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1.
Nucleic Acids Res ; 50(21): 12543-12557, 2022 11 28.
Artigo em Inglês | MEDLINE | ID: mdl-36454022

RESUMO

Several basic leucine zipper (bZIP) transcription factors have accessory motifs in their DNA-binding domains, such as the CNC motif of CNC family or the EHR motif of small Maf (sMaf) proteins. CNC family proteins heterodimerize with sMaf proteins to recognize CNC-sMaf binding DNA elements (CsMBEs) in competition with sMaf homodimers, but the functional role of the CNC motif remains elusive. In this study, we report the crystal structures of Nrf2/NFE2L2, a CNC family protein regulating anti-stress transcriptional responses, in a complex with MafG and CsMBE. The CNC motif restricts the conformations of crucial Arg residues in the basic region, which form extensive contact with the DNA backbone phosphates. Accordingly, the Nrf2-MafG heterodimer has approximately a 200-fold stronger affinity for CsMBE than canonical bZIP proteins, such as AP-1 proteins. The high DNA affinity of the CNC-sMaf heterodimer may allow it to compete with the sMaf homodimer on target genes without being perturbed by other low-affinity bZIP proteins with similar sequence specificity.


Assuntos
Regulação da Expressão Gênica , Fator 2 Relacionado a NF-E2 , Fator 2 Relacionado a NF-E2/genética , Fator 2 Relacionado a NF-E2/metabolismo , Fatores de Transcrição de Zíper de Leucina Básica/metabolismo , DNA/genética
2.
Angew Chem Int Ed Engl ; : e202409973, 2024 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-38837490

RESUMO

Prenylation of peptides is widely observed in the secondary metabolites of diverse organisms, granting peptides unique chemical properties distinct from proteinogenic amino acids. Discovery of prenylated peptide agents has largely relied on isolation or genome mining of naturally occurring molecules. To devise a platform technology for de novo discovery of artificial prenylated peptides targeting a protein of choice, here we have integrated the thioether-macrocyclic peptide (teMP) library construction/selection technology, so-called RaPID (Random nonstandard Peptides Integrated Discovery) system, with a Trp-C3-prenyltransferase KgpF involved in the biosynthesis of a prenylated natural product. This unique enzyme exhibited remarkably broad substrate tolerance, capable of modifying various Trp-containing teMPs to install a prenylated residue with tricyclic constrained structure. We constructed a vast library of prenylated teMPs and subjected it to in vitro selection against a phosphoglycerate mutase. This selection platform has led to the identification of a pseudo-natural prenylated teMP inhibiting the target enzyme with an IC50 of 30 nM. Importantly, the prenylation was essential for the inhibitory activity, enhanced serum stability, and cellular uptake of the peptide, highlighting the benefits of peptide prenylation. This work showcases the de novo discovery platform for pseudo-natural prenylated peptides, which is readily applicable to other drug targets.

3.
J Am Chem Soc ; 144(44): 20332-20341, 2022 11 09.
Artigo em Inglês | MEDLINE | ID: mdl-36282922

RESUMO

Bioengineering of ribosomally synthesized and post-translationally modified peptides (RiPPs) is an emerging approach to explore the diversity of pseudo-natural product structures for drug discovery purposes. However, despite the initial advances in this area, bioactivity reprogramming of multienzyme RiPP biosynthetic pathways remains a major challenge. Here, we report a platform for de novo discovery of functional thiopeptides based on reengineered biosynthesis of lactazole A, a RiPP natural product assembled by five biosynthetic enzymes. The platform combines in vitro biosynthesis of lactazole-like thiopeptides and mRNA display to prepare and screen large (≥1012) combinatorial libraries of pseudo-natural products. We demonstrate the utility of the developed protocols in an affinity selection against Traf2- and NCK-interacting kinase (TNIK), a protein involved in several cancers, which yielded a plethora of candidate thiopeptides. Of the 11 synthesized compounds, 9 had high affinities for the target kinase (best KD = 1.2 nM) and 10 inhibited its enzymatic activity (best Ki = 3 nM). X-ray structural analysis of the TNIK/thiopeptide interaction revealed the unique mode of substrate-competitive inhibition exhibited by two of the discovered compounds. The thiopeptides internalized to the cytosol of HEK293H cells as efficiently as the known cell-penetrating peptide Tat (4-6 µM). Accordingly, the most potent compound, TP15, inhibited TNIK in HCT116 cells. Altogether, our platform enables the exploration of pseudo-natural thiopeptides with favorable pharmacological properties in drug discovery applications.


Assuntos
Produtos Biológicos , Produtos Biológicos/farmacologia , Produtos Biológicos/metabolismo , Processamento de Proteína Pós-Traducional , Peptídeos/química , Vias Biossintéticas , Descoberta de Drogas
4.
Antimicrob Agents Chemother ; 66(3): e0208121, 2022 03 15.
Artigo em Inglês | MEDLINE | ID: mdl-35041512

RESUMO

Limited data are available on breakthrough fungemia, defined as fungemia that develops on administration of antifungal agents, in patients with hematological disorders. We reviewed the medical and microbiological records of adult patients with hematological diseases who had breakthrough fungemia between January 2008 and July 2019 at Toranomon Hospital and Toranomon Hospital Kajigaya in Japan. A total of 121 cases of breakthrough fungemia were identified. Of the 121 involved patients, 83, 11, 5, and 22 were receiving micafungin, voriconazole, itraconazole, and liposomal amphotericin B, respectively, when the breakthrough occurred. Of the 121 causative breakthrough fungal strains, 96 were Candida species, and the rest were 13 cases of Trichosporon species, 7 of Fusarium species, 2 of Rhodotorula mucilaginosa, and 1 each of Cryptococcus neoformans, Exophiala dermatitidis, and Magnusiomyces capitatus. The crude 14-day mortality rate of breakthrough fungemia was 36%. Significant independent factors associated with the crude 14-day mortality rate were age of ≥60 years (P = 0.011), chronic renal failure (P = 0.0087), septic shock (P < 0.0001), steroid administration (P = 0.0085), and liposomal amphotericin B breakthrough fungemia (P = 0.0011). An absolute neutrophil count of >500/µL was significantly more common in candidemia in the multivariate analysis (P = 0.0065), neutropenia and nonallogeneic hematopoietic stem cell transplants were significantly more common in Trichosporon fungemia (P = 0.036 and P = 0.033, respectively), and voriconazole breakthrough fungemia and neutropenia were significantly more common in Fusarium fungemia (P = 0.016 and P = 0.016, respectively). The epidemiological and clinical characteristics of breakthrough fungemia of patients with hematological disorders were demonstrated. Some useful factors to predict candidemia, Trichosporon fungemia, and Fusarium fungemia were identified.


Assuntos
Candidemia , Cryptococcus neoformans , Fungemia , Fusarium , Doenças Hematológicas , Trichosporon , Adulto , Antifúngicos/uso terapêutico , Candida , Candidemia/tratamento farmacológico , Fungemia/tratamento farmacológico , Fungemia/microbiologia , Doenças Hematológicas/complicações , Doenças Hematológicas/tratamento farmacológico , Humanos , Pessoa de Meia-Idade
5.
J Am Chem Soc ; 143(44): 18481-18489, 2021 11 10.
Artigo em Inglês | MEDLINE | ID: mdl-34723512

RESUMO

Cyclotides are plant-derived peptides with complex structures shaped by their head-to-tail cyclic backbone and cystine knot core. These structural features underpin the native bioactivities of cyclotides, as well as their beneficial properties as pharmaceutical leads, including high proteolytic stability and cell permeability. However, their inherent structural complexity presents a challenge for cyclotide engineering, particularly for accessing libraries of sufficient chemical diversity to design potent and selective cyclotide variants. Here, we report a strategy using mRNA display enabling us to select potent cyclotide-based FXIIa inhibitors from a library comprising more than 1012 members based on the cyclotide scaffold of Momordica cochinchinensis trypsin inhibitor-II (MCoTI-II). The most potent and selective inhibitor, cMCoFx1, has a pM inhibitory constant toward FXIIa with greater than three orders of magnitude selectivity over related serine proteases, realizing specific inhibition of the intrinsic coagulation pathway. The cocrystal structure of cMCoFx1 and FXIIa revealed interactions at several positions across the contact interface that conveyed high affinity binding, highlighting that such cyclotides are attractive cystine knot scaffolds for therapeutic development.


Assuntos
Proteínas Sanguíneas/farmacologia , Ciclotídeos/farmacologia , Fator XIIa/metabolismo , Proteínas Sanguíneas/química , Ciclotídeos/química , Fator XIIa/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos
6.
Clin Infect Dis ; 67(4): 573-578, 2018 08 01.
Artigo em Inglês | MEDLINE | ID: mdl-29462291

RESUMO

Background: Previous studies suggest that Helicobacter cinaedi can cause recurrent bacteremia. In this study, we elucidated the risk factors for recurrent H. cinaedi bacteremia and explored the efficacy of selective digestive decontamination (SDD) as a preventive measure. Methods: We retrospectively reviewed the medical records of patients with H. cinaedi bacteremia between March 2009 and December 2016 at 2 Japanese hospitals. Results: We identified 168 patients with H. cinaedi bacteremia. Bacteremia recurred in 34 patients. The 100-day cumulative incidence rate of recurrent bacteremia was 18.7%. In univariate analysis of factors associated with recurrent bacteremia, anticancer chemotherapy (hazard ratio [HR], 3.75; 95% confidence interval [CI], 1.86-7.58; P < .001), systemic steroids (HR, 3.79; 95% CI, 1.70-8.45; P = .0011), and hematological malignancy (HR, 3.18; 95% CI, 1.64-6.19; P < .001) were detected. Multivariate analysis indicated that anticancer chemotherapy (HR, 2.47; 95% CI, 1.19-5.12; P = .015) and systemic steroids (HR, 2.40; 95% CI, 1.03-5.61; P = .044) were the independent risk factors. Of the 168 patients, 47 received SDD. According to Gray's test, SDD might have reduced the rate of recurrence but this was not statistically significant (HR, 0.46; 95% CI, 0.18-1.18; P = .11). However, in a proportional hazard modeling analysis, SDD reduced the rate of recurrence (HR, 0.36; 95% CI, 0.13-1.00; P = .050). Conclusions: The 100-day cumulative incidence of recurrent H. cinaedi bacteremia was 18.7%. Anticancer chemotherapy and systemic steroids were independent risk factors for recurrent bacteremia. SDD is a potential strategy for reducing the recurrence.


Assuntos
Antibacterianos/uso terapêutico , Bacteriemia/prevenção & controle , Infecções por Helicobacter/prevenção & controle , Canamicina/uso terapêutico , Prevenção Secundária/métodos , Adulto , Idoso , Idoso de 80 Anos ou mais , Descontaminação , Feminino , Trato Gastrointestinal/microbiologia , Helicobacter/efeitos dos fármacos , Infecções por Helicobacter/tratamento farmacológico , Humanos , Incidência , Masculino , Prontuários Médicos , Pessoa de Meia-Idade , Recidiva , Estudos Retrospectivos , Fatores de Risco
7.
Helicobacter ; 23(1)2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-29218758

RESUMO

BACKGROUND: The route of Helicobacter cinaedi bacteremia has not yet been clarified. Although bacterial translocation from the intestinal tract into the circulation has been suggested, it has not been demonstrated thus far. The objective of this study was to investigate the port of entry of this bacterium. MATERIAL AND METHODS: We conducted a retrospective study on patients with H. cinaedi bacteremia between March 2009 and May 2013. Records of patients in whom H. cinaedi was detected in both blood and stool cultures were extracted. H. cinaedi was identified using gyrB-targeted PCR. Pulse-field gel electrophoresis was used to investigate the consistency of the genotypes. RESULTS: Seventy-one patients were diagnosed with H. cinaedi bacteremia during the study period. H. cinaedi was detected in both blood and stool samples of 21 patients. Pulse-field gel electrophoresis was used to investigate the consistency of the genotypes in 18 evaluable strains (from 9 patients). The pulse-field gel electrophoresis patterns of the stool- and blood-derived strains of H. cinaedi were consistent among all 9 patients. Most of the 9 patients analyzed were immunocompromised and being treated with anticancer drugs or steroids, which suggests reduced intestinal immunity. CONCLUSIONS: This is the first study to demonstrate that bacterial translocation from the intestinal tract could represent one route of H. cinaedi bacteremia.


Assuntos
Bacteriemia/microbiologia , Translocação Bacteriana/fisiologia , Infecções por Helicobacter/microbiologia , Helicobacter/isolamento & purificação , Intestinos/microbiologia , Adulto , Idoso , Proteínas de Bactérias/genética , DNA Girase/genética , Fezes/microbiologia , Feminino , Helicobacter/genética , Infecções por Helicobacter/sangue , Humanos , Hospedeiro Imunocomprometido , Japão , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Estudos Retrospectivos
8.
Nat Commun ; 15(1): 2580, 2024 Mar 22.
Artigo em Inglês | MEDLINE | ID: mdl-38519511

RESUMO

Histone H2B monoubiquitination (at Lys120 in humans) regulates transcription elongation and DNA repair. In humans, H2B monoubiquitination is catalyzed by the heterodimeric Bre1 complex composed of Bre1A/RNF20 and Bre1B/RNF40. The Bre1 proteins generally function as tumor suppressors, while in certain cancers, they facilitate cancer cell proliferation. To obtain structural insights of H2BK120 ubiquitination and its regulation, we report the cryo-electron microscopy structure of the human Bre1 complex bound to the nucleosome. The two RING domains of Bre1A and Bre1B recognize the acidic patch and the nucleosomal DNA phosphates around SHL 6.0-6.5, which are ideally located to recruit the E2 enzyme and ubiquitin for H2BK120-specific ubiquitination. Mutational experiments suggest that the two RING domains bind in two orientations and that ubiquitination occurs when Bre1A binds to the acidic patch. Our results provide insights into the H2BK120-specific ubiquitination by the Bre1 proteins and suggest that H2B monoubiquitination can be regulated by nuclesomal DNA flexibility.


Assuntos
Neoplasias , Nucleossomos , Humanos , Microscopia Crioeletrônica , DNA/metabolismo , Histonas/metabolismo , Ubiquitina-Proteína Ligases/metabolismo , Ubiquitinação
9.
Nat Commun ; 12(1): 6605, 2021 11 15.
Artigo em Inglês | MEDLINE | ID: mdl-34782608

RESUMO

Dimethylated histone H3 Lys36 (H3K36me2) regulates gene expression, and aberrant H3K36me2 upregulation, resulting from either the overexpression or point mutation of the dimethyltransferase NSD2, is found in various cancers. Here we report the cryo-electron microscopy structure of NSD2 bound to the nucleosome. Nucleosomal DNA is partially unwrapped, facilitating NSD2 access to H3K36. NSD2 interacts with DNA and H2A along with H3. The NSD2 autoinhibitory loop changes its conformation upon nucleosome binding to accommodate H3 in its substrate-binding cleft. Kinetic analysis revealed that two oncogenic mutations, E1099K and T1150A, increase NSD2 catalytic turnover. Molecular dynamics simulations suggested that in both mutants, the autoinhibitory loop adopts an open state that can accommodate H3 more often than the wild-type. We propose that E1099K and T1150A destabilize the interactions that keep the autoinhibitory loop closed, thereby enhancing catalytic turnover. Our analyses guide the development of specific inhibitors of NSD2.


Assuntos
Carcinogênese/genética , Carcinogênese/metabolismo , Metilação de DNA , Histona-Lisina N-Metiltransferase/metabolismo , Histonas/metabolismo , Proteínas Repressoras/metabolismo , Microscopia Crioeletrônica , Epigenômica , Histona-Lisina N-Metiltransferase/química , Histona-Lisina N-Metiltransferase/genética , Histonas/química , Histonas/genética , Humanos , Cinética , Metilação , Simulação de Dinâmica Molecular , Neoplasias/genética , Neoplasias/metabolismo , Oncogenes , Proteínas Repressoras/química , Proteínas Repressoras/genética
10.
Jpn J Infect Dis ; 74(4): 325-332, 2021 Jul 21.
Artigo em Inglês | MEDLINE | ID: mdl-33390432

RESUMO

Studies describing reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay-based infection control strategies (LAMP-based ICSs) for coronavirus disease 2019 (COVID-19) are limited. We reviewed the medical records of cases in which RT-LAMP was performed. Standard ICSs and LAMP-based ICSs were implemented during the study period. The strategies were intended to impose longer periods of infection control precautions (ICPs) for specific patients, such as those with a history of exposure to COVID-19 patients and/or bilateral ground glass opacities (bGGO) on chest computed tomography (CT). Of 212 patients, which included 13 confirmed COVID-19 patients in the diagnostic cohort, exposure to COVID-19 patients (P <0.0001) and chest CT bGGO (P = 0.0022) were identified as significant predictors of COVID-19. In the 173 hospitalized patients in which the results of the first RT-LAMP were negative, the duration of ICPs was significantly longer in patients with exposure to COVID-19 and/or a high clinical index of suspicion and patients with bGGO than in the remaining patients (P = 0.00046 and P = 0.0067, respectively). Additionally, no confirmed COVID-19 cases indicating nosocomial spread occurred during the study period. Establishing a comprehensive system that combines rational LAMP-based ICSs with standard ICSs might be useful for preventing nosocomial spread.


Assuntos
Teste para COVID-19/métodos , COVID-19/diagnóstico , COVID-19/virologia , Controle de Infecções/métodos , Transcrição Reversa/genética , SARS-CoV-2/genética , Adulto , Técnicas de Laboratório Clínico/métodos , Feminino , Hospitais , Humanos , Masculino , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Amplificação de Ácido Nucleico/métodos , RNA Viral/genética , Sensibilidade e Especificidade , Tóquio , Adulto Jovem
11.
Diagn Microbiol Infect Dis ; 96(3): 114964, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31884027

RESUMO

We examined the applicability of Matrix-assisted laser desorption ionization-time of flight mass spectrometry using 54 Helicobacter cinaedi isolates from humans. In all 54 isolates, MALDI-TOF MS detected H. cinaedi as the best match organism. Our findings suggest that MALDI TOF-MS can be used effectively to identify H. cinaedi.


Assuntos
Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Hemocultura , DNA Bacteriano/química , Helicobacter/genética , Helicobacter/isolamento & purificação , Infecções por Helicobacter/microbiologia , Humanos
12.
Pflugers Arch ; 459(1): 93-103, 2009 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-19641936

RESUMO

Macrophages are distributed in all peripheral tissues and play a critical role in the first line of the innate immune defenses against bacterial infection by phagocytosis of bacterial pathogens through the macrophage scavenger receptor 1 (MSR1). Within tissues, the partial pressure of oxygen (pO2) decreases depending on the distance of cells from the closest O2-supplying blood vessel. However, it is not clear how the expression of MSR1 in macrophages is regulated by low pO2. On the other hand, hypoxia-inducible factor (HIF)-1alpha is well known to control hypoxic responses through regulation of hypoxia-inducible genes. Therefore, we investigated the effects of hypoxia and HIF-1alpha on MSR1 expression and function in the macrophage cell line RAW264. Exposure to 1% O2 or treatment with the hypoxia-mimetic agent cobalt chloride (CoCl2) significantly suppressed the expression of MSR1 mRNA, accompanied by a markedly increase in levels of nuclear HIF-1alpha protein. The overexpression of HIF-1alpha in RAW264 cells suppressed the expression of MSR1 mRNA and protein, transcriptional activity of the MSR1 gene, and phagocytic capacity against the Gram-positive bacteria Listeria monocytogenes. The suppression of MSR1 mRNA by hypoxia or CoCl2 was inhibited by YC-1, an inhibitor of HIF-1alpha, or by the depletion of HIF-1alpha expression by small interference RNA. These results indicate that hypoxia transcriptionally suppresses MSR1 expression through HIF-1alpha.


Assuntos
Hipóxia Celular/fisiologia , Regulação da Expressão Gênica , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Macrófagos/metabolismo , Receptores Depuradores Classe A/biossíntese , Animais , Antimutagênicos/farmacologia , Western Blotting , Linhagem Celular , Cobalto/farmacologia , Expressão Gênica , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Oxigênio , Pressão Parcial , RNA Mensageiro/análise , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transfecção
13.
Int J Infect Dis ; 73: 60-62, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-29879525

RESUMO

Tuberculosis screening was performed for a healthy asymptomatic woman to determine whether she had been infected with active genital tuberculosis via sexual intercourse with her husband who had epididymal tuberculosis. Vaginal swab culture yielded Mycobacterium tuberculosis. Furthermore, whole genome sequencing revealed that the two causative isolates were genetically identical. This appears to be the first report on the sexual transmission of genital tuberculosis from a man to an asymptomatic woman, detected by active screening for genital tuberculosis and molecular analysis, including whole genome sequencing. Active screening for genital tuberculosis in the female partner should be considered soon after diagnosis of male genital tuberculosis, even when the female partner is asymptomatic.


Assuntos
Doenças Bacterianas Sexualmente Transmissíveis/diagnóstico , Tuberculose dos Genitais Femininos/diagnóstico , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Comportamento Sexual , Cônjuges , Tuberculose dos Genitais Femininos/transmissão
14.
Intern Med ; 57(7): 1045-1048, 2018 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-29269681

RESUMO

A 60-year-old man developed pneumonia after undergoing autologous peripheral blood stem cell transplantation for diffuse large-B cell lymphoma. A urinary antigen test and sputum culture were both negative for Legionella pneumophila; however, a sputum sample that was examined by loop-mediated isothermal amplification (LAMP) was positive for Legionella spp. On admission, the results of blood culturing using a BACTEC system were negative for 7 days. However, L. pneumophila serogroup 5 was detected in a blood subculture using WYOα medium. The patient was successfully treated with a fluoroquinolone-based regimen. LAMP is useful for the diagnosis of Legionella spp.


Assuntos
Bacteriemia/genética , Fluoroquinolonas/uso terapêutico , Legionella pneumophila/classificação , Doença dos Legionários/diagnóstico , Doença dos Legionários/tratamento farmacológico , Técnicas de Amplificação de Ácido Nucleico , Pneumonia/diagnóstico , Aminoacridinas , Humanos , Linfoma/terapia , Masculino , Pessoa de Meia-Idade , Neutropenia/diagnóstico , Neutropenia/microbiologia , Transplante de Células-Tronco de Sangue Periférico/efeitos adversos , Pneumonia/diagnóstico por imagem , Pneumonia/microbiologia , Sorogrupo , Escarro/microbiologia , Resultado do Tratamento
15.
Int J Infect Dis ; 58: 18-21, 2017 May.
Artigo em Inglês | MEDLINE | ID: mdl-28257816

RESUMO

OBJECTIVES: The aim of this study was to evaluate the in vitro effects and clinical efficacies of trimethoprim-sulfamethoxazole (SXT) combined with other antimicrobial agents against Stenotrophomonas maltophilia. METHODS: In vitro analysis was conducted on 89 S. maltophilia strains isolated from blood and the respiratory tract between June 2012 and October 2014. Levofloxacin (LVX), ticarcillin-clavulanic acid (TIM), and minocycline (MIN) were selected for an examination of their effects when individually combined with SXT by the checkerboard method. In addition, 29 S. maltophilia bacteremia cases were reviewed and the clinical efficacies of SXT-based combination therapies were analyzed. RESULTS: SXT+LVX showed synergy in 21, no interactions in 61, and antagonism in 7. SXT+TIM showed synergy in 71, and no interactions in 18. SXT+MIN showed synergy in 10, and no interactions in 79. The review of clinical data indicated that a combination of SXT+fluoroquinolone was not associated with improved prognosis compared with monotherapy. CONCLUSIONS: The in vitro data indicated that SXT+TIM had beneficial microbiological effects and was not antagonistic. Our in vitro and clinical data analyses do not support the routine use of SXT+fluoroquinolone combination therapy for S. maltophilia infection.


Assuntos
Antibacterianos/uso terapêutico , Infecções por Bactérias Gram-Negativas/tratamento farmacológico , Neoplasias/complicações , Stenotrophomonas maltophilia , Combinação Trimetoprima e Sulfametoxazol/uso terapêutico , Idoso , Bacteriemia/tratamento farmacológico , Ácidos Clavulânicos , Quimioterapia Combinada , Feminino , Fluoroquinolonas/uso terapêutico , Infecções por Bactérias Gram-Negativas/complicações , Infecções por Bactérias Gram-Negativas/microbiologia , Humanos , Levofloxacino/uso terapêutico , Masculino , Pessoa de Meia-Idade , Minociclina/uso terapêutico , Neoplasias/tratamento farmacológico , Stenotrophomonas maltophilia/isolamento & purificação , Ticarcilina , Resultado do Tratamento
16.
J Nutr Sci Vitaminol (Tokyo) ; 51(1): 27-33, 2005 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-15915665

RESUMO

The objective of this study is to clarify the difference in susceptibility to protease digestion by kiwifruit juice between collagen domains under different conditions. In addition, the effect of pre-treatment with kiwifruit juice on collagen in meat during cooking processes was examined. Kiwifruit juice can degrade denatured collagen, but it can not cleave the triple helical domain of collagen. Thus, kiwifruit juice does not have collagenase activity. On the other hand, the cross-linked subunits of acid-soluble collagen were converted to monomeric subunits by kiwifruit juice treatment at acidic pH, suggesting that the globular domains, in which cross-links preferentially occur, can be degraded by kiwifruit juice. The pre-treatment with kiwifruit juice significantly decreased the shear force of connective tissue in comparison with other pre-treatments without protease activity, but inversely increased the liberation of collagen-related peptides in the outer solution by heating processes at 50 and 70 degrees C or by a shorter heating time at 100 degrees C. This can be explained by the protease-mediated degradation of globular domains. However, this effect was not observed with a prolonged heating period at 100 degrees C, and the liberation of collagen-related peptides by pre-treatment with kiwifruit juice at 100 degrees C was less than that at 70 degrees C for all heating periods. Thus, it can be suggested that the pre-treatment with kiwifruit juice might be useful in meat softening under vacuum-cooking and grilling, but not under stewing.


Assuntos
Actinidia/enzimologia , Colágeno/metabolismo , Frutas/enzimologia , Carne/análise , Peptídeo Hidrolases/metabolismo , Animais , Bebidas , Bovinos , Colágeno/química , Tecido Conjuntivo/metabolismo , Culinária , Temperatura Alta , Desnaturação Proteica , Estrutura Secundária de Proteína , Solubilidade
17.
J Clin Microbiol ; 45(9): 2853-7, 2007 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-17596362

RESUMO

Helicobacter cinaedi has being recognized as an important human pathogen which causes bloodstream infections. Although the first case of bacteremia with this pathogen in Japan was reported in 2003, the true prevalence of H. cinaedi as a pathogen of bloodstream infections in this country is not yet known. Therefore, the aim of our study was to assess the incidence of bacteremia with H. cinaedi in Japan. We conducted a prospective, multicenter analysis in 13 hospitals during 6 months in Tokyo, Japan. Among positive blood cultures from 1 October 2003 to 31 March 2004, isolates suspected of being Helicobacter species were studied for further microbial identification. Identification of the organisms was based on their biochemical traits and the results of molecular analysis of their 16S rRNA gene sequences. A total of 16,743 blood culture samples were obtained during the study period, and 2,718 samples (17.7%) yielded positive culture results for coagulase-negative staphylococci. Among nine isolates suspected to be Helicobacter species, six isolates were finally identified as H. cinaedi. The positivity rate for H. cinaedi in blood culture was 0.06% of total blood samples and 0.22% of blood samples with any positive culture results. All patients with bacteremia with H. cinaedi were found to have no human immunodeficiency virus (HIV) infection, but many of them had complications with either malignancy, renal failure, or a history of surgical operation. Therefore, our results suggest that bacteremia with H. cinaedi is rare but can occur in compromised hosts other than those with HIV infection in Japan.


Assuntos
Bacteriemia/epidemiologia , Bacteriemia/microbiologia , Infecções por Helicobacter/epidemiologia , Helicobacter/isolamento & purificação , Adolescente , Adulto , Idoso , Técnicas de Tipagem Bacteriana , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Feminino , Infecções por HIV/complicações , Helicobacter/classificação , Helicobacter/genética , Hospitais , Humanos , Incidência , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Neoplasias/complicações , Filogenia , Complicações Pós-Operatórias/microbiologia , Estudos Prospectivos , RNA Ribossômico 16S/genética , Insuficiência Renal/complicações , Análise de Sequência de DNA , Tóquio/epidemiologia
18.
Neurochem Res ; 27(6): 497-500, 2002 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-12199154

RESUMO

A timed profile of glutathione oxidation and reactive nitrogen species during reperfusion after cerebral ischemia in rat was obtained. Dialysate was collected every 25 min from a microdialysis probe inserted into the cerebral cortex before and after cerebral ischemia. NO2-, NO3-, and reduced and oxidized glutathione (GSH, GSSG) were detected by high-performance liquid chromatography. GSH and GSSG increased and reached a peak: 3408 +/- 1710% (mean +/- SE) at 25 min of reperfusion (P < 0.0001) and 329 +/- 104% at 50 min of reperfusion (P = 0.06), respectively. Oxidation ratio decreased from 0.82 +/- 0.04 to 0.42 +/- 0.07 (P < 0.0001) at 25 min of reperfusion. NO3- levels significantly decreased (68.3 +/- 9.1%) (P < 0.01) during ischemia and remained lower than the control value during reperfusion. NO2- levels did not significantly change. These data suggest that GSH releases during early phase of reperfusion and that its rapid oxidation contributes to prevent an increase in reactive nitrogen species.


Assuntos
Isquemia Encefálica/metabolismo , Glutationa/metabolismo , Nitrogênio/metabolismo , Traumatismo por Reperfusão/metabolismo , Animais , Oxirredução , Ratos , Ratos Wistar
19.
Microbiol Immunol ; 46(1): 51-4, 2002.
Artigo em Inglês | MEDLINE | ID: mdl-11911189

RESUMO

Legionella antigen detection kits for diagnosing legionellosis from urine have become widely used, but basic information about reactivity of the kits to non-serogroup (SG) 1 L. pneumophila and other Legionella species remains incomplete. We evaluated Biotest EIA and the most recently developed Binax NOW by using in-vitro extracted antigens of 22 L. pneumophila SG 1 to 15 strains and of 27 other Legionella species. Both kits showed excellent sensitivity to L pneumophila SG 1 antigens, but reacted to different sets of non-SG I L. pneumophila with different sensitivity. No cross-reactivity was observed to Legionella species other than L. pneumophila.


Assuntos
Antígenos de Bactérias/imunologia , Antígenos de Bactérias/urina , Imunoensaio , Legionella/imunologia , Legionelose/diagnóstico , Antígenos de Bactérias/isolamento & purificação , Reações Cruzadas , Humanos , Técnicas Imunoenzimáticas , Legionella/classificação , Legionella pneumophila/classificação , Legionella pneumophila/imunologia , Doença dos Legionários/diagnóstico , Kit de Reagentes para Diagnóstico , Sensibilidade e Especificidade , Sorotipagem
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