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1.
Artigo em Inglês | MEDLINE | ID: mdl-38501552

RESUMO

Accessible Summary What is known on the subject? Functioning is one of the most affected areas in schizophrenia. Social, occupational and personal domains are affected, and these deficits are responsible for a major part of the disability associated with the disorder. There are several instruments to measure functioning, but the HoNOS provides a wide assessment of impairment in 12 areas of functioning. What does the paper add to existing knowledge? The Spanish version of the HoNOS shows good properties in terms of reliability and validity for use in schizophrenia patients. Although some authors divide the scale according to proposed underlying dimensions, in schizophrenia this division may not be appropriate. What are the implications for practice? A reliable and easy-to-use measure of impairment in different areas of functioning is useful for optimizing the treatment and rehabilitation of patients with schizophrenia. ABSTRACT: INTRODUCTION: The HoNOS scale was designed for the assessment of psychosocial impairment in various domains. While it is widely used in psychiatric settings, it has not been validated in Spanish for use in patients with schizophrenia. AIM: To examine the psychometric properties of the Spanish version of the HoNOS scale in a sample of schizophrenia patients. METHOD: A total of 194 individuals aged 18 to 65 with schizophrenia spectrum diagnoses were evaluated using the HoNOS. Illness severity and level of functioning were also assessed. RESULTS: The HoNOS showed moderate internal consistency, good inter-observer reliability and good test-retest reliability. Factor analysis revealed an internal structure consisting of four factors, with item distribution differing from the theoretical dimensions proposed for the original scale. DISCUSSION: The Spanish version of the HoNOS scale is a reliable and valid instrument for assessing psychosocial impairment in individuals diagnosed with schizophrenia spectrum disorders. However, further research is needed to determine its internal structure more accurately. IMPLICATIONS FOR PRACTICE: The HoNOS scale provides researchers and clinicians with a valid measure of impairment in twelve different domains, which can facilitate and guide the treatment of schizophrenia patients.

2.
Rev Esp Enferm Dig ; 115(2): 99, 2023 02.
Artigo em Inglês | MEDLINE | ID: mdl-35748467

RESUMO

Neuroendocrine tumors (NETs) or carcinoids represent a small percentage of gastrointestinal neoplasms (2%). The ileum (41.8%), rectum (27.4%) and stomach (8.7%) are the most common locations. We present an unusual case of NET due to its origin in the ampulla of Vater, as an extremely rare cause of biliary obstruction. A bibliographic review of the current recommendations of management and treatment for this case, different from other locations of the gastrointestinal tract is carried out. The curative treatment of choice for early-stage Vater ampulla NETs is Whipple surgery with lymphadenectomy due to Its high rate of lymphatic and metastatic dissemination, regardless of size and histological grade, although there are no updated official guidelines.


Assuntos
Ampola Hepatopancreática , Tumor Carcinoide , Neoplasias do Ducto Colédoco , Tumores Neuroendócrinos , Humanos , Tumores Neuroendócrinos/cirurgia , Tumores Neuroendócrinos/patologia , Ampola Hepatopancreática/cirurgia , Ampola Hepatopancreática/patologia , Neoplasias do Ducto Colédoco/cirurgia , Neoplasias do Ducto Colédoco/patologia , Tumor Carcinoide/cirurgia , Tumor Carcinoide/patologia , Pancreaticoduodenectomia
4.
Rev. esp. enferm. dig ; 112(12): 903-908, dic. 2020. tab, graf
Artigo em Espanhol | IBECS | ID: ibc-200577

RESUMO

INTRODUCCIÓN: la biopsia hepática percutánea es un procedimiento necesario para el diagnóstico de hepatopatías no exento de complicaciones y con malestar psicológico para el paciente. OBJETIVO: determinar el perfil de seguridad del propofol en la biopsia hepática percutánea, las complicaciones de la técnica per se y la satisfacción de los pacientes tras su realización. MÉTODOS: estudio observacional retrospectivo mediante recogida de datos de tolerancia y complicaciones en pacientes sometidos a biopsia hepática bajo sedación profunda con propofol. Valoración de la calidad y satisfacción percibida por los pacientes mediante una encuesta transversal. RESULTADOS: incluimos 97 pacientes con una dosis media de propofol de 170,46 mg. De las complicaciones derivadas de la sedación, se registraron seis desaturaciones leves (6,2 %) resueltas con maniobras posturales (50 %) y parada de la bomba de propofol (50 %) y once episodios de hipotensión (11,3 %) resueltos de forma espontánea (82,82 %) o fluidoterapia (18,18 %). De las complicaciones derivadas de la técnica, se registraron tres casos de dolor precoz (3,1 %) y uno tardío (1,03 %), todos resueltos con 1 g de paracetamol intravenoso. Todos los pacientes iniciaron tolerancia oral y fueron dados de alta a las 24 horas del procedimiento sin necesidad de analgesia ambulatoria. La satisfacción general, así como el malestar psicológico fueron evaluados como muy buenos/excelentes en el 100 % de los pacientes. DISCUSIÓN: el propofol presenta un adecuado perfil de seguridad en la biopsia hepática y mantiene el éxito de la prueba, con buena tolerancia de la misma por el paciente. Consideramos posible ampliar la utilidad de la sedación con propofol a este procedimiento


No disponible


Assuntos
Humanos , Feminino , Pessoa de Meia-Idade , Fígado/patologia , Hepatopatias/patologia , Sedação Profunda/métodos , Propofol/administração & dosagem , Resultado do Tratamento , Biópsia , Hepatopatias/diagnóstico , Biópsia por Agulha/métodos , Hepatopatias/psicologia , Estudos Transversais , Acetaminofen/administração & dosagem , Satisfação do Paciente , Estudos Retrospectivos , Inquéritos e Questionários
5.
Rev. esp. enferm. dig ; 112(11): 854-859, nov. 2020. tab
Artigo em Espanhol | IBECS | ID: ibc-198770

RESUMO

INTRODUCCIÓN: el uso del consentimiento informado es necesario en los procedimientos invasivos como documento garantizador de la relación sanitaria ética y de la seguridad del paciente. OBJETIVO: analizar si se poseen y utilizan los documentos de consentimiento informado para paracentesis en los centros sanitarios, así como obtener algunos datos de interés sobre la técnica. MATERIAL Y MÉTODO: realizamos un estudio observacional descriptivo mediante una encuesta transversal on-line difundida por redes sociales, destinada a los especialistas y residentes de aparato digestivo durante diciembre de 2019. RESULTADOS: incluimos 203 encuestas anónimas (55,2 % adjuntos y 44,8 % residentes) de 74 centros sanitarios de 34 provincias españolas. Noventa encuestados (44,3 %) tenían dicho documento en sus centros; de estos, 29 (32,2 %) lo entregaban siempre; 31 (34,4 %), algunas veces; y 21 (23,3 %), nunca. Setenta y dos profesionales (35,5 %) contestaron no tenerlo y 41 (20,5 %), ser desconocedores; de entre ellos, 77 (68,1 %) consideraban necesaria su creación, 31 (27,4 %) no lo creían así y cinco (4,4 %) no contestaron. En cuanto a la técnica, 173 facultativos (85,2 %) realizan la punción bajo visión directa y 30 (14,8 %), ecoguiada en la mayoría de las ocasiones. Ciento nueve (53,7 %) aplican siempre anestésico local, 80 (39,4 %) lo aplican en algunas ocasiones y 14 (6,9 %) no lo utilizan. Ciento sesenta y siete encuestados (82,3 %) consideraron que es una técnica sencilla, frente a 36 (17,7 %) que la consideran de complejidad intermedia. En cuanto al riesgo, 150 (73,5 %) lo consideran bajo y 52 (25,6 %), medio. Noventa y nueve de ellos (48,8 %) refieren haber tenido complicaciones menores y 37 (18,2 %), mayores. CONCLUSIONES: la paracentesis es una técnica habitual en los servicios de digestivo y, a pesar de ser considerada sencilla y segura, entraña complicaciones. Por ello consideramos necesaria la formación reglada en esta técnica, así como la creación, difusión y utilización de los consentimientos informados dada la importante variabilidad intra e interhospitalaria que presenta


No disponible


Assuntos
Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Consentimento Livre e Esclarecido/estatística & dados numéricos , Paracentese/ética , Pesquisas sobre Atenção à Saúde/estatística & dados numéricos , Estudos Transversais , Internet , Corpo Clínico Hospitalar/estatística & dados numéricos , Padrões de Prática Médica , Espanha
6.
Rev Esp Enferm Dig ; 112(12): 903-908, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33118358

RESUMO

INTRODUCTION: the percutaneous hepatic biopsy is a necessary procedure for the diagnosis of liver diseases which can cause complications and psychological discomfort for the patient. AIMS: to determine the safety profile of propofol in percutaneous hepatic biopsy, the complications of the technique per se and patients satisfaction once completed. METHODS: a retrospective observational study was performed via the acquisition of data of tolerance and perceived quality by the patients using a transversal survey. RESULTS: ninety-seven patients were included with an average propofol dose of 170.46 mg. Of the complications resulting from the sedation, there were six slight desaturations (6.2 %) resolved with a forehead maneuver (50 %) or cessation of the propofol infusion pump (50 %) and eleven hypotension episodes (11.3 %) resolved without intervention (82.82 %) or with fluid replacement (18.18 %). Of the complications resulting from the technique, there were three cases of early-onset pain (3.1 %) and one delayed (1.03 %); all were resolved with 1 g of intravenous paracetamol. All patients were discharged with oral tolerance and without the need for analgesia 24 hours after the procedure. General satisfaction, as well as psychological discomfort, were evaluated as "very good/excellent" in 100 % of the patients. DISCUSSION: propofol demonstrated a favorable safety profile in hepatic biopsy, aiding in the ultimate success of the procedure and tolerance for the patient. We propose the expansion of the use of sedation with propofol to this procedure.


Assuntos
Propofol , Biópsia , Sedação Consciente , Humanos , Hipnóticos e Sedativos/efeitos adversos , Dor , Propofol/efeitos adversos
7.
Rev Esp Enferm Dig ; 112(11): 854-859, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-33054303

RESUMO

INTRODUCTION: informed consent is necessary for invasive procedures as a document that guarantees the ethical health relationship and patient safety. AIMS: to analyze whether we have and use informed consent documents for paracentesis in our hospitals and to obtain data on the technique. METHODS: a descriptive observational study was performed during December 2019, via a cross-sectional survey disseminated through social networks, aimed at specialists and residents of gastroenterology. RESULTS: two hundred and three anonymous surveys were included (55.2 % gastroenterologist and 44.8 % residents) from 74 hospitals in 34 Spanish provinces. Ninety respondents (44.3 %) stated that they had the document in their centers. Of these, 29 (32.2 %) always provided it, 31 (34.4 %) provided it sometimes and 21 (23.3 %) never. Seventy-two professionals (35.5 %) answered that they did not have it and 41 (20.5 %) selected "unknown". Of these, 77 (68.1 %) considered it was necessary to create this document, 31 (27.4 %) did not think it was necessary and five (4.4 %) did not answer. With regards to the technique, 173 (85.2 %) performed paracentesis under direct visualization and 30 (14.8 %) were eco-guided on most occasions. One hundred and nine (53.7 %) always applied local anesthetic, 80 (39.4 %) sometimes and 14 (6.9 %) did not. One hundred and sixty-seven respondents (82.3 %) considered it to be a simple technique versus 36 (17.7 %) who thought that it was of intermediate complexity. In terms of risk, 150 (73.5 %) considered it was low and 52 (25.6 %), medium. Ninety-nine (48.8 %) experienced minor complications and 37 (18.2 %) experienced major complications. CONCLUSIONS: paracentesis is a common technique in digestive services and could be associated with complications, even though it is considered to be simple and safe. Due to the important intra- and inter-hospital variability that this technique presents, we consider standardized training in this technique is necessary, as well as the creation, spread and use of informed consents.


Assuntos
Consentimento Livre e Esclarecido , Paracentese , Estudos Transversais , Humanos , Inquéritos e Questionários
8.
Extremophiles ; 23(5): 587-597, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31250111

RESUMO

Reactive oxygen species and nitrogen species (ROS and RNS), produced in a wide range of physiological process even under low oxygen availability, are among the main stressors found in the environment. Strategies developed to combat them constitute key features in bacterial adaptability and survival. Pseudomonas extremaustralis is a metabolic versatile and stress resistant Antarctic bacterium, able to grow under different oxygen conditions. The present work explores the effect of oxidative stress under low oxygen conditions in P. extremaustralis, by combining RNA deep sequencing analysis and physiological studies. Cells grown under microaerobiosis exhibited more oxidative damage in macromolecules and lower survival rates than under aerobiosis. RNA-seq analysis showed an up-regulation of genes related with oxidative stress response, flagella, chemotaxis and biofilm formation while chaperones and cytochromes were down-regulated. Microaerobic cultures exposed to H2O2 also displayed a hyper-flagellated phenotype coupled with a high motility behavior. Moreover, cells that were subjected to oxidative stress presented increased biofilm formation. Altogether, our results suggest that a higher motile behavior and augmented capacity to form biofilm structures could work in addition to well-known antioxidant enzymes and non-enzymatic ROS scavenging mechanisms to cope with oxidative stress at low oxygen tensions.


Assuntos
Quimiotaxia , Flagelos/metabolismo , Estresse Oxidativo , Pseudomonas/metabolismo , Transcriptoma , Biofilmes , Genes Bacterianos , Oxigênio/metabolismo , Pseudomonas/genética , Pseudomonas/fisiologia
9.
J Ind Microbiol Biotechnol ; 45(1): 15-23, 2018 01.
Artigo em Inglês | MEDLINE | ID: mdl-29116430

RESUMO

Diesel fuel is one of the most important sources of hydrocarbon contamination worldwide. Its composition consists of a complex mixture of n-alkanes, branched alkanes and aromatic compounds. Hydrocarbon degradation in Pseudomonas species has been mostly studied under aerobic conditions; however, a dynamic spectrum of oxygen availability can be found in the environment. Pseudomonas extremaustralis, an Antarctic bacterium isolated from a pristine environment, is able to degrade diesel fuel and presents a wide microaerophilic metabolism. In this work RNA-deep sequence experiments were analyzed comparing the expression profile in aerobic and microaerophilic cultures. Interestingly, genes involved in alkane degradation, including alkB, were over-expressed in micro-aerobiosis in absence of hydrocarbon compounds. In minimal media supplemented with diesel fuel, n-alkanes degradation (C13-C19) after 7 days was observed under low oxygen conditions but not in aerobiosis. In-silico analysis of the alkB promoter zone showed a putative binding sequence for the anaerobic global regulator, Anr. Our results indicate that some diesel fuel components can be utilized as sole carbon source under microaerophilic conditions for cell maintenance or slow growth in a Pseudomonas species and this metabolism could represent an adaptive advantage in polluted environments.


Assuntos
Alcanos/metabolismo , Gasolina , Pseudomonas/metabolismo , Aerobiose , Biodegradação Ambiental , Citocromo P-450 CYP4A/genética , Citocromo P-450 CYP4A/metabolismo , Pseudomonas/enzimologia , Pseudomonas/genética , Transcriptoma
10.
J Crohns Colitis ; 11(12): 1430-1439, 2017 Dec 04.
Artigo em Inglês | MEDLINE | ID: mdl-28981652

RESUMO

BACKGROUND AND AIMS: This study sought to determine the prevalence of malnutrition in patients with inflammatory bowel disease, to analyse the dietary beliefs and behaviours of these patients, to study their body composition, to evaluate their muscular strength and to identify the factors associated with malnutrition in these patients. METHODS: This was a prospective, multicentre study. Crohn's disease and ulcerative colitis patients from 30 Spanish centres, from the outpatient clinics, were included. A questionnaire of 11 items was applied to obtain data from patients' dietary behaviour and beliefs. Patients who accepted were evaluated to assess their nutritional status using Subjective Global Assessment and body mass index. Body composition was evaluated through bioelectrical impedance. RESULTS: A total of 1271 patients were included [51% women, median age 45 years, 60% Crohn's disease]. Of these, 333 patients underwent the nutritional evaluation. A total of 77% of patients declared that they avoided some foods to prevent disease relapse. Eighty-six per cent of patients avoided some foods when they had disease activity because of fear of worsening the flare. Sixty-seven per cent of patients modified their dietary habits after disease diagnosis. The prevalence of malnutrition was 16% [95% confidence interval = 12-20%]. In the multivariate analysis, history of abdominal surgery, active disease and avoidance of some foods during flares were associated with higher risk of malnutrition. CONCLUSIONS: The prevalence of malnutrition in inflammatory bowel disease patients was high. We identified some predictive factors of malnutrition. Most of the patients had self-imposed food restrictions, based on their beliefs.


Assuntos
Dieta , Comportamento Alimentar , Conhecimentos, Atitudes e Prática em Saúde , Doenças Inflamatórias Intestinais/fisiopatologia , Desnutrição/epidemiologia , Desnutrição/fisiopatologia , Estado Nutricional , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Composição Corporal , Índice de Massa Corporal , Doença de Crohn , Impedância Elétrica , Feminino , Alimentos , Força da Mão , Humanos , Doenças Inflamatórias Intestinais/complicações , Masculino , Pessoa de Meia-Idade , Avaliação Nutricional , Prevalência , Estudos Prospectivos , Fatores de Risco , Espanha/epidemiologia , Exacerbação dos Sintomas , Adulto Jovem
11.
Environ Microbiol ; 19(2): 645-658, 2017 02.
Artigo em Inglês | MEDLINE | ID: mdl-27768818

RESUMO

Pseudomonas putida strains are generally recognized as solvent tolerant, exhibiting varied sensitivity to organic solvents. Pan-genome analysis has revealed that 30% of genes belong to the core-genome of Pseudomonas. Accessory and unique genes confer high degree of adaptability and capabilities for the degradation and synthesis of a wide range of chemicals. For the use of these microbes in bioremediation and biocatalysis, it is critical to understand the mechanisms underlying these phenotypic differences. In this study, RNA-seq analysis compared the short- and long-term responses of the toluene-sensitive KT2440 strain and the highly tolerant DOT-T1E strain. The sensitive strain activates a larger number of genes in a higher magnitude than DOT-T1E. This is expected because KT2440 bears one toluene tolerant pump, while DOT-T1E encodes three of these pumps. Both strains activate membrane modifications to reduce toluene membrane permeability. The KT2440 strain activates the TCA cycle to generate energy, while avoiding energy-intensive processes such as flagellar biosynthesis. This suggests that KT2440 responds to toluene by focusing on survival mechanisms. The DOT-T1E strain activates toluene degradation pathways, using toluene as source of energy. Among the unique genes encoded by DOT-T1E is a 70 kb island composed of genes of unknown function induced in response to toluene.


Assuntos
Regulação Bacteriana da Expressão Gênica/fisiologia , Pseudomonas putida/efeitos dos fármacos , Pseudomonas putida/metabolismo , Solventes/farmacologia , Tolueno/farmacologia , Transcriptoma , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Biodegradação Ambiental , Pseudomonas putida/genética
12.
PLoS One ; 10(12): e0145353, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26671564

RESUMO

Temperature is one of the most important factors for bacterial growth and development. Cold environments are widely distributed on earth, and psychrotolerant and psychrophilic microorganisms have developed different adaptation strategies to cope with the stress derived from low temperatures. Pseudomonas extremaustralis is an Antarctic bacterium able to grow under low temperatures and to produce high amounts of polyhydroxyalkanoates (PHAs). In this work, we analyzed the genome-wide transcriptome by RNA deep-sequencing technology of early exponential cultures of P. extremaustralis growing in LB (Luria Broth) supplemented with sodium octanoate to favor PHA accumulation at 8°C and 30°C. We found that genes involved in primary metabolism, including tricarboxylic acid cycle (TCA) related genes, as well as cytochromes and amino acid metabolism coding genes, were repressed at low temperature. Among up-regulated genes, those coding for transcriptional regulatory and signal transduction proteins were over-represented at cold conditions. Remarkably, we found that genes involved in ethanol oxidation, exaA, exaB and exaC, encoding a pyrroloquinoline quinone (PQQ)-dependent ethanol dehydrogenase, the cytochrome c550 and an aldehyde dehydrogenase respectively, were up-regulated. Along with RNA-seq experiments, analysis of mutant strains for pqqB (PQQ biosynthesis protein B) and exaA were carried out. We found that the exaA and pqqB genes are essential for growth under low temperature in LB supplemented with sodium octanoate. Additionally, p-rosaniline assay measurements showed the presence of alcohol dehydrogenase activity at both 8°C and 30°C, while the activity was abolished in a pqqB mutant strain. These results together with the detection of ethanol by gas chromatography in P. extremaustralis cultures grown at 8°C support the conclusion that this pathway is important under cold conditions. The obtained results have led to the identification of novel components involved in cold adaptation mechanisms in this bacterium, suggesting for the first time a role of the ethanol oxidation pathway for bacterial growth at low temperatures.


Assuntos
Temperatura Baixa , Etanol/metabolismo , Perfilação da Expressão Gênica , Genes Bacterianos , Pseudomonas/genética , Álcool Desidrogenase/metabolismo , Regiões Antárticas , Regulação Bacteriana da Expressão Gênica , Fases de Leitura Aberta/genética , Oxirredução , Pseudomonas/crescimento & desenvolvimento , Software , Regulação para Cima/genética
13.
Environ Microbiol ; 17(9): 3251-62, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-25581266

RESUMO

Multi-drug resistant bacteria are a major threat to humanity, especially because the current battery of known antibiotics is not sufficient to combat infections produced by these microbes. Therefore, the study of how current antibiotics act and how bacteria defend themselves against antibiotics is of critical importance. Pseudomonas putida DOT-T1E exhibits an impressive array of RND efflux pumps, which confer this microorganism high resistance to organic solvents and antibiotics that would kill most other microorganisms. We have chosen DOT-T1E as a model microbe to study the microbial responses to a wide battery of antibiotics (chloramphenicol, rifampicin, tetracycline, ciprofloxacin, ampicillin, kanamycin, spectinomycin and gentamicin). Ribonucleic acid sequencing (RNA)-seq analyses revealed that each antibiotic provokes a unique transcriptional response profile in DOT-T1E. While many of the genes identified were related to known antibiotic targets, others were unrelated or encoded hypothetical proteins. These results indicate that our knowledge of antibiotic resistance mechanisms is still partial. We also identified 138 new small RNAs (sRNAs) in DOT-T1E, dramatically adding to the 16 that have been previously described. Importantly, our results reveal that a correlation exists between the expression of messenger RNA and sRNA, indicating that some of these sRNAs are likely involved in fine tuning the expression of antibiotic resistance genes. Taken together, these findings open new frontiers in the fight against multi-drug resistant bacteria and point to the potential use of sRNAs as novel antimicrobial targets.


Assuntos
Antibacterianos/farmacologia , Farmacorresistência Bacteriana Múltipla/genética , Pseudomonas putida/efeitos dos fármacos , Pequeno RNA não Traduzido/genética , Transcrição Gênica/efeitos dos fármacos , Pseudomonas putida/genética , RNA Mensageiro/genética , Solventes/farmacologia
14.
Environ Microbiol Rep ; 7(2): 227-36, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25394275

RESUMO

RNA sequencing (RNA-seq) has revealed several hundreds of previously undetected small RNAs (sRNAs) in all bacterial species investigated, including strains of Pseudomonas aeruginosa, Pseudomonas putida and Pseudomonas syringae. Nonetheless, only little is known about the extent of conservation of expressed sRNAs across strains and species. In this study, we have used RNA-seq to identify sRNAs in P. putida DOT-T1E and Pseudomonas extremaustralis 14-3b. This is the first strain of P. extremaustralis and the second strain of P. putida to have their transcriptomes analysed for sRNAs, and we identify the presence of around 150 novel sRNAs in each strain. Furthermore, we provide a comparison based on sequence conservation of all the sRNAs detected by RNA-seq in the Pseudomonas species investigated so far. Our results show that the extent of sRNA conservation across different species is very limited. In addition, when comparing the sRNAs expressed in different strains of the same species, we observe that numerous sRNAs exhibit a strain-specific expression pattern. These results support the idea that the evolution of most bacterial sRNAs is rapid, which limits the extent of both interspecies and intraspecies conservation.


Assuntos
Pseudomonas/genética , RNA Bacteriano/genética , Pequeno RNA não Traduzido/genética , Sequência Conservada , Perfilação da Expressão Gênica , Variação Genética , Pseudomonas/classificação , RNA Bacteriano/análise , Pequeno RNA não Traduzido/análise , Homologia de Sequência do Ácido Nucleico
15.
BMC Genomics ; 15: 783, 2014 Sep 11.
Artigo em Inglês | MEDLINE | ID: mdl-25213728

RESUMO

BACKGROUND: RNA sequencing technologies reveal that bacteria express RNA molecules other than mRNA, rRNA or tRNA. During the last years genome-wide bacterial transcriptomes have been shown to comprise intergenic RNA, antisense RNA, and untranslated regions, all capable of performing diverse regulatory functions. RESULTS: In this study we used RNA-seq to identify 232 antisense RNAs (asRNAs) in the opportunistic pathogen Pseudomonas aeruginosa grown under 13 different conditions. The conditions studied include exponential and stationary growth as well as osmotic, oxidative and antibiotic stress. We found a significant overrepresentation of asRNAs that are transcribed opposite to genes involved in cell division and in cell wall, lipopolysaccharide (LPS), and capsule biosynthesis, most likely reflecting the conditions used in this study. A substantial number of asRNAs significantly changed their expression under osmotic, oxidative and antibiotic stress, suggesting that asRNAs may play regulatory roles during these conditions. We also made a comparison between the asRNAs detected in this study in P. aeruginosa PAO1 with the asRNAs detected in two previous studies in P. aeruginosa PA14, and found that the extent of overlap between the studies is very limited. CONCLUSIONS: RNA-seq experiments are revealing hundreds of novel transcripts in all bacterial genomes investigated. The comparison between independent studies that used RNA-seq to detect novel asRNAs in P. aeruginosa shows that the overlap between the results reported is very narrow. It is necessary to address how reproducibility of these kind of studies should be reported in order to avoid misleading conclusions when comparing data generated by non-identical methods.


Assuntos
Pseudomonas aeruginosa/genética , RNA Antissenso/genética , RNA Bacteriano/genética , Antibacterianos/farmacologia , Parede Celular/genética , Regulação Bacteriana da Expressão Gênica , Oxirredução , Pseudomonas aeruginosa/crescimento & desenvolvimento , Análise de Sequência de RNA , Estresse Fisiológico
16.
mBio ; 5(5): e01592-14, 2014 Sep 16.
Artigo em Inglês | MEDLINE | ID: mdl-25227464

RESUMO

UNLABELLED: Microbial population polymorphisms are commonly observed in natural environments, including long-term infected hosts. However, the underlying processes promoting and stabilizing diversity are difficult to unravel and are not well understood. Here, we use chronic infection of cystic fibrosis airways by the opportunistic pathogen Pseudomonas aeruginosa as a system for investigating bacterial diversification processes during the course of infection. We analyze clonal bacterial isolates sampled during a 32-year period and map temporal and spatial variations in population diversity to different infection sites within the infected host. We show that the ancestral infecting strain diverged into distinct sublineages, each with their own functional and genomic signatures and rates of adaptation, immediately after initial colonization. The sublineages coexisted in the host for decades, suggesting rapid evolution of stable population polymorphisms. Critically, the observed generation and maintenance of population diversity was the result of partitioning of the sublineages into physically separated niches in the CF airway. The results reveal a complex within-host population structure not previously realized and provide evidence that the heterogeneity of the highly structured and complex host environment promotes the evolution and long-term stability of pathogen population diversity during infection. IMPORTANCE: Within-host pathogen evolution and diversification during the course of chronic infections is of importance in relation to therapeutic intervention strategies, yet our understanding of these processes is limited. Here, we investigate intraclonal population diversity in P. aeruginosa during chronic airway infections in cystic fibrosis patients. We show the evolution of a diverse population structure immediately after initial colonization, with divergence into multiple distinct sublineages that coexisted for decades and occupied distinct niches. Our results suggest that the spatial heterogeneity in CF airways plays a major role in relation to the generation and maintenance of population diversity and emphasize that a single isolate in sputum may not represent the entire pathogen population in the infected individual. A more complete understanding of the evolution of distinct clonal variants and their distribution in different niches could have positive implications for efficient therapy.


Assuntos
Adaptação Fisiológica/genética , Evolução Molecular , Interações Hospedeiro-Patógeno/genética , Pseudomonas aeruginosa/genética , Clonagem Molecular , Fibrose Cística/microbiologia , Meio Ambiente , Genes Bacterianos , Genética Populacional , Genômica , Humanos , Fenótipo , Filogenia , Polimorfismo de Nucleotídeo Único , Infecções por Pseudomonas/microbiologia , Pseudomonas aeruginosa/classificação , Pseudomonas aeruginosa/isolamento & purificação , Percepção de Quorum/fisiologia , Sistema Respiratório , Análise de Sequência de DNA , Escarro/microbiologia
17.
Methods Mol Biol ; 1149: 433-56, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24818924

RESUMO

Small regulatory RNAs (sRNAs) in bacteria are known to modulate gene expression and control a variety of processes including metabolic reactions, stress responses, and pathogenesis in response to environmental signals. A method to identify bacterial sRNAs on a genome-wide scale based on RNA sequencing (RNA-seq) is described that involves the preparation and analysis of three different sequencing libraries. As a significant number of unique sRNAs are identified in each library, the libraries can be used either alone or in combination to increase the number of sRNAs identified. The approach may be applied to identify sRNAs in any bacterium under different growth and stress conditions.


Assuntos
Pseudomonas aeruginosa/genética , RNA Bacteriano/genética , Análise de Sequência de RNA/métodos , Pareamento de Bases/genética , Sequência de Bases , Desoxirribonuclease I/metabolismo , Biblioteca Gênica , Pseudomonas aeruginosa/crescimento & desenvolvimento , Pirofosfatases/metabolismo , RNA Bacteriano/isolamento & purificação , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Transcriptoma/genética
18.
Environ Microbiol ; 14(8): 2006-16, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22533370

RESUMO

Bacterial small regulatory RNAs (sRNAs) function in post-transcriptional control of gene expression and control a variety of processes including metabolic reactions, stress responses and pathogenesis in response to environmental signals. A variety of approaches have been used previously to identify 44 sRNAs in the opportunistic human pathogen Pseudomonas aeruginosa. In this work, RNA sequencing (RNA-seq) is used to identify novel transcripts in P.aeruginosa involving a combination of three different sequencing libraries. Almost all known sRNAs and over 500 novel intergenic sRNAs are identified with this approach. Although the use of three libraries increased the number of novel transcripts identified, there were significant differences in the subset of transcripts detected in each library, underscoring the importance of library preparation strategy and relative sRNA abundance for successful sRNA detection. Nearly 90% of the novel sRNAs have no orthologous bacterial sequences outside of P.aeruginosa, supporting a limited degree of sequence conservation and rapid evolution of sRNAs at the species level. We anticipate that the data will be useful for the study of regulatory sRNAs in bacteria and that the approach described here may be applied to identify sRNAs in any bacterium under different growth and stress conditions.


Assuntos
Genoma Bacteriano , Pseudomonas aeruginosa/genética , RNA Bacteriano/genética , Sequência Conservada/genética , Regulação Bacteriana da Expressão Gênica , Biblioteca Gênica , Pseudomonas aeruginosa/metabolismo , RNA Bacteriano/metabolismo , Reprodutibilidade dos Testes , Análise de Sequência de RNA
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