76 research outputs found
Characterization of the GacS/GacA system in the virulence regulation in Pseudomonas savastanoi.
The two-component regulatory system GacS/GacA is one of the main mechanisms for global regulation in bacteria. GacS/GacA is a highly conserved system that has been studied in many pathogenic bacteria. However, its characterization has been mainly focused on pathogenic bacteria of herbaceous plants. Despite previous works have reported that GacS/GacA regulates the expression of virulence factors, its role in virulence varies among different species and strains. The aim of this work was the identification of virulence factors regulated by the GacS/GacA system in the model bacterium Pseudomonas savastanoi pv. savastanoi (Psv), causal agent of olive knot disease. To this end, we generated a gacA deletion mutant in the strain Psv NCPPB 3335, whose transcriptomic profile was further analyzed using a massive RNA sequencing (RNA-seq) strategy. The bioinformatic analysis of RNA-seq data showed that the Psv GacS/GacA system regulates a large number of genes, including some virulence factors already described, such as those related to the type III secretion system, the biosynthesis of phytohormones and the catabolism of aromatic compounds, among others. In addition, small Rsm-type RNAs and regulatory proteins (RsmA) were identified in the regulatory cascade of the GacS/GacA system. Finally, the involvement of some of the virulence factors of Psv NCPP 3335 were further studied through different phenotypic assays, such as plant virulence assays, induction of hypersensitive response, leaf adhesion tests and translocation of type III effectors. Results obtained in this work indicate that GacS/GacA system presents a role in regulation of virulence factors of Psv NCPPB 3335.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech
Secreted citrate serves as iron carrier for the marine pathogen photobacterium damselae subsp damselae
Photobacterium damselae subsp damselae (Pdd) is a Vibrionaceae that has a wide
pathogenic potential against many marine animals and also against humans. Some
strains of this bacterium acquire iron through the siderophore vibrioferrin. However, there
are virulent strains that do not produce vibrioferrin, but they still give a strong positive
reaction in the CAS test for siderophore production. In an in silico search on the genome
sequences of this type of strains we could not find any ORF which could be related
to a siderophore system. To identify genes that could encode a siderophore-mediated
iron acquisition system we used a mini-Tn10 transposon random mutagenesis approach.
From more than 1,400 mutants examined, we could isolate a mutant (BP53) that showed
a strong CAS reaction independently of the iron levels of the medium. In this mutant the
transposon was inserted into the idh gene, which encodes an isocitrate dehydrogenase
that participates in the tricarboxylic acid cycle. The mutant did not show any growth
impairment in rich or minimal media, but it accumulated a noticeable amount of citrate
(around 7 mM) in the culture medium, irrespective of the iron levels. The parental strain
accumulated citrate, but in an iron-regulated fashion, being citrate levels 5–6 times higher
under iron restricted conditions. In addition, a null mutant deficient in citrate synthase
showed an impairment for growth at high concentrations of iron chelators, and showed
almost no reaction in the CAS test. Chemical analysis by liquid chromatography of the
iron-restricted culture supernatants resulted in a CAS-positive fraction with biological
activity as siderophore. HPLC purification of that fraction yielded a pure compound which
was identified as citrate from its MS and NMR spectral data. Although the production
of another citrate-based compound with siderophore activity cannot be ruled out, our
results suggest that Pdd secretes endogenous citrate and use it for iron scavenging
from the cell environmentThis work was supported by grants AGL2012-39274-C02-01/02
and AGL2015-63740-C2-1/2-R (AEI/FEDER, EU) from the State
Agency for Research (AEI) of Spain, and co-funded by the
FEDER Programme from the European Union. The support
of Xunta de Galicia (Spain) with grant GRC-2014/007 is also
acknowledgedS
Implicaciones de la inteligencia emocional en la salud y el bienestar emocional
In this article, we have tried to group the results from different researches related to EI and physic and mental health. One of the conclusions we have is that EI perceived through report (TMMS, etc.) as well as EI executed (MSCEIT) are related to people’s health and welfare. Likewise, the average of MSCEIT grades for different clinic groups of people (schizophrenia, AIDS, Depression, etc.) and non - clinic groups of people(university students, students, etc), give us a vision of psychopathology- normal continuum, being the highest grades in EI of MSCEIT the ones which are positively related to personal health and stable emotional state.En este artículo hemos intentado agrupar los resultados de distintas investigaciones en relación a la IE y la salud física y mental. Una de las conclusiones que obtenemos es que tanto la IE percibida mediante autoinforme (TMMS, etc.) como la IE de ejecución (MSCEIT) están relacionadas con la salud y el bienestar de las personas. Del mismo modo, las medias de las puntuaciones globales del MSCEIT para diferentes poblaciones clínicas (Esquizofrenia, SIDA, Depresión, etc.) y no-clínicas (universitarios, estudiantes, etc.), nos dan una visión del continuo normalidad-psicopatología, siendo las puntuaciones más altas en IE del MSCEIT las que se asocian positivamente a salud personal y estado emocional estable
An efficient reduction of azide to amine: a new methodology to synthesize ethyl 7-amino-1-ethyl-6,8-difluoroquinolone-3- carboxylate and its spectroscopic characterization
Most of the quinolone antibacterial research has
been focused on the functionality at C-7 position where the
nature of substituents is responsible for antibacterial
spectrum, potency, bioavailability, and side effects of the
quinolones. Then, a 7-amino-fluoroquinolone could be the
starting point of a wide variety of potentially useful compounds like tetracyclic and tricyclic quinolones or secondary amines with side chain derivatives. This attracted
our attention to synthesize a 7-azide-fluoroquinolone,
which could be converted to amine performing a photochemical reaction using CuI as catalyst. FT-IR and H1
NMR spectra of the final product, ethyl 7-amino-1-ethyl6,8-difluoroquinolone-3-carboxylate, suggests the formation of dimers, a feature already observed in norfloxacin
Dysregulation of the endocannabinoid signaling system in the cerebellum and brainstem in a transgenic mouse model of spinocerebellar ataxia type-3.
Spinocerebellar ataxia type-3 (SCA-3) is a rare disease but it is the most frequent type within the autosomal dominant inherited ataxias. The disease lacks an effective treatment to alleviate major symptoms and to modify disease progression. Our recent findings that endocannabinoid receptors and enzymes are significantly altered in the post-mortem cerebellum of patients affected by autosomal-dominant hereditary ataxias suggest that targeting the endocannabinoid signaling system may be a promising therapeutic option. Our goal was to investigate the status of the endocannabinoid signaling system in a transgenic mouse model of SCA-3, in the two CNS structures most affected in this disease cerebellum and brainstem. These animals exhibited progressive motor incoordination, imbalance, abnormal gait, muscle weakness, and dystonia, in parallel to reduced in vivo brain glucose metabolism, deterioration of specific neuron subsets located in the dentate nucleus and pontine nuclei, small changes in microglial morphology, and reduction in glial glutamate transporters. Concerning the endocannabinoid signaling, our data indicated no changes in CB2 receptors. By contrast, CB1 receptors increased in the Purkinje cell layer, in particular in terminals of basket cells, but they were reduced in the dentate nucleus. We also measured the levels of endocannabinoid lipids and found reductions in anandamide and oleoylethanolamide in the brainstem. These changes correlated with an increase in the FAAH enzyme in the brainstem, which also occurred in some cerebellar areas, whereas other endocannabinoid-related enzymes were not altered. Collectively, our results in SCA-3 mutant mice confirm a possible dysregulation in the endocannabinoid system in the most important brain structures affected in this type of ataxia, suggesting that a pharmacological manipulation addressed to correct these changes could be a promising option in SCA-3.This study has been supported by MICINN (SAF2009-11847 and SAF2015-68580-C2-1-R), CIBERNED (CB06/05/0089), "Fundacion Eugenio Rodriguez Pascual" and the Research and Education Component of the Advancing a Healthier Wisconsin Endowment at the Medical College of Wisconsin. Carmen Rodriguez-Cueto was a predoctoral fellow supported by FPI Program-Ministry of Science. Authors are indebted to Yolanda Garcia-Movellan for administrative support
Fatty liver and fibrosis in glycine N-methyltransferase knockout mice is prevented by nicotinamide
Deletion of glycine N-methyltransferase (GNMT) in mice, the main gene involved in liver S-adenosylmethionine (SAMe) catabolism, leads to the hepatic accumulation of this molecule and the development of fatty liver and fibrosis. To demonstrate that the excess of hepatic SAMe is the main agent contributing to liver disease in GNMT-KO mice, we treated 1.5-month old GNMT-KO mice for 6 weeks with nicotinamide (NAM), a substrate of the enzyme NAM N-methyltransferase. NAM administration markedly reduced hepatic SAMe content, prevented DNA-hypermethylation and normalized the expression of critical genes involved in fatty acid metabolism, oxidative stress, inflammation, cell proliferation, and apoptosis. More important, NAM treatment prevented the development of fatty liver and fibrosis in GNMT-KO mice. Because GNMT expression is down-regulated in patients with cirrhosis and there are subjects with GNMT mutations who have spontaneous liver disease, the clinical implication of the present findings is obvious at least with respect to these latter individuals. Especially since NAM has been used for many years to treat a broad spectrum of diseases including pellagra and diabetes without significant side effects, it should be considered in subjects with GNMT mutations.ConclusionsThese results indicate that the anomalous accumulation of SAMe in GNMT-KO mice can be corrected by NAM treatment leading to the normalization of the expression of many genes involved in fatty acid metabolism, oxidative stress, inflammation, cell proliferation and apoptosis, and to the reversion of the appearance of the pathologic phenotype
Técnicas basadas en las tecnologías de Bioréplicas impresas en 3D y Realidad Aumentada de alteraciones anatómicas y disfunciones fisiológicas para la mejora docente en las asignaturas de Anatomía y Fisiología
Memoria final de la CONVOCATORIA DE PROYECTOS DE INNOVACIÓN Y BUENAS PRÁCTICAS DOCENTES (PIBD Fase I), 2020/2021.
Este proyecto tiene como objetivos revelar los factores que ayudan a integrar el binomio enseñanza-aprendizaje empleando las tecnologías de realidad aumentada (RA) y el modelaje de bioréplicas por renderización (a través de segmentación, fotogrametría o diseño asistido por ordenador), preferiblemente a través de imágenes digitales de pacientes reales o colaboradores, y/o deposición fundida (FDM).
Los entornos de aprendizaje elegidos fueron las asignaturas del departamento de fisiología en el Campus de Ceuta ("Anatomía Humana" y "Fisiología I"), impartidas en el grado de enfermería de la Universidad de Granada. Junto con las clases tradicionales, en prácticas y seminarios se complementó la docencia tradicional con la tecnología emergente para comprobar los resultados que podrían tener en el aprendizaje del discente, además de comprobar su apreciación ante la utilidad percibida y satisfacción. Las encuestas utilizadas fueron de tipo Likert, con valoraciones del 1 al 7, siendo el valor de 1 el más desfavorable. El modelo de aceptación tecnológica (TAM) se pasó a los discentes antes y después de la experiencia. Solo la encuesta de motivación de materiales de instrucción (IMMS) se pasó al finalizar el curso. Además, se valoró el conocimiento mediante una prueba de evaluación al finalizar las asignaturas.
Los resultados mostraron que ambas tecnologías eran igual de eficientes, mejorando el aprendizaje de las materias. Además, estas técnicas permite que los discentes de primer año de grado puedan correlacionar los conceptos teóricos con modelos visuales reales. De esa manera profundizan en conceptos difíciles de explicar de manera tradicional, al no permitir su visualización y creación de una imagen mental, y por otro lado porque no tendrían acceso hasta último año de grado, situación donde ocurre un acercamiento real a los pacientes del hospital.
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This project aims to reveal the factors that help to integrate the teaching-learning binomial using augmented reality (AR) technologies and the modeling of bioreplicas by rendering (through segmentation, photogrammetry or computer-aided design), preferably through of digital images of real patients or collaborators, and/or fused deposition (FDM).
The chosen learning environments were the subjects of the physiology department at the Ceuta Campus ("Human Anatomy" and "Physiology I"), taught in the nursing degree at the University of Granada. Along with traditional classes, practices and seminars, traditional teaching was complemented with emerging technology to verify the results that they could have in the student's learning, in addition to checking their appreciation of the perceived usefulness and satisfaction. The surveys used were of the Likert type, with ratings from 1 to 7, with a value of 1 being the most unfavorable. The technology acceptance model (TAM) was passed to the students before and after the experience. Only the Instructional Materials Motivation Survey (IMMS) was passed at the end of the course. In addition, knowledge was assessed through an evaluation test at the end of the subjects.
The results showed that both technologies were equally efficient, improving the learning of the subjects. In addition, these techniques allow first-year students to correlate theoretical concepts with real visual models. In this way, they delve into concepts that are difficult to explain in a traditional way, by not allowing their visualization and creation of a mental image, and on the other hand because they would not have access until the last year of degree, a situation where a real approach to hospital patients occurs.Departamento de Fisiología
Glycemic markers and relation with arterial stiffness in Caucasian subjects of the MARK study
[EN]BACKGROUND:
Effect of prediabetes and normal glucose on arterial stiffness remains controversial. The primary aim of this study was to investigate the relationship of fasting plasma glucose (FPG), postprandial glucose (PG) and glycosylated haemoglobin (HbA1c) with brachial-ankle pulse wave velocity (baPWV) and cardio-ankle vascular index (CAVI) in Caucasian adults. The secondary aim was to analyse this relationship by glycaemic status.
METHODS:
Cross-sectional study. Setting: Primary care. Participants: 2,233 subjects, 35-74 years. Measures: FPG (mg/dL) and HbA1c (%) of all subjects were measured using standard automated enzymatic methods. PG (mg/dL) was self-measured at home two hours after meals (breakfast, lunch and dinner) for one day using an Accu-chek ® glucometer. CAVI was measured using a VaSera VS-1500® device (Fukuda Denshi), and baPWV was calculated using a validated equation.
RESULTS:
CAVI and baPWV values were significantly higher in subjects with diabetes mellitus than in glucose normal and prediabetes groups (p<0.001). FPG, PG and HbA1c were positively associated with CAVI and baPWV. The β regression coefficient for: HbA1c was 0.112 (CI 95% 0.068 to 0.155) with CAVI, 0.266 (CI 95% 0.172 to 0.359) with baPWV; for PG was 0.006 (CI 95% 0.004 to 0.009 and for FPG was 0.005 (CI 95% 0.002 to 0.008) with baPWV; and for PG was 0.002 (CI 95% 0.001 to 0.003) and 0.003 (CI 95% 0.002 to 0.004) with CAVI (p<0.01 in all cases). When analysing by hyperglycaemic status, FPG, PG and HbA1c were positively associated with CAVI and baPWV in subjects with type 2 diabetes mellitus.
CONCLUSION:
FPG, PG and HbA1c show a positive association with CAVI and baPWV, in Caucasian adults with intermediate cardiovascular risk factors. When analysing by hyperglycaemic status, the association is only maintained in subjects with type 2 diabetes mellitus
Manejo anetésico de Hematoma Subdural bilateral en paciente anciano a propósito de un caso.
El tratamiento quirúrgico y el manejo anestésico de los hematomas subdurales bilaterales en el adulto mayor es un tema complejo y controversial por la elevada mortalidad que lo acompaña. El paciente anciano portador de hematoma subdural bilateral representa un reto para el anestesiólogo. La disponibilidad de drogas intravenosas potentes, agentes inhalatorios con propiedades fisicoquímicas únicas, resultan fundamentales para estos pacientes. El objetivo de este reporte de caso es dar a conocer el desenlace clínico en el paciente anciano con hematoma subdural bilateral y la importancia del manejo neuroanestesico en estas situaciones. Se comenta el caso de un paciente masculino de 81 años de edad sometido a drenaje de hematoma subdural bilateral; para lo cual se administra anestesia general balanceada con propofol (en la inducción), sevorane y remifentanil (en el mantenimiento), con buen control hemodinámico, y exitosa recuperación. El manejo anestésico de un paciente anciano neuroquirurgico se complica cuando el anestesiólogo no está familiarizado con los parámetros hemodinámicos, mismos que son de vital importancia para mantener un adecuado funcionamiento cardiovascular, tomando importante relevancia la presión de perfusión cerebral. Este tópico reviste gran importancia debido al elevado porcentaje de pacientes ancianos neuroquirurgicos y por la elevada morbi-mortalidad, siendo la anestesia general balanceada la técnica de elección en este caso en particular. Se concluyó que a pasar de los riesgos que implica el acto quirúrgico, la edad y comorbilidades se conserva la presión de perfusión cerebral, es de vital importancia para llevar a cabo un manejo anestésico adecuado y una pronta recuperación neurológica
Binding of Natural and Synthetic Polyphenols to Human Dihydrofolate Reductase
Dihydrofolate reductase (DHFR) is the subject of intensive investigation since it appears to be the primary target enzyme for antifolate drugs. Fluorescence quenching experiments show that the ester bond-containing tea polyphenols (-)-epigallocatechin gallate (EGCG) and (-)-epicatechin gallate (ECG) are potent inhibitors of DHFR with dissociation constants (KD)of 0.9 and 1.8 μM, respectively, while polyphenols lacking the ester bound gallate moiety [e.g., (-)-epigallocatechin (EGC) and (-)-epicatechin (EC)] did not bind to this enzyme. To avoid stability and bioavailability problems associated with tea catechins we synthesized a methylated derivative of ECG (3-O-(3,4,5-trimethoxybenzoyl)-(-)-epicatechin; TMECG), which effectively binds to DHFR (KD = 2.1 μM). In alkaline solution, TMECG generates a stable quinone methide product that strongly binds to the enzyme with a KD of 8.2 nM. Quercetin glucuronides also bind to DHFR but its effective binding was highly dependent of the sugar residue, with quercetin-3-xyloside being the stronger inhibitor of the enzyme with a KD of 0.6 μM. The finding that natural polyphenols are good inhibitors of human DHFR could explain the epidemiological data on their prophylactic effects for certain forms of cancer and open a possibility for the use of natural and synthetic polyphenols in cancer chemotherapy
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