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1.
Plant Cell Physiol ; 60(10): 2180-2192, 2019 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-31198965

RESUMO

A putative silent gene of the freshwater cyanobacterium Synechococcus elongatus strain PCC 7942, encoding a small protein with two transmembrane helices, was named nrtS, since its overexpression from an inducible promoter conferred nitrate uptake activity on the nitrate transport-less NA4 mutant of S. elongatus. Homologs of nrtS, encoding proteins of 67-118 amino acid residues, are present in a limited number of eubacteria including mostly cyanobacteria and proteobacteria, but some others, e.g. the actinobacteria of the Mycobacterium tuberculosis complex, also have the gene. When expressed in NA4, the nrtS homolog of the γ-proteobacterium Marinomonas mediterranea took up nitrate with higher affinity for the substrate as compared with the S. elongatus NrtS (Km of 0.49 mM vs. 2.5 mM). Among the 61 bacterial species carrying the nrtS homolog, the marine cyanobacterium Synechococcus sp. strain PCC 7002 is unique in having two nrtS genes (nrtS1 and nrtS2) located in tandem on the chromosome. Coexpression of the two genes in NA4 resulted in nitrate uptake with a Km (NO3-) of 0.15 mM, while expression of either of the two resulted in low-affinity nitrate uptake activity with Km values of >3 mM, indicating that NrtS1 and NrtS2 form a heteromeric transporter complex. The heteromeric transporter was shown to transport nitrite as well. A Synechococcus sp. strain PCC 7002 mutant defective in the nitrate transporter (NrtP) showed a residual activity of nitrate uptake, which was ascribed to the NrtS proteins. Blue-native PAGE and immunoblotting analysis suggested a hexameric structure for the NrtS proteins.


Assuntos
Proteínas de Transporte de Ânions/genética , Nitratos/metabolismo , Nitritos/metabolismo , Synechococcus/genética , Sequência de Aminoácidos , Proteínas de Transporte de Ânions/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Transporte Biológico , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Mutação , Transportadores de Nitrato , Alinhamento de Sequência , Synechococcus/metabolismo
2.
Nucleic Acids Res ; 45(D1): D551-D554, 2017 01 04.
Artigo em Inglês | MEDLINE | ID: mdl-27899668

RESUMO

The first ever cyanobacterial genome sequence was determined two decades ago and CyanoBase (http://genome.microbedb.jp/cyanobase), the first database for cyanobacteria was simultaneously developed to allow this genomic information to be used more efficiently. Since then, CyanoBase has constantly been extended and has received several updates. Here, we describe a new large-scale update of the database, which coincides with its 20th anniversary. We have expanded the number of cyanobacterial genomic sequences from 39 to 376 species, which consists of 86 complete and 290 draft genomes. We have also optimized the user interface for large genomic data to include the use of semantic web technologies and JBrowse and have extended community-based reannotation resources through the re-annotation of Synechocystis sp. PCC 6803 by the cyanobacterial research community. These updates have markedly improved CyanoBase, providing cyanobacterial genome annotations as references for cyanobacterial research.


Assuntos
Cianobactérias/genética , Bases de Dados Genéticas , Genoma Bacteriano , Genômica/métodos , Biologia Computacional/métodos , Navegador
3.
Plant Cell Physiol ; 56(8): 1608-15, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-26063393

RESUMO

Most organisms capable of oxygenic photosynthesis have an aas gene encoding an acyl-acyl carrier protein synthetase (Aas), which activates free fatty acids (FFAs) via esterification to acyl carrier protein. Cyanobacterial aas mutants are often used for studies aimed at photosynthetic production of biofuels because the mutation leads to intracellular accumulation of FFAs and their secretion into the external medium, but the physiological significance of the production of FFAs and their recycling involving Aas has remained unclear. Using an aas-deficient mutant of Synechococcus elongatus strain PCC 7942, we show here that remodeling of membrane lipids is activated by high-intensity light and that the recycling of FFAs is essential for acclimation to high-light conditions. Unlike wild-type cells, the mutant cells could not increase their growth rate as the light intensity was increased from 50 to 400 µmol photons m(-2) s(-1), and the high-light-grown mutant cells accumulated FFAs and the lysolipids derived from all the four major classes of membrane lipids, revealing high-light-induced lipid deacylation. The high-light-grown mutant cells showed much lower PSII activity and Chl contents as compared with the wild-type cells or low-light-grown mutant cells. The loss of Aas accelerated photodamage of PSII but did not affect the repair process of PSII, indicating that PSII is destabilized in the mutant. Thus, Aas is essential for acclimation of the cyanobacterium to high-light conditions. The relevance of the present finding s to biofuel production using cyanobacteria is discussed.


Assuntos
Carbono-Enxofre Ligases/metabolismo , Synechococcus/enzimologia , Aclimatação , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Carbono-Enxofre Ligases/genética , Ácidos Graxos não Esterificados/metabolismo , Luz , Lipídeos de Membrana/metabolismo , Mutação , Fotossíntese/fisiologia , Fotossíntese/efeitos da radiação , Complexo de Proteína do Fotossistema II/fisiologia , Complexo de Proteína do Fotossistema II/efeitos da radiação , Synechococcus/genética , Synechococcus/fisiologia , Synechococcus/efeitos da radiação
4.
Plant Cell Physiol ; 56(12): 2467-77, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26468506

RESUMO

An RND (resistance-nodulation-division)-type transporter having the capacity to export free fatty acids (FFAs) was identified in the cyanobacterium Synechococcus elongatus strain PCC 7942 during characterization of a mutant strain engineered to produce FFAs. The basic strategy for construction of the FFA-producing mutant was a commonly used one, involving inactivation of the endogenous acyl-acyl carrier protein synthetase gene (aas) and introduction of a foreign thioesterase gene ('tesA), but a nitrate transport mutant NA3 was used as the parental strain to achieve slow, nitrate-limited growth in batch cultures. Also, a nitrogen-regulated promoter PnirA was used to drive 'tesA to maximize thioesterase expression during the nitrate-limited growth. The resulting mutant (dAS2T) was, however, incapable of growth under the conditions of nitrate limitation, presumably due to toxicity associated with FFA overproduction. Incubation of the mutant culture under the non-permissive conditions allowed for isolation of a pseudorevertant (dAS2T-pr1) capable of growth on nitrate. Genome sequence and gene expression analyses of this strain suggested that expression of an RND-type efflux system had rescued growth on nitrate. Targeted inactivation of the RND-type transporter genes in the wild-type strain resulted in loss of tolerance to exogenously added FFAs including capric, lauric, myristic, oleic and linolenic acids. Overexpression of the genes in dAS2T, on the other hand, enhanced FFA excretion and cell growth in nitrate-containing medium, verifying that the genes encode an efflux pump for FFAs. These results demonstrate the importance of the efflux system in efficient FFA production using genetically engineered cyanobacteria.


Assuntos
Proteínas de Bactérias/metabolismo , Ácidos Graxos não Esterificados/metabolismo , Synechococcus/metabolismo , Sequência de Bases , Transporte Biológico , Genes de Plantas , Mutação/genética , Nitratos/metabolismo , Filogenia , Synechococcus/genética , Synechococcus/crescimento & desenvolvimento
5.
J Infect Chemother ; 21(5): 340-5, 2015 May.
Artigo em Inglês | MEDLINE | ID: mdl-25727286

RESUMO

Worldwide, the most important concern in the treatment of sexually transmitted infections is the increase in antimicrobial resistant Neisseria gonorrhoeae strains including resistance to cephalosporins, penicillins, fluoroquinolones or macrolides. To investigate the trends of antimicrobial susceptibility among N. gonorrhoeae strains isolated from male patients with urethritis, a Japanese surveillance committee conducted the second nationwide surveillance study. Urethral discharge was collected from male patients with urethritis at 26 medical facilities from March 2012 to January 2013. Of the 151 specimens, 103 N. gonorrhoeae strains were tested for susceptibility to 20 antimicrobial agents. None of the strains was resistant to ceftriaxone, but the minimum inhibitory concentration (MIC) 90% of ceftriaxone increased to 0.125 µg/ml, and 11 (10.7%) strains were considered less susceptible with an MIC of 0.125 µg/ml. There were 11 strains resistant to cefixime, and the MICs of these strains were 0.5 µg/ml. The distributions of the MICs of fluoroquinolones, such as ciprofloxacin, levofloxacin and tosufloxacin, were bimodal. Sitafloxacin, a fluoroquinolone, showed strong activity against all strains, including strains resistant to other three fluoroquinolones, such as ciprofloxacin, levofloxacin and tosufloxacin. The azithromycin MICs in 2 strains were 1 µg/ml.


Assuntos
Antibacterianos/farmacologia , Farmacorresistência Bacteriana , Neisseria gonorrhoeae/efeitos dos fármacos , Vigilância da População , Uretrite/microbiologia , Adolescente , Adulto , Idoso , Azitromicina/farmacologia , Cefixima/farmacologia , Ceftriaxona/farmacologia , Fluoroquinolonas/farmacologia , Humanos , Japão , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Penicilinas/farmacologia , Adulto Jovem
6.
Plant Cell Physiol ; 55(7): 1311-24, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-24904028

RESUMO

Some cyanobacterial genomes encode an integral membrane protein of the HPP family, which exhibited nitrite transport activity when expressed in the nitrite transport-less NA4 mutant of the cyanobacterium Synechococcus elongatus strain PCC 7942. AT5G62720 and AT3G47980 were found to encode Arabidopsis homologs of the cyanobacterial protein. The product of AT5G62720 was localized to the chloroplast envelope membrane and was shown to confer nitrite uptake activity on the NA4 mutant when expressed with an N-terminally truncated transit peptide or as a fusion with the N-terminal region of the cyanobacterial HPP family protein. Kinetic analyses showed that the Arabidopsis protein has much higher affinity for nitrite (K(m) = 13 µM) than the cyanobacterial protein (K(m) = 150 µM). Illuminated chloroplasts isolated from the mutant lines of AT5G62720 showed much lower activity of nitrite uptake than the chloroplasts isolated from the wild-type Col-0 plants, while the chloroplasts of the mutants of AT1G68570 (AtNPF3.1), the gene previously reported to encode a plastid nitrite transporter AtNitr1, showed wild-type levels of nitrite uptake activity. AT3G47980 was expressed in roots but not in shoots. It has a putative transit peptide similar to that of AT5G62720 and its fusion with the N-terminal region of the cyanobacterial HPP protein showed low but significant activity of nitrite transport in the cyanobacterial cell. Transcription of AT5G62720 (AtNITR2;1) and AT3G47980 (AtNITR2;2) was stimulated by nitrate under the control of the NIN-like proteins, suggesting that the HPP proteins represent nitrate-inducible components of the nitrite transport system of plastids.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Proteínas de Bactérias/metabolismo , Proteínas de Membrana Transportadoras/metabolismo , Nitritos/metabolismo , Synechococcus/metabolismo , Sequência de Aminoácidos , Arabidopsis/citologia , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Proteínas de Bactérias/genética , Proteínas de Cloroplastos/genética , Proteínas de Cloroplastos/metabolismo , Cloroplastos/metabolismo , Genes Reporter , Proteínas de Membrana Transportadoras/genética , Dados de Sequência Molecular , Mutação , Nitratos/metabolismo , Especificidade de Órgãos , Raízes de Plantas/citologia , Raízes de Plantas/genética , Raízes de Plantas/metabolismo , Brotos de Planta/citologia , Brotos de Planta/genética , Brotos de Planta/metabolismo , Plantas Geneticamente Modificadas , Proteômica , Proteínas Recombinantes de Fusão , Plântula/citologia , Plântula/genética , Plântula/metabolismo , Alinhamento de Sequência , Synechococcus/citologia , Synechococcus/genética
7.
Plant Cell Physiol ; 55(2): 281-92, 2014 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-24319077

RESUMO

Elevated CO2 has been reported to stimulate plant growth under nitrogen-sufficient conditions, but the effects of CO2 on growth in a constantly nitrogen-limited state, which is relevant to most natural habitats of plants, remain unclear. Here, we maintained Arabidopsis seedlings under such conditions by growing a mutant with reduced nitrate uptake activity on a medium containing nitrate as the sole nitrogen source. Under nitrogen-sufficient conditions (i.e. in the presence of ammonium), growth of shoots and roots of both the wild type (WT) and the mutant was increased approximately 2-fold by elevated CO2. Growth stimulation of shoots and roots by elevated CO2 was observed in the WT growing with nitrate as the sole nitrogen source, but in the mutant grown with nitrate, the high-CO2 conditions stimulated only the growth of roots. In the mutant, elevated CO2 caused well-known symptoms of nitrogen-starved plants, including decreased shoot/root ratio, reduced nitrate content and accumulation of anthocyanin, but also had an increased Chl content in the shoot, which was contradictory to the known effect of nitrogen depletion. A high-CO2-responsive change specific to the mutant was not observed in the levels of the major metabolites, although CO2 responses were observed in the WT and the mutant. These results indicated that elevated CO2 causes nitrogen limitation in the seedlings grown with a constantly limited supply of nitrogen, but the Chl content and the root biomass of the plant increase to enhance the activities of both photosynthesis and nitrogen uptake, while maintaining normal metabolism and response to high CO2.


Assuntos
Arabidopsis/fisiologia , Dióxido de Carbono/farmacologia , Metaboloma , Nitrogênio/deficiência , Compostos de Amônio/metabolismo , Antocianinas/metabolismo , Arabidopsis/efeitos dos fármacos , Arabidopsis/crescimento & desenvolvimento , Biomassa , Dióxido de Carbono/metabolismo , Clorofila/metabolismo , Técnicas de Inativação de Genes , Mutação , Nitratos/metabolismo , Nitrogênio/metabolismo , Fotossíntese , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/fisiologia , Brotos de Planta/efeitos dos fármacos , Brotos de Planta/crescimento & desenvolvimento , Brotos de Planta/fisiologia , Plântula/efeitos dos fármacos , Plântula/crescimento & desenvolvimento , Plântula/fisiologia , Solo
8.
Photosynth Res ; 121(2-3): 151-7, 2014 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-24585024

RESUMO

The carboxylase activities of crude carboxysome preparations obtained from the wild-type Synechococcus elongatus strain PCC 7942 strain and the mutant defective in the carboxysomal carbonic anhydrase (CA) were compared. The carboxylation reaction required high concentrations of bicarbonate and was not even saturated at 50 mM bicarbonate. With the initial concentrations of 50 mM and 25 mM for bicarbonate and ribulose-1,5-bisphosphate (RuBP), respectively, the initial rate of RuBP carboxylation by the mutant carboxysome (0.22 µmol mg(-1) protein min(-1)) was only 30 % of that observed for the wild-type carboxysomes (0.71 µmol mg(-1) protein min(-1)), indicating the importance of the presence of CA in efficient catalysis by ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco). While the mutant defective in the ccmLMNO genes, which lacks the carboxysome structure, could grow under aeration with 2 % (v/v) CO2 in air, the mutant defective in ccaA as well as ccmLMNO required 5 % (v/v) CO2 for growth, indicating that the cytoplasmically localized CcaA helped utilization of CO2 by the cytoplasmically localized Rubisco by counteracting the action of the CO2 hydration mechanism. The results predict that overexpression of Rubisco would hardly enhance CO2 fixation by the cyanobacterium at CO2 levels lower than 5 %, unless Rubisco is properly organized into carboxysomes.


Assuntos
Anidrases Carbônicas/metabolismo , Synechococcus/enzimologia , Dióxido de Carbono/metabolismo , Ribulose-Bifosfato Carboxilase/metabolismo , Ribulosefosfatos/metabolismo
9.
Genes Genet Syst ; 992024 Apr 18.
Artigo em Inglês | MEDLINE | ID: mdl-38447993

RESUMO

The budding yeast Saccharomyces cerevisiae is an excellent model organism for studying chromatin regulation with high-resolution genome-wide analyses. Since newly generated genome-wide data are often compared with publicly available datasets, expanding our dataset repertoire will be beneficial for the field. Information on transcription start sites (TSSs) determined at base pair resolution is essential for elucidating mechanisms of transcription and related chromatin regulation, yet no datasets that cover two different cell types are available. Here, we present a CAGE (cap analysis of gene expression) dataset for a-cells and α-cells grown in defined and rich media. Cell type-specific genes were differentially expressed as expected, ensuring the reliability of the data. Some of the differentially expressed TSSs were medium-specific or detected due to unrecognized chromosome rearrangement. By comparing the CAGE data with a high-resolution nucleosome map, major TSSs were primarily found in +1 nucleosomes, with a peak approximately 30 bp from the promoter-proximal end of the nucleosome. The dataset is available at DDBJ/GEA.


Assuntos
Estudo de Associação Genômica Ampla , Nucleossomos , Reprodutibilidade dos Testes , Cromatina/metabolismo , Saccharomyces cerevisiae/genética
10.
Plant Cell Physiol ; 54(9): 1504-14, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23811238

RESUMO

Among the known functions of the P(II) protein (the glnB gene product) in the cyanobacterium Synechococcus elongatus, negative regulation of the activity of PipX, a transcriptional co-activator of the NtcA regulon, has been thought to be essential for cell viability, because all the P(II)-less mutants thus far constructed carry spontaneous mutations in pipX. PipX is thus deduced to be a toxic protein, but its toxicity has not been clearly defined because of the lack of P(II)-deficient mutants carrying wild-type pipX. In this study, we developed a method to construct a targeted P(II)-less mutant of S. elongatus without the pipX mutation and determined the contribution of PipX to the detrimental effects of P(II) deficiency. Growth defects of the mutant were severe under nitrogen-replete conditions, i.e. in the presence of ammonium, but were also apparent under nitrogen-limited conditions. Genetic analyses indicated that the growth impairment observed under the nitrogen-limited conditions is largely due to the toxicity of PipX. Some of the phenotypes observed under the nitrogen-replete conditions, including reduced pigmentation and death of most of the cells after transfer from nitrogen-limited conditions to nitrogen-replete conditions, were ascribed to the toxicity of PipX, but inactivation of pipX only partially rescued the growth defect observed in the presence of ammonium, indicating the presence of an as yet unknown P(II) function(s) required for normal growth. Effects of ammonium addition on the nitrite uptake activity of the glnB mutant revealed a new function for P(II) in regulation of the activity of the ABC-type cyanate/nitrite transporter.


Assuntos
Proteínas de Bactérias/genética , Mutação , Proteínas PII Reguladoras de Nitrogênio/genética , Synechococcus/genética , Transportadores de Cassetes de Ligação de ATP/genética , Transportadores de Cassetes de Ligação de ATP/metabolismo , Sequência de Aminoácidos , Compostos de Amônio/metabolismo , Compostos de Amônio/farmacologia , Proteínas de Bactérias/metabolismo , Sequência de Bases , Divisão Celular/efeitos dos fármacos , Divisão Celular/genética , Cianatos/metabolismo , Dados de Sequência Molecular , Nitrato Redutase/genética , Nitrato Redutase/metabolismo , Nitritos/metabolismo , Nitrogênio/metabolismo , Nitrogênio/farmacologia , Proteínas PII Reguladoras de Nitrogênio/metabolismo , Espectinomicina/farmacologia , Synechococcus/efeitos dos fármacos , Synechococcus/metabolismo
11.
Sex Transm Infect ; 89(6): 528-30, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23349337

RESUMO

OBJECTIVES: Trichomonas vaginalis is one of the pathogens causing sexually transmitted infections. This microorganism is a common pathogen among women, but its significance as a cause of morbidity among men remains uncertain. We sought to determine the prevalence and morbidity of T. vaginalis infection in Japanese men with and without urethritis. METHODS: We examined urine specimens from 215 men with urethritis and 98 men without urethritis for the presence of urethral T. vaginalis by PCR assay. RESULTS: Only four patients-one with gonococcal urethritis, one with non-gonococcal chlamydial urethritis, one with non-gonococcal non-chlamydial urethritis and one without urethritis-were positive for T. vaginalis. The prevalence of T. vaginalis was 1.4% in men with urethritis and 1.0% in men without urethritis. A possible relation between the appearance of T. vaginalis and clinical symptoms was not confirmed. CONCLUSIONS: In the present study, the incidence of urethral T. vaginalis infection appears to be rare in Japanese men with or without urethritis, and T. vaginalis may be an uncommon pathogen in male urethritis in Japan.


Assuntos
Tricomoníase/epidemiologia , Tricomoníase/patologia , Trichomonas vaginalis/isolamento & purificação , Uretra/parasitologia , Adolescente , Adulto , Coinfecção/parasitologia , Coinfecção/patologia , DNA de Protozoário/genética , DNA de Protozoário/isolamento & purificação , Humanos , Incidência , Japão/epidemiologia , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Prevalência , Trichomonas vaginalis/genética , Uretrite/parasitologia , Uretrite/patologia , Urina/parasitologia , Adulto Jovem
12.
J Infect Chemother ; 19(5): 941-5, 2013 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-23749142

RESUMO

To clarify the clinical efficacy of STFX for patients with non-gonococcal urethritis (NGU), including chlamydial urethritis and Mycoplasma genitalium-positive urethritis, this study included male patients with NGU who were 20 years old or older. The pathogens, including Chlamydia trachomatis, M. genitalium and Ureaplasma urealyticum, were detected by nucleic acid amplification tests and the patients were treated with sitafloxacin 100 mg twice daily for 7 days. Microbiological and clinical efficacies were assessed for the patients with NGU posttreatment. Among the 208 patients enrolled in this study, data for a total of 118 patients could be analyzed. The median age was 32 (20-61) years. The median duration from the completion of treatment to the second visit was 21 (14-42) days. There were 68 pathogen-positive NGU cases and 50 with NGU without any microbial detection. Microbiological cure was achieved in 95.6% of the pathogen-positive NGU patients. Total clinical cure was achieved in 91.3% (105/115). In this study, STFX was able to eradicate 95.7% of C. trachomatis, 93.8% of M. genitalium and 100% of U. urealyticum. The results of our clinical research indicate that the STFX treatment regimen should become a standard regimen recommended for patients with NGU. In addition, this regimen is recommended for patients with M. genitalium-positive NGU.


Assuntos
Antibacterianos/administração & dosagem , Fluoroquinolonas/administração & dosagem , Infecções por Bactérias Gram-Negativas/tratamento farmacológico , Uretrite/tratamento farmacológico , Adulto , Antibacterianos/efeitos adversos , Chlamydia trachomatis/efeitos dos fármacos , Chlamydia trachomatis/isolamento & purificação , Fluoroquinolonas/efeitos adversos , Infecções por Bactérias Gram-Negativas/microbiologia , Humanos , Masculino , Pessoa de Meia-Idade , Mycoplasma genitalium/efeitos dos fármacos , Mycoplasma genitalium/isolamento & purificação , Estudos Prospectivos , Ureaplasma urealyticum/efeitos dos fármacos , Ureaplasma urealyticum/isolamento & purificação , Uretrite/microbiologia , Adulto Jovem
13.
J Infect Chemother ; 19(4): 571-8, 2013 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-23813093

RESUMO

Neisseria gonorrhoeae is one of the most important pathogens causing sexually transmitted infection, and strains that are resistant to several antimicrobials are increasing. To investigate the trends of antimicrobial susceptibility among N. gonorrhoeae strains isolated from male patients with urethritis, a Japanese surveillance committee conducted the first nationwide surveillance. The urethral discharge was collected from male patients with urethritis at 51 medical facilities from April 2009 to October 2010. Of the 156 specimens, 83 N. gonorrhoeae strains were tested for susceptibility to 18 antimicrobial agents. The prevalence of ß-lactamase-producing strains and chromosomally mediated resistant strains were 7.2 % and 16.5 %, respectively. None of the strains was resistant to ceftriaxone, but the minimum inhibitory concentration (MIC) of ceftriaxone for 7 strains (8.4 %) was 0.125 µg/ml. One strain was resistant to cefixime (MIC 0.5 µg/ml). The MICs of fluoroquinolones, such as ciprofloxacin, levofloxacin, and tosufloxacin, showed a bimodal distribution. The MIC of sitafloxacin was lower than those of the three fluoroquinolones listed here, and it was found that the antimicrobial activity of sitafloxacin was stronger than that of the fluoroquinolones. The MIC of azithromycin in 2 strains was 2 µg/ml, but no high-level resistance to macrolides was detected.


Assuntos
Antibacterianos/farmacologia , Neisseria gonorrhoeae/efeitos dos fármacos , Uretrite/epidemiologia , Uretrite/microbiologia , Adolescente , Adulto , Idoso , Farmacorresistência Bacteriana , Humanos , Japão/epidemiologia , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Neisseria gonorrhoeae/isolamento & purificação , Prevalência , Vigilância em Saúde Pública
14.
Hinyokika Kiyo ; 59(12): 785-9, 2013 Dec.
Artigo em Japonês | MEDLINE | ID: mdl-24419010

RESUMO

A 58-year-old man from Brazil was followed as an outpatient with asymptomatic macroglobulinemia and idiopathic thrombocytopenic purpura (ITP). Abdominal enhanced computed tomographic (CT) scan for elevated liver enzymes revealed a left renal tumor. The tumor was in the middle outer left kidney, measured 18 mm in diameter, was discovered in its early phase, and appeared half exophytic. After investigations, the patient was diagnosed with left renal cell carcinoma associated with ITP. His preoperative platelet count was 10,000/µl ; five days of intravenous gamma globulin therapy with high-dose dexamethasone increased the platelet count to 76,000/µl just before operation. Laparoscopic left partial nephrectomy was performed successfully using the retroperitoneal approach. The renal artery was clamped and the tumor excised with an adequate margin. Renal parenchymal repair was completed using running sutures. Ischemia time was 16 minutes. There was no severe oozing of blood intraoperatively. The platelet count decreased to 15,000/µl on postoperative day three (POD 3), and there was oozing of blood around the retroperitoneal drain tube. The bleeding stopped after administration of platelet transfusion. The patient was discharged on POD 9. The histopathological diagnosis was clear cell carcinoma, and surgical margins were negative.


Assuntos
Carcinoma de Células Renais/cirurgia , Neoplasias Renais/cirurgia , Laparoscopia , Nefrectomia/métodos , Púrpura Trombocitopênica Idiopática/complicações , Humanos , Masculino , Pessoa de Meia-Idade , Púrpura Trombocitopênica Idiopática/sangue
15.
IEEE/ACM Trans Comput Biol Bioinform ; 20(5): 2970-2980, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37079418

RESUMO

Three-dimensional (3D) reconstruction for cryogenic electron microscopy (cryo-EM) often falls into an ill-posed problem owing to several uncertainties in observations, including noise. To reduce excessive degree of freedom and avoid overfitting, the structural symmetry is often used as a powerful constraint. In the case of the helix, the entire 3D structure is determined by the subunit 3D structure and two helical parameters. There is no analytical method to simultaneously obtain both of the subunit structure and helical parameters. A common approach is to employ an iterative reconstruction in which the two optimizations are performed alternately. However, iterative reconstruction does not necessarily converge when a heuristic objective function is used for each optimization step. Also, the obtained 3D reconstruction highly depends on the initial guess of the 3D structure and the helical parameters. Herein, we propose a method for estimating the 3D structure and helical parameters that also performs an iterative optimization; however, the objective function for each step is derived from a single objective function to make the algorithm convergent and less sensitive to the initial guess. Finally, we evaluated the effectiveness of the proposed method by testing it on cryo-EM images, which were challenging to reconstruct using conventional methods.


Assuntos
Algoritmos , Imageamento Tridimensional , Microscopia Crioeletrônica/métodos , Imageamento Tridimensional/métodos , Teorema de Bayes , Processamento de Imagem Assistida por Computador/métodos
16.
J Exp Bot ; 62(4): 1411-24, 2011 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-21282331

RESUMO

Nitrate assimilation by cyanobacteria is inhibited by the presence of ammonium in the growth medium. Both nitrate uptake and transcription of the nitrate assimilatory genes are regulated. The major intracellular signal for the regulation is, however, not ammonium or glutamine, but 2-oxoglutarate (2-OG), whose concentration changes according to the change in cellular C/N balance. When nitrogen is limiting growth, accumulation of 2-OG activates the transcription factor NtcA to induce transcription of the nitrate assimilation genes. Ammonium inhibits transcription by quickly depleting the 2-OG pool through its metabolism via the glutamine synthetase/glutamate synthase cycle. The P(II) protein inhibits the ABC-type nitrate transporter, and also nitrate reductase in some strains, by an unknown mechanism(s) when the cellular 2-OG level is low. Upon nitrogen limitation, 2-OG binds to P(II) to prevent the protein from inhibiting nitrate assimilation. A pathway-specific transcriptional regulator NtcB activates the nitrate assimilation genes in response to nitrite, either added to the medium or generated intracellularly by nitrate reduction. It plays an important role in selective activation of the nitrate assimilation pathway during growth under a limited supply of nitrate. P(II) was recently shown to regulate the activity of NtcA negatively by binding to PipX, a small coactivator protein of NtcA. On the basis of accumulating genome information from a variety of cyanobacteria and the molecular genetic data obtained from the representative strains, common features and group- or species-specific characteristics of the response of cyanobacteria to nitrogen is summarized and discussed in terms of ecophysiological significance.


Assuntos
Cianobactérias/metabolismo , Nitratos/metabolismo , Transdução de Sinais , Proteínas de Transporte de Ânions/genética , Proteínas de Transporte de Ânions/metabolismo , Proteínas de Transporte de Ânions/fisiologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Sequência de Bases , Cianobactérias/genética , Regulação Bacteriana da Expressão Gênica , Ácidos Cetoglutáricos/metabolismo , Modelos Genéticos , Dados de Sequência Molecular , Transportadores de Nitrato , Regiões Promotoras Genéticas , Compostos de Amônio Quaternário/metabolismo , Alinhamento de Sequência , Fatores de Transcrição/metabolismo
17.
J Infect Chemother ; 17(6): 821-4, 2011 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-21710162

RESUMO

Many recent studies have shown that Mycoplasma genitalium is among the pathogens responsible for Chlamydia trachomatis-negative nongonococcal urethritis (NGU). A single 1-g dose of azithromycin (AZM) has been recommended for the treatment of NGU, including M. genitalium-positive NGU, irrespective of whether it is positive or negative for Chlamydia trachomatis. The purpose of this study was to determine the minimal inhibitory concentrations of AZM against Mycoplasma genitalium strains, and to assess its clinical efficacy against Mycoplasma genitalium-positive NGU. Seven Mycoplasma genitalium strains were obtained from the American Type Culture Collection, and susceptibility testing of seven antimicrobial agents was performed using a broth microdilution method. Thirty men with M. genitalium-positive NGU were enrolled in this study and treated with a single 1-g dose of AZM. AZM and clarithromycin (CAM) were highly active against M. genitalium strains. Fluoroquinolone activities were moderate, and of the three fluoroquinolones tested, gatifloxacin (GFLX) and sparfloxacin (SPFX) were more active than levofloxacin (LVFX). In 25 of 30 (83.3%) men treated with a single 1-g dose of AZM, M. genitalium was eradicated from first-void urine samples, as determined by polymerase chain reaction. AZM was highly active against M. genitalium, and a single 1-g dose of AZM for M. genitalium-positive NGU was tolerated in Japan. These findings may be helpful in establishing optimal treatment for M. genitalium-positive NGU.


Assuntos
Antibacterianos/farmacologia , Azitromicina/farmacologia , Infecções por Mycoplasma/tratamento farmacológico , Mycoplasma genitalium/efeitos dos fármacos , Uretrite/tratamento farmacológico , Antibacterianos/uso terapêutico , Azitromicina/uso terapêutico , Humanos , Masculino , Testes de Sensibilidade Microbiana , Infecções por Mycoplasma/microbiologia , Mycoplasma genitalium/isolamento & purificação , Uretrite/microbiologia
18.
Int J Urol ; 18(1): 72-5, 2011 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-21198939

RESUMO

Previous studies have shown that lower prostate-specific antigen (PSA) levels in obese men might decrease the sensitivity of prostate cancer screening, leading to delayed diagnosis and unfavorable prognosis. We examined whether the effect of obesity is important in prostate cancer screening of Japanese men, who have a low prevalence of obesity. We analyzed 19,294 male subjects from a large cohort of Toyota Motor Corporation (TMC) employees (aged > 50 years, serum PSA level ≤ 4.0 ng/mL) who underwent physical examinations from August 2006 to December 2009. The relationship between PSA level and obesity-related factors was analyzed by simple and multiple regression analysis. The relationships between six body mass index (BMI) categories, and PSA level and PSA mass (PSA concentration × plasma volume) were analyzed. PSA level decreased significantly with increasing BMI, but the coefficient of determination was very low. Mean PSA values decreased from 1.02 to 0.85 ng/mL as BMI increased from underweight (BMI <18.5) to morbidly obese (BMI >35). However, PSA mass peaked in the overweight category and was slightly reduced with increasing BMI. On multiple regression analysis, PSA level was influenced by age, diastolic blood pressure and high-density lipoprotein as well as BMI. We found an inverse but weak relationship between PSA level and BMI. Obesity seems to have very limited influence on prostate cancer screening in this population. Nonetheless, when considering indications for prostatic biopsy in obese men, we should be aware that the hemodilution effect might reduce PSA levels.


Assuntos
Obesidade/sangue , Antígeno Prostático Específico/sangue , Adulto , Idoso , Povo Asiático , Índice de Massa Corporal , Estudos de Coortes , Humanos , Japão , Masculino , Programas de Rastreamento , Pessoa de Meia-Idade , Neoplasias da Próstata/diagnóstico , Análise de Regressão
19.
J Urol ; 184(3): 851-8; quiz 1235, 2010 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-20643433

RESUMO

PURPOSE: The emergence and spread of Neisseria gonorrhoeae with resistance to oral antibiotics have led to difficulty in treating gonorrhea. We review drug resistance in N. gonorrhoeae with a particular emphasis on resistance to fluoroquinolones, cefixime and azithromycin. MATERIALS AND METHODS: Literature selected from peer reviewed journals listed in MEDLINE(R)/PubMed(R) from 1943 to 2009 and from resources cited in those articles was reviewed comprehensively. RESULTS: Due to the spread of fluoroquinolone resistant N. gonorrhoeae fluoroquinolones are no longer recommended for the treatment of gonorrhea. The emergence of N. gonorrhoeae with a mosaic penicillin-binding protein 2 associated with oral cephalosporin resistance has threatened cefixime treatment for gonorrhea. Emergence of N. gonorrhoeae with high level resistance to azithromycin has also been documented. However, injectable antibiotics (sepctinomycin and ceftriaxone) retain their activity against N. gonorrhoeae. To monitor drug resistance in N. gonorrhoeae several national and international programs have become functional. CONCLUSIONS: Oral regimens for the treatment of gonorrhea are limited. At present to our knowledge ceftriaxone is the most reliable and available agent for the treatment of gonorrhea. To prevent the further emergence and international spread of drug resistance, and allow for the selection of appropriate treatments, a comprehensive global program is needed including surveillance for drug resistance in N. gonorrhoeae and collection of patient epidemiological data. Clinicians should effectively treat patients with gonorrhea, always being conscious of local trends of drug resistance in N. gonorrhoeae, and should perform culture and antimicrobial susceptibility testing in those with persistent gonorrhea after treatment.


Assuntos
Azitromicina/farmacologia , Cefixima/farmacologia , Fluoroquinolonas/farmacologia , Neisseria gonorrhoeae/efeitos dos fármacos , Farmacorresistência Bacteriana , Gonorreia/tratamento farmacológico , Humanos , Vigilância da População , Uretrite/tratamento farmacológico , Uretrite/microbiologia
20.
Hinyokika Kiyo ; 56(4): 225-8, 2010 Apr.
Artigo em Japonês | MEDLINE | ID: mdl-20448447

RESUMO

Carcinoid tumors are low-grade malignant tumors that arise from neuroendocrine cells. Primary renal carcinoid tumors are extremely uncommon. A 63-year-old woman presented with a right abdominal mass and fever. Abdominal computed tomography demonstrated a mass in the right kidney; the mass measured 120 mm in diameter and showed hemorrhage. The patient underwent an uneventful right radical nephrectomy, and histological appearance was typical of carcinoid tumor. Immunohistochemistry demonstrated strong cytoplasmic labeling for neuron-specific enolase and synaptophysin. Additional examinations of the gastrointestinal tract did not show any evidence of carcinoid tumors. The patient remains free from disease recurrence at 8 months after the operation. The prognosis for primary renal carcinoid tumor is relatively optimistic. Complete surgical excision is the only recommended treatment for localized renal carcinoid tumor.


Assuntos
Tumor Carcinoide/cirurgia , Hemorragia/complicações , Nefropatias/complicações , Neoplasias Renais/cirurgia , Tumor Carcinoide/complicações , Tumor Carcinoide/diagnóstico , Tumor Carcinoide/patologia , Feminino , Humanos , Neoplasias Renais/complicações , Neoplasias Renais/diagnóstico , Neoplasias Renais/patologia , Imageamento por Ressonância Magnética , Pessoa de Meia-Idade , Tomografia Computadorizada por Raios X , Resultado do Tratamento
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