Among 61 patients admitted to our hospital because of bacterial pneumonia during the last six years, we investigated several risk factors influencing delayed resolution of pneumonia shadows, such as age, extent of the lesion, period of fever and sputum production after antibiotic therapy, WBC, CRP and PaO2 values on admission, the period of increased values of WBC and CRP, the presence or absence of etiological pathogens, the period from the appearance of symptoms to admission, and the period of drug therapy. In our series, the period from appearance of symptoms to admission (r=0.62) and that of increased WBC (r=0.35) significantly correlated with the period of pneumonia shadows to the time of their resolution. Similar results were obtained from the multiple variate analysis of various factors. These results suggest that the duration period (nontherapy period) of inflammation closely associated with leukocytosis plays a crucial role in the delayed resolution of pneumonia shadows.
To clarify the role of local production (exohepatic) of complement on primary host defense mechanisms against microbial infections in the host who decreased the amount of complement in serum, the kinetics of the complement production by the complement-producing cells in exohepatic tissue was examined by measuring the amount of Clq, subcomponent of the first complement component, in cultured supernatant of the monolayer of peritoneal macrophages (PMφ) collected from 5, 15, 35, and 48 weeks-old female mice of NZB/W Fl (B/W Fl). The C lq production of PMφ in B/W Fl mice showed remarkable decrease at 15 weeks-old. After that, however, the C lq producibility of PMφ recovered gradually and, at mice 48 weeks old, the complement produced finally exceeded the amount observed in mice 5 weeks-old, contrariwise the Clq values in serum were significantly lowering in the same aged mice. The increased production of Clq of PMφ was observed in both 35 and 48 weeks-old mice, of which the value corresponded with increase of anti-nuclear antibody titer in serum and of the amount of protein in urine. On female mice of ddY (control), the amount of C lq production of PMφ in 5 weeks-old mice was two-fold higher than that of 5 weeks-old B/WF1 mice. But at 15 weeks, the production showed a 1/2 decrease in that of mice 5 weeks-old and the decreased values were kept to 35 weeks-old. The serum C lq values in ddY mice showed lowering only during a period of 5 to 35 weeks-old, and was increased more than the values observed in 5 weeks-old mice at 48 weeks-old. These results suggest that the increasing complement producibility of the producing cells in exohepatic tissue may play an important role in primary defense against microbial infections in host decreasing complement values in the serum.
We investigated the clinical and laboratory data of 215 hospitalized patients (mean age were 76.9±12.1) to analyze both the characteristics of senile UTI and the influence of the way of urination. UTI was present in 121 of 1897 patients (6.4%), 95 of whom (78.5%) were female. Comparison of the parameters between non-infected and infected patients were as follows: body temperature was 36.57±0.64°C vs. 37.49±0.77°C; WBC, 5410±2040/μl vs. 7260±3230/μl; CRP, 1.2±2.4 mg/dl vs. 3.5±3.4mg/dl; mean class of urinary RBC, 0-1/hpf vs. 3-5/hpf; and mean class of urinary WBC, 5-10/hpfvs. 30-50/hpf. All parameters were significantly elevated (p<0.001) in the patients with UTI. The rate of detection of causative bacteria was 88.7%; with 14.8% Escherichia coli, 12.8% Providencia species, 9.6% Enterococci, and 8.7% Pseudomonas aeruginosa. Patients with UTI were divided into three groups according to their method of urination: normal urination, use of diapers and catheterization. Body temperature (≥37.5°C) was 2.8%, 10.1% and 34.9%; WBC (≥9.000), 2.7%, 6.1%, and 14.3%; CRP, 16.9%, 36.1% and 51.1%; urine RBC (≥6-10/hpf), 8.4%, 7.1% and 36.1%; urine WBC (≥15-30/hpf), 20.4%, 44.4% and 76.9%, respectively. There was a significant difference (p<0.05-0.001) between all parameters except for urine RBC between the normal-urination patients and diaper using patients. This investigation suggested that the use of diapers was a risk factor for UTI in elderly patients.
A polymerase chain reaction (PCR) procedure for detection of Ureaplasma urealyticum was developed. A set of oligonucreotides based on sequences within the 16S ribosomal RNA gene from U. urealyticum were used as extension primers for the PCR. A DNA fragment of 397 by was amplified by the PCR, when U.urealyticum DNA was template for the PCR. No amplified product was detected from other bacterial DNA including those of Mycoplasma genus. The amplified DNA fragment of 397by was detected on agarose gel electrophoresis, when DNA of≥102 cells of U. urealyticum per PCR was used as template for the PCR. Thus, the PCR procedure was shown to be a simple, rapid and specific method for detection of U. urealyticum and could be applied to detection of U. urealyticum from clinical specimens.
Two Shigella strains (89-141 and 89-11) isolated from the stool of patients returning from abroad (both from India) in Tokyo in 1989 showed an atypical serologic reaction of agglutination with only polyvalent antiserum to S. flexneri prepared commercially. These strains had the typical biochemical characteristics of S. flexneri and were biochemically identical. Both strains were positive for Sereny test in guinea pig eye and cell-invasion test in HeLa cells. The strains also had virulence-plasmid encoding outer membrane proteins, indicating pathogenicity. The results of antigenic analyses showed that the strains were serologically identical to each other and gave significant cross-reactions with S. flexneri variant Y that has only group 3, 4 factor antigen. However, the results of reciprocal absorption tests showed that the 0 antigen of these strains was not identical to S. flexneri variant Y, and they were confirmed to have an additional type-specific antigen which is not included among the known S. flexneri type antigen I-VI or provisional type 88-893 which were proposed by us in 1992. Futhermore, one stock strain (TSHSO8) formally identified as S. flexneri variant X has this type antigen, suggesting that the new antigen can be classified into two subtypes by combination of group factor antigen. Strain 89-141 is designated as the test strain for this new type antigen of S. flexneri.
The α-streptococci, consisted of normal oral flora mainly, with inhibitory activity against pathogenic microbes in healthy individuals was investigated by group A Streptococcus (indicator strain 6-22 nonmucoid T-12). Rate of α-streptococci with inhibitory activity against group A Streptococcus was increased as aging, and the rate in pre-school children was higher than that in school children. These results suggested that more than 90% of the tested α-streptococci with strong inhibitory activities (S. salivarius) against indicator strain had inhibitory activities against group A Streptococcus (mucoid T-6), H. influenzae, S. pneumoniae, group C Streptococcus, and from 40% to 70% of the tested strains had also inhibitory activities against other pathogens. As there were many strains of a-streptococci with inhibitory activities against pathogens, that usually detected in the upper respiratory infection, the problem on the strains in the future will explain significance of the defense mechanism against upper respiratory infection and this can be applied clinically.
Soil samples from the sugar cane fields and muscovado samples from the manufacturing processes at sugar manufactories in many parts of Okinawa Prefecture were collected, mainly in 1988 and 1989, and examined for Clostridium botulinum and Clostridium tetani. Of 290 soil samples 21 (7.2%) were positive for C. botulinum. Four (1.4%) and 17 (5.9%) contained types E and C respectively. C. botulinum type E was demonstrated in the North and South areas of Okinawa main island. Type A and B were not demonstrated from soil samples in Okinawa Prefecture. Of 53 samples of the manufactoring processes of muscovado 5 (9.4%) were positive for C. botulinumtype C. Type A, B and E were not found from muscovado samples in Okinawa Prefecture. Of 290 soil samples 54 (18.6%) were positive for C. tetani. Among the 53 muscovado samples from the manufactoring processes of the sugar cane, 10 (18.9%) were positive for C. tetani.
An alkaline-phosphatase-labelled synthetic oligonucleotide probe was applied to detect the structural gene of cholera enterotoxin (ctx). This DNA probe has a complementary base sequence to 30 base of the CT-A subunit. This method was, for the first time, applied to diagnosis of a diarrheal patient. The probe detected ctx rapidly and simply as compared with reversed passive latex agglutination (RPLA) and Beads-ELISA. The cfu minimal dose for detection with the probe was about 106-7/ml. This method can be easily performed in any clinical laboratory because the procedure is safe, simple and rapid (it can be completed within about 3 hours).
In vitro proliferative response and gamma-interferon (IFN-γ) production to Pneumocystis cariniiantigen were investigated in peripheral blood mononuclear cells (PBMC) from healthy volunteers and cord blood mononuclear cells (CBMC) from umbilical cords after delivery. Twenty three of 25 samples of PBMC had strong proliferative response, and 15 of 25 samples of PBMC produced IFN-γ. In the samples showing strong proliferative response, CD4 lymphocytes expressed interleukin 2 receptors. None of the 3 CBMC samples had proliferative response nor production of IFN-γ. When CD4 lymphocytes were depleted in PBMC, the proliferative response was markedly decreased and IFN-yproduction was abolished. In conclusion, the lymphocytes of healthy adults are activated and produce IFN-γ by stimulation with P.carinii antigen, and these two response are both induced by CD4 lymphocytes.
The interactions between aerobic and anaerobic bacteria in the development of anaerobic bacterial pneumonia were studied by introducing Bacteroides fragilis and Escherichia coli alone or in combination into guinea pigs by tracheal infusion. The lung lesions induced by B. fragilis were mainly located near the pleura, unlike those induced by E. coli, and were accompanied by pneumonia, lung abscess. and uleuritis. The lung lesions produced by mixed infection with B. fragilis (109 cfu) and E. coli (107 cfu) were significantly more severe than those induced by either microbe alone, and the redox potentials at the foci of inflammation were markedly reduced (max: 330 mV). Analysis of lung lesions after treatment with aztreonam and clindamycin and neutrophil phagocytic activity suggested that E. coli was primarily responsible for the lung lesions and that B. fragilis promoted the accompanying inflammation, resulting in increased pathogenicity of the mixed infections.
We studied the epidemiology of enterovirus infection in Aichi Prefecture from 1985to 1989. We examined the age distribution of aseptic meningitis patients (AM) and exanthematous disease patients (Ex) and a seroepidemiological study of echovirus type 7 (E7), E9, E18 and group A coxsackievirus type 9 (CA9), was performed. The results is as follows: 1) E7, E9, E18 and CA9 were isolated from AM and Ex but E6, Ell and group B Coxsackie viruses (CB) were isolated in fewer cases from Ex. 2) The AM was consistently increased from June to August. Whereas the Ex was seen in all seasons but a slight increase was noted between June to July, and enteroviruses (EV) isolation wasi ncreased in this season. 3) The AM occurred in 0 year olds and 4 year olds whereas the Ex was seen in 0 to 1 years. EV was mainly isolated from 0-1 year olds. 4) The relationship of clinical manifestations and age was very clear in E9 and E18, a higher proportion of children at 1 years or under were those of the Ex and most children of the latter part of 4 years were those of the AM. The Ex had the same results with E7 and CA9 but AM was increased in 0 years and 4 year olds. 5) We studied the age distributions of neutralizing antibodies against E7, E9, E18 and CA9. The positive rate of neutralizing antibody after prevalence rose between 2-5 years of age. There were few patients among the 2 to 3 year olds but the neutralizing antibody was raised in this age. I considered that reason the enteroviruses infected mainly the 2 to 6 year olds showing no clinical symptoms where as some of the 3 year olds had aseptic meningitis and some under 1 year had symptoms of exanthematous diseases.
We developed two PCR methods, which amplify bovine tuberculous MPB70 gene and mycobacterial 16S rRNA gene, for detection of tubercle bacilli and mycobacteria in sputum, respectively.A mong 27 Mycobacterium species and 57 species of 30 genuses other than Mycobacterium, only M.tuberculosis (TB) complex, i.e., M. tuberculosis, M. bovis, M. africanum, M. microti showed DNA amplification by PCR for MPB70, and amplification of 16S rRNA gene were observed specific in Mycobacterium species. A combination of these PCR abilities were available to differentiate the TB complex and nontuberculous mycobacteria (NTM). We investigated the correlation between these methods and conventional methods with 311 sputa that were suspected mycobacteriosis. The PCR method could detect 12 cases of TB complex and 4 cases of NTM in 17 specimens, which were positive by conventional methods, but could not for one specimen. Among 294 specimens that were negative with conventional methods, the PCR method detected 13 and 8 cases of TB complex and NTM, respectively. These results were confirmed by commercial tuberculous specific DNA probe or investigation of the clinical background of the patients. On the other hand, 273 specimens showed negative result either PCR nor conventional methods. The PCR method did not detect tuberculous DNA in normal 197 sputa, which were not suspected mycobacteriosis. These results indicate that each one of these PCR methods is highly specific to TB complex or Mycobacterium species. We concluded that these PCR methods are useful and advanced methods for rapid and direct detection of tubelculosis and mycobacteriosis.
A 58-year-old male was referred to our division because of antibiotic-resistant pneumonia. A chest X-ray film revealed severe pneumonia over the left lung field but laboratory data showed no leukocytosis. Transbronchial lung biopsy findings showed the evidence consistent with organizing pneumonia. One-month prednisolone therapy produced a disappearance of the pneumonia shadow, but a giant bulla was found at the same site. It was considered that it was necessary to treat this case with a combination of effectve antibiotics and steroids in the early phase of the disease.