RESUMO
BACKGROUND: Couple relationship distress is common and associated with poor physical, psychological, and relational outcomes for both partners. Emotionally Focused Therapy for couples (EFT) is a short-term structured approach based on attachment theory that integrates a humanistic, experiential approach to restructuring emotional experience and a systemic structural approach to restructuring interactions. This model has been shown to be an effective treatment for couple distress. The supporting research, however, has only been conducted with English-speaking couples. Despite Spanish being the second-most spoken language and meaningful cultural differences between English- and Spanish-speaking countries, the efficacy of EFT has not been examined in this cultural context. This study will examine the efficacy of EFT in this particular context and advance the understanding of potential mechanisms of change. METHODS: We will use a multicenter randomized wait-list controlled design to examine the efficacy of EFT in a Spanish-speaking sample of moderately distressed couples. One hundred forty individuals in 70 couples in Argentina, Costa Rica, Guatemala, Mexico, and Spain will be randomly assigned to receive 19-21 sessions of EFT or be placed on a waitlist. Outcomes on a range of relational and individual mental health variables will be assessed prior to random assignment, throughout treatment, and at the conclusion of treatment. Primary outcomes will include dyadic adjustment, couple satisfaction, and attachment. Secondary variables, such as loneliness, parenting, affective communication, and sexual satisfaction, will be included as potential mediators of the treatment effect. Couples in the treatment group will also be assessed at 3-, 6-, 12-, 18-, and 24-month follow-ups. Process variables such as the therapeutic alliance will also be assessed routinely in couples assigned to the treatment group. Couples in the waitlist will receive a psycho-educational program based on EFT after completing the study. DISCUSSION: This study will be the first RCT of Emotionally Focused Therapy in a Spanish-speaking context. The results of the study will inform researchers interested in whether treatments developed and tested in the USA and Canada can be effective in differing cultural contexts. It may also point researchers and clinicians to areas where cultural adaptation is needed to improve efficacy. TRIAL REGISTRATION: ClinicalTrials.gov NCT04277325. Registered on February 20, 2020.
Assuntos
Terapia de Casal , Terapia Focada em Emoções , Humanos , Terapia de Casal/métodos , Idioma , México , Espanha , Argentina , Costa Rica , Guatemala , Ensaios Clínicos Controlados Aleatórios como Assunto , Estudos Multicêntricos como AssuntoRESUMO
Preeclampsia, a disorder with a heterogeneous physiopathology, can be attributed to maternal, fetal, and/or placental factors. Long non-coding RNAs (lncRNAs) refer to a class of non-coding RNAs, the essential regulators of biological processes; their differential expression has been associated with the pathogenesis of multiple diseases. The study aimed to identify lncRNAs, expressed in the placentas and plasma of patients who presented with preeclampsia, as potential putative biomarkers of the disease. In silico analysis was performed to determine lncRNAs differentially expressed in the placentas of patients with preeclampsia, using a previously published RNA-Seq dataset. Seven placentas and maternal plasma samples collected at delivery from preterm preeclamptic patients (≤37 gestational weeks of gestation), and controls were used to validate the expression of lncRNAs by qRT-PCR. Six lncRNAs were validated and differentially expressed (p < 0.05) in the preeclampsia and control placentas: UCA1 and HCG4 were found upregulated, and LOC101927355, LINC00551, PART1, and NRAD1 downregulated. Two of these lncRNAs, HCG4 and LOC101927355, were also detected in maternal plasma, the latter showing a significant decrease (p = 0.03) in preeclamptic patients compared to the control group. In silico analyses showed the cytoplasmic location of LOC101927355, which suggests a role in post-transcriptional gene regulation. The detection of LOC101927355 in the placenta and plasma opens new possibilities for understanding the pathogenesis of preeclampsia and for its potential use as a biomarker.
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Estimado director: Hemos leído con interés el artículo Prevalencia del síndrome metabólico en la población dos consultorios del Policlínico Primero de Enero, de los autores Rivero Sabournin y otros.1 Dicho trabajo demuestra el potencial investigativo en la Atención Primaria de Salud, así como la pertinencia de este escenario en los estudios epidemiológicos. Para el desarrollo de la investigación sus autores se basan en los criterios de síndrome metabólico (SM) del National Cholesterol Education Program Adult Treatment Panel III (NCEP-ATP III),1 aunque resulta imprescindible evaluar los criterios de la Clasificación Consensuada o Armonizada de Alberti y otros del año 2009,2 los cuales constituyen la guía más importante para la evaluación por parte del médico de cabecera de los pacientes que pudieran padecer de SM. Otro elemento importante en esta investigación es que se resalta la relación SM y envejecimiento, aunque no se argumenta cuáles pudieran ser los nexos entre ellos. Actualmente se considera que sea el endotelio (END) y la disfunción endotelial (DISF) la vía común de cada una de estas alteraciones como lo han propuesto varios autores: niveles elevados de ácidos grasos,3 envejecimiento,4 oxidación de LDL,5 hiperglucemia,6 niveles séricos de adipoquinas7,8 y las especies reactivas del oxígeno.9 Para un abordaje de esta relación SM-E-END sería necesario analizar que la base fisiopatológica del SM es la insulinorresistencia (IR) y que, precisamente, el endotelio tiene una función importante, donde la insulina (INS) logra desempeñar su función.8 En el endotelio se produce el factor de crecimiento similar a la INS (IGF, según sus siglas en inglés)9 y es donde se encuentran proteínas de membrana que sirven de transportadores a la INS. Una vez que se instaura el daño endotelial o la disfunción endotelial (DE), la pérdida...(AU)
Assuntos
Humanos , Masculino , Feminino , Idoso , Idoso de 80 Anos ou mais , Atenção Primária à Saúde , Envelhecimento/fisiologia , Síndrome Metabólica/epidemiologiaRESUMO
Hemos leído con gran atención el artículo de los autores González Rey y otros, titulado: Disfunción endotelial en una etapa precoz del diagnóstico de hipertensión arterial. Resulta muy interesante el tratamiento de un tema básico de gran interés en la clínica a través del uso de biomarcadores casi siempre a la disposición de nuestros profesionales de la salud en los diferentes niveles de atención como resulta ser el caso de la microalbuminuria.1 El endotelio resulta cada vez de mayor interés para investigadores y médicos de asistencia, pues es el punto de confluencia de las enfermedades vasculares, metabólicas y neurodegenerativas, y es el primer eslabón en el desarrollo de la aterosclerosis. Se conoce que los diferentes factores involucrados en la activación y daño endotelial como las altas concentraciones de ácido úrico,2 los niveles elevados de ácidos grasos,3 el envejecimiento4 y la hiperglicemia,5 son los mismos que contribuyen a posteriori con el desarrollo y las complicaciones de la placa de ateroma. Vale destacar el aporte...(AU)
Assuntos
Humanos , Endotélio/fisiopatologia , Hipertensão Essencial/epidemiologiaRESUMO
We characterized the antimicrobial resistance of 70 Escherichia coli isolates obtained from patients with a urinary tract infection (UTI) from 8 public hospitals in Peru. Resistance profiles were identified using the automated MicroScan® system. A standard polymerase chain reaction was used for the detection of the bla TEM, bla CTX-M, bla SHV and bla PER genes. The 65.7% (46/70) of the isolates presented a multidrug-resistant phenotype and 55.7% (39/70) were extended-spectrum beta-lactamases producers. High levels of resistance were detected for ampicillin (77,1%), ciprofloxacin (74,3%), trimethoprim/sulfamethoxazole (62,9%), cefepime (57,1%), and cefuroxime (57,1%). The bla TEM gene was the most frequent (31,4%), followed by bla CTX-M (18,6%) and bla SHV (2,9%) genes. These results show high resistance levels to antimicrobials of clinical use in E. coli isolates from hospital UTI patients in Peru.
Se caracterizó la resistencia antimicrobiana de 70 aislados de Escherichia coli obtenidos de pacientes con infección del tracto urinario (ITU) provenientes de ocho hospitales públicos en el Perú. Los perfiles de resistencia fueron identificados mediante el uso del sistema automatizado MicroScan®. Se utilizó una reacción en cadena de la polimerasa convencional para la detección de los genes bla TEM, bla CTX-M, bla SHV y bla PER. El 65,7% (46/70) de los aislados presentó un fenotipo multidrogorresistente y el 55,7% (39/70) fue identificado como productores de betalactamasas de espectro extendido. Se detectaron altos niveles de resistencia para ampicilina (77,1%), ciprofloxacina (74,3%), trimetoprim/sulfametoxazol (62,9%), cefepime (57,1%) y cefuroxima (57,1%). El gen bla TEM fue el más frecuente con un 31,4%, seguido por bla CTX-M (18,6%) y bla SHV (2,9%). Los resultados evidencian altos niveles de resistencia a antimicrobianos de importancia clínica en aislados de E. coli de pacientes con ITU en el Perú.
Assuntos
Infecções por Escherichia coli , Escherichia coli Uropatogênica , Antibacterianos/farmacologia , Farmacorresistência Bacteriana/genética , Infecções por Escherichia coli/tratamento farmacológico , Hospitais Públicos , Humanos , Testes de Sensibilidade Microbiana , Peru , Escherichia coli Uropatogênica/genética , beta-Lactamases/genéticaRESUMO
RESUMEN Se caracterizó la resistencia antimicrobiana de 70 aislados de Escherichia coli obtenidos de pacientes con infección del tracto urinario (ITU) provenientes de ocho hospitales públicos en el Perú. Los perfiles de resistencia fueron identificados mediante el uso del sistema automatizado MicroScan®. Se utilizó una reacción en cadena de la polimerasa convencional para la detección de los genes bla TEM, bla CTX-M, bla SHV y bla PER. El 65,7% (46/70) de los aislados presentó un fenotipo multidrogorresistente y el 55,7% (39/70) fue identificado como productores de betalactamasas de espectro extendido. Se detectaron altos niveles de resistencia para ampicilina (77,1%), ciprofloxacina (74,3%), trimetoprim/sulfametoxazol (62,9%), cefepime (57,1%) y cefuroxima (57,1%). El gen bla TEM fue el más frecuente con un 31,4%, seguido por bla CTX-M (18,6%) y bla SHV (2,9%). Los resultados evidencian altos niveles de resistencia a antimicrobianos de importancia clínica en aislados de E. coli de pacientes con ITU en el Perú.
ABSTRACT We characterized the antimicrobial resistance of 70 Escherichia coli isolates obtained from patients with a urinary tract infection (UTI) from 8 public hospitals in Peru. Resistance profiles were identified using the automated MicroScan® system. A standard polymerase chain reaction was used for the detection of the bla TEM, bla CTX-M, bla SHV and bla PER genes. The 65.7% (46/70) of the isolates presented a multidrug-resistant phenotype and 55.7% (39/70) were extended-spectrum beta-lactamases producers. High levels of resistance were detected for ampicillin (77,1%), ciprofloxacin (74,3%), trimethoprim/sulfamethoxazole (62,9%), cefepime (57,1%), and cefuroxime (57,1%). The bla TEM gene was the most frequent (31,4%), followed by bla CTX-M (18,6%) and bla SHV (2,9%) genes. These results show high resistance levels to antimicrobials of clinical use in E. coli isolates from hospital UTI patients in Peru.
Assuntos
Humanos , Masculino , Peru , Infecções Urinárias , Reação em Cadeia da Polimerase , Farmacorresistência Bacteriana , Escherichia coli , Hospitais Públicos , Pacientes , Doenças Urológicas , Resistência beta-Lactâmica , Escherichia coli UropatogênicaRESUMO
RESUMEN Se caracterizó la resistencia antimicrobiana de 70 aislados de Escherichia coli obtenidos de pacientes con infección del tracto urinario (ITU) provenientes de ocho hospitales públicos en el Perú. Los perfiles de resistencia fueron identificados mediante el uso del sistema automatizado MicroScan®. Se utilizó una reacción en cadena de la polimerasa convencional para la detección de los genes bla TEM, bla CTX-M, bla SHV y bla PER. El 65,7% (46/70) de los aislados presentó un fenotipo multidrogorresistente y el 55,7% (39/70) fue identificado como productores de betalactamasas de espectro extendido. Se detectaron altos niveles de resistencia para ampicilina (77,1%), ciprofloxacina (74,3%), trimetoprim/sulfametoxazol (62,9%), cefepime (57,1%) y cefuroxima (57,1%). El gen bla TEM fue el más frecuente con un 31,4%, seguido por bla CTX-M (18,6%) y bla SHV (2,9%). Los resultados evidencian altos niveles de resistencia a antimicrobianos de importancia clínica en aislados de E. coli de pacientes con ITU en el Perú.
ABSTRACT We characterized the antimicrobial resistance of 70 Escherichia coli isolates obtained from patients with a urinary tract infection (UTI) from 8 public hospitals in Peru. Resistance profiles were identified using the automated MicroScan® system. A standard polymerase chain reaction was used for the detection of the bla TEM, bla CTX-M, bla SHV and bla PER genes. The 65.7% (46/70) of the isolates presented a multidrug-resistant phenotype and 55.7% (39/70) were extended-spectrum beta-lactamases producers. High levels of resistance were detected for ampicillin (77,1%), ciprofloxacin (74,3%), trimethoprim/sulfamethoxazole (62,9%), cefepime (57,1%), and cefuroxime (57,1%). The bla TEM gene was the most frequent (31,4%), followed by bla CTX-M (18,6%) and bla SHV (2,9%) genes. These results show high resistance levels to antimicrobials of clinical use in E. coli isolates from hospital UTI patients in Peru.
Assuntos
Humanos , Masculino , Feminino , Resistência beta-Lactâmica , Escherichia coli , Hospitais Públicos , Infecções , Pacientes , Peru , Infecções Urinárias , Doenças Urológicas , beta-Lactamases , Farmacorresistência BacterianaRESUMO
El acretismo placentario se define como la invasión anómala de la placenta al miometrio, denominado percretismo en su forma más severa. En la actualidad se describe la coexistencia de placenta previa con cicatriz de cesárea, como el principal factor de riesgo. Siendo esta última variable la que explica su incidencia al alza en los últimos años. El pronóstico depende mayoritariamente del grado de adherencia, del diagnóstico prenatal y del adecuado manejo multidisciplinario. Con el objetivo de mostrar una presentación poco frecuente de percretismno placentario se presenta a continuación un caso clínico sin diagnóstico prenatal con requerimiento de cirugía de urgencia y su respectivo outcome.
Placental acretism is defined as an abnormal invasion of the placenta to the myometrium, it's most severe presentation being placenta percreta. The main risk factor for this disease is the coexistence of placenta previa and previous cesarean section. Its incidence has been progressively rising, mainly because of the increase in cesarean sections. Extent of adherence, prenatal vs intra surgery diagnosis, and multidisciplinary management are accountable for the prognosis of placental acretism. A case report with no prenatal diagnosis, which required emergency surgery, and its outcome is presented.
Assuntos
Humanos , Feminino , Gravidez , Adulto , Placenta Acreta/cirurgia , Placenta Acreta/diagnóstico , Hemoperitônio/etiologia , Ruptura Uterina , Resultado da Gravidez , EmergênciasRESUMO
Background During pregnancy, there is an increase in the amount of extracellular vesicles, especially placental exosomes, in maternal plasma. Aim To isolate and characterize extracellular vesicles from blood during the three trimesters of pregnancy and to evaluate their capacity to identify patients at risk of developing gestational diabetes. Material and Methods A case-control study was conducted in a cohort of 50 pregnant women with plasma samples taken in each trimester. Six women who developed gestational diabetes were paired with three healthy controls per case (a total of 19). Clinical characteristics were recorded at first prenatal appointment, and blood samples were obtained during the first, second and third trimesters. Extracellular vesicles were isolated from plasma by the commercial kit, ExoQuick™. Nanoparticle tracking analysis, was used to characterize the obtained extracellular vesicles. Results The total concentration of extracellular particles isolated from maternal plasma increased along with gestational age. The size of the extracellular vesicles obtained in the first trimester of pregnancy was very similar between groups (144 ± 37 nm for controls and 143 ± 34 nm for patients with gestational diabetes mellitus). Moreover, the concentration of extracellular vesicles collected in the first trimester, was significantly higher in patients who developed gestational diabetes mellitus later in pregnancy compared to normoglycemic pregnant women (7.94 x 10 8 and 5.15 x 10 8 , p = 0.03). Conclusions Our results provide an insight into the potential capacity of first trimester plasma extracellular vesicles as early biomarkers for the prediction of gestational diabetes mellitus.
Assuntos
Humanos , Feminino , Gravidez , Adulto , Diabetes Gestacional/sangue , Vesículas Extracelulares/metabolismo , Biomarcadores/sangue , Estudos de Casos e Controles , Valor Preditivo dos Testes , Estudos Prospectivos , Sensibilidade e Especificidade , Diabetes Gestacional/diagnósticoRESUMO
RESUMEN El cáncer de mama puede originar de forma muy excepcional metástasis en órganos genitales. Estas suelen presentarse de forma asintomática siendo su diagnóstico generalmente tardío. En el estudio de la enfermedad diseminada la tomografía de emisión de positrones con fluordeoxiglucosa asociada a la tomografía computada (FDG PET/CT) juega un rol importante, sin embargo presenta limitantes al momento de diferenciar entre lesiones secundarias, primarias y neoplasias benignas que pueden tener elevada captación de fluordeoxiglucosa (1). Por lo expuesto anteriormente el diagnóstico de certeza solo puede realizarse mediante el estudio anatomopatológico. El caso presentado a continuación abarca uno de los posibles diagnósticos diferenciales con FDG PET/CT que evidencia un incremento en la captación de fluordeoxiglucosa.
ABSTRACT Breast cancer rarely metastasize on pelvic organs and its diagnosis is often delayed due to the asymptomatic nature of this disease. Fluorodeoxyglucose Positron Emission Tomography/Computed Tomography (FDG PET/CT) is a useful method for early diagnosis. However, may present a false positive diagnosis from benign lesions that have an increased uptake of glucose. We present a case in which a patient who had breast cancer had an increase glucose uptake in FDG PET/CT from a benign lesion.
Assuntos
Humanos , Feminino , Pessoa de Meia-Idade , Neoplasias da Mama/complicações , Neoplasias da Mama/diagnóstico , Leiomioma/patologia , Neoplasias Uterinas/patologia , Diagnóstico Diferencial , Tomografia por Emissão de Pósitrons combinada à Tomografia ComputadorizadaRESUMO
Intrauterine devices (IUDs) are among the most efficient and widely used contraceptive methods available. Removal is recommended after a certain time or in some cases when adverse effects are observed. A considerable number of patients have nonvisible guides or "lost threads" on speculum examinations, hindering the extraction. In this article, we report a consecutive series of 254 nonpregnant patients referred to our center after 1 or more failed attempts at IUD removal. We describe a novel ultrasound-guided approach, using a laparoscopic forceps to safely and effectively remove IUDs in nonpregnant patients.
Assuntos
Remoção de Dispositivo/métodos , Dispositivos Intrauterinos , Ultrassonografia de Intervenção/métodos , Útero/diagnóstico por imagem , Adulto , Endoscopia , Feminino , Humanos , Estudos Retrospectivos , Instrumentos CirúrgicosRESUMO
Background During pregnancy, there is an increase in the amount of extracellular vesicles, especially placental exosomes, in maternal plasma. Aim To isolate and characterize extracellular vesicles from blood during the three trimesters of pregnancy and to evaluate their capacity to identify patients at risk of developing gestational diabetes. Material and Methods A case-control study was conducted in a cohort of 50 pregnant women with plasma samples taken in each trimester. Six women who developed gestational diabetes were paired with three healthy controls per case (a total of 19). Clinical characteristics were recorded at first prenatal appointment, and blood samples were obtained during the first, second and third trimesters. Extracellular vesicles were isolated from plasma by the commercial kit, ExoQuick™. Nanoparticle tracking analysis, was used to characterize the obtained extracellular vesicles. Results The total concentration of extracellular particles isolated from maternal plasma increased along with gestational age. The size of the extracellular vesicles obtained in the first trimester of pregnancy was very similar between groups (144 ± 37 nm for controls and 143 ± 34 nm for patients with gestational diabetes mellitus). Moreover, the concentration of extracellular vesicles collected in the first trimester, was significantly higher in patients who developed gestational diabetes mellitus later in pregnancy compared to normoglycemic pregnant women (7.94 x 10 8 and 5.15 x 10 8 , p = 0.03). Conclusions Our results provide an insight into the potential capacity of first trimester plasma extracellular vesicles as early biomarkers for the prediction of gestational diabetes mellitus.
Assuntos
Diabetes Gestacional/sangue , Vesículas Extracelulares/metabolismo , Adulto , Biomarcadores/sangue , Estudos de Casos e Controles , Diabetes Gestacional/diagnóstico , Feminino , Humanos , Valor Preditivo dos Testes , Gravidez , Estudos Prospectivos , Sensibilidade e EspecificidadeRESUMO
Objective: To assess the level of happiness and satisfaction in the life and medical practice of dermatologists in Mexico. Method: A descriptive study (online survey) was conducted focused on practicing dermatologists in our country. Questions included demographic characteristics, the Pemberton happiness index (with local validation) and questions that assessed the degree of personal satisfaction. Descriptive statistics were used to obtain the central tendency and dispersion. Measures of central tendency and dispersion were performed; to compare categorical variables, contingency tables for chi-square test were used and when comparing quantitative variables with normal distribution, Student's t t-test was used. Results: 219 surveys were included, 72.6% female and 27.4% male, with an average age of 45.6 and an average of 16 years of medical practice. Most of them (64.8%) graduate from Mexico City; 93% were very satisfied with the specialty and 98.6% of them would choose the same once again, the most important reason is to encompass medical and surgical areas. The level of happiness by using the Pemberton scale was "high" (mode: 9.11; standard deviation: 1.73). Conclusions: This first study in Latin America on this subject in dermatologists showed high levels of satisfaction and happiness in both professional and personal areas.
Objetivo: Conocer la felicidad y la satisfacción laboral de los dermatólogos en México. Método: Estudio descriptivo, mediante encuesta en línea dirigida a dermatólogos especialistas, graduados y con ejercicio en México. Se incluyeron elementos demográficos, el Índice de Felicidad de Pemberton (con validación local) y preguntas para evaluar la satisfacción personal. Se realizaron medidas de tendencia central y dispersión; para comparar las variables categóricas, se utilizaron tablas de contingencia para ji al cuadrado, y al comparar variables cuantitativas con distribución normal se utilizó la prueba t de Student. Resultados: Se incluyeron 219 encuestados, el 72.6% mujeres y el 27.4% hombres, con una media de edad de 45.6 años y un promedio de 16 años de ejercicio profesional. La mayoría (64.8%) eran egresados de la Ciudad de México y el 75.3% estaban muy satisfechos con su escuela de formación. Un 93% se manifestó muy satisfecho con su especialidad y el 98.6% volvería a escogerla; el mayor motivo sería abarcar ámbitos médicos y quirúrgicos. El grado de felicidad según el Índice de Pemberton fue «alto¼ (media: 9.11; desviación estándar: 1.73). Conclusión: Este primer estudio en América Latina sobre este aspecto en dermatólogos, muestra unos altos índices de satisfacción y felicidad, tanto en el área profesional como en la personal.
Assuntos
Dermatologistas/psicologia , Dermatologia , Felicidade , Satisfação no Emprego , Satisfação Pessoal , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
Janibacter sp. strain R02 (BNM 560) was isolated in our laboratory from an Antarctic soil sample. A remarkable trait of the strain was its high lipolytic activity, detected in Rhodamine-olive oil supplemented plates. Supernatants of Janibacter sp. R02 displayed superb activity on transesterification of acyl glycerols, thus being a good candidate for lipase prospection. Considering the lack of information concerning lipases of the genus Janibacter, we focused on the identification, cloning, expression and characterization of the extracellular lipases of this strain. By means of sequence alignment and clustering of consensus nucleotide sequences, a DNA fragment of 1272bp was amplified, cloned and expressed in E. coli. The resulting recombinant enzyme, named LipJ2, showed preference for short to medium chain-length substrates, and displayed maximum activity at 80°C and pH 8-9, being strongly activated by a mixture of Na+ and K+. The enzyme presented an outstanding stability regarding both pH and temperature. Bioinformatics analysis of the amino acid sequence of LipJ2 revealed the presence of a consensus catalytic triad and a canonical pentapeptide. However, two additional rare motifs were found in LipJ2: an SXXL ß-lactamase motif and two putative Y-type oxyanion holes (YAP). Although some of the previous features could allow assigning LipJ2 to the bacterial lipase families VIII or X, the phylogenetic analysis showed that LipJ2 clusters apart from other members of known lipase families, indicating that the newly isolated Janibacter esterase LipJ2 would be the first characterized member of a new family of bacterial lipases.
Assuntos
Actinobacteria/enzimologia , Actinobacteria/genética , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Lipase/genética , Lipase/metabolismo , Sequência de Aminoácidos , Proteínas de Bactérias/classificação , Clonagem Molecular , Sequência Conservada , Estabilidade Enzimática , Escherichia coli/enzimologia , Escherichia coli/genética , Esterases/classificação , Esterases/genética , Esterases/metabolismo , Expressão Gênica , Genes Bacterianos , Temperatura Alta , Concentração de Íons de Hidrogênio , Cinética , Lipase/classificação , Modelos Moleculares , FilogeniaRESUMO
Several Pseudomonas sp. CR611 Lip I.3 mutants with overall increased activity and a shift towards longer chain substrates were constructed. Substitution of residues Y29 and W310 by smaller amino acids provided increased activity on C18-substrates. Residues G152 and S154, modified to study their influence on interfacial activation, displayed a five and eleven fold increased activity.
Assuntos
Lipase/química , Lipase/metabolismo , Mutagênese , Pseudomonas/enzimologia , Lipase/genética , Modelos Moleculares , Mutação , Conformação Proteica , Especificidade por SubstratoRESUMO
A new xylanolytic strain, Paenibacillus favisporus CC02-N2, was isolated from sugarcane plantation fields in Brazil. The strain had a xylan-degrading system with multiple enzymes, one of which, xylanase Xyn30A, was identified and characterized. The enzyme is a single-domain xylanase belonging to family 30 of the glycosyl hydrolases (GH30). Xyn30A shows high activity on glucuronoxylans, with a Vmax of 267.2 U mg⻹, a Km of 4.0 mg/ml, and a kcat of 13,333 min⻹ on beechwood xylan, but it does not hydrolyze arabinoxylans. The three-dimensional structure of Xyn30A consists of a common (ß/α)8 barrel linked to a side-chain-associated ß-structure, similar to previously characterized GH30 xylanases. The hydrolysis products from glucuronoxylan were methylglucuronic-acid-substituted xylooligomers (acidic xylooligosaccharides). The enzyme bound to insoluble xylan but not to crystalline cellulose. Our results suggest a specific role for Xyn30A in xylan biodegradation in natural habitats. The enzyme is a good candidate for the production of tailored xylooligosaccharides for use in the food industry and in the biotechnological transformation of biomass.
Assuntos
Proteínas de Bactérias/metabolismo , Paenibacillus/enzimologia , Microbiologia do Solo , Xilosidases/metabolismo , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Brasil , Clonagem Molecular , Estabilidade Enzimática , Cinética , Dados de Sequência Molecular , Paenibacillus/química , Paenibacillus/genética , Paenibacillus/isolamento & purificação , Especificidade por Substrato , Xilosidases/química , Xilosidases/genéticaRESUMO
The gene for a Geobacillus thermoleovorans CCR11 thermostable lipase was recovered by PCR and cloned. Four genetic constructions were designed and successfully expressed in E. coli: (i) the lipase structural gene (lipCCR11) in the PinPoint Xa vector; (ii) the lipase structural gene (lipACCR11) in the pET-28a(+) vector; (iii) the lipase structural gene minus the signal peptide (lipMatCCR11) in the pET-3b vector; and (iv) the lipase structural gene plus its own promoter (lipProCCR11) in the pGEM-T cloning vector. The lipase gene sequence analysis showed an open reading frame of 1,212 nucleotides coding for a mature lipase of 382 residues (40 kDa) plus a 22 residues signal peptide. Expression under T7 and T7lac promoter resulted in a 40- and 36-fold increase in lipolytic activity with respect to the original strain lipase. All recombinant lipases showed an optimal activity at pH 9.0, but variations were found in the temperature for maximum activity and the substrate specificity among them and when compared with the parental strain lipase, especially in the recombinant lipases that contained fusion tags. Therefore, it is important to find the appropriate expression system able to attain a high concentration of the recombinant lipase without compromising the proper folding of the protein.
Assuntos
Bacillaceae/enzimologia , Lipase/química , Proteínas Recombinantes/química , Sequência de Aminoácidos , Concentração de Íons de Hidrogênio , Lipase/genética , Lipase/metabolismo , Conformação Proteica , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Análise de Sequência de DNA , TemperaturaRESUMO
Objetivo: Establecer la relación entre la colección diagnosticada ecográficamente en la cúpula vaginal posterior a una histerectomía vaginal y las complicaciones que puedan devenir. Lugar: Servicio de Ginecología del Pabellón 5-II del Departamento de Ginecología y Obstetricia del Hospital Arzobispo Loayza entre agosto del 2001 hasta marzo del 2002. Material y métodos: estudio con una muestra de 153 pacientes, con indicaciones para la cirugía electiva de histerectomía vaginal y que al tercer día del postoperatorio se le realizaba una ecografía pélvica, para determinar la existencia de alguna colección en la cúpula. De acuerdo a los diversos análisis estadísticos se consideró significancia estadística a un p menor 0,05. Resultados: en las pacientes estudiadas, la edad y la gesta promedio fue de 56 años y 7 respectivamente; se presentaron 40,5 por ciento de complicaciones, siendo las dos terceras partes de índole infecciosa con predominio de las infecciones urinarias atribuidas a la permanencia de la sonda vesical por más de 72 horas y el resto relacionados a infección y hematoma de cúpula, en menor medida se presentaron fallas en la técnica operatoria. Discusión: es notable la asociación entre la presencia de la colección en la cúpula con los procesos infecciosos y complicaciones de la misma, siendo estadísticamente significativa con p menor 0,00 IC95 por ciento RR=77,5; indicando un aumento de 77 veces el riesgo de tener una complicación a ese nivel. Los pocos reportes en la literatura mundial difieren de lo encontrado por nosotros. Podríamos afirmar que la presencia de colección en la cúpula vaginal después de una histerectomía vaginal es un claro indicador de una complicación mediata a ese nivel.