Neutrophil's CL (CL-index) in were assayed in 18 patients with chronic lower tract infection (13 cases in the exacerbated phase and five cases in the stable phase) three times per case: before, during and after chemotherapy. Before the initiation of chemotherapy, the CL-index? N (neutrophil numbers × CL-index) in patients with exacerbation remained higher than in those without exacerbation. At the exacerbated phase, the CL-index of nine patients and the CL-index · N of 10 patients fell lower than those obtained before treatment. AT the stable phase, the CL-index and the CL-index · N did not change remarkably during and after chemotherapy.

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Polymerase chain reaction (PCR) with nested primer pairs was used to diagnose Tsutsugamushi disease and identify the Rickettsia tsutsugamushi serotype. The primer pairs used for PCR were designed on the basis of the nucleotide sequence of the gene that encodes the 56-kDa antigen. Five serovariants, the Gilliam, Karp, Kato, Kawasaki, and Kuroki strains of Rickettsia tsutsugamushi were detected and identified by nested PCR. The serotypes of patients registered during 1990 to 1992 in Kanagawa Prefecture were identified by nested PCR. Sixty percentage of patients showed Kawasaki types, 20% Karp types, and 20% Kuroki types. This result suggested that the recent Tsutsugamushi disease were mostly caused by Kawasaki types in Kanagawa Prefecture.

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We evaluated the usefulness of urine sediment from male gonorrhea patients for detecting gonococcal antigens by enzyme immunoassay, Gonozyme^® (Abbott Laboratories). Experiments for dilution rate of urine sediment with buffer provided in a kit indicated that dilution of sediment at 1: 4-6 would be practical in performing Gonozyme. All urine sediment specimens from 18 gonorrhea patients showed positive by Gonozyme, and 68, except for 3, culture-negative control urine sediments, including 8 sediments from patients with chlamydial urethlitis, were negative. Based on comparing results of Gonozyme with those of culture, sensitivities and specificity were 88.9-100% and 95.6%, respectively. During storage of urine sediment at 5°C for 7 days, absorbance values were stable. Therefore, urine sediment could be preserved or transported. We conclude that urine sediment is useful for diagnosis of gonococcal urethritis in males by Gonozyme.

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A clinical assessment of fungal infection in hepatobiliary and pancreatic diseases during 1975 and 1991 was made and 25 cases of systemic mycosis were noted. Among 25 cases there were 20 liver diseases (hepatocellular carcinoma 12, liver cirrhosis 5, fulminant hepatitis 2, polyarteriris nodosa 1), 2 cases of gallbladder cancer and 3 cases of pancreatic cancer. The fungus was consisted of 14 cases (56%) of Candida, 9 cases of Aspergillus (36%), and 2 cases of Cryptococcus (8%).Fungal infection was most frequent in the lung (8 cases) and esophagus (6 cases), but rarely in the stomach, lyrnphnode, liver, thyroid, kidney and gallbladder. Generalized fungus infection was noted in four cases (16%).
Fatal fungal infection was complicated in liver cirrhosis (2 cases), fulminant hepatitis (one case), gallbladder cancer (one case) and cystadenocarcinoma of the pancreas (one case). In five fatal cases three cases of Aspergillus pneumonia and two cases of Candida septicemia were included. Glucocorticoid was used in 13 cases (52%) and anti-cancer drugs was administered in two cases (12%). However, in 9 cases (36%) without treatment of glucocorticoid or anti-cancer drug fungal infection was detected. In conclusion, there is a possibility of fungal infection in grave hepatic diseases and empirical administration of anti-fungal agent may be necessary.

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In this clinical study we tried to find out the clinical significance of positive blood cultures between Jan. 1984 and Dec. 1989 in the Nagasaki Rosai Hospital. In this period the association between the isolated bacteria from blood cultures (398 specimens: 17.9%) and clinical background were evaluated. Each year the positive cultures were seen in about 18-20%. The distribution of bacteria showed increase percentage of the gram-positive organisms such as coagulase-negative Staphylococcus. Gram-negative organisms in included P. aeruginosa, A. calcoaceticus and E. coli. A. calcoaceticus was frequently found from the dept. of neurosurgery. Fungus, mostly Candida, was isolated in about 10-20%. The specimens were provided from department of internal medicine (62.9%) and neurosurgery (22.8%)
An analysis of 213 episodes of 188 bacteremic cases was made. The onset of bacteremia had two peaks, 24 h or 1 month, after admission. The primary infected sites were the urinary tract (26.7%), respiratory system (14.5%) and central nervous system (6.1%). The central venous catheter was an important infected site. The major underlying diseases included acute cerebrovascular diseases, cardiac failure due to valvular heart diseases or acute myocardial infarction and/or iatrogenic immunocompromised state. The major complications were disseminated intravascular coagulation (DIC) (24.4%) and multiple organ failure (MOF) (8.9%). Half of the DIC episodes was caused by grampositive cocci such as CNS, E. faecalis and MRSA. 68.4% of MOF episodes was associated with gram-negative bacteria. While the remainder was caused by gram-positive cocci including CNS and MRSA.
The comparison between the effective group and the poor group of antimicrobial chemoterapy revealed that ixn the poor group E. faecalis, A. calcoaceticus and Candida were major pathogens and the respiratory infection and central venous catheter were also major risk factors of chemotherapy for bacteremia. We recommend minocycline to be included in the combination treatment when we considered infection of the CNS where S. aureus and A. calcoaceticus as a causative organism of bacteremia.

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A retrospective evaluation of the status of soil-transmitted helminthic infection in Nepal during the period from 1985 to 1992 was conducted at the Parasitology Laboratory, Department of Pathology, TU Teaching Hospital in Kathmandu. An average of 6, 537 faecal samples were examined each year for the presence of various types of intestinal parasites. The annual rate of positivity of these samples for soiltransmitted helminthiasis ranged from 18.0 to 36.6%. The annual incidence decreased every successive calender year in both adults and children, irrespective of sex. Among the various types of helminths, the most common was Ascaris lumbricoides (roundworm), followed by hookworm. The incidence of A. lumbricoides remained constant throughout the study period, while that of other parasites markedly decreased each year.

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Anaerobic cultures were performed on various kinds of specimens obtained from 67 patients with respiratory infections. The infections consisted of 33 cases of pneumonia, 14 cases of lung abscess, 6 cases of pyothorax, 9 cases of chronic lower respiratory tract infection, and 5 cases of infection of pulmonary cyst. Specimens included bronchoscopic specimens obtained using a single tube or protected catheter brush (PCB), transtracheal aspirate, percutaneous lung aspirate, aspirate using a suction tube through the trachostomy or intubation tube, and pleural fluid. Anaerobes were isolated in 80% of pyothorax, 43% of lung abscess, and 15% of pneumonia (42% of aspiration pneumonia), respectively. Anaerobes isolated frequently in respiratory infections included Prevotella spp., Fusobacterium spp., and Peptostreptococcus spp. In regard to the type of specimen, bronchial aspirate obtained by bronchoscopy was useful in examination of pneumonia, and was useful as percutaneous lung aspirate in examination of lung abscess. But there was discrepancy of culture results between specimens obtained by different methods in the same patient. To confirm anaerobes as pathogens in respiratory infections, it is necessary to select an appropriate method to obtain specimens and to interpret the culture results comprehensively.

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A multicenter clinical study was carried out to investigate the clinical efficacy icacy of SY5555 from viewpoint of its MIC breakpoint for Enterococcus faecalis.
SY5555 was administered in dosages of 150 mg×3/day and 300 mg×3/day, for 7 days, to patients with complicated urinary tract infections of moderate severity (no indwelling catheter). Based primarily on the bacteriological efficacy (eradication) as the criterion for evaluation, the MIC breakpoint of SY5555 was investigated.
SY5555 showed potent antimicrobial activity towards E. faecalis, and its MIC₈₀ value for the clinical isolates of this bacterial species was 1.56μg/ml. Analysis was carried out to determine the extent to which this in vitro activity is corroborated by the drug's clinical efficacy in eradicating E. faecalis
1. Experimental study
In a study employing an experimental model of complicated bladder infections of moderate severity, it was judged that the MIC breakpoints of SY5555 administered at 150 mg×3/day and 300mg×3/day were 2μg/ml and 8μg/ml.
2. Clinical study
SY5555 was administered at 150 mg×3/day (7 days) to six patients and at 300 mg×3/day (7 days) to 32 patients, and the clinical efficacy of these treatments was investigated.
2.1. Investigation of the clinical MIC breakpoint showed it to be 12.5μg/ml for both the 150 mg×3/day and 300 mg×3/day regimens. However, since the number of cases in this study was small, analysis was performed of the data from a large number of cases by including the cases registered in a nation-wide Phase II study of SY5555. That analysis showed the clinical MIC breakpoint for the two dosages to be 3.13μg/ml and 6.25μg/ml. Thus, this analysis of large number of cases yielded clinical MIC breakpoint that are in agreement with the abovementioned experimental MIC breakpoints.
2.2. In the evaluation of the efficacy rate using the UTI drug efficacy evaluation criteria, cases rated as “good” or “excellent” numbered five of the six patients in the 150 mg×3/day treatment group and 25 of the 32 (78.1%) patients in the 300 mg×3/day treatment group, or 30/33 (78.9%) cases overall. These efficacy rates are about the same as the 82.0% reported for the nation-wide Phase II study.
2.3. Each of the strains of E. faecalis isolated from the patients was found to be eradicated by the SY5555 treatment: four strains in the 150 mg dosage group (the MIC of each was 3.13μg/ml or less) and 18 strains in the 300 mg dosage group (the MIC of each was 6.25μg/ml or less). Moreover, in both dosage groups, E. faecalis was not found to recur after the SY5555 treatment.
3. Conclusions
In consideration of the above findings in the investigations of treatment of patients with complicated urinary tract infections of moderate severity, the following conclusions can be drawn.
3.1. In antimicrobial treatment employing SY5555 in a regimen consisting of 300 mg×3/day for 7 days, the clinical breakpoint is in the range of 6.25-12.5μg/ml.
3.2. In paticular, as would be surmised from the fact that the MIC₈₀ value of SY5555 for E. faecalis isolated is 1.56μg/ml, this drug shows quite strong efficacy in the eradication of E. faecalis from cases of complicated urinary tract infection.

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During a 12-years period between September, 1980 and November, 1992, hemolytic streptococci (group A, B, C, & G) in throats of healthy school children in Osaka were examined every other month. The results were summerized as follows.
1. 5, 023 strains of hemolytic streptococci were detected from 11, 647 specimens, 43.1%. Among them, 2, 395 strains (20.6%) belonged to group A, 1, 647 (14.4%) to group B, 767 (6.6%) to group G and 187 (1.6%) to group C.
2. In the first half of the research, group A streptococci were detected predominantly and the last half, group B generally. Both this lower rate of group A to the total strains (47.7%) and this higher rate of group B (33.3%) were due to the small number of lower school grade children from whom group A streptococci are often detected and also due to application of the selective enrichment medium.
3. Group A streptococci were classified T-type. The most common serotype was T-1, and 638 strains (26.6%) were detected, followed by T-12, 377 (15.7%), T-6, 210 (8.8%) and T-13, 203 (8.5%). The dominant serotype was exchanged annually, but only T-1 was the most common serotype for 4 years (1983-1986).
4. In group B, the most common serotype was Ia, and 524 strains (31.3%) were detected, followed by III, 417 (24.9%), Ib, 164 (9.8%), III/R, 130 (7.8%) and Ia/c, 122 (7.3%). Annual changes of serotype were as follows; Ia was dominant from 1980 to 1988, III from 1989 to 1990 and NT6 in 1991-1992 generally.

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During a 12-years period between 1980 and 1992, 5, 023 strains of hemolytic streptococci were detected from throats of school children, and described in part I. Among them, the following strains their susceptibility were tested to 14 kinds of antibiotics; 1, 511 strains of group A, 1, 038 of group B, 125 of group C, 553 of group G.
1. No resistant strains against both penicillins (PCG, AMPC, ABPC, ACPC) and cephems (CER, CET, CCL, CEX) could be found. Strains of group B were less susceptible than the others to penicillins and cephems.
2. Some resistant strains were found against macrolides (EM, OL & JM), 5.9-8.6%. These resistant strains belonged many to group A and a few to group B & G. Since 1983, the gradual decrease of the resistant strains was noted though few were found after 1986.
3. To TC a number of resistant strains were detected in group A, B, C & G through this study, (12.5-48.4%).
4. To CP some resistant strains were found from 1980 to 1985 among group A, B & G, 3.0-5.1%. Since 1986, the sharp decrease of the resistant strains was noted.
5. Multiresistant strains to TC, CP and macrolides were found since 1980 to 1982. They were found in many of group A and a few of group B and G, but after 1986 decreased sharply.
6. Most of the multiresistant strains of group A belonged to serotype T-12 and group B to serotype Ia. Ia/c, III and III/R.
7. Among the strains of group A type T-13, there were many resistant strains to TC. The rate of resistant occupied over 60% from 1980 to 1986, but decreased slightly thereafter.

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Studies on β-hemolytic streptococci have been carried out in 2 institutions for handicapped preschool children during the last nine-year period from 1985 to 1993. These studies were based on continuous throat cultures, 4 to 12 times a year, from 244 children including Down's syndrome and mentally retarded children in K-institution (K), and from 134 including cerebral palsy and other cerebrospinal disorders in H-institution (H). A significant difference (p<0.05) was demonstrated in the isolation rates between K (11.1%) and H (6.4%).
In K, isolation rates of β-hemolytic streptococci ranged from 0 to 43.5%, showing a higher rate in winter (16.5%) than in summer (6.6%), and elevated-isolation rates of homologous types were demonstrate four times during the study period; i. e., type T12 in '87, type T28 in '88, type T1 in '90 and '92.
In H, on the other hand, no higher rates (as to group A) were observed in winter or summer.
In relation to disorders of children, the rates were 12.6% in Down's syndrome (mean age 2.7y), 10.0% in mentally retarded children (3.9y) and 6.4% in cerebral palsy or others with cerebrospinal disorders (3.3y). Down's syndrome, the youngest group, revealed the highest isolation rate of β-hemolytic streptococci.
In only two cases isolated hemologous types detected were over two times continuously.

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Fibrinogen binding activities in culture supernates of group A streptococci were detected by dot blot assay using horseradish peroxidase conjugated human fibrinogen. Various intensities in colored dots were seen in 63 of 70 strains isolated from pharyngitis patients in 1992. Strong binding activities in all of 16 strains were partially sensitive to trypsin digestion. The binding activities in the concentrated culture supernate of M-type 4 strain SS91 were analysed by SDS-PAGE and Western blotting. Intense colored bands and faint ones were seen in the position corresponding to 89K, 66K, 59K, 49K, 42KDa and 77K, 53K, 51K, 44KDa, respectively. Only one band at 42KDa remained after trypsin digestion.

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It is known that the neonatal group B streptococcal disease in most cases is acquired by contamination during passeage through the birth canal at the time of delivery. Therefore, to investigate the influence of colonization of group B streptococci in the female genital tract by age, we carried out the isolation of group B streptococci from the urine of healthy children of school age group, and those age groups were as follow: kindergarten primary school students, junior high school students, high school students and women over twenty years of age.
The isolation rates of group B streptococci from the urine of the healthy persons by each group were as follow: 12.5-18.2% in 93 females and 0-0.3% in 90 males in kindergarten aged 3-5 years, 6.4-25.3% in 285 females and 0.3-3.2% males in primary school aged 6-11 years, 11.8-19.1% in 183 females and 0-7.1% in 233 males in junior high school aged 12-14 years, 16.4-23.7% in 416 females and 2.6-7.1% in 114 males in high school aged 15-17 years, and 35.0-50.0% in adult females aged 20-60 years.
Isolation rates according to sex were 2.2% in 756 males to 17.5% in 977 females of aged 3-17 years, and there was a clearly highly significant difference between males and females (p<0.01).
Group B streptococci isolated from urine of 1, 818 persons were 220 strains (12.1%), and the 146 (66.4%) in all strains could be typed by the serological test. The most frequently isolated type and number of the types distribution in each school age group were as follows: type Ia, type Ib/c and two types others in kindergarteners, type Ia and seven types others in primary school students, type Ia/c and seven types others in junior high school students, type NT/R and ten types others in high school students.

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The structural gene for type 3 M protein of Streptococcus pyogenes, which consists of an N-terminal variable portion and C-terminal conserved repeat regions, has been cloned by the polymerase chain reaction (PCR) with two primers (K-1 and K-2). They were selected from the best conserved region of the leader sequences and of the C-terminal portion near the Hexapeptide (LPSTGE) sequence found in different M proteins. From the nucleotide sequence of the product, 1645 base pairs were determined, including 32 amino acids of the leader sequences, the complete N-terminal variable region and the conserved C repeat regions. Analysis of the deduced amino acids of the sequence revealed the existence of two major repeat regions, the B and C repeat regions. Comparison of the C-repeat regions among M3 and other M proteins showed them to be more than 90% identical. The two B repeat blocks in M3 protein are also similar to those in M12 protein. Predictive secondary structure analysis of M3 protein reveals a strong alpha-helical potential. The algorithm also shows that the beta-sheet and turn potential for region 23-42 in M3 protein are similar to those for region 28-50 in M12 protein. The results indicate that M3 protein is closely related to M12 protein.

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The Lancefield's serotyping system of Streptococcus pyogenes is based on the M protein. The typing sera. used for this system are prepared in rabbits immunized with whole organisms of specific serotypes. To remove cross reactive antibodies, the sera should be extensively absorbed with organisms of selected heterologous serotypes. In this study, the possibility of using monoclonal antibody (MAb) as a type specific reagent was discussed.
MAb which specifically reacted to M type 12 protein of S. pyogenes were produced by cell fusion. This specific MAb reacted to hot acid extracted M type 12 protein in ELISA but didn't react in agglutination and precipitation. Latex beads were sensitized with MAb and examined by the coagglutination method. The latex reagent could detect a very small amount of M type 12 protein, so it could be used for M typing of S. pyogenes which produced a small amount of M 12 protein.

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For prevention of group B streptococcal (GBS) infection in neonates, GBS carriers of pregnant women should be detected. Procedures for detection of carriers were needed to get higher positive rate and to be handled with easy. Urine samples were shown to detect higher rates than vaginal smear samples in pregnant women.
GBS in the urine were adsorbed to charcoal granula for easier transporting. These adsorbed charcoal were tested as to whether they could be preserved with different temperatures and days. Urine with 10¹ CFU/ml of GBS added could be detected after 7 days at emperatures of 4, 25, 37, 42°C. The cultures of this procedure with the cultures of urine sediment of pregnant women were compared. Detecting rates of this procedure and urine sediment were 177 (26.2%) and 136 (20.1%) of 676 pregnant women, respectively.
These data suggested that adsorbed charcoal culture was useful with higher positive rates for detection of the GBS carriers in pregnant women.

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