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1.
Nihon Hinyokika Gakkai Zasshi ; 108(1): 24-29, 2017.
Artigo em Japonês | MEDLINE | ID: mdl-29367505

RESUMO

(Objectives) We report the clinical features about polymicrobial bacteria detection cases in the uncomplicated urinary tract infection of the premenopausal woman from the voided midstream urine culture. (Methods) We retrospectively reviewed the premenopausal woman from 18-49 years patients visited Sendai City Hospital from April, 2006 to December, 2014, diagnosed uncomplicated cystitis or uncomplicated pyelonephritis. We analyzed for 375 specimens from the voided midstream urine culture. (Results) Among 375 specimens, the urine culture-positive for uropathogens were 211 specimens. The monomicrobial bacterial were detected in 184 specimens (87.2%) and polymicrobial bacterial specimens were 27 specimens (12.8%). The most combination group was the caused bacteria and periurethral microorganisms in 20 specimens (74.1%). Then 6 periurethral microorganisms specimens (22.2%), the caused bacteria were only 1 specimen was overlapped (3.7%). The case of urinary tract infections recurrence or revealed voiding dysfunction that need periodic treatment were more prevalent in the polymicrobial than the monomicrobial group (22.2% vs 9.8%, p=0.043). (Conclusions) When polymicrobial bacteria were detected in uncomplicated urinary tract infection in premenopausal woman, it was confirmed that there were the most combinations of caused bacteria and periurethral microorganisms. In these cases, treatment intended for only the caused bacteria. A risk of the infection recurrence and voiding dysfunction were statistically significant higher rate in the polymicrobial bacteria detection cases, and it might be necessary to consider that search to complicated urinary tract infection.


Assuntos
Bacteriúria/microbiologia , Escherichia coli/isolamento & purificação , Klebsiella pneumoniae/isolamento & purificação , Pré-Menopausa , Proteus mirabilis/isolamento & purificação , Infecções Urinárias/microbiologia , Adolescente , Adulto , Corynebacterium/isolamento & purificação , Feminino , Humanos , Pessoa de Meia-Idade , Staphylococcus/isolamento & purificação , Staphylococcus saprophyticus/isolamento & purificação , Infecções Urinárias/complicações , Transtornos Urinários/etiologia , Transtornos Urinários/microbiologia , Adulto Jovem
2.
J Infect Chemother ; 21(11): 828-30, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-26188420

RESUMO

Parvimonas micra was renamed species as within Gram-positive anaerobic cocci and rarely causes severe infections in healthy people. We report the first confirmed case of spondylodiscitis with epidural abscess caused by P. micra in a healthy women. The patient has a pain in low back and anterior left thigh. Magnetic resonance imaging and computed tomography detected the affected lesion at the L2 and L3 vertebral bodies. All isolates from the surgical and needle biopsy specimens were identified as P. micra by 16S rRNA and MALDI-TOF. In this case, P. micra showed high sensitivity to antimicrobial therapy. She was successfully treated with debridement and sulbactam/ampicillin, followed by oral metronidazole for a total of 10 weeks. The causative microorganisms of spondylodiscitis are not often identified, especially anaerobic bacteria tend to be underestimated. On the other hand, antimicrobial therapy for spondylodiscitis is usually prolonged. Accordingly, we emphasize the importance of performing accurate identification including anaerobic bacteria.


Assuntos
Discite , Abscesso Epidural , Firmicutes , Discite/diagnóstico , Discite/microbiologia , Discite/fisiopatologia , Abscesso Epidural/diagnóstico , Abscesso Epidural/microbiologia , Abscesso Epidural/fisiopatologia , Feminino , Humanos , Pessoa de Meia-Idade
3.
J Infect Chemother ; 21(2): 138-40, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25240269

RESUMO

Culture-negative endocarditis remains a diagnostic and therapeutic challenge despite recent medical advances. Streptococcus tigurinus, a novel member of the Streptococcus mitis group, was first identified in Zurich. S. tigurinus possesses virulence determinants and causes invasive infections. We report a case of culture-negative endocarditis with serious complications due to S. tigurinus, which was identified by 16S ribosomal RNA gene sequence analysis of excised valve tissue specimens. This technique is useful for identification of the causative microorganism in patients with culture-negative endocarditis and may facilitate early diagnosis and appropriate antimicrobial treatment.


Assuntos
Endocardite Bacteriana/microbiologia , Infecções Estreptocócicas/microbiologia , Streptococcus/genética , Adulto , DNA Bacteriano/genética , Valvas Cardíacas/microbiologia , Valvas Cardíacas/cirurgia , Humanos , Masculino , RNA Ribossômico 16S/genética , Streptococcus/isolamento & purificação
4.
J Infect Chemother ; 21(10): 729-36, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26271590

RESUMO

Molecular characterization of community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) is generally conducted referred to staphylococcal cassette chromosome mec (SCCmec) type IV or V. CA-MRSA is now a cause of concern since such strains have been isolated not only from individuals in a community but also from patients in healthcare settings. The aim of this study was to analyze microbiological and molecular epidemiological features of CA-MRSA strains at a Japanese tertiary care hospital using PCR based-open reading frame typing (POT). This technique allows for molecular classification into CA-MRSA (POT-CA) and hospital-associated (HA-) MRSA (POT-HA) with clonal discrimination. Clinical MRSA isolates obtained from consecutive patients between October 1, 2012 and September 30, 2013 at the hospital were analyzed in combination with the clinical definition for CA-MRSA by the Centers for Disease Control and Prevention and POT. Of 219 isolates (76 clonal groups), 64 (29.3%) were clinical-HA/POT-CA isolates (22 clonal groups). Some clones of them accumulated in this hospital and might be involved in nosocomial transmission. Virulent factors of the isolates were analyzed, and only one (1.6%) Panton-Valentine leukocidin gene positive isolate but no arginine catabolic mobile element genes positive isolate were found in clinical-HA/POT-CA. Additionally, clinical-HA/POT-CA isolates showed higher antimicrobial susceptibility than clinical-HA/POT-HA, especially to minocycline, doxycycline, and amikacin. The most frequent genotype of molecular CA-MRSA was multi-locus sequence type 5-SCCmecIV, previously not detected in Japan. Although CA-MRSA at this hospital showed low virulence and higher antimicrobial susceptibility, the risk of nosocomial infection from them should be recognized, requiring stricter infection control measures.


Assuntos
Staphylococcus aureus Resistente à Meticilina , Infecções Estafilocócicas/microbiologia , Infecções Comunitárias Adquiridas , Humanos , Japão/epidemiologia , Staphylococcus aureus Resistente à Meticilina/genética , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Staphylococcus aureus Resistente à Meticilina/metabolismo , Staphylococcus aureus Resistente à Meticilina/patogenicidade , Epidemiologia Molecular , Reação em Cadeia da Polimerase , Infecções Estafilocócicas/epidemiologia , Centros de Atenção Terciária , Virulência
5.
J Clin Microbiol ; 52(3): 911-5, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24403303

RESUMO

Acinetobacter baumannii is generally the most frequently isolated Acinetobacter species. Sequence analysis techniques allow reliable identification of Acinetobacter isolates at the species level. Forty-eight clinical isolates of Acinetobacter spp. were obtained from blood cultures at Tohoku University Hospital. These isolates were identified at the species level by partial sequencing of the RNA polymerase ß-subunit (rpoB), 16S rRNA, and gyrB genes. Then further characterization was done by using the PCR for detection of OXA-type ß-lactamase gene clusters, metallo-ß-lactamases, and carO genes. Pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing were also performed. The most frequent isolate was Acinetobacter soli (27.1%). Six of the 13 A. soli isolates were carbapenem nonsusceptible, and all of these isolates produced IMP-1. PFGE revealed that the 13 A. soli isolates were divided into 8 clusters. This study demonstrated that A. soli accounted for a high proportion of Acinetobacter isolates causing bacteremia at a Japanese tertiary hospital. Non-A. baumannii species were identified more frequently than A. baumannii and carbapenem-nonsusceptible isolates were found among the non-A. baumannii strains. These results emphasize the importance of performing epidemiological investigations of Acinetobacter species.


Assuntos
Infecções por Acinetobacter/epidemiologia , Infecções por Acinetobacter/microbiologia , Acinetobacter/isolamento & purificação , Bacteriemia/epidemiologia , Bacteriemia/microbiologia , Infecção Hospitalar/epidemiologia , Infecção Hospitalar/microbiologia , Acinetobacter/classificação , Proteínas da Membrana Bacteriana Externa/genética , Sangue/microbiologia , Análise por Conglomerados , DNA Girase/genética , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , RNA Polimerases Dirigidas por DNA/genética , Eletroforese em Gel de Campo Pulsado , Genótipo , Humanos , Japão/epidemiologia , Tipagem de Sequências Multilocus , Filogenia , Prevalência , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Centros de Atenção Terciária , beta-Lactamases/genética
6.
J Infect Chemother ; 20(8): 498-501, 2014 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-24861538

RESUMO

In cooperation with the Miyagi prefectural government, we conducted a survey of the management of sanitation at evacuation centers and the health of the evacuees by visiting 324 evacuation centers at two weeks after the 2011 Great East Japan Earthquake. The facilities often used as evacuation centers were community centers (36%), schools (32.7%) and Nursing homes (10.2%). It was more difficult to maintain a distance of at least 1 m between evacuees at the evacuation centers with a larger number of residents. At evacuation centers where the water supply was not restored, hygienic handling of food and the hand hygiene of the cooks were less than adequate. Among evacuation centers with ≤50 evacuees, there was a significant difference in the prevalence rate of digestive symptoms between the centers with and without persons in charge of health matters (0.3% vs. 2.1%, respectively, p < 0.001). The following three factors had an important influence on the level of sanitation at evacuation centers and the health of evacuees: 1) the size of the evacuation center, 2) the status of the water supply, and 3) the allocation of persons in charge of health matters. Given that adjusting the number of evacuees to fit the size of the evacuation center and prompt restoration of the water supply are difficult to achieve immediately after an earthquake, promptly placing persons in charge of health matters at evacuation centers is a practicable and effective measure, and allocation of at least one such person per 50 evacuees is desirable.


Assuntos
Doenças Transmissíveis/epidemiologia , Terremotos/história , Abrigo de Emergência/estatística & dados numéricos , Saneamento/estatística & dados numéricos , História do Século XXI , Humanos , Japão/epidemiologia , Fatores de Risco
7.
Tohoku J Exp Med ; 234(2): 111-6, 2014 10.
Artigo em Inglês | MEDLINE | ID: mdl-25224029

RESUMO

Susceptible healthcare personnel (HCP) are at high risk for acquiring and transmitting measles, mumps, rubella, and varicella (MMRV). Presumptive evidence of immunity to MMRV is recommended for HCP. The aim of this investigation was to examine the seroprevalence of MMRV in Japanese HCP and the association with history or vaccination in terms of occupational safety. To improve infection control at our hospital, we also assessed their immune status by implementing prevaccination antibody screening and an immunization program with postvaccination serological testing. We implemented seroprevalence surveys on MMRV antibodies among 243 newly and 2,664 previously hired HCP in a Japanese tertiary care hospital. Self-administered questionnaires about history of MMRV and vaccination with or without written documentation were completed for newly hired HCP. Prevaccination and postvaccination serological tests were performed using virus-specific IgG enzyme-linked immunosorbent assays. Indeed, only a few HCP accurately remembered or had written records of their disease or vaccination history. After our immunization program was implemented, the seropositivity rate reached levels as high as ~98% for measles, rubella, and varicella, and increased to ~80% for mumps. Our program was cost-effective, and no severe adverse reactions were reported. The prevaccination antibody screening for HCP would be helpful, given the lack of written vaccination records or documented disease history, and is also useful for the prevention of adverse reactions associated with unnecessary vaccination. It is important for infection control practitioners to comprehend the immune status of HCP against MMRV, and then provide an appropriate immunization program for susceptible HCP.


Assuntos
Vacina contra Varicela/uso terapêutico , Varicela/prevenção & controle , Pessoal de Saúde , Vacina contra Sarampo-Caxumba-Rubéola/uso terapêutico , Sarampo/prevenção & controle , Caxumba/prevenção & controle , Rubéola (Sarampo Alemão)/prevenção & controle , Adulto , Idoso , Anticorpos Antivirais/sangue , Vacina contra Varicela/efeitos adversos , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Programas de Imunização , Japão , Masculino , Vacina contra Sarampo-Caxumba-Rubéola/efeitos adversos , Pessoa de Meia-Idade , Estudos Soroepidemiológicos , Centros de Atenção Terciária , Vacinação , Adulto Jovem
8.
Emerg Infect Dis ; 19(5): 799-801, 2013 May.
Artigo em Inglês | MEDLINE | ID: mdl-23648069

RESUMO

Tuberculosis was diagnosed in a person who had stayed in a shelter after the 2011 Great East Japan Earthquake. A contact investigation showed that the prevalence of latent tuberculosis infection among other evacuees at the shelter was 20%. Our report underscores the importance of tuberculosis prevention and control after natural disasters.


Assuntos
Terremotos , Tuberculose Latente/epidemiologia , Mycobacterium tuberculosis/isolamento & purificação , Tuberculose Pulmonar/epidemiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Desastres , Feminino , Humanos , Japão/epidemiologia , Tuberculose Latente/diagnóstico , Tuberculose Latente/transmissão , Masculino , Pessoa de Meia-Idade , Habitação Popular , Tuberculose Pulmonar/diagnóstico , Tuberculose Pulmonar/transmissão
9.
Rinsho Byori ; 61(12): 1160-5, 2013 Dec.
Artigo em Japonês | MEDLINE | ID: mdl-24605551

RESUMO

On 11 March 2011, an earthquake measuring 9.0 on the Richter scale occurred off the northeast coast of Honshu Island, Japan, produced a devastating tsunami that destroyed many towns and villages near the coast in Iwate, Miyagi, and Fukushima prefectures. Miyagi Prefecture was the area most severely devastated by the tsunami, with extensive loss of life and property; hundreds of thousands of people lost their houses and were forced to move to evacuation areas. In the days and weeks following devastating natural disasters, the threat of infectious disease outbreak is high. We initiated cooperation and support activities in terms of infection control at evacuation centers in the aftermath of the disaster. For example, we assessed sanitary and infectious risk factors in evacuation centers, in collaboration with Miyagi Prefectural Government and public health centers in the devastated area, to prevent the transmission of infectious diseases among evacuees. We also supported the control of two outbreaks of influenza A, which occurred in different centers in Miyagi Prefecture in the early period after the disaster. Both outbreaks subsided without any complicated or fatal cases of influenza as a result of the prompt implementation of a systemic approach with a bundle of control measures.


Assuntos
Desastres , Surtos de Doenças/prevenção & controle , Terremotos , Controle de Infecções , Equipe de Assistência ao Paciente , Humanos , Controle de Infecções/métodos , Japão , Tsunamis
10.
Clin Infect Dis ; 54(1): e5-7, 2012 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-21976468

RESUMO

We describe 2 post-tsunami outbreaks of influenza A in evacuation centers in Miyagi Prefecture, Japan, in 2011. Although containment of the outbreak was challenging in the evacuation settings, prompt implementation of a systemic approach with a bundle of control measures was important to control the influenza outbreaks.


Assuntos
Surtos de Doenças , Influenza Humana/epidemiologia , Tsunamis , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Pré-Escolar , Controle de Doenças Transmissíveis/métodos , Feminino , Humanos , Japão/epidemiologia , Masculino , Pessoa de Meia-Idade , Adulto Jovem
11.
J Antimicrob Chemother ; 67(7): 1623-6, 2012 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-22447879

RESUMO

OBJECTIVES: Acinetobacter baumannii presents a clinical challenge when it is non-susceptible to carbapenems. The prevalence of carbapenem-non-susceptible A. baumannii in Japan is unclear, as previous studies have been limited in scope. We investigated the spread of carbapenem-non-susceptible A. baumannii in Japan and performed a comparison with findings from overseas. METHODS: A total of 305 non-duplicate clinical isolates of Acinetobacter spp. from 176 medical facilities in all geographical regions of Japan were tested for susceptibility to antimicrobial agents by the agar dilution method. Isolates with MICs of imipenem ≥ 4 mg/L underwent PCR analysis of OXA-type ß-lactamase gene clusters and metallo-ß-lactamase genes. These isolates were further analysed by sequencing of OXA-type ß-lactamases and by multilocus sequence typing (MLST). RESULTS: Fifty-five of the 305 clinical isolates had MICs of imipenem ≥ 4 mg/L. The OXA-51-like carbapenemase gene was detected in 52 of these 55 isolates. Within the OXA-51-like gene cluster, OXA-66 was found in 43 (82.7%) of the 52 isolates. MLST identified the following sequence types (STs): ST74, ST76, ST92, ST106, ST188 and ST195 in 2 (3.8%), 2 (3.8%), 40 (76.9%), 5 (9.6%), 2 (3.8%) and 1 (1.9%) of the isolates, respectively. In particular, ST92 was found in 31 (91.2%) of the 34 A. baumannii isolates with MICs of imipenem ≥ 16 mg/L. CONCLUSIONS: This is the first report on the molecular epidemiology of A. baumannii with MICs of imipenem ≥ 4 mg/L in Japan. OXA-66 and ST92 were dominant among these isolates.


Assuntos
Infecções por Acinetobacter/epidemiologia , Infecções por Acinetobacter/microbiologia , Acinetobacter baumannii/classificação , Acinetobacter baumannii/efeitos dos fármacos , Antibacterianos/farmacologia , Carbapenêmicos/farmacologia , Resistência beta-Lactâmica , Acinetobacter baumannii/genética , Acinetobacter baumannii/isolamento & purificação , DNA Bacteriano/química , DNA Bacteriano/genética , Genótipo , Humanos , Japão/epidemiologia , Testes de Sensibilidade Microbiana , Epidemiologia Molecular , Tipagem de Sequências Multilocus
12.
Int Immunol ; 23(2): 97-108, 2011 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-21172897

RESUMO

Invariant NK T (iNKT) cells are known to play a critical role in the regulation of inflammatory responses in various clinical settings. In the present study, we assessed the contribution of iNKT cells to the development of acute lung injury (ALI), which was caused by intra-tracheal administration of LPS. Jα18 gene-disrupted mice lacking these cells underwent neutrophilic inflammatory responses in lungs at an equivalent level as control mice. Next, mice were sensitized intra-tracheally with α-galactosylceramide, an activator of iNKT cells, followed by challenge with LPS. In this model, mice showed severe lung injury, and all mice were killed within 72 h after LPS injection. IFN-γ and tumor necrosis factor (TNF)-α were strikingly elevated in the lungs of these mice. Administration of neutralizing mAb against IFN-γ and TNF-α attenuated lung injury in a histopathological analysis and improved their survival rate. Flow cytometric analysis revealed that IFN-γ was expressed in NK cells, iNKT cells and also Gr-1(dull+)Ly-6C(+) monocytes and TNF-α was detected mainly in Gr-1(bright+)Ly-6G(+) neutrophils and Gr-1(dull+)Ly-6C(+) monocytes. Otherwise, in mice treated with LPS alone, IFN-γ was not detected in the lungs and Gr-1(bright+)Ly-6G(+) neutrophil was a main cellular source of TNF-α production. Anti-Gr-1 mAb resulted in the attenuation of ALI and decrease in the level of these cytokines. These results indicated that activation of iNKT cells led to striking exacerbation of ALI caused by LPS and that Gr-1(+) monocytes were recruited in the lungs with expressing IFN-γ and TNF-α and played an important role in the development of these responses.


Assuntos
Lesão Pulmonar Aguda/fisiopatologia , Interferon gama/imunologia , Lipopolissacarídeos/farmacologia , Monócitos/imunologia , Células T Matadoras Naturais/imunologia , Receptores de Quimiocinas/imunologia , Fator de Necrose Tumoral alfa/imunologia , Lesão Pulmonar Aguda/induzido quimicamente , Lesão Pulmonar Aguda/imunologia , Animais , Pulmão/efeitos dos fármacos , Pulmão/imunologia , Ativação Linfocitária , Camundongos , Camundongos Knockout , Camundongos Transgênicos
13.
J Infect Chemother ; 18(3): 347-51, 2012 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-22116462

RESUMO

It is not clear whether antipseudomonal agents can kill cell-associated bacteria within a short time. Madin-Darby canine kidney (MDCK) and A549 cells were infected with Pseudomonas aeruginosa ATCC 27853 and PAO1 and the bactericidal activity of ceftazidime, imipenem, meropenem, gentamicin, and ciprofloxacin against the organisms was investigated. In both MDCK and A549 cells, ß-lactams could not kill epithelial cell-associated bacteria within 2 h. Gentamicin at concentrations ≤32 µg/ml killed more than 99% of epithelial cell-associated bacteria. Ciprofloxacin at 0.5 µg/ml killed more than 99.9% of MDCK cell-associated bacteria. Ciprofloxacin has the strongest and most rapid bactericidal activity against epithelial cell-associated bacteria, which may be explained by the combination of potent in-vitro bactericidal activity and high penetration ability into epithelial cells.


Assuntos
Antibacterianos/farmacologia , Células Epiteliais/microbiologia , Testes de Sensibilidade Microbiana/métodos , Pseudomonas aeruginosa/efeitos dos fármacos , Animais , Linhagem Celular Transformada , Linhagem Celular Tumoral , Ciprofloxacina/farmacologia , Cães , Gentamicinas/farmacologia , Humanos , Imipenem/farmacologia , Células Madin Darby de Rim Canino
14.
Artigo em Japonês | MEDLINE | ID: mdl-23547484

RESUMO

On 11 March 2011, an earthquake measuring 9.0 on the Richter scale off the northeast coast of Honshu Island, Japan, produced a devastating tsunami that destroyed many towns and villages near the coast in Iwate, Miyagi, and Fukushima prefectures. Miyagi Prefecture was the area most severely devastated by the tsunami, with extensive loss of life and property; hundreds of thousands of people lost their houses and were forced to move to evacuation areas. In the days and weeks following devastating natural disasters, the threat of infectious disease outbreak is high. Rapid diagnostic tests can be performed at or near the site of patient care and the tests were very useful in this disaster, because they enabled us to manage patients appropriately in the settings where medical resources were limited. Here we report actual cases where the rapid diagnostic tests for infectious diseases were useful in the patient management.


Assuntos
Terremotos , Infecções/diagnóstico , Humanos , Japão , Tsunamis
15.
J Antimicrob Chemother ; 66(10): 2255-62, 2011 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-21733965

RESUMO

OBJECTIVES: Extended-spectrum ß-lactamases (ESBLs) have become a problem among AmpC-producing Enterobacteriaceae and the emergence of concomitant quinolone resistance in ß-lactamase-producing isolates poses a global threat. In this study we investigated the prevalence and regional variation of ESBLs in Japanese clinical isolates of Citrobacter spp. and analysed plasmid-mediated quinolone resistance (PMQR) determinants in ESBL-producing Citrobacter spp. METHODS: A total of 348 clinical isolates of Citrobacter spp. collected throughout Japan were studied. Screening and the boronic acid disc test were performed to detect ESBLs in Citrobacter spp. with chromosomal AmpC ß-lactamases. PCR and sequencing were done to identify ESBL and PMQR genes. For ESBL-producing Citrobacter spp., PFGE was performed using the SfiI restriction enzyme. RESULTS: The number of ESBL-producing isolates confirmed phenotypically was 67 (19.3%). The prevalence of ESBL-producing Citrobacter koseri was significantly higher (32.1%) than that of ESBL-producing Citrobacter freundii (4.6%) (P < 0.01). Moreover, the prevalence of ESBLs was notably higher among C. koseri from southern Japan (60.0%). CTX-M-2 was predominant in C. koseri. Of the ESBL-producing C. koseri analysed, 23.2% possessed PMQR determinants, and there was a significant association between qnrB4 and bla(SHV-12). The 57 ESBL-producing Citrobacter spp. possessing bla(CTX-M), bla(SHV) or bla(TEM) were divided into 18 unique PFGE types. CONCLUSIONS: This is the first report about the prevalence of PMQR determinants among ESBL-producing Citrobacter spp. from Japan. Our data suggest that ESBLs and PMQR determinants are spreading among C. koseri in Japan.


Assuntos
Citrobacter/enzimologia , Citrobacter/genética , Farmacorresistência Bacteriana Múltipla/genética , beta-Lactamases/genética , Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Sequência de Bases , Citrobacter/efeitos dos fármacos , Citrobacter/isolamento & purificação , Infecções por Enterobacteriaceae/tratamento farmacológico , Infecções por Enterobacteriaceae/epidemiologia , Infecções por Enterobacteriaceae/microbiologia , Humanos , Japão , Testes de Sensibilidade Microbiana , Quinolonas/farmacologia , Análise de Sequência de DNA , beta-Lactamases/metabolismo
16.
J Antimicrob Chemother ; 66(3): 494-8, 2011 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-21172783

RESUMO

OBJECTIVES: Streptococcus pyogenes causes various diseases in humans. While the prevalence of fluoroquinolone-resistant S. pyogenes isolates has been increasing since 2000 in the USA and Europe, it has remained very low in Japan. We isolated a fluoroquinolone-resistant S. pyogenes strain and analysed its genetics. METHODS: TU-296, a strain of S. pyogenes resistant to levofloxacin (MIC 16 mg/L), was isolated from the throat of a patient in their thirties with pharyngitis in autumn 2007. We carried out susceptibility tests for various antimicrobial agents and PCR analysis of the genes gyrA, gyrB, parC and parE in the quinolone resistance-determining region, followed by sequencing of the PCR products to find mutation(s) and the resulting amino acid substitution(s). We then sequenced the PCR product of the emm gene and determined the emm genotype. RESULTS: S. pyogenes TU-296 was found to have the following mutations and amino acid substitutions: adenine 476 to cytosine in gyrA and cytosine 367 to thymine in parC, resulting in Glu-85→Ala in GyrA and Ser-79→Phe in ParC. The genotype of the isolate was emm11. CONCLUSIONS: Amino acid substitutions in fluoroquinolone-resistant S. pyogenes have already been reported from Europe and the USA, including Ser-81→Phe or Tyr and Met-99→Leu in GyrA, as well as Ser-79→Phe, Tyr or Ala and others in ParC. Numerous point mutations were found in parC and parE of S. pyogenes TU-296. In addition, a new amino acid substitution was detected (Glu-85→Ala in GyrA). To our knowledge, there have been no previous reports of this substitution in a clinical isolate of S. pyogenes.


Assuntos
Substituição de Aminoácidos/genética , Antibacterianos/farmacologia , Farmacorresistência Bacteriana , Fluoroquinolonas/farmacologia , Mutação Puntual , Streptococcus pyogenes/efeitos dos fármacos , Streptococcus pyogenes/genética , Adulto , Antígenos de Bactérias/genética , Proteínas da Membrana Bacteriana Externa/genética , Proteínas de Transporte/genética , DNA Girase/genética , DNA Topoisomerase IV/genética , DNA Bacteriano/química , DNA Bacteriano/genética , Humanos , Japão , Testes de Sensibilidade Microbiana , Faringite/microbiologia , Faringe/microbiologia , Reação em Cadeia da Polimerase , Análise de Sequência de DNA , Infecções Estreptocócicas/microbiologia , Streptococcus pyogenes/isolamento & purificação
17.
Tohoku J Exp Med ; 220(1): 15-9, 2010 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-20046047

RESUMO

Pseudomonas (P.) aeruginosa is a major opportunistic pathogen especially in immunocompromised patients. To evaluate the invasiveness of respiratory pathogens, we developed monolayer culture systems and examined the degree of invasion by P. aeruginosa and invasive Salmonella (S.) typhimurium strains using human respiratory cell lines: A549 (derived from lung cancer), BEAS-2B (normal bronchial epithelium), and Calu-3 (pleural effusion of a patient with adenocarcinoma of the lung). Cells were seeded into filter units containing 0.33 cm(2) filter membranes with 3.0 microm pores, and were incubated at 37 degrees C under 5% CO(2) for 4-10 days. By monitoring the trans-monolayer electrical resistance (TER), we judged that BEAS-2B cells (TER values: 436.2 +/- 16.8 to 628.8 +/- 66.3 Omega cm(2)) and Calu-3 cells (TER values: 490.5 +/- 25.2 to 547.8 +/- 21.6 Omega cm(2)) formed monolayers with tight junctions, but not A549 cells. On day 8 of culture, monolayer cultures were infected with bacteria, and the number of microorganisms penetrating into the basolateral medium was counted. Wild-type P. aeruginosa PAO1 (PAO1 WT) and S. typhimurium SL1344 were detected in the basolateral medium of BEAS-2B monolayer system by 3 h after inoculation, while only P. aeruginosa PAO1 WT was detected in the basolateral medium of Calu-3 monolayer, indicating poor invasiveness of S. typhimurium SL1344 in the Calu-3 system. These findings suggest that BEAS-2B or Calu-3 monolayer system could be useful for evaluating the invasiveness of respiratory pathogens. Because of the difference in bacterial invasiveness, we may need to choose a suitable cell system for each target pathogen.


Assuntos
Bactérias/patogenicidade , Técnicas de Cultura de Células/métodos , Células Epiteliais/citologia , Células Epiteliais/microbiologia , Sistema Respiratório/citologia , Sistema Respiratório/microbiologia , Animais , Linhagem Celular , Separação Celular , Cães , Impedância Elétrica , Humanos
19.
Antimicrob Agents Chemother ; 53(10): 4225-30, 2009 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-19651910

RESUMO

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 at < or = 0.5 microg/ml (susceptible). A disk diffusion breakpoint for piperacillin-tazobactam of > or = 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.


Assuntos
Ampicilina/farmacologia , Antibacterianos/farmacologia , Haemophilus influenzae/efeitos dos fármacos , beta-Lactamases/farmacologia , Combinação Amoxicilina e Clavulanato de Potássio , Ciprofloxacina/farmacologia , Farmacorresistência Bacteriana Múltipla/genética , Haemophilus influenzae/genética , Testes de Sensibilidade Microbiana , Mutação
20.
Tohoku J Exp Med ; 217(2): 117-21, 2009 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-19212104

RESUMO

(1-3)-beta-D-glucan (BDG) is a cell-wall polysaccharide component found in most fungi. The measurement of BDG is a useful diagnostic marker for invasive fungal infections. However, it is well known that interfering substances can result in false positive reactions. We encountered a patient who underwent lung transplantation and presented with highly elevated BDG values, despite having no evidence of invasive fungal infection. We therefore hypothesized that elevated BDG values were originated from the gauze products used during surgery. While it is known that gauze products contain BDG, there have been no previous reports to quantitatively correlate amount of gauze usage and BDG levels. In this study, we extracted BDG from various gauze products and measured BDG to better understand the degree of which gauze contributes to elevated BDG values. Six types of commonly used surgical gauze products were selected for our study. Each of the surgical gauze was immersed in sterile, purified water for up to 120 minutes. At set intervals, BDG values in the water extracts were measured. Purified water samples without gauze were used as negative controls (< 4 pg/ml). After 120-minute extraction, BDG levels varied greatly depending on gauze products, ranging from 11.7 pg/ml to 6612 pg/ml. The gauze made of lyocell, which is a fiber produced from wood pulp cellulose, yielded the lowest levels of BDG, and probably would not cause false positive for fungal infections. There is a need for the development of a gauze product that does not contribute to elevated BDG values.


Assuntos
Curativos Oclusivos , beta-Glucanas/análise , Humanos , Fatores de Tempo
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