miXAFS: a program for X-ray absorption fine-structure data analysis.

A new program called miXAFS for the analysis of X-ray absorption fine-structure (XAFS) data is presented. miXAFS can analyze the XAFS functions simultaneously for all measured X-ray absorption edges of the constituent elements in a sample under the constraints for the structural parameters over the edges. The program provides a surface plot of the R-factor as a function of two structural parameters, which is useful to validate the optimized structural parameters. The structural parameters can be obtained from the XAFS data in a few steps using the setting file and batch process. The program, which is coded in MATLAB and freely available, runs on Macintosh and Windows operating systems. It has a graphical user interface and loads experimental data and XAFS functions in a variety of ASCII data formats.

Calphostin C-mediated translocation and integration of Bax into mitochondria induces cytochrome c release before mitochondrial dysfunction.

Calphostin C-mediated apoptosis in glioma cells was reported previously to be associated with down-regulation of Bcl-2 and Bcl-xL. In this study, we report that 100 nM calphostin C also induces translocation and integration of monomeric Bax into mitochondrial membrane, followed by cytochrome c release into cytosol and subsequent decrease of mitochondrial inner membrane potential (DeltaPsim) before activation of caspase-3. The integration of monomeric Bax was associated with acquirement of alkali-resistance. The translocated monomeric Bax was partly homodimerized after cytochrome c release and decrease of DeltaPsim. The translocation and homodimerization of Bax, cytochrome c release, and decrease of DeltaPsim were not blocked by 100 microM z-VAD.fmk, a pan-caspase inhibitor, but the homodimerization of Bax and decrease of DeltaPsim were inhibited by 10 microM oligomycin, a mitochondrial F0F1-ATPase inhibitor. Therefore, it would be assumed that mitochondrial release of cytochrome c results from translocation and integration of Bax and is independent of permeability transition of mitochondria and caspase activation, representing a critical step in calphostin C-induced cell death.

Proteasome inhibitors induce Fas-mediated apoptosis by c-Myc accumulation and subsequent induction of FasL message in human glioma cells.

Proteasome inhibitors were shown previously to induce mitochondria-independent and caspase-3-dependent apoptosis in human glioma cell lines by unknown mechanisms. Here, we showed that treatment with proteasome inhibitors, lactacystin or acetyl-leucinyl-leucinyl-norleucinal, led to elevation of the steady-state c-Myc protein but not c-myc mRNA, suggesting the accumulation of c-Myc protein by proteasome inhibitors. In addition, the marked association of c-Myc protein with ubiquitin by treatment with proteasome inhibitors indicated the involvement of proteasome in c-Myc proteolysis and the stabilization of c-Myc protein by proteasome inhibitors in vivo. The expression of Fas (also termed CD95 or APO-1) mRNA, if analyzed by reverse transcriptase polymerase chain reaction assay, was found to occur constitutively, and increased slightly by the treatment with proteasome inhibitors. In contrast, the expression of Fas ligand (FasL) mRNA was markedly induced temporarily before the activation of caspase-3 by the treatment. Agonistic anti-Fas antibody (CH11) induced apoptotic cell death, suggesting the presence of a functional Fas receptor. In addition, proteasome inhibitor-induced apoptosis was prevented by the addition of antagonistic anti-FasL antibody (4A5) or z-IETD.fmk, a potent inhibitor of caspase-8, indicating the involvement of the Fas receptor-ligand apoptotic signaling system in proteasome inhibitor-mediated apoptosis. Thus, it is suggested that proteasome inhibitors cause the accumulation of c-Myc protein which induces transiently FasL message to stimulate the Fas receptor-ligand apoptotic signaling pathway.

Proteasome inhibitors induce mitochondria-independent apoptosis in human glioma cells.

The proteasome inhibitors lactacystin and AcLLNal induced p53-independent apoptosis in two human glioma cell lines, and the apoptosis was accompanied by up-regulation of immunoreactive wild-type p53, p21Waf1, Mdm2, and p27Kip1. Pretreatment with cycloheximide decreased the induction of cell death independently of p53 protein status, suggesting that the up-regulation of short-lived proteins is associated with proteasome inhibitor-induced apoptosis. Caspase-3-like proteases were activated in the proteasome inhibitor-mediated apoptosis, and the induction of cell death was inhibited more effectively in the presence of z-VAD.fmk than in the presence of Ac-DEVD.fmk, suggesting that caspases other than caspase-3 are involved. Nonetheless, there were no significant alterations in levels of immunoreactive Bcl-2, Bcl-X(L), Bax, Bad, and Bak, nor any evidence of cytochrome c release into cytosol and dissipation of delta(psi)m. Thus, the proteasome inhibitor-induced apoptosis is mediated by a mitochondria-independent mechanism, and the once activated caspase-3 does not cause the cytochrome c release and the delta(psi)m disruption.

A clinical study of fluconazole for the treatment of deep mycoses.

A multicenter clinical study of fluconazole was conducted at 41 hospital sites in Japan. Fluconazole was administered orally or intravenously at daily doses of 50 to 400 mg to 199 patients with deep-seated mycoses. Clinical efficacy was evaluable in 125 of these patients. Most cases were complicated with serious underlying diseases such as cancer, leukemia, or AIDS. Clinical cures were achieved in 56 (87.5%) of 64 cases of candidiasis, in 11 (68.8%) of 16 cases of cryptococcosis, in 19 (44.2%) of 43 cases of aspergillosis, and in one case each (100%) of mucormycosis and fungemia due to an unspecified yeast. Eradication rates of causative fungi were 87.9% in Candida spp., 62.5% in Cryptococcus neoformans, and 52.2% in Aspergillus spp. Side effects were observed in 13 cases, with an incidence rate of 6.5%. In most cases fluconazole was well tolerated. Changes in laboratory test values due to the drug were reported in 35 patients with an incidence rate of 17.6%. The changes were minor and transient; primarily increases in liver enzyme. Fluconazole is a useful antifungal agent for the treatment of systemic deep-seated mycoses.

Inhibition by 5'-methylthioadenosine of cell growth and tyrosine kinase activity stimulated by fibroblast growth factor receptor in human gliomas.

Stimulation of three human glioma cell lines with basic fibroblast growth factor (bFGF) led to the enhancement of cell growth and the rapid tyrosine phosphorylation of cellular proteins, including major substrates of 90 kD. A methyltransferase inhibitor, 5'-methylthioadenosine (MTA), inhibited dose dependently the bFGF-stimulated cell growth and protein tyrosine phosphorylation in glioma cells by blocking both receptor autophosphorylation and substrate phosphorylation, as shown by immunoblotting with antiphosphotyrosine antibodies and cross-linking bFGF to receptors. The antiproliferative activity of MTA correlated quantitatively with its potency as an inhibitor of bFGF-stimulated protein tyrosine kinase activity. The methyltransferase inhibitor MTA had no effect on either epidermal growth factor- or platelet-derived growth factor-stimulated protein tyrosine phosphorylation in glioma cells, but inhibited specifically bFGF-stimulated protein tyrosine kinase activity. The concentration of MTA required for inhibition of protein methylation correlated well with the concentration required for inhibition of bFGF-stimulated cell growth and protein tyrosine phosphorylation. Because MTA had no effect on numbers and dissociation constants of high- and low-affinity bFGF receptors, the inhibition of bFGF-stimulated bFGF receptor tyrosine kinase activity is not likely to be the result of a reduction in bFGF receptor and bFGF binding capacity. In fact, MTA delayed and reduced the internalization and nuclear translocation of bFGF, and the internalized bFGF was submitted to a limited proteolysis that converted it to lower molecular peptides whose presence remained for at least 22 hours. The effect of MTA on bFGF-stimulated tyrosine phosphorylation was immediate and readily reversible.

Apoptosis of human glioma cells in response to calphostin C, a specific protein kinase C inhibitor.

Calphostin C acts at the regulatory domain as a highly selective inhibitor of protein kinase C (PKC), and staurosporine acts at the catalytic domain as a nonspecific PKC inhibitor. The authors investigated the capacity of calphostin C and staurosporine to promote apoptotic fragmentation of DNA in four human glioma cell lines. The exposure of glioma cell lines to 100 nM calphostin C for 2 to 8 hours induced a decrease in particulate PKC activities and exposure for 16 to 24 hours produced a concentration-dependent increase in internucleosomal DNA cleavage on agarose gel electrophoresis. In addition, the human glioma cells showed the classic morphological features of apoptosis: cell shrinkage, nuclear condensation, and the formation of apoptotic bodies. A 24-hour exposure to staurosporine failed to induce internucleosomal DNA fragmentation at concentrations generally used to achieve maximum inhibition of enzyme activity (50 nM) but promoted fragmentation at considerably higher concentration (more than 200 nM). Deoxyribonucleic acid fragments obtained from cells exposed to 100 nM calphostin C for 16 to 24 hours possessed predominantly 5'-phosphate termini, consistent with the action of a Ca++/Mg(++)-dependent endonuclease. Northern and Western blot analyses revealed that the exposure to 100 nM calphostin C for 4 hours failed to alter bcl-2 transcript and protein, but exposure for more than 8 hours decreased the amount of bcl-2 transcript and protein. Together, these observations suggest that calphostin C is capable of inducing apoptotic DNA fragmentation and cell death in a highly concentration dependent manner in human glioma cells and that the apoptosis is closely associated with the decrease in transcription and translation of bcl-2.

Synthesis of (+)- and (-)-nojirimycin and their 1-deoxy derivatives from myo-inositol.

The conversion of the naturally abundant cyclitol, myo-inositol (4), into (+)-nojirimycin (1a), its enantiomer (1b), and their 1-deoxy analogues (2a and 2b) is described. Biological assay of 2a, 2b, and the bisulfite adducts of 1a and 1b (3a and 3b) showed that the compounds having the unnatural L-gluco configuration (2b and 3b) possess moderate-to-high inhibitory activity against almond beta-D-glucosidase and bovine liver beta-D-galactosidase.

Lung abscess: analysis of 66 cases from 1979 to 1991.

Sixty-six patients with bacterial lung abscess were treated between 1979 and 1991 in our hospital. Among these patients, death occurred in one of the 42 cases of community-acquired infection (mortality rate: 2.4%) and in 16 of the 24 cases of nosocomial infection (mortality rate: 66.7%). Of all 66 cases, 55 were culture-positive, and the etiologic agents isolated from 24 of the culture-positive cases were found to be anaerobic bacteria. The most common aerobes isolated from the foci were identified as Staphylococcus aureus, Klebsiella spp. and Pseudomonas aeruginosa, while the most common anaerobes were Bacteroides spp., Peptostreptococcus, Fusobacterium spp., microaerophilic Streptococcus and Veillonella. The mortality was higher in the cases with P. aeruginosa, Klebsiella spp. and Candida spp. than in those with other bacteria. The prognosis of lung abscess patients proved to depend on the presence of underlying diseases and on superinfection with aerobes.

Periodic fever compatible with familial Mediterranean fever.

A 55-year-old male presented with a recurrent fever of over 38 degrees C, occurring at irregular intervals 1-6 times a month with chest, back or abdominal pain. After admission to our hospital, we found the following characteristics: 1) the febrile attacks were accompanied by obvious inflammatory findings and pleuritis or peritonitis; 2) the patient's elder sister had a similar periodic fever; and 3) there were no apparent causative factors responsible for his symptoms. Therefore, we diagnosed this as a case compatible with familial Mediterranean fever. The febrile attacks have been completely suppressed by daily colchicine. This is the seventh case of familial Mediterranean fever reported in Japan.

Effect of enrichment of infusion solutions with branched chain amino acids in parenteral nutrition of rats.

The effect of enrichment of the branched chain amino acids (BCAAs) leucine, isoleucine and valine on total parenteral nutrition was studied in rats. Experimental infusion solutions with a sufficient, marginal or deficient level of glucose contained either the conventional amino acid composition (22.6% BCAAs) or a BCAA-enriched amino acid composition (36% BCAAs). Rats were infused with experimental solutions for 4 days and several parameters of protein metabolism were evaluated in various tissues. Under conditions of sufficient energy supply, BCAA-enriched and conventional groups showed similar body weight gains and muscle protein degradations as measured by urinary 3-methylhistidine excretion. Polysome profiles in the liver and gastrocnemius muscle of the BCAA-enriched group were more heavily aggregated than those of the conventional group. Under the conditions of marginal or deficient energy supply, beneficial effects of BCAA enrichment over the conventional amino acid composition became more evident in terms of better body weight retention, higher RNA/DNA ratio and heavier polysome profile in both liver and muscle, and reduced protein catabolism in muscle. The present study suggests that enrichment of BCAAs, particularly valine and isoleucine, may be useful for nutritional support under hypercatabolic or stressed conditions.

[Asporogenic anaerobic empyema--clinical and bacteriological investigations of 31 patients with anaerobic empyema].

The author reviewed the records of 31 patients with asporogenic anaerobic empyema mostly seen in the wards of Internal Medicine, Juntendo University Hospital during the 27 years between 1961 and 1988, and obtained the following results. 1. There were 25 males and 8 females with an average age of 57.8 and 51.0 y/o (range, 25 to 79 y/o), respectively males more than forty years old occupied 74.2 percent of all cases. 28 patients (90%) had underlying conditions. 2. The cases of mixed infections with anaerobes and aerobes were only 22.6%. 3. The isolated bacteria were microaerophilic streptococcus, Bacteroides spp., Peptostreptococcus spp., Fusobacterium spp. etc. in this order. 4. There were no relationships between anaerobic infections with or without aerobes and putrid odor of pleural effusion. 5. Bacteroides spp. were isolated most in the group with putrid pleural effusion, however, they were not isolated in the group without putrid pleural effusion at all. This fact suggests that there is an intimate relationship between putrid odor and Bacteroides spp. 6. There was no deceased case which pleural effusion had been drained sufficiently with open or closed drainage. It suggests that sufficient drainage is the most important in therapeutic procedures of asporogenic anaerobic empyema.

[Clinical studies of intravenous drip infusion of micronomicin. Internal field infections].

Micronomicin sulfate (MCR) by intravenous drip infusion for internal field infections was studied in 30 cooperative research institutions and the following results were obtained. Clinical efficacy of MCR on respiratory tract infections was 79.4% for pneumonia, 51.5% for chronic bronchitis and 81.0% for respiratory tract infections associated with bronchiectasis, respectively, with a total effectiveness rate of 70.9%. MCR was effective on urinary tract infections with a rate of 85% and on septicemia with a rate of 80%. There were no cases of clinical symptoms seemed to be adverse reactions. Abnormal laboratory test values were noted in 13 out of 167 cases (7.8%), but all of them were transient without severe reactions observed. It is fully expected that MCR intravenous drip infusion for internal field infection is clinically effective.

[Clinical trials of netilmicin against respiratory tract infection].

Netilmicin, a new semisynthetic aminoglycoside antibiotic recently developed by Shering Co., was injected to 5 patients with respiratory tract infection (2 cases of diffuse panbronchiolitis, 2 cases of bronchiectasis and 1 case of bronchial asthma with infection). A daily dose of netilmicin was 200 mg by intramuscular injection and duration of netilmicin therapy was for 7 to 14 days. Clinical response to netilmicin therapy of respiratory tract infection was good in 3 cases, fair in 1 case and poor in 1 case. No subjective and objective findings considered as the side effect was observed and laboratory examinations showed no abnormality. It may be concluded from the above clinical results that netilmicin may be effective for respiratory tract infections and further study may prove its efficacy.

[Susceptibility of bacteria isolated from patients with lower respiratory tract infections to antibiotics (1987)].

Since 1981, in cooperation with research institutions across the nation, Ikemoto, et al. have been collecting clinical isolates from patients with respiratory tract infections and conducting an annual retrospective survey of patients' background factors and of isolated strains and their sensitivities to various antibacterial agents and antibiotics. In the period from October, 1987 to September, 1988, 17 institutions participated in the survey and a total of 706 strains which were demonstrated to be causative organisms were isolated from 562 patients with respiratory tract infections. Strains were mostly isolated from the sputum. The taxonomic breakdown of these strains was: Staphylococcus aureus (69 strains), Streptococcus pneumoniae (120), Haemophilus influenzae (170), Mucoid-producing Pseudomonas aeruginosa (42), Non-mucoid-producing P. aeruginosa (87), Escherichia coli (11), Klebsiella pneumoniae (35), Brahamella catarrharis (72), etc. Of these strains, 629 were used to determine MICs of various antibacterial agents and antibiotics for susceptibility analyses. Relationships between patient backgrounds and diagnoses and between infections diseases and causative organisms were also investigated. Most of the major causative organisms, such as H. influenzae and P. aeruginosa, showed no substantial changes from previous years, with regard to their sensitivities to antibiotic agent, but S. aureus, particularly methicillin/cephem-resistant strains of S. aureus (MCRSA) showed somewhat lower sensitivity to beta-lactams, and as in recent years, to ofloxacin, a new quinolone drug, as well. Regarding background factors of patients, the age distribution was heavily concentrated in age brackets of 50 years and older, thus patients in these age group accounted for 75.2% of all the patients, which was comparable to 73.5% in 1985 and 77.9% in 1986. Among infections encountered, bacterial pneumonia was most frequent at 28.3%, followed by chronic bronchitis (27.2%) and bronchiectasis (16.0%). Bacterial pneumonia was actually the most frequent, throughout the entire age groups accounting for 34.3% of patients up to 29 years, 26.6% in the group of 30-69 years and 30.7% in patients aged 70 years and older. Chronic bronchitis was next most frequent and accounted for 20.0%, 26.4% and 30.7% among the three age groups, respectively. Breaking down clinical isolates by diagnosis, H. influenzae, S. pneumoniae and P. aeruginosa were isolated frequently from most of the infectious diseases.(ABSTRACT TRUNCATED AT 400 WORDS)

[Clinical study of miconazole in deep-seated fungal infections].

We administered miconazole to patients with deep-seated fungal infections, to investigate into the clinical and mycological responses to the drug and also into its clinical safety. We also looked into the responses of amphotericin B (AMPH), flucytosine (5-FC) and the combination of the 2 drugs to deep mycoses in the past 10 years, for retrospective comparison of the findings achieved in the treatment with miconazole. A clinical response rate of 85% (29/34 patients) was achieved in the treatment of deep-seated fungal infections with miconazole. Mycologically, a fungus eradication rate of 79% (22/28 patients) and a fungus decrease rate of 11% (3/28) were achieved with the miconazole treatment, comparison of the response to miconazole alone and that to AMPH alone revealed that the former was significantly preferable. Side effects of miconazole were observed in 23% of the treated patients (15/66). Statistical analysis of the incidence of side effects of miconazole and that of AMPH showed that the former was significantly lower.

[Susceptibility of bacteria isolated from the patients with lower respiratory tract infections to antibiotics (1984)].

Bacterial isolates from patients with pulmonary infections have been collected over the last 4 years in collaboration with investigators at 14 hospitals in various parts of Japan to study isolation frequency of pathogens from patients and drug susceptibilities of these isolates. Possible causative pathogens mainly isolated from sputum of patients with lower respiratory tract infections were collected during a period from September 1984 to March 1985. We first determined types of respiratory diseases and found that, between 1981 and 1983, 57.9 approximately 64.5% of the examined diseases were chronic respiratory infections such as chronic bronchitis, chronic bronchiolitis and bronchiectasis, and that these infections including diffuse panbronchiolitis accounted for 63.1% in 1984. Bacterial pneumonia was found to be 24.8% in 1981, but it was 11.0% in 1983 and 15.1% in 1984. These results seemed to reflect decreases in the occurrence of bacterial pneumonia in young population. We then investigated the correlations between these infections and isolates and found that distributions of causative organisms of chronic bronchitis and bronchiectasis during the 4 years were similar while the detection rate of Staphylococcus aureus from bacterial pneumonia increased in 1982 and 1983, and that of Gram-positive organisms such as Enterococcus faecalis rose in 1984. Branhamella catarrhalis was considered to be a non-pathogenic organism normally harbored in the upper respiratory tract. Recently, however, respiratory infections caused by this organism have been reported by some investigators. In our research also, this Gram-negative diplococcus was isolated as a causative organism of respiratory infections as 6 strains were found in 1983 and 29 strains in 1984, hence an increase was observed.

[Susceptibility of bacteria isolated from patients with lower respiratory tract infections to antibiotics (1985)].

Collaborated studies on species of respiratory tract infection (RTI)-related organisms for their identification and drug susceptibilities have been carried out since 1981 at about 20 centers in Japan. On this occasion, the data obtained between 1982 and 1985 were reanalyzed to determine whether or not drug susceptibilities differed depending upon diseases, from which the organisms were isolated. The results summarized below were obtained in this study. 1. Among Staphylococcus aureus, Streptococcus pneumoniae, Haemophilus influenzae, Klebsiella pneumoniae and Pseudomonas aeruginosa examined, differences in drug susceptibilities according to different diseases were found among S. aureus and also mucoid strains of P. aeruginosa. 2. Susceptibility to beta-lactam antibiotics was definitely lower in S. aureus strains isolated from pneumonia than in those isolated from chronic bronchitis and bronchiectasia. 3. The isolation frequency of methicillin-and cefazolin-resistant strains of S. aureus was 30.3% and 25.9%, respectively, and was especially high among strains isolated from pneumonia. The antibiotic potency of minocycline against S. aureus, including methicillin resistant S. aureus, was the strongest among 9 drugs examined; S. aureus maintained relatively sufficient sensitivity to dicloxacillin among beta-lactam antibiotics. 4. Mucoid producing strains of P. aeruginosa isolated from chronic bronchitis had slightly lower drug susceptibility than those isolated from bronchiectasia. 5. When drug susceptibilities of H. influenzae were compared among groups separated according to diseases using MIC50, MIC80 and MIC90 as indicators, there were no clear differences. The isolation frequency of ampicillin (ABPC)-resistant strains, however, was clearly different among diseases; namely, resistant strains were the most and the least frequently isolated from chronic bronchitis and from pneumonia, respectively. In addition, the drug susceptibility of H. influenzae isolated in 1985 was analyzed in relation to the production of beta-lactamase. As a result, it was suspected that some factors, other than beta-lactamase, participated in the mechanism of ABPC-resistance. 6. These results suggest that drugs to be used for the chemotherapy of RTI should be selected considering the fact that drug susceptibilities of the pathogens differ, even among the same species, according to diseases.

[Susceptibility of bacteria isolated from lower respiratory tract infections to antibiotics (1982)].

Two hundred seventy-six bacterial strains were isolated as possible causative pathogens mainly from sputum in 248 patients with lower respiratory tract infections at 12 medical institutions in various parts of Japan during the period from September 1982 to March 1983. Of these, 272 isolates including 28 Staphylococcus aureus strains, 38 Streptococcus pneumoniae strains, 107 Haemophilus influenzae strains, 68 Pseudomonas aeruginosa strains, 17 Klebsiella pneumoniae strains, 9 Escherichia coli strains and 5 strains of other species were tested in vitro for MICs of various antibiotics, and their drug sensitivity distributions determined. Data were also analyzed for distribution of cases by clinical entities, age and sex, interrelations between the types of infections and the species and frequency of isolation of organisms, and relations of the antimicrobial regimens at collection of clinical specimens to the species and frequency of isolation of the organisms. It engenders great interest that there was a significant increase in frequency of S. aureus isolation within 7 days after antibiotic therapy, compared to pretreatment isolation frequency, in the 1982 series. This seems to deserve further investigation in detail. The H. influenzae strains isolated with the highest frequency in 1981 and those in 1982 were examined as to susceptibility to several representative antibiotics, with interdrug comparisons: ABPC vs. SBPC, CTM vs. CMZ, and CMX vs. LMOX. The isolates demonstrated high degrees of susceptibility to these drugs and there was no conspicuous change in bacterial sensitivity to the drugs.

[Susceptibility of bacteria isolated from lower respiratory tract infections to antibiotics (1983)].

Bacterial isolates from the patients with pulmonary infections have been collected over these 3 years, in collaboration with investigators at 13 hospitals in various parts of Japan for the study on frequency of isolation of pathogens among the patients and their drug susceptibilities. Possible causative pathogens mainly isolated from sputum in patients with lower respiratory tract infections during period from September 1983 to March 1984, were collected. The frequency of bacterial isolates from the sputum and their drug susceptibility on H. influenzae among the various pathogens are discussed. The kinds of bacterial species isolated from the patients with respiratory tract infections associated with chronic bronchitis, chronic capillary bronchitis or bronchiectasis and their relative frequency of isolation were as follows; Total numbers of bacterial isolates collected from various hospitals were 220 strains in 1981, 168 strains 1982 and 258 strains in 1983. H. influenzae was always isolated with highest frequency of 50.5% in 1981, 45.8% in 1982 and 40.7% in 1983, and followed by P. aeruginosa (24.1%), S. aureus (8.2%), S. pneumoniae (7.3%) and K. pneumoniae (5.9%) and E. coli (4.1%) in 1981; P. aeruginosa (30.4%), S. pneumoniae (11.9%), S. aureus (4.8%), K. pneumoniae (3.0%) and E. coli (1.8%) in 1982; P. aeruginosa (26.7%), S. pneumoniae (10.1%), S. aureus (7.8%), K. pneumoniae (3.5%) and E. coli (3.5%) in 1983. The drug susceptibility test of H. influenzae to ampicillin (ABPC), piperacillin (PIPC), mezlocillin (MZPC), sulbenicillin (SBPC), gentamicin (GM), amikacin (AMK), cefotiam (CTM), cefmetazole (CMZ), cefoperazone (CPZ), cefotaxime (CTX), ceftizoxime (CZX), cefmenoxime (CMX) and latamoxef (LMOX) was done by using agar micro-broth dilution methods. H. influenzae was most markedly susceptible to cephems of the third generation, especially to CMX, CZX and CTX by which about 95% of H. influenzae tested were inhibited the growth under the concentration with less than or equal to 0.10 microgram/ml of the drugs. Furthermore, annual changes in susceptibility of H. influenzae to various antibiotics was analyzed over the period from 1981 to 1983. The frequency of S. pneumoniae isolated from patients with chronic bronchitis or bronchiectasis was about 10% in this survey. However, in the case of respiratory infections associated with bacterial pneumonia, the frequencies were as follows; 24.8% (77 cases/311 cases) in 1981, 17.7% (44 cases/248 cases) in 1982, and 11.0% (39 cases/355 cases) in 1983. The frequency of isolation of S. pneumoniae decreased every year.(ABSTRACT TRUNCATED AT 400 WORDS)