RESUMEN
BACKGROUND: Urinary tract infections (UTIs) are common bacterial infections, primarily caused by uropathogenic Escherichia coli (UPEC), leading to significant health issues and economic burden. Although antibiotics have been effective in treating UPEC infections, the rise of antibiotic-resistant strains hinders their efficacy. Hence, identifying novel bacterial targets for new antimicrobial approaches is crucial. Bacterial factors required for maintaining the full virulence of UPEC are the potential target. MepM, an endopeptidase in E. coli, is involved in the biogenesis of peptidoglycan, a major structure of bacterial envelope. Given that the bacterial envelope confronts the hostile host environment during infections, MepM's function could be crucial for UPEC's virulence. This study aims to explore the role of MepM in UPEC pathogenesis. RESULTS: MepM deficiency significantly impacted UPEC's survival in urine and within macrophages. Moreover, the deficiency hindered the bacillary-to-filamentous shape switch which is known for aiding UPEC in evading phagocytosis during infections. Additionally, UPEC motility was downregulated due to MepM deficiency. As a result, the mepM mutant displayed notably reduced fitness in causing UTIs in the mouse model compared to wild-type UPEC. CONCLUSIONS: This study provides the first evidence of the vital role of peptidoglycan endopeptidase MepM in UPEC's full virulence for causing UTIs. MepM's contribution to UPEC pathogenesis may stem from its critical role in maintaining the ability to resist urine- and immune cell-mediated killing, facilitating the morphological switch, and sustaining motility. Thus, MepM is a promising candidate target for novel antimicrobial strategies.
Asunto(s)
Infecciones por Escherichia coli , Infecciones Urinarias , Escherichia coli Uropatógena , Infecciones Urinarias/microbiología , Escherichia coli Uropatógena/genética , Escherichia coli Uropatógena/patogenicidad , Escherichia coli Uropatógena/enzimología , Escherichia coli Uropatógena/efectos de los fármacos , Animales , Ratones , Infecciones por Escherichia coli/microbiología , Virulencia , Endopeptidasas/genética , Endopeptidasas/metabolismo , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/metabolismo , Femenino , Peptidoglicano/metabolismo , Macrófagos/microbiología , Macrófagos/inmunología , Humanos , Modelos Animales de EnfermedadRESUMEN
INTRODUCTION: Carbon dioxide-dependent Proteus mirabilis has been isolated from clinical specimens. It is not clear whether mutations in carbonic anhydrase are responsible for the carbon dioxide dependence of P. mirabilis. The pathogenicity of carbon dioxide-dependent P. mirabilis also remains unclear. The purpose of this study was to determine the cause carbon dioxide dependence of P. mirabilis and its pathogenicity. METHODS: The DNA sequence of can encoding carbonic anhydrase of a carbon dioxide-dependent P. mirabilis small colony variant (SCV) isolate was analyzed. To confirm that impaired carbonic anhydrase activity is responsible for the formation of the carbon dioxide-dependent SCV phenotype of P. mirabilis, we performed complementation experiments using plasmids with intact can. Additionally, mouse infection experiments were performed to confirm the change in virulence due to the mutation of carbonic anhydrase. RESULTS: We found that the can gene of the carbon dioxide-dependent P. mirabilis SCV isolate showed had a frameshift mutation with a deletion of 1 bp (c. 173delC). The can of P. mirabilis encodes carbonic anhydrase was also found to function in Escherichia coli. The cause of the carbon dioxide-dependent SCV phenotype of P. mirabilis was an abnormality in carbonic anhydrase. Nevertheless, no changes were observed in virulence due to the mutation of carbonic anhydrase in mouse infection experiments. CONCLUSIONS: The can gene is essential for the growth of P. mirabilis in ambient air. The mechanisms underlying this fitness advantage in terms of infection warrant further investigation.
Asunto(s)
Dióxido de Carbono , Anhidrasas Carbónicas , Infecciones por Proteus , Proteus mirabilis , Proteus mirabilis/genética , Proteus mirabilis/patogenicidad , Proteus mirabilis/aislamiento & purificación , Dióxido de Carbono/metabolismo , Animales , Ratones , Infecciones por Proteus/microbiología , Anhidrasas Carbónicas/genética , Anhidrasas Carbónicas/metabolismo , Humanos , Virulencia/genética , Femenino , Mutación del Sistema de LecturaRESUMEN
A man in his 50s was diagnosed with right upper lobe non-small-cell lung cancer (cT3N1M0, stage â ¢A) on bronchoscopy. The tumor was located at the right hilum and was bordered extensively on the pulmonary artery. We observed significant tumor shrinkage (ycT1bN1M0, stage â ¡B), following three cycles of systemic chemotherapy combined with an immune checkpoint inhibitor and performed right upper sleeve lobectomy + ND2a-2 via thoracotomy for radical resection. Postoperative histopathological examination showed no residual tumor cells, and the patient was deemed to have a histopathologic complete response. Currently, the patient is being followed up without adjuvant chemotherapy. Several recent studies have reported the usefulness of systemic chemotherapy combined with immune checkpoint inhibitor administration as preoperative induction chemotherapy. However, the role of adjuvant immunotherapy in patients with a histopathologic complete response remains unclear, and careful treatment decision-making is important.
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Carcinoma de Pulmón de Células no Pequeñas , Neoplasias Pulmonares , Masculino , Humanos , Carcinoma de Pulmón de Células no Pequeñas/tratamiento farmacológico , Carcinoma de Pulmón de Células no Pequeñas/cirugía , Carcinoma de Pulmón de Células no Pequeñas/patología , Neoplasias Pulmonares/tratamiento farmacológico , Neoplasias Pulmonares/cirugía , Neoplasias Pulmonares/patología , Inhibidores de Puntos de Control Inmunológico/uso terapéutico , Pulmón/patología , Neumonectomía/efectos adversosRESUMEN
BACKGROUND: Localized pleural adhesion (LPA) evaluation in the apical region is difficult even with four-dimensional ultra-low-dose computed tomography (4D-ULDCT) in the supine position due to smaller pleural movements. PURPOSE: To assess usability of 4D-ULDCT in the lateral decubitus (LD) position for LPA detection in the apical region. MATERIAL AND METHODS: Forty-seven patients underwent 4D-ULDCT of a single respiration cycle with 16-cm coverage of body axis in supine and LD positions with the affected lung uppermost. Intraoperative thoracoscopic findings confirmed LPA presence. A pleural point and a corresponding point on costal outer edge were placed in identical axial planes at end-inspiration. Pleuro-chest wall distance between two points (PCD) was calculated at each respiratory phase. In the affected lung, average change in amount of PCD (PCDACA) was compared between patients with and without LPA in total and two sub-groups (non-COPD and COPD, non-emphysematous and emphysematous patients) in supine and non-dependent (ND) LD positions. Receiver operating characteristic (ROC) curve analysis was performed to determine optimal thresholds in PCDACA for differentiating patients with LPA from those without. RESULTS: In COPD/emphysematous patients and total population, PCDACA with LPA was smaller than in those without in the supine and NDLD positions for overall, lateral, and dorsal regions. For the lateral region in COPD patients, area under ROC curve (AUC) increased from supine (0.64) to NDLD position (0.81). For the dorsal region in emphysematous patients, AUC increased from supine (0.76) to NDLD position (0.96). CONCLUSION: 4D-ULDCT in LD position may be useful for LPA detection in apical regions for COPD and/or emphysematous patients.
Asunto(s)
Tomografía Computarizada Cuatridimensional , Posicionamiento del Paciente , Enfermedades Pleurales/diagnóstico por imagen , Anciano , Anciano de 80 o más Años , Femenino , Tomografía Computarizada Cuatridimensional/métodos , Humanos , Pulmón/diagnóstico por imagen , Masculino , Persona de Mediana Edad , Dosis de Radiación , Adherencias TisularesRESUMEN
A 73 years old male patient with the past history of kidney transplantation was admitted to our hospital for treatment of coronavirus disease 2019 (COVID-19) pneumonia. On the 25th day after the onset of symptoms when his condition was improving, he suddenly developed pneumothorax. Chest tube drainage was performed and connected the tube to the drainage device using a high efficiency particulate air (HEPA) filter. Because of the improvement of infection, the HEPA filter was removed from the drainage device on day 28. Chest tube drainage was continued by day 35, and he was discharged and introduced home oxygen therapy on day 51.
Asunto(s)
COVID-19 , Neumotórax , Anciano , Tubos Torácicos , Drenaje , Humanos , Masculino , Neumotórax/diagnóstico por imagen , Neumotórax/cirugía , SARS-CoV-2RESUMEN
A carbon dioxide-dependent small-colony variant of Escherichia coli SH4888 was isolated from blood cultures of a patient with cholangitis. To date, little is known regarding the molecular mechanisms leading to formation of carbon dioxide-dependent phenotypes in clinical isolates, but abnormalities in the carbonic anhydrase are thought to cause carbon dioxide autotrophy. In this study DNA sequence analysis of the carbonic anhydrase-encoding can locus in the carbon dioxide-dependent E. coli SH4888 revealed that the isolate had a 325-bp deletion spanning from the 3'-terminal region of can to the 3'-terminal region of hpt, which encodes a hypoxanthine phosphoribosyltransferase. To confirm that the carbon dioxide-dependent SCV phenotype of E. coli SH4888 was due to the can mutation, we performed a complementation test with a plasmid carrying an intact can that restored the normal phenotype. However, E. coli SH4888 had increased virulence compared to the can-complemented E. coli SH4888 in a murine infection model. In conclusion, these data confirm that impaired carbonic anhydrase function can cause a carbon dioxide-dependent SCV phenotype in E. coli SH4888 and provides a fitness advantage in terms of infection.
Asunto(s)
Dióxido de Carbono/metabolismo , Anhidrasas Carbónicas/genética , Escherichia coli/genética , Escherichia coli/metabolismo , Anciano , Animales , Cultivo de Sangre , Colangitis/microbiología , Farmacorresistencia Bacteriana , Genes Bacterianos , Prueba de Complementación Genética , Humanos , Masculino , Ratones , Ratones Endogámicos BALB C , Viabilidad Microbiana/genética , Mutación , Análisis de Secuencia de ADNRESUMEN
BACKGROUND: Extraintestinal pathogenic E. coli (ExPEC) is a common gram-negative organism causing various infections, including urinary tract infections (UTIs), bacteremia, and neonatal meningitis. The cjrABC-senB gene cluster of E. coli contributes to ExPEC virulence in the mouse model of UTIs. Consistently, the distribution of cjrABC-senB is epidemiologically associated with human UTIs caused by E. coli. cjrABC-senB, which has previously been proposed to encode an iron uptake system, may facilitate ExPEC survival in the iron availability-restricted urinary tract. Given that the bloodstream is also an iron limited environment to invading bacteria, the pathogenic role of cjrABC-senB in ExPEC bacteremia, however, remains to be investigated. METHODS: The ability of ExPEC RS218 strains with and without cjrABC-senB to survive in the mouse bloodstream and human serum was evaluated. Subsequently, the role of this gene cluster in the ExPEC interaction with the complement system was evaluated. Finally, the distribution of cjrABC-senB in human clinical E. coli isolates was determined by PCR. The frequency of cjrABC-senB in bacteremia isolates that were not associated with UTIs (non-UTI bacteremia isolates) was compared with that in UTI-associated isolates and fecal isolates. RESULTS: Expression of cjrABC-senB attenuated the survival of RS218 in the mouse bloodstream and human serum. The cjrABC-senB-harboring strains triggered enhanced classical- and alternative-complement pathway activation and became more vulnerable to complement-mediated killing in serum. cjrA was identified as the major gene responsible for the attenuated serum survival. Expressing cjrABC-senB and cjrA increased bacterial susceptibility to detergent and induced periplasmic protein leakage, suggesting that the expression of these genes compromises the integrity of the outer membrane of ExPEC. In addition, the frequency of cjrABC-senB in non-UTI bacteremia isolates was significantly lower than that in UTI-associated isolates, while the frequencies in non-UTI bacteremia isolates and fecal isolates showed no significant difference. Consistently, this epidemiological investigation suggests that cjrABC-senB does not contribute to E. coli bacteremia in humans. CONCLUSION: The contribution of cjrABC-senB to the pathogenesis of ExPEC is niche dependent and contradictory because the genes facilitate ExPEC UTIs but hinder bacteremia. The contradictory niche-dependent characteristic may benefit the development of novel strategies against E. coli-caused infections.
Asunto(s)
Bacteriemia/microbiología , Activación de Complemento , Infecciones por Escherichia coli/microbiología , Proteínas de Escherichia coli/metabolismo , Escherichia coli Patógena Extraintestinal/fisiología , Genes Bacterianos , Familia de Multigenes , Animales , Escherichia coli Patógena Extraintestinal/genética , Ratones , Ratones Endogámicos BALB CRESUMEN
BACKGROUND: Extraintestinal pathogenic E. coli (ExPEC) remains one of the most prevalent bacterial pathogens that cause extraintestinal infections, including neonatal meningitis, septicemia, and urinary tract (UT) infections (UTIs). Antibiotic therapy has been the conventional treatment for such infections, but its efficacy has decreased due to the emergence of antibiotic-resistant bacteria. Identification and characterization of bacterial factors that contribute to the severity of infection would facilitate the development of novel therapeutic strategies. The ExPEC periplasmic protease Prc contributes to the pathogen's ability to evade complement-mediated killing in the serum. Here, we further investigated the role of the Prc protease in ExPEC-induced UTIs and the underlying mechanism. METHODS: The uropathogenic role of Prc was determined in a mouse model of UTIs. Using global quantitative proteomic analyses, we revealed that the expression of FliC and other outer membrane-associated proteins was altered by Prc deficiency. Comparative transcriptome analyses identified that Prc deficiency affected expression of the flagellar regulon and genes that are regulated by five extracytoplasmic signaling systems. RESULTS: A mutant ExPEC with a prc deletion was attenuated in bladder and kidney colonization. Global quantitative proteomic analyses of the prc mutant and wild-type ExPEC strains revealed significantly reduced flagellum expression in the absence of Prc, consequently impairing bacterial motility. The prc deletion triggered downregulation of the flhDC operon encoding the master transcriptional regulator of flagellum biogenesis. Overexpressing flhDC restored the prc mutant's motility and ability to colonize the UT, suggesting that the impaired motility is responsible for attenuated UT colonization of the mutant. Further comparative transcriptome analyses revealed that Prc deficiency activated the σE and RcsCDB signaling pathways. These pathways were responsible for the diminished flhDC expression. Finally, the activation of the RcsCDB system was attributed to the intracellular accumulation of a known Prc substrate Spr in the prc mutant. Spr is a peptidoglycan hydrolase and its accumulation destabilizes the bacterial envelope. CONCLUSIONS: We demonstrated for the first time that Prc is essential for full ExPEC virulence in UTIs. Our results collectively support the idea that Prc is essential for bacterial envelope integrity, thus explaining how Prc deficiency results in an attenuated ExPEC.
Asunto(s)
Endopeptidasas/genética , Infecciones por Escherichia coli/genética , Proteínas de Escherichia coli/genética , Escherichia coli Patógena Extraintestinal/genética , Flagelina/genética , Infecciones Urinarias/genética , Animales , Farmacorresistencia Bacteriana/genética , Infecciones por Escherichia coli/microbiología , Infecciones por Escherichia coli/patología , Escherichia coli Patógena Extraintestinal/patogenicidad , Regulación Bacteriana de la Expresión Génica/genética , Humanos , Ratones , Proteómica , Transducción de Señal/genética , Infecciones Urinarias/microbiología , Infecciones Urinarias/patología , Escherichia coli Uropatógena/genética , Escherichia coli Uropatógena/patogenicidad , Factores de Virulencia/genéticaRESUMEN
Nuss procedure for pediatric patients with pectus excavatum has been practiced worldwide, including in Japan, due to the simple procedure and has a high therapeutic effect. Because it is usually performed under thoracoscopy to secure the safety, it is performed not only by pediatric or plastic surgeons but also by general thoracic surgeons. On the other hand, a risk of infection must always be considered in this method in which a foreign metal bar is used. In particular, when the skin barrier mechanism is declining due to skin diseases such as atopic dermatitis, the risk of infection of the implant may increase. The present case was an 8-year-old male with a history of atopic dermatitis. He underwent thoracoscopic Nuss procedure. Although there was no problem during his hospitalization, the bar was exposed from the skin on the 58th postoperative day with the infection triggered, and the unexpected early bar removal was performed on the 66th postoperative day. We report this case with some literature review.
Asunto(s)
Dermatitis Atópica , Tórax en Embudo , Infecciones , Niño , Humanos , Japón , Masculino , Procedimientos Quirúrgicos Mínimamente Invasivos , Estudios Retrospectivos , ToracoscopíaRESUMEN
Wall teichoic acids (WTAs) are anionic polymers that are covalently linked to peptidoglycan and play important roles in cell shape determination, cell division, autolysis, pathogenesis and antibiotic resistance in Gram-positive bacteria. In Bacillus subtilis, WTA is synthesized in the cytoplasm and translocated by an ABC transporter, TagGH. In this study, we found that the transmembrane segment of TagH is required for WTA transport under high temperatures. Cells expressing TagH302-FL (a construct fused to the 6×FLAG tag after the transmembrane segment, which lacks the C-terminal extracellular domain) grew normally at high temperatures, similar to those expressing the full-length TagH-FL fusion. In contrast, cells expressing TagH275-FL, which lacks both the transmembrane segment and the extracellular domain, exhibited a temperature-sensitive phenotype at temperatures above 49 °C and a growth defect at 50 °C. Interestingly, this growth defect was dissolved by an additional incubation at 37 °C. A similar temperature-sensitive phenotype was observed in cells expressing an N-terminal 6×FLAG tag fusion of TagH275. Immunofluorescence microscopy (IFM) indicated that TagG and TagH are localized on the cytoplasmic membrane in a patch-like manner. In addition, the C-terminal-truncated forms, TagH275-FL and TagH302-FL, were localized in similar patch-like patterns at 37 °C; only foci for TagH275-FL were remarkably reduced at high temperatures. Moreover, cell surface decoration with WTA was considerably reduced in cells harbouring TagH275-FL at high temperature, supporting the results of IFM observation. These results suggest that the transmembrane segment of TagH plays an important role in WTA export at high temperatures.
Asunto(s)
Transportadoras de Casetes de Unión a ATP/metabolismo , Bacillus subtilis/fisiología , Proteínas Bacterianas/metabolismo , Transporte Biológico/genética , Pared Celular/metabolismo , Respuesta al Choque Térmico/fisiología , Hidrolasas/metabolismo , Ácidos Teicoicos/metabolismo , Transportadoras de Casetes de Unión a ATP/química , Transportadoras de Casetes de Unión a ATP/genética , Bacillus subtilis/genética , Bacillus subtilis/crecimiento & desarrollo , Bacillus subtilis/metabolismo , Proteínas Bacterianas/química , Proteínas Bacterianas/genética , Membrana Celular/metabolismo , Eliminación de Gen , Hidrolasas/química , Hidrolasas/genética , Dominios Proteicos , Proteínas Recombinantes de Fusión/química , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/metabolismoRESUMEN
Bacterial cells are covered with peptidoglycan (PG) layer(s), serving as the cellular exoskeleton. The PG sacculus changes its shape during cell growth, and thus both the synthesis and disassembly of PG are important for cell proliferation. In Bacillus subtilis, four dl-endopeptidases (DLEPases; LytE, LytF, CwlO and CwlS) are involved in the maintenance of cell morphology. The lytE cwlO double mutant exhibits synthetic lethality and defective cell elongation, while the lytE lytF cwlS triple mutant exhibits defective cell separation, albeit with septum formation. LytE is involved in both cell separation and elongation. We propose that DLEPases have varied roles in cell separation and elongation. To determine these roles, the catalytic domain of LytE was substituted with another catalytic domain that digests the other bonds in PG. By using the chimeric enzymes, we assessed the suppression of the synthetic lethality by the cell elongation defect and the disruption of chain morphology by the cell separation defect. All the constructed chimeric enzymes suppressed the cell separation defect, restoring the chain morphology. Digestion at any position of PG broke the linkage between two daughter cells, releasing them from each other. However, only d,d-endopeptidases suppressed the lack of DLEPase in the lytE cwlO double mutant. This indicated that the release of tension on the expanding PG sacculus is not the sole essential function of DLEPases. Considering that the structure of the digested PG is important for cell elongation, the digested product might be reused in the growth process in some way.
Asunto(s)
Bacillus subtilis/crecimiento & desarrollo , Bacillus subtilis/metabolismo , Proteínas Bacterianas/metabolismo , Endopeptidasas/metabolismo , Peptidoglicano/metabolismo , Bacillus subtilis/citología , Bacillus subtilis/enzimología , Proteínas Bacterianas/química , Proteínas Bacterianas/genética , Dominio Catalítico/genética , División Celular , Pared Celular/metabolismo , Endopeptidasas/química , Endopeptidasas/genética , Peptidoglicano/biosíntesis , Proteínas Recombinantes de Fusión/análisis , Proteínas Recombinantes de Fusión/metabolismo , Mutaciones Letales SintéticasRESUMEN
Non-C5 -units terpenoids (norisoprenoids) with an acetonyl group are widely distributed in nature. However, studies on the biosynthesis of norisoprenoids are scarce. Now, the C33 norisoprenoid, (all-E)-farnesylfarnesylacetone, was identified from Bacillus spp. and it was elucidated for the first time that superoxide mediates the cleavage of menaquinones (vitaminâ K) to form norisoprenoids in saponification treatment. From inâ vivo experiments using gene-disrupted Bacillus subtilis strains targeted for enzymes responsible for menaquinone biosynthesis and for superoxide dismutase, it was suggested that the non-enzymatic cleavage (autoxidation) of menaquinone with superoxide resulted in norisoprenoid synthesis in Bacillus cells. Furthermore, the bioactive norisoprenoids, farnesylacetone and phytone, were produced in Bacillus cells by this novel synthesis system.
Asunto(s)
Bacillus/química , Superóxidos/química , Terpenos/metabolismo , Bacillus/citología , Bacillus/metabolismo , Estructura Molecular , Terpenos/químicaRESUMEN
BACKGROUND: An attenuated mutant (designated NY303) of Vibrio vulnificus, which causes serious wound infection and septicemia in humans, was isolated fortuitously from a clinical strain YJ016. This mutant was defective in cytotoxicity, migration on soft agar and virulence in the mouse. The purpose of this study was to map the mutation in this attenuated mutant and further explore how the gene thus identified is involved in virulence. METHODS: The whole genome sequence of mutant NY303 determined by next-generation sequencing was compared with that of strain YJ016 to map the mutations. By isolating and characterizing the specific gene-knockout mutants, the gene associated with the phenotype of mutant NY303 was identified. This gene encodes a global regulator, Lrp. A mutant, YH01, deficient in Lrp was isolated and examined in vitro, in vivo and ex vivo to find the affected virulence mechanisms. The target genes of Lrp were further identified by comparing the transcriptomes, which were determined by RNA-seq, of strain YJ016 and mutant YH01. The promoters bound by Lrp were identified by genome footprinting-sequencing, and those related with virulence were further examined by electrophoretic mobility shift assay. RESULTS: A mutation in lrp was shown to be associated with the reduced cytotoxicity, chemotaxis and virulence of mutant NY303. Mutant YH01 exhibited a phenotype resembling that of mutant NY303, and was defective in colonization in the mouse and growth in mouse serum, but not the antiphagocytosis ability. 596 and 95 genes were down- and up-regulated, respectively, in mutant YH01. Many of the genes involved in secretion of the MARTX cytotoxin, chemotaxis and iron-acquisition were down-regulated in mutant YH01. The lrp gene, which was shown to be negatively autoregulated, and 7 down-regulated virulence-associated genes were bound by Lrp in their promoters. A 14-bp consensus sequence, mkCrTTkwAyTsTG, putatively recognized by Lrp was identified in the promoters of these genes. CONCLUSIONS: Lrp is a global regulator involved in regulation of cytotoxicity, chemotaxis and iron-acquisition in V. vulnificus. Down-regulation of many of the genes associated with these properties may be responsible, at least partly, for loss of virulence in mutant NY303.
Asunto(s)
Proteínas Bacterianas/genética , Regulación hacia Abajo , Proteína Reguladora de Respuesta a la Leucina/genética , Mutación , Vibrio vulnificus/genética , Vibrio vulnificus/patogenicidad , Virulencia/genética , Animales , Proteínas Bacterianas/metabolismo , Proteína Reguladora de Respuesta a la Leucina/metabolismo , Ratones , Ratones Endogámicos C3H , Enfermedades de los Roedores/microbiología , Vibriosis/microbiología , Vibrio vulnificus/fisiologíaRESUMEN
We treated a 72-year-old Japanese female with sustained high fever and overall body exhaustion. An infectious liver cyst and right lung pneumonia were suspected causes. Hepatic cystectomy and various antibiotics did not resolve symptoms. Pneumonia exacerbation and ascitic fluid retention, left lumbar spinal osteomyelitis, and peri-gastric lymph node abscess penetrating the stomach were observed. Mycobacterium avium was identified in sputum, ascites, vertebral body abscess puncture specimen, and pus mucus secretion in the stomach. We diagnosed a disseminated nontuberculous mycobacterial infection. She seemed immunocompetent, without signs of AIDS or hematological malignancy. Serum anti-IFN-γ autoantibodies tested positive and were suspected to be involved in the illness onset.
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Autoanticuerpos/sangre , Interferón gamma/inmunología , Infecciones por Mycobacterium no Tuberculosas/inmunología , Anciano , Femenino , Humanos , Infecciones por Mycobacterium no Tuberculosas/tratamiento farmacológicoRESUMEN
Type 1 fimbriae are filamentous structures on Escherichia coli. These structures are important adherence factors. Because binding to the host cells is the first step of infection, type 1 fimbria is an important virulence factor of pathogenic E. coli. Expression of type 1 fimbria is regulated by a phase variation in which each individual bacterium can alternate between fimbriated (phase-ON) and nonfimbriated (phase-OFF) states. The phase variation is regulated by the flipping of the 314-bp fimS fragment, which contains the promoter driving the expression of the genes required for the synthesis of type 1 fimbria. Thus, the bacterial proteins able to interact with fimS are likely to be involved in regulating the expression of type 1 fimbria. To identify novel type 1 fimbria-regulating factors, we used an E. coli K12 proteome chip to screen for the bacterial factors able to interact with a 602-bp DNA fragment containing fimS and its adjacent regions. The Spr protein was identified by the proteome chip-based screening and further confirmed to be able to interact with fimS by electrophoretic mobility shift assay. Deletion of spr in the neonatal meningitis E. coli strain RS218 significantly increased the ratio of the bacterial colonies that contained the type 1 fimbria phase-ON cells on agar plates. In addition, Spr interfered with the interactions of fimS with the site-specific recombinases, FimB and FimE, which are responsible for mediating the flipping of fimS. These results suggest that Spr is involved in the regulation of type 1 fimbria expression through direct interaction with the invertible element fimS. These findings facilitate our understanding of the regulation of type 1 fimbria.
Asunto(s)
Proteínas Bacterianas/biosíntesis , Fimbrias Bacterianas/genética , Proteoma/genética , Factores de Virulencia/biosíntesis , ADN Nucleotidiltransferasas/biosíntesis , Escherichia coli K12/genética , Regulación Bacteriana de la Expresión Génica , Regiones Promotoras Genéticas , Proteoma/biosíntesis , Recombinación Genética , Factores de Virulencia/genéticaRESUMEN
Bacillus subtilis LytF plays a principal role in cell separation through its localization at the septa and poles on the vegetative cell surface. In this study, we found that a mutation in a major lipoteichoic acid (LTA) synthase gene--ltaS--results in a considerable reduction in the σ(D)-dependent transcription of lytF. The lytF transcription was also reduced in mutants that affected glycolipid anchor biosynthesis. Immunofluorescence microscopy revealed that both the numbers of cells expressing LytF and the LytF foci in these mutants were decreased. In addition, the transcriptional activity of lytF was almost abolished in the double (ltaS yfnI), triple (ltaS yfnI yqgS), and quadruple (ltaS yfnI yqgS yvgJ) mutants during vegetative growth. Cell separation defects in these mutants were partially restored with artificial expression of LytF. Interestingly, when lytF transcription was induced in the ltaS single or multiple mutants, LytF was localized not only at the septum, but also along the sidewall. The amounts of LytF bound to cell wall in the single (ltaS) and double (ltaS yfnI) mutants gradually increased as compared with that in the WT strain, and those in the triple (ltaS yfnI yqgS) and quadruple mutants were almost similar to that in the double mutant. Moreover, reduction of the lytF transcription and chained cell morphology in the ltaS mutant were completely restored with artificial induction of the yqgS gene. These results strongly suggest that LTA influences the temporal, σ(D)-dependent transcription of lytF and is an additional inhibitory component to the vegetative cell separation enzyme LytF.
Asunto(s)
Bacillus subtilis/química , Bacillus subtilis/enzimología , Proteínas Bacterianas/análisis , Endopeptidasas/análisis , Regulación Bacteriana de la Expresión Génica , Bacillus subtilis/genética , Bacillus subtilis/fisiología , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , División Celular , Endopeptidasas/genética , Microscopía Fluorescente , Factor sigma/metabolismo , Transcripción GenéticaRESUMEN
The Gram-positive bacterium Bacillus subtilis has a thick cell wall. The cell wall contains various proteins, both for secretion and for peptidoglycan (PG) maintenance. Penicillin-binding proteins for PG synthesis, PG hydrolases (autolysins), and regulator proteins for the autolysins are the known components of the PG maintenance system. YqgA was identified as an abundant protein attached to the cell wall of B. subtilis through a proteomics analysis. The YqgA protein was localized at cell division sites during the transition period between the exponential and the stationary phases. YqgA localization was affected by mutations in the dl-endopeptidases (DLEPases), which are the autolysins involved in cell morphogenesis. Furthermore, yqgA mutations on a background of defective DLEPases led to delays in cell growth and cell morphological changes. These results demonstrate that yqgA is genetically related to the genes encoding DLEPases involved in cell morphogenesis.
Asunto(s)
Bacillus subtilis/citología , Bacillus subtilis/genética , Proteínas Bacterianas/genética , Pared Celular/metabolismo , Endopeptidasas/genética , Homología de Secuencia de Ácido Nucleico , Bacillus subtilis/enzimología , Bacillus subtilis/metabolismo , Proteínas Bacterianas/metabolismo , Proliferación Celular , Regulación Bacteriana de la Expresión Génica , Espacio Intracelular/metabolismo , Mutación , Transporte de ProteínasRESUMEN
OBJECTIVE: To evaluate the heat effects of radiofrequency ablation (RFA) on normal bone by mechanical testing, MRI, and histology. MATERIALS AND METHODS: The institutional animal care and use committee approved the animal study. Thirty-two adult Japanese white rabbits were included in our study. Bone biopsy needles were inserted from the distal end of the right (RFA side) and the left (control side) femurs using a fluoroscopic guide. A 17-gauge internally cooled RFA electrode with a 2-cm active tip was inserted through the needle to the right femur, and RFA was performed for 12 min using a 200-W generator. Animals were divided into four groups and 8 animals from each group were euthanized on day 1, day 7, day 30, and day 60 after RFA. MRI was performed prior to euthanasia. Three-point bending test was performed to measure flexural strength. Student's t test was used to evaluate for significant differences between RFA and control side for each group. Femurs underwent histological examination by hematoxylin and eosin staining after the bending test. RESULTS: MRI showed a high-intensity rim around the bone on T2WI. Three-point bending test showed no statistically significant differences (P < 0.05) between the RFA and the control side in any of the groups. Histologically, osteocytes of cortical bone showed cell death, but the lamellar structure was preserved in all groups and bone remodeling was observed. CONCLUSION: Heat by RFA did not change normal bone strength within 2 months, despite the heat effects in the cortical bone and cell death.
Asunto(s)
Ablación por Catéter/métodos , Fémur/fisiopatología , Fémur/cirugía , Animales , Módulo de Elasticidad , Fémur/patología , Calor , Imagen por Resonancia Magnética , Conejos , Estrés Mecánico , Resistencia a la Tracción/fisiologíaRESUMEN
We report a rare case of pulmonary metastasis of invasive thymoma, with endobronchial polypoid growth causing hemosputum in a 77-year-old man. The patient had been admitted 8 years earlier for the treatment of invasive thymoma and had undergone extended thymo-thymectomy through a mid-sternotomy, followed by a course of radiotherapy. Pulmonary metastases developed 3 years after surgery, for which the patient received several courses of chemotherapy; however, the tumor continued to progress gradually. He presented at our emergency unit within 4 years of completion of the chemotherapy, with sudden massive hemoptysis. We performed endotracheal intubation to prevent suffocation and bronchoscopic examination revealed that a tumor and blood clots had obstructed the left main bronchus. We performed bronchial arterial embolization and endoscopic electrosurgery to resect the tumor, then occluded the responsible bronchus with an endobronchial Watanabe spigot to prevent further endobronchial polypoid growth and bronchial hemorrhage from the invasive thymoma.
Asunto(s)
Neoplasias de los Bronquios/patología , Neoplasias Pulmonares/secundario , Pólipos/patología , Timoma/secundario , Neoplasias del Timo/patología , Anciano , Arterias Bronquiales , Neoplasias de los Bronquios/terapia , Terapia Combinada , Electrocirugia/métodos , Embolización Terapéutica , Endoscopía , Hemoptisis/etiología , Humanos , Neoplasias Pulmonares/terapia , Masculino , Invasividad Neoplásica , Pólipos/terapia , Esputo , Timectomía , Timoma/complicaciones , Timoma/cirugía , Timoma/terapia , Neoplasias del Timo/complicaciones , Neoplasias del Timo/cirugía , Resultado del TratamientoRESUMEN
We reported a case of surgically resected double bronchogenic cysts within the anterior mediastinum. An anterior mediastinal tumor had been found at medical examination 6 years ago in a 66-year-old man, but has been followed up without treatment. After the treatment of another disease, he was referred to our hospital for evaluation of the mediastinal tumor. A chest computed tomography showed 2 anterior mediastinal nodules. Nodules in the thymus were resected with video-assisted thoracic surgery. The tumors were both pathologically diagnosed as bronchogenic cysts.