[New Concept and Approach to Rapid Diagnosis for Infectious Diseases]
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Journal Article↗
Biomedical subjects
N Yamane
Publications and source records attributed to N Yamane.
[Evaluation of Fluo-Card Milleri to Identify the Isolates of Streptococcus milleri Group: Comparative Identication with Referral Fluorogenic Phenotypic Dierentiation]
Clinical isolates which belong to the "Streptococcus milleri" group were identied by the referral uorogenic phenotypic tests described by Whiley et al. [J. Clin. Microbiol., 28: 1497-1501 (1990)] and a rapid, commercially available test kits; Fluo-Card Milleri (KEY Scientific Products, Round Rock, Tex., U.S.A.) to the species level. Of 218 clinical isolates included, 196 (89.9%) were correctly identied by the Fluo-Card Milleri when compared with the reference identications. Ten isolates (4.6%) of S. constellatus resulted in the "unidentied" due to the negative interpretations for all the three enzymatic reactions. A total of twelve isolates (5.5%); five of S. anginosus, five of S. constellatus, and two of S. intermedius, were misidentied. The levels of agreement were 95.7% for S. anginosus, 91.3% for S. intermedius, and 92.6% for S. constellatus when the unidentied results were excluded.
Journal Article↗
[New clinical tests that physicians should know for diagnosis of infectious diseases].
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Bacteria↗
Florid reactive follicular hyperplasia in elderly patients. A clinicopathological study of 23 cases.
Florid reactive follicular hyperplasia (FRFH) of the enlarged lymph node in elderly patients requiring biopsy is a relatively uncommon phenomenon as compared with younger age groups. We experienced 23 patients, aged 60 years or more, from whom the biopsied lymph node specimens histologically showed inappropriate FRFH for their age, in the period between 1982 and 1996. These cases were morphologically subdivided into three groups, FRFH with interfollicular plasmacytosis, that with progressive transformation of germinal center, and FRFH without additional specific findings. FRFH with interfollicular plasmacytosis were observed in 11 cases, all of whom were accompanied with several immunological abnormalities (six with rheumatoid arthritis, three with multicentric Castleman's disease and one each with myoepithelial sialoadenitis and autoimmune hemolytic anemia). Three men with uncertain etiology exhibited an unusual histology of progressive transformed germinal centers which were clinically characterized by a bulky neck mass. Among the nine cases with nonspecific FRFH, only four had a specific etiology (one each with adult onset Still's disease, chronic sinusitis, Epstein-Barr virus infection and infectious lateral cervical cyst), while the other five with unknown etiology showed abnormal laboratory findings suggestive of an abnormal humoral immune response, i.e. hypergammaglobulinemia and seropositivities for some autoantibodies. None of our patients developed malignant lymphoma during the follow-up period. Of note, 16 (70%) of the 23 cases were found to be associated with various types of imbalances of the immune system, some of which appeared to be currently ill-defined as clinicopathological entities that were simply categorized as autoimmune disease.
Aged↗
Occurrence of monocytoid B-cells in reactive lymph node lesions.
Benign monocytoid B-cells are a peculiar subset of B-cells. They are closely related to marginal zone B-lymphocytes, show cytological diversity and may be recognized in a variety of reactive lymph node conditions. To analyze the incidence, cytological spectrum and phenotypic features of benign monocytoid B-cells, we investigated a series of 301 consecutively biopsied and unselected cases of reactive lymph node change from 1988 and 1995. A monocytoid B-cell reaction was identified in 46 (15%) cases and could be cytologically subclassified into two groups: 31 (67%) cases with common-type cells and 15 (33%) cases with large, transformed cells, according to the description by Plank et al. [19]. These reactions were regularly associated with follicular hyperplasia (95%) and were part of an epithelioid cell response in 24 cases (50%). Immunohistologically, both types of benign monocytoid B-cells were negative for bcl-2 protein expression, which was in contrast to the bcl-2 positive reaction in marginal zone B-lymphocytes and their neoplastic counterpart in monocytoid B-cell lymphoma. An association of Epstein-Barr virus (EBV) with monocytoid B-cells was investigated by in situ-hybridization. EBV genomes were detected in five (15%) of 31 cases tested. In each of these five cases, positive cells were represented in both high and low numbers. The morphologic features of the EBV-positive cells were not consistent with monocytoid B-cells, but rather with medium-sized to large lymphoid cells. It appeared that the occurrence of monocytoid B-cell reaction in reactive lymph node lesions was not related to EBV infection in the majority of cases.
Adolescent↗
Clinical evaluation of the ASTY colorimetric microdilution panel for antifungal susceptibility testing.
A method using a commercially prepared colorimetric microdilution panel (ASTY; Kyokuto Pharmaceutical Industrial Co., Ltd.) was compared in four different laboratories with the National Committee for Clinical Laboratory Standards (NCCLS) reference microdilution method by testing 802 clinical isolates of Candida spp. (C. albicans, C. glabrata, C. tropicalis, C. parapsilosis, C. krusei, C. lusitaniae, C. guilliermondii, C. lipolytica, C. rugosa, and C. zeylanoides) against amphotericin B, 5-fluorocytosine (5FC), fluconazole, and itraconazole. Reference MIC endpoints were established after 48 h of incubation, and ASTY endpoints were established after 24 and 48 h of incubation. ASTY endpoints were determined to be the time at which the color of the first well changed from red (indicating growth) to purple (indicating growth inhibition) or blue (indicating no growth). Excellent agreement (within 2 dilutions) between the reference and colorimetric MICs was observed. Overall agreement was 93% at 24 h and 96% at 48 h. Agreement ranged from 90% with itraconazole and 5FC to 96% with amphotericin B at 24 h and from 92% with itraconazole to 99% with amphotericin B and 5FC at 48 h. The ASTY colorimetric microdilution panel method appears to be comparable to the NCCLS reference method for testing the susceptibilities of Candida spp. to a variety of antifungal agents.
Amphotericin B↗
[Enhanced induction of apoptosis of human colorectal cancer cells after preoperative treatment with 5-fluorouracil its relationship to DNA ploidy pattern].
We examined the relationship between apoptosis induced by 5-fluorouracil (5-FU) that was given preoperatively to colorectal cancer patients and DNA ploidy pattern, and investigated the cell cycle changes, and the expression of Ki-67. Twenty-nine patients with advanced colorectal cancer were divided into four groups, 3 days, 5 days, 7 days, and 10 days. Groups received continuous intravenous 5-FU at 500 mg/body/day preoperatively. Then, patients were divided into two groups by DNA ploidy pattern, diploid(D) and aneuploid(A). Apoptotic cells were stained by the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) method. The expression of Ki-67 was examined by immunohistochemical staining. We used flow cytometry (FCM) for analysis of cell cycle distribution. Apoptosis of cancer cells was mostly increased in 7 days 5-FU administration in both D and A groups. The expression of Ki-67 was reduced according to the prolongation of the term of 5-FU administration in both D and A groups. We assessed S-phase fraction (SPF) to evaluate the cell cycle changes by 5-FU. Tumor samples of all patients after injection of 5-FU showed S-phase accumulation. The ratio of SPF (after 5-FU/before 5-FU) was the highest in the 5-day 5-FU administration group in both D and A groups. We concluded that apoptosis and S-phase accumulation were increased, and proliferative activity was decreased by preoperative 5-FU administration in colorectal cancer patients. However, there was no clear correlation between DNA ploidy pattern and these changes.
Adenocarcinoma↗
[Determination of pyrazinamide susceptibility for Mycobacterium tuberculosis by use of Middlebrook culture media and comparison with results of pyrazinamidase test].
We developed a new test method to determine pyrazinamide (PZA) susceptibility for Mycobacterium tuberculosis in an acidified Middlebrook 7H9 broth (pH6.0), and evaluated in comparison with the agar proportion method of the National Committee for Clinical Laboratory Standards (NCCLS) M24-T and with pyrazinamidase assay. The test method is based on a culture in 4 ml of the modified Middlebrook 7H9 broth containing 100, 200 and 400 micrograms PZA/ml, respectively. First, the cell suspension was adjusted to a McFarland #1 turbidity, and then diluted 1:10. After mixing, 0.1 ml of the diluted cell suspension was inoculated and incubated at 36 +/- 1 degrees C in an ambient air. After 7 day-incubation, the test broth was read in comparison with the growth control. When a significant growth at 100 micrograms PZA/ml or an attenuated growth at 100 micrograms PZA/ml but a significant growth at 400 micrograms PZA/ml were observed, the test isolate was interpreted as being PZA-resistant. When PZA-susceptible and PZA-resistant ATCC reference strains were repeatedly tested, the results obtained were highly precise and accurate. A total of 65 clinical isolates were tested, the results indicating 95.4% of agreements with the agar proportion method and 90.8% with pyrazinamidase assay. There found six discrepant results of 13 resistant isolates; three were susceptible by the agar proportion and all the six were positive by pyrazinamidase assay. Accordingly, we can conclude that, in place of radiometric Bactec System, our newly developed test method is an accurate, practical, rapid and nonradiometric alternative to determine PZA susceptibility for M. tuberculosis in clinical mycobacteriology laboratories.
Amidohydrolases↗
[Evaluation of a newly developed broth microdilution test method to determine minimum inhibitory concentrations (MICs) of antimicrobial agents for mycobacteria].
We developed a new broth microdilution antimycobacterial susceptibility test for determination of minimum inhibitory concentration (MICs) using an air-dried microplate containing serially diluted antimicrobial agents and the modified Middlebrook 7H9 broth. The eight agents included were streptomycin (SM), isoniazid (INH), rifampicin (RFP), ethambutol (EB), kanamycin (KM), levofloxacin (LVFX), sparfloxacin (SPFX) and clarithromycin (CAM). Serial dilutions of the agents (128 micrograms/ml to 0.125 micrograms/ml) were prepared in microplates, and were reconstituted by inoculation of 0.2ml of cell suspensions (approximately 3 x 10(5) cells/ml). The test plates were incubated at 36 degrees C in 5% CO2, and the growth endpoints were read visually after 5-day, 7-day and 10-day incubations. Four ATCC reference strains, Mycobacterium tuberculosis, M. avium, M. kansasii and M. intracellulare, were repeatedly tested at three sites. Of 480 determination against eight agents, 455 (94.8%), 470 (97.9%) and 455 (94.8%) of the MICs read after 5-day, 7-day and 10-day incubations fell within 3log2 dilutions, respectively. The MICs gradually elevated during the incubation, however those of 7-day incubation were highly precise and easily determined. A total of 160 clinical isolates of M. tuberculosis and 114 of nontuberculous mycobacteria were tested against eight agents. As for the primary drugs (SM, INH, RFP and EB), most isolates of M. tuberculosis were highly susceptible with MIC90, < or = micrograms/ml. Both LVFX and SPFX were also active. The MICs against nontuberculous mycobacteria distributed in a wide range, and the activities of RFP, LVFX, SPFX and CAM were more potent. These results demonstrate this newly developed test method to be a practical, rapid, quantitative and nonradiometric alternative for the determination of MICs in clinical mycobacteriology laboratories.
Antitubercular Agents↗
[Blood culture: gold standard for definitive diagnosis of bacterial and fungal infections--from the laboratory aspect].
Blood culture has been long recognized as a gold standard for definitive diagnosis of bacterial and fungal infections worldwide. However, due to the lack of this recognition, particularly among physicians, very few number of blood cultures have been tested in microbiology laboratories throughout Japan. Positive results from blood cultures would have a great impact on clinical decisions; initiation of antibiotic chemotherapy, change to effective chemotherapy, and more frequently, change to less expensive chemotherapy. The "Rules of Three"; nearly all instances of bacteremia will be recognized after the incubation of three blood cultures for three days, originally proposed by Young et al. is still applicable to the most clinical instances, and will increase the value of blood cultures. The key to assuring greater clinical impact from blood cultures is rapidity to detect positive by the laboratory. Several automated blood culture systems are now commercially available, and will enable us to detect positive cultures significantly earlier than before.
Bacterial Infections↗
[Characteristics and outcome of stroke patients discharged from hospitals in an urban area of Japan].
The purpose of this study was to obtain the overall characteristics and outcome in stroke patients in an urban area of Japan, in order to develop some strategies for a community-based care system for improvement of the prognosis and activities of daily life in stroke patients. The present study, based on 4 hospitals in Yao city in Osaka, showed results as follows: 1) The number of stroke patients discharged from 4 hospitals during a recent one year period was 377 (216 men and 161 women). 2) The proportion of patients who died was 27%. Of the living patients, the proportions of patients who had motor paralysis or speech disturbance at discharge were 54% and 27%, respectively. 3) The proportion of patients who were to return to their home was 77% for all living patients. The proportion of patients who were not able to walk by themselves, or needed assistance in at least one of the activities of daily life was 31% and 28%, respectively, for all living patients who were to return to their home. This study showed that stroke registry from major hospitals in an urban area is useful for obtaining the overall characteristics of stroke patients in the area and promote community-wide programs in the prevention and home care of stroke.
Activities of Daily Living↗
Systemic lupus erythematosus (SLE) lymphadenopathy presenting with histopathologic features of Castleman' disease: a clinicopathologic study of five cases.
Lymph node enlargement is common in active systemic lupus erythematosus (SLE), a disease characterized by well defined clinical criteria. Although numerous reports have described the characteristic histology of SLE lymphadenopathy to include necrotizing lesions and hematoxylin bodies, no detailed description has examined the histopathologic features that are similar to Castleman's disease (CD) in SLE patients. In this report, we describe the clinicopathologic findings of CD-like peripheral lymphadenopathy, which was identified in five (26%) of 19 SLE patients. These five patients were all female with an age range of 24 to 44 years, and four of them presented with multicentric lymphadenopathy. They also had systemic symptoms and abnormal laboratory findings, indicating active disease, although two patients had not fulfilled the diagnostic criteria of SLE at the initial disease. The size of the enlarged lymph nodes seldom exceeded 2.0 cm in diameter, and biopsies revealed histopathologic features similar to CD, of intermediate type in three patients and hyaline vascular type in two according to the classification of Flendrig [7]. Immunohistochemical studies demonstrated polyclonal plasma cell populations in all five cases. Epstein-Barr virus genomes were detected in the small lymphocytes of two of the three cases examined by in situ hybridization studies. Recently, the histopathologic findings of CD have been associated with a disrupted immune response, and the present data suggest that SLE should be listed as one of the diseases showing the histopathologic features similar to CD.
Adult↗
[Effects of globulin preparations on phagocytic killing of oral streptococci].
The phagocytic bacterial activity on Streptococcus intermedius and Streptococcus oralis, clinical isolates from patients suffering from severe oral surgical infections, was evaluated by using sera and neutrophils of patients who had been treated with polyethylene glycal treated human normal immunoglobulin (Venoglobulin-IH). The phagocytic function of the neutrophils against the two strains, approximately 10 cells/neutrophil, was unrelated to the administration of Venoglobulin-IH. The bactericidal function of the serum and neutrophils on the third day following Venoglobulin-IH administration, on the other hand, rose to 10 times that of the sera and neutrophils of patients with mild infections. The agglutinin titer of Venoglobulin-IH against S. intermedius and S. oralis were 32 and 16 times, respectively, indicating an evident rise in these titers against the clinical isolates in comparison with the reference strains of the same organisms. These findings indicated that the oral streptococci that are isolated from patients with grave infections are associated with an antigenicity that is different from the normal bacterial flora; and these organisms are more labile to phagocytosis by neutrophils that were obtained from patients who have been treated with Venoglobulin-IH.
Administration, Oral↗
[Multisite evaluation of a colorimetric broth microdilution antifungal susceptibility testing].
A new colorimetric microdilution antifungal susceptibility testing using an air-dried microplate containing serially diluted antifungal agents and an oxidation-reduction dye, sodium resazurin, as a color indicator to detect the growth of yeasts was evaluated at multiple sites. Six ATCC reference strains described in the National Committee for Clinical Laboratory Standards (NCCLS) M27-T, two quality control and four reference strains, were repeatedly tested against five antifungal agents, amphotericin B(AMPH), flucytosine (5-FC), fluconazole (FCZ), miconazole (MCZ) and itraconazole (ITZ) at 17 different laboratories. Of 1,521 MIC determinations, 1,393 (91.6%) were within the acceptable limits described in the M27-T, ranging from 23.8% to 100%. However, significant discrepancies from the M27-T limits were noted in some combinations, such as FCZ against C. albicans ATCC 90028. In addition, the MIC limits for MCZ and ITZ against the six reference strains were determined. A total of 1,645 clinical isolates of yeasts were tested against five antifungal agents. Most isolates of C. albicans were highly susceptible to the five agents with MIC90 ranging from 0.03 microgram/ml (MCZ) to 1.0 microgram/ml (AMPH). The MICs against AMPH and 5-FC were tightly clustered, except with C. krusei and Trichosporon spp. Whereas, the distribution of MICs for FCZ, MCZ and ITZ had a wide range from < or = 0.03 microgram/ml to > 64 micrograms/ml. From these results, it can be concluded that our colorimetric broth microdilution antifungal susceptibility test is reliable, easy-to-perform and a suitable alternative for the determination of the MICs in clinical microbiology laboratories.
Amphotericin B↗
[Laboratory-evaluation of antimicrobial susceptibility testings to detect vancomycin-resistant enterococci].
The emergence and dissemination of vancomycin-resistant enterococci (VRE) emphasizes the need for laboratories to be able to correctly detect them. The study described was conducted to evaluate the test methods presently available in Japan to discriminate between the isolates of VRE and those susceptible (VSE). Among the phenotypic test methods evaluated, an agar screening method which utilized 8 micrograms per ml of vancomycin in Mueller-Hinton agar plate appeared to have a sufficient accuracy. When 23 isolates of vanA positive, 31 of vanB positive, 4 of both positive and 60 of both negative were tested, the sensitivity and specificity to detect VRE were estimated to be 98.3% and 100%, respectively. Also, all the VRE isolates were interpreted as being resistant or intermediate by the E test recently approved in Japan, when the results were read after 48 hr-incubation. Whereas, two disk diffusion tests, Showa disk and NCCLS-based Sensi-disc, were evaluated, but both methods failed to discriminate between VRE and VSE, in particular, between the isolates with vanB positive and negative. The automated system, Vitek GPS-TA produced high frequencies of very major errors; 8.7% for vanA positives and 58% for vanB positives. A total of 1,214 enterococcal isolates from multisite laboratories in Japan, comprising 7 different species, were first tested onto agar screening test plates, but none of isolates represented phenotypic vancomycin resistance. With these results, it can be recommended to detect VRE in clinical microbiology laboratories as follows: First, all the enterococcal isolates will be tested onto the agar screening plates or by the E test. Then, if the isolate is interpreted as being resistant or intermediate, the laboratory should confirm whether it is positive for vanA or vanB by polymerase chain reaction (PCR) specified.
Anti-Bacterial Agents↗
[Determination of in vitro synergy when amphotericin B is combined with various antimicrobial agents against yeasts by using a colorimetric microdilution checkerboard].
We determined in vitro interactions when amphoteric B (AMPH) is combined with various antimicrobial agents against yeast by using a colorimetric microdilution transfer plate technique, principally based on the current National Committee for Clinical Laboratory Standards (NCCLS) M27-T. An oxidation-reduction dye, sodium resazurin, was used as a color indicator to detect the growth of yeasts, and a standard two-dimensional, two-agent microdilution checkerboard in RPMI 1640 was employed to determine in vitro interactions; synergistic, indifferent or antagonistic. The study included 125 clinical isolates of Candida species and nine reference strains of American Type Culture Collection described in M27-T. Among the 34 antimicrobial agents first tested, polymyxin B (PL), rifampicin (RFP), tetracycline (TC) and erythromycin (EM) showed significant synergism. The fractional inhibitory concentration (FIC) indices of the respective agents were; PL 0.16 to 0.51 (mean, 0.315), RFP 0.13 to 0.56 (0.255), TC 0.06 to 0.75 (0.353) and EM 0.27 to 1.0 (0.550). RFP was the most potent agent, 118 of 125 clinical isolates (94.4%) showing synergism (FIC, < or = 0.5). With these results, we can conclude that several antibacterial agents are potentially effective when combined with AMPH against yeasts, probably due to alteration in the permeability barrier of the surface membrane by AMPH. Antifungal synergism may be promising for more effective, and less toxic therapy, and thus in vivo study will be necessary to determine their clinical significance.
Amphotericin B↗
[Inactivated influenza virus vaccine: the status quo and several new approaches for future application].
Inactivated influenza vaccines were first developed in the 1930s and then, more efficient methods of virus purification and disruption have led to less toxic vaccine, e.g. HA-split vaccine. A program of annual immunization to schoolchildren had been recommended in Japan since 1960. This assumed that the high incidence of influenza morbidity among schoolchildren might play a role in extending influenza in general population. However, due to the lack of detailed data, the public had bee reluctant to accept, especially for healthy children. Accordingly, the Japanese government changed its policy to vaccinate on demand, which resulted in a marked decrease of vaccinees. Several alternative approaches are in progress to produce more efficient vaccines. DNA vaccine, plasmid DNA encoding influenza peptides, seems promising for future application. However, as it will take more years for clinical evaluation, it is urgent to design a protocol to estimate cost-effectiveness of the presently available inactivated vaccine.
Child↗
[Clinical microbiology--main simple instruments and reagents in the field of clinical microbiology].
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Fluorescent Antibody Technique↗