originating from gastro mucous membrane respectively in a Bainiku-ekisu concentration of 0.156%, 0.313%, 0.625% and 0.9%, and measured the level of MIC (minimum inhibiting concentration of development). As a result, out of ten H.p. strains four of them presented strong bacteriosterile effects in a concentration of less than 0.156%, and six of them, in a concentration of less than 0.313%. Furthermore, in order to measure the bacteriosterile effects against H.p. in the stomach we measured the quantity of bacteria in 0 minutes, five minutes, and ten minutes after mixing ten H.p. strains suspended in physiological salt solution with a Bainiku-ekisu solution of 0.3% and 0.9% dissolved in aseptic physiological salt solution. As a result, within five minutes after mixing, every one of the ten H.p. strains was observed to present strong bacteriosterile effects. These results suggest that Bainiku-ekisu can be considered as a fool likely to prevent the development of H.p. as well as the future possibility of making clinical applications to H.p. infection.