Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 631
Filtrar
Mais filtros

Tipo de documento
Intervalo de ano de publicação
1.
Actas Dermosifiliogr ; 115(1): 36-47, 2024 Jan.
Artigo em Inglês, Espanhol | MEDLINE | ID: mdl-37678633

RESUMO

BACKGROUND: Several studies support the hypothesis that scabies is on the rise in Spain. There are also concerns about the possible development of resistance to treatment and an increase in atypical presentations. The aims of this study were to describe the demographic and clinical characteristics of patients with scabies seen by dermatologists in Spain, to identify the possible emergence of atypical forms of scabies, and to explore the frequency of treatment failures and associated risk factors. METHODS: We conducted an observational, cross-sectional, multicenter study of data collected prospectively in April and May 2023 using the CLINI-AEDVp platform created by the Spanish Academy of Dermatology and Venereology (AEDV). RESULTS: Participating dermatologists from 31 hospitals in 15 of Spain's autonomous communities recorded 186 cases of active scabies (51% in women) during the study period. A diagnostic certainty level of A, B or C as per the International Alliance for the Control of Scabies Consensus Criteria was required for diagnosis. Overall, 92% of patients had typical scabies and 66% had already been treated with a scabicide for the current episode. Of the treated patients, only 36% had received and completed adequate treatment (including the simultaneous treatment of all household members) and 50% had not received clear written recommendations. CONCLUSIONS: In a high proportion of the cases of scabies studied, the patient had already received treatment. In those cases, we observed several remediable shortcomings that could explain why some of these treatments had failed. Remedying these deficiencies should lead to better control of scabies and an improved assessment of the actual effectiveness of currently available scabicides.


Assuntos
Escabiose , Humanos , Feminino , Escabiose/tratamento farmacológico , Escabiose/epidemiologia , Espanha/epidemiologia , Estudos Transversais , Falha de Tratamento , Academias e Institutos
2.
Actas Dermosifiliogr ; 115(1): T36-T47, 2024 Jan.
Artigo em Inglês, Espanhol | MEDLINE | ID: mdl-37923079

RESUMO

BACKGROUND: Several studies support the hypothesis that scabies is on the rise in Spain. There are also concerns about the possible development of resistance to treatment and an increase in atypical presentations. The aims of this study were to describe the demographic and clinical characteristics of patients with scabies seen by dermatologists in Spain, to identify the possible emergence of atypical forms of scabies, and to explore the frequency of treatment failures and associated risk factors. METHODS: We conducted an observational, cross-sectional, multicenter study of data collected prospectively in April and May 2023 using the CLINI-AEDVp platform created by the Spanish Academy of Dermatology and Venereology (AEDV). RESULTS: Participating dermatologists from 31 hospitals in 15 of Spain's autonomous communities recorded 186 cases of active scabies (51% in women) during the study period. A diagnostic certainty level of A, B or C as per the International Alliance for the Control of Scabies Consensus Criteria was required for diagnosis. Overall, 92% of patients had clinical features of classic scabies and 66% had already been treated with a scabicide for the current episode. Of the treated patients, only 36% had received and completed adequate treatment (including the simultaneous treatment of all household members) and 50% had not received clear written recommendations. CONCLUSIONS: In a high proportion of scabies cases, the patient has already received treatment. In those cases, we observe several remediable shortcomings that could explain why some of these treatments fail. Remedying these deficiencies should lead to better control of scabies and an improved assessment of the actual effectiveness of currently available scabicides.


Assuntos
Escabiose , Humanos , Feminino , Escabiose/diagnóstico , Escabiose/tratamento farmacológico , Escabiose/epidemiologia , Espanha/epidemiologia , Estudos Transversais , Falha de Tratamento , Academias e Institutos
3.
Metab Eng ; 75: 78-90, 2023 01.
Artigo em Inglês | MEDLINE | ID: mdl-36368470

RESUMO

Conversion of CO2 to value-added products presents an opportunity to reduce GHG emissions while generating revenue. Formate, which can be generated by the electrochemical reduction of CO2, has been proposed as a promising intermediate compound for microbial upgrading. Here we present progress towards improving the soil bacterium Cupriavidus necator H16, which is capable of growing on formate as its sole source of carbon and energy using the Calvin-Benson-Bassham (CBB) cycle, as a host for formate utilization. Using adaptive laboratory evolution, we generated several isolates that exhibited faster growth rates on formate. The genomes of these isolates were sequenced, and resulting mutations were systematically reintroduced by metabolic engineering, to identify those that improved growth. The metabolic impact of several mutations was investigated further using RNA-seq transcriptomics. We found that deletion of a transcriptional regulator implicated in quorum sensing, PhcA, reduced expression of several operons and led to improved growth on formate. Growth was also improved by deleting large genomic regions present on the extrachromosomal megaplasmid pHG1, particularly two hydrogenase operons and the megaplasmid CBB operon, one of two copies present in the genome. Based on these findings, we generated a rationally engineered ΔphcA and megaplasmid-deficient strain that exhibited a 24% faster maximum growth rate on formate. Moreover, this strain achieved a 7% growth rate improvement on succinate and a 19% increase on fructose, demonstrating the broad utility of microbial genome reduction. This strain has the potential to serve as an improved microbial chassis for biological conversion of formate to value-added products.


Assuntos
Cupriavidus necator , Cupriavidus necator/genética , Cupriavidus necator/metabolismo , Dióxido de Carbono/metabolismo , Óperon , Carbono/metabolismo , Formiatos/metabolismo
4.
Pediatr Cardiol ; 44(6): 1285-1292, 2023 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-37147525

RESUMO

The implantation of percutaneous balloon expandable valves in native or patched right ventricular outflow tracts (nRVOT) is a challenging technique due to the diversity of anatomies and shapes, the large sizes, and the distensibility of the nRVOT, for which specific techniques have been developed. We present a single center experience with balloon expandable percutaneous pulmonary valves in nRVOT, describing the techniques used, complications observed, and a short-mid term follow-up.. This is a single center descriptive study of patients who underwent a percutaneous pulmonary valve implantation in a nRVOT with a balloon expandable pulmonary valve in our center between September 2012 and June 2022.. We implanted successfully 45 valves in 46 patients (20 Sapien and 25 Melody). Tetralogy of Fallot or pulmonary atresia with VSD were the main congenital heart disease (n = 32). All were pre-stented, 18 in a one step procedure. We used a Dryseal sheath in 13/21 Sapien. In 6 patients we used the anchoring technique, 5 with a very large nRVOT and one pyramidal nRVOT. In the 3.5 year follow-up 7 patients developed endocarditis and 3 required a valve redilation, no fractures were observed. PPVI of native RVOT with balloon expandable valves is feasible in a number of selected anatomies, including large or pyramidal nRVOT, using specific techniques, (presenting, LPA anchoring).


Assuntos
Cardiopatias Congênitas , Implante de Prótese de Valva Cardíaca , Próteses Valvulares Cardíacas , Insuficiência da Valva Pulmonar , Valva Pulmonar , Humanos , Valva Pulmonar/cirurgia , Implante de Prótese de Valva Cardíaca/métodos , Próteses Valvulares Cardíacas/efeitos adversos , Resultado do Tratamento , Cardiopatias Congênitas/cirurgia , Cateterismo Cardíaco/métodos , Desenho de Prótese , Estudos Retrospectivos
5.
Encephale ; 49(3): 211-218, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35331466

RESUMO

OBJECTIVES: Behavior problems are one of the most common reasons for seeking mental health services in pediatric populations. The objectives are to evaluate the effects of the EQUIPE program (Étude Québécoise d'Intervention pour les Parents d'Enfants avec des problèmes de comportement) and to analyze the impact of the severity of behavior problems and of parental characteristics. METHODS: This program was translated from the Community Parent Education Program. The effects of EQUIPE, as compared to a control group, were evaluated by using Child Behavior Checklist and Parent Stress Index questionnaires before (T0) and after the intervention program (T1), and at 6 (T2) and 12months (T3) follow-up visits. RESULTS: In total, 533 participants were enrolled in intervention (n=465) (with "severe" or "mild" subgroups according to CBCL-T score) and a control group (n=68). In the two groups, the results showed a statistically significant decrease in Child Behavior Checklist and Parent Stress Index total scores at T1, T2 and T3, with the exception of Child Behavior Checklist total scores in the control group at T2. In the intervention group Child Behavior Checklist total scores were significantly higher in the "severe"; which was not the case for Parent Stress Index total scores at T2 and T3. DISCUSSION: Socioeconomic characteristics, family details, parental medical history and the age of the children appeared to influence changes in Child Behavior Checklist and Parent Stress Index total scores. CONCLUSION: The EQUIPE program is an effective intervention for reducing behavior problems and parents' stress in a French-Canadian population.


Assuntos
Serviços de Saúde Mental , Pais , Humanos , Criança , Canadá , Pais/psicologia , Relações Mãe-Filho , Comportamento Infantil/psicologia
6.
Med Oral Patol Oral Cir Bucal ; 28(5): e404-e411, 2023 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-37330967

RESUMO

BACKGROUND: Persistent periapical lesions (PPL) are the result of pulpar necrosis induced by bacterial infection resulting in bone degradation and culminating with the loss of dental piece. Pathological changes in the peripapice are associated with the presence of free radicals. The transcription factor Nrf2 is the main regulator of the endogenous antioxidant response against oxidative stress and has been implicated in the regulation of osteoclastogenesis.The aim is to determine the oxidative condition in samples from patients with Persistent Periapical Injuries as a detonating factor of tissue damage. MATERIAL AND METHODS: An observational, descriptive, cross-sectional study was carried out in samples with PPL (cases) and samples by removal of third molars (controls) obtained in the clinic of the specialty in endodontics, University of Guadalajara. Samples were submitted to histological staining with Hematoxylin-Eosin, lipoperoxide analysis, Superoxide Dismutase (SOD), Glutathione-Peroxidase (GPx) and Catalase (CAT) activities were determined by immunoenzymatic assays and NrF2 by Western Blot analysis. RESULTS: Samples from PPL patients histologically showed an increased presence of lymphocytes, plasma cells, and eosinophils, as well as a decrease in extracellular matrix proteins and fibroblast cells. There was a rise in lipid peroxidation, GPx and SOD activities, but an important decline (36%) in Catalase activity was observed (p<0.005); finally, NrF2-protein was diminished at 10.41%. All comparisons were between cases vs controls. CONCLUSIONS: The alterations in antioxidants endogenous NrF2-controlled are related to osseous destruction in patients with PPL.


Assuntos
Antioxidantes , Fator 2 Relacionado a NF-E2 , Catalase/metabolismo , Fator 2 Relacionado a NF-E2/metabolismo , Estudos Transversais , Antioxidantes/metabolismo , Superóxido Dismutase/metabolismo , Glutationa Peroxidase/metabolismo
7.
J Struct Biol ; 214(3): 107872, 2022 09.
Artigo em Inglês | MEDLINE | ID: mdl-35660516

RESUMO

Image processing in cryogenic electron tomography (cryoET) is currently at a similar state as Single Particle Analysis (SPA) in cryogenic electron microscopy (cryoEM) was a few years ago. Its data processing workflows are far from being well defined and the user experience is still not smooth. Moreover, file formats of different software packages and their associated metadata are not standardized, mainly since different packages are developed by different groups, focusing on different steps of the data processing pipeline. The Scipion framework, originally developed for SPA (de la Rosa-Trevín et al., 2016), has a generic python workflow engine that gives it the versatility to be extended to other fields, as demonstrated for model building (Martínez et al., 2020). In this article, we provide an extension of Scipion based on a set of tomography plugins (referred to as ScipionTomo hereafter), with a similar purpose: to allow users to be focused on the data processing and analysis instead of having to deal with multiple software installation issues and the inconvenience of switching from one to another, converting metadata files, managing possible incompatibilities, scripting (writing a simple program in a language that the computer must convert to machine language each time the program is run), etcetera. Additionally, having all the software available in an integrated platform allows comparing the results of different algorithms trying to solve the same problem. In this way, the commonalities and differences between estimated parameters shed light on which results can be more trusted than others. ScipionTomo is developed by a collaborative multidisciplinary team composed of Scipion team engineers, structural biologists, and in some cases, the developers whose software packages have been integrated. It is open to anyone in the field willing to contribute to this project. The result is a framework extension that combines the acquired knowledge of Scipion developers in close collaboration with third-party developers, and the on-demand design of functionalities requested by beta testers applying this solution to actual biological problems.


Assuntos
Tomografia com Microscopia Eletrônica , Software , Algoritmos , Microscopia Crioeletrônica/métodos , Processamento de Imagem Assistida por Computador/métodos , Reprodutibilidade dos Testes
8.
Faraday Discuss ; 240(0): 210-227, 2022 11 08.
Artigo em Inglês | MEDLINE | ID: mdl-35861059

RESUMO

The number of maps deposited in public databases (Electron Microscopy Data Bank, EMDB) determined by cryo-electron microscopy has quickly grown in recent years. With this rapid growth, it is critical to guarantee their quality. So far, map validation has primarily focused on the agreement between maps and models. From the image processing perspective, the validation has been mostly restricted to using two half-maps and the measurement of their internal consistency. In this article, we suggest that map validation can be taken much further from the point of view of image processing if 2D classes, particles, angles, coordinates, defoci, and micrographs are also provided. We present a progressive validation scheme that qualifies a result validation status from 0 to 5 and offers three optional qualifiers (A, W, and O) that can be added. The simplest validation state is 0, while the most complete would be 5AWO. This scheme has been implemented in a website https://biocomp.cnb.csic.es/EMValidationService/ to which reconstructed maps and their ESI can be uploaded.


Assuntos
Processamento de Imagem Assistida por Computador , Microscopia Crioeletrônica/métodos , Microscopia Eletrônica
9.
BMC Psychiatry ; 22(1): 614, 2022 09 19.
Artigo em Inglês | MEDLINE | ID: mdl-36123659

RESUMO

BACKGROUND: Police officers are increasingly required to respond to incidents involving psychiatric patients. However, few studies have assessed whether the attitude of police officers depends on prior knowledge of their specific psychiatric diagnosis. Our aim was to analyze the effects of psychiatric diagnosis on the behavior of police officers. METHODS: We utilized the Attribution Questionnaire adapted to the police context to examine the attitudes of 927 officers of the Spanish National Police Force towards persons diagnosed with either schizophrenia or depressive disorder playing the role of somebody in need of assistance, a victim of a crime, a witness, or a suspect in a criminal case. Different socio-demographic variables were also collected. RESULTS: Compared to attitudes to individuals with a known psychiatric diagnosis, police officers expressed increased willingness to help psychiatric patients and increased sympathy and attributing to them less responsibility for their actions. They also showed increased feelings of avoidance, reported a greater perception of danger and a greater need for isolation and involuntary treatment. This was especially so in the case of schizophrenia. Stigmatizing attitudes were less apparent when the person was a woman, a veteran officer, or someone with a history of work experience. CONCLUSIONS: Police officers may hold certain stigmatizing attitudes towards persons with mental illness, particularly schizophrenia, that require special attention, as they may negatively affect police action. We found several factors associated with the persistence of these stigmatizing attitudes among police officers that may guide us when implementing training programs for promoting attitude change, especially at the beginning of an officer's professional career.


Assuntos
Transtornos Mentais , Esquizofrenia , Atitude , Feminino , Humanos , Transtornos Mentais/diagnóstico , Transtornos Mentais/psicologia , Polícia , Esquizofrenia/diagnóstico , Inquéritos e Questionários
10.
Hum Genet ; 140(12): 1665-1678, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34448047

RESUMO

Syndromic retinal diseases (SRDs) are a group of complex inherited systemic disorders, with challenging molecular underpinnings and clinical management. Our main goal is to improve clinical and molecular SRDs diagnosis, by applying a structured phenotypic ontology and next-generation sequencing (NGS)-based pipelines. A prospective and retrospective cohort study was performed on 100 probands with an a priori diagnosis of non-Usher SRDs, using available clinical data, including Human Phenotype Ontology annotation, and further classification into seven clinical categories (ciliopathies, specific syndromes and five others). Retrospective molecular diagnosis was assessed using different molecular and bioinformatic methods depending on availability. Subsequently, uncharacterized probands were prospectively screened using other NGS approaches to extend the number of analyzed genes. After phenotypic classification, ciliopathies were the most common SRD (35%). A global characterization rate of 52% was obtained, with six cases incompletely characterized for a gene that partially explained the phenotype. An improved characterization rate was achieved addressing prospective cases (83%) and well-recognizable syndrome (62%) subgroups. The 27% of the fully characterized cases were reclassified into a different clinical category after identification of the disease-causing gene. Clinical-exome sequencing is the most appropriate first-tier approach for prospective cases, whereas whole-exome sequencing and bioinformatic reanalysis increases the diagnosis of uncharacterized retrospective cases to 45%, mostly those with unspecific symptoms. Our study describes a comprehensive approach to SRDs in daily clinical practice and the importance of thorough clinical assessment and selection of the most appropriate molecular test to be used to solve these complex cases and elucidate novel associations.


Assuntos
Oftalmopatias Hereditárias/diagnóstico , Ontologia Genética , Sequenciamento de Nucleotídeos em Larga Escala , Doenças Retinianas/diagnóstico , Ciliopatias/genética , Estudos de Coortes , Oftalmopatias Hereditárias/genética , Feminino , Estudos de Associação Genética , Testes Genéticos , Humanos , Masculino , Técnicas de Diagnóstico Molecular , Mutação , Fenótipo , Estudos Prospectivos , Doenças Retinianas/genética , Estudos Retrospectivos , Síndrome
11.
Clin Exp Dermatol ; 46(6): 1097-1101, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-33713349

RESUMO

Azathioprine is an immunosuppressant drug used in many dermatological and nondermatological pathologies. Azathioprine hypersensitivity syndrome (AHS) is a rare idiosyncratic reaction that is not related to dose or thiopurine methyltransferase activity. Up to half of cases of AHS can present with variable cutaneous manifestations besides fever, malaise and other systemic symptoms. It is important to be aware of AHS, as continuance or reintroduction of the drug can led to multiorgan failure and cardiovascular collapse.


Assuntos
Azatioprina/efeitos adversos , Síndrome de Hipersensibilidade a Medicamentos/etiologia , Síndrome de Hipersensibilidade a Medicamentos/patologia , Imunossupressores/efeitos adversos , Pele/patologia , Diagnóstico Diferencial , Edema/patologia , Humanos , Masculino , Pessoa de Meia-Idade , Neutrófilos/patologia
12.
Aging Clin Exp Res ; 32(6): 1043-1048, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30989508

RESUMO

BACKGROUND: Health professionals commonly use gait speed in the evaluation of functional status in older people. However, only a limited number of studies have assessed gait speed in the absence of disorders of gait, using confounding factors and exclusion criteria coming from studies conducted in younger people. Our study aims to analyse which factors are associated with gait speed in older people with normal clinical gait. METHODS: An observational cross-sectional study was conducted in 119 community-dwelling residents without relevant comorbidities (Charlson index < 2), preserved function (Barthel > 85) and normal gait by visual exploration. Exclusion criteria included suffering from any illness that could modify the characteristics of gait, terminal status or the presence of an acute medical illness in the past 3 months. We used a stepwise linear regression of several variables (sociodemographic characteristics, cognition, body composition, drugs, falls, sarcopenia, frailty and physical activity) on 6-metre gait speed. RESULTS: The mean age was 78 years (range 70-96 years) and 71.4% were women. Variables that remained associated with gait speed in the multivariate final model were age (B = - 0.020, p < 0.001); gender (B = - 0.184, p < 0.001); waist-to-height ratio (B = - 0.834, p = 0.002); number of falls (B = - 0.049, p = 0.003) and the number of Fried's frailty criteria (B = - 0.064, p = 0.019). CONCLUSION: Falls, frailty and the waist-to-height ratio modify gait speed in older people with normal gait. Studies analysing the potential effect of several factors on gait speed should consider them as confounding factors.


Assuntos
Velocidade de Caminhada , Acidentes por Quedas , Idoso , Idoso de 80 Anos ou mais , Cognição , Estudos Transversais , Exercício Físico , Feminino , Idoso Fragilizado , Fragilidade , Humanos , Vida Independente , Masculino , Sarcopenia/fisiopatologia
13.
Immun Ageing ; 16: 14, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31312227

RESUMO

BACKGROUND: Seasonal influenza virus infection is a significant cause of morbimortality in the elderly. However, there is poor vaccine efficacy in this population due to immunosenescence. We aimed to explore several homeostatic parameters in the elderly that could impact influenza vaccine responsiveness. METHODS: Subjects (> 60 years old) who were vaccinated against influenza virus were included, and the vaccine response was measured by a haemagglutination inhibition (HAI) test. At baseline, peripheral CD4 and CD8 T-cells were phenotypically characterized. Thymic function and the levels of different inflammation-related biomarkers, including Lipopolysaccharide Binding Protein (LBP) and anti-cytomegalovirus (CMV) IgG antibodies, were also measured. RESULTS: Influenza vaccine non-responders showed a tendency of higher frequency of regulatory T-cells (Tregs) before vaccination than responders (1.49 [1.08-1.85] vs. 1.12 [0.94-1.63], respectively, p = 0.061), as well as higher expression of the proliferation marker Ki67 in Tregs and different CD4 and CD8 T-cell maturational subsets. The levels of inflammation-related biomarkers correlated with the frequencies of different proliferating T-cell subsets and with thymic function (e.g., thymic function with D-dimers, r = - 0.442, p = 0.001). CONCLUSIONS: Age-related homeostatic dysregulation involving the proliferation of CD4 and CD8 T-cell subsets, including Tregs, was related to a limited responsiveness to influenza vaccination and a higher inflammatory status in a cohort of elderly people.

14.
Int J Obes (Lond) ; 42(4): 618-624, 2018 04.
Artigo em Inglês | MEDLINE | ID: mdl-29158541

RESUMO

BACKGROUND/OBJECTIVES: We aimed to evaluate mitochondrial biogenesis (MB), structure, metabolism and dysfunction in abdominal adipose tissue from male pediatric patients with obesity. SUBJECTS/METHODS: Samples were collected from five children with obesity (percentile ⩾95) and five eutrophic boys (percentile ⩾5/⩽85) (8-12 years old) following parental informed consent. We analyzed the expression of key genes involved in MB (sirtuin-1 (SIRT1), peroxisome proliferator-activated receptor-γ (PPARγ), PPARγ coactivator-1α (PGC1α), nuclear respiratory factors 1 and 2 (NRF1, NRF2) and mitochondrial transcription factor A (TFAM) and surrogates for mitochondrial function/structure/metabolism (porin, TOMM20, complex I and V, UCP1, UCP2, SIRT3, SOD2) by western blot. Citrate synthase (CS), complex I (CI) activity, adenosine triphosphate (ATP) levels, mitochondrial DNA (mtDNA) content and oxidative stress end points were also determined. RESULTS: Most MB proteins were significantly decreased in samples from children with obesity except complex I, V and superoxide dismutase-2 (SOD2). Similarly, CS and CI activity showed a significant reduction, as well as ATP levels and mtDNA content. PPARγ, PGC1α, complex I and V and SOD2 were hyperacetylated compared with lean samples. Concurrently, in samples from children with obesity, we found decreased SOD2 activity and redox state imbalance highlighted by decreased reduced glutathione/oxidized glutathione (GSH/GSSG) ratio and significant increases in protein carbonylation. CONCLUSIONS: Adipose tissue from children with obesity demonstrates a dysregulation of key modulators of MB and organelle structure, and displays hyperacetylation of key proteins and altered expression of upstream regulators of cell metabolism.


Assuntos
Tecido Adiposo/fisiopatologia , Mitocôndrias/fisiologia , Biogênese de Organelas , Obesidade Infantil/fisiopatologia , Acetilação , Tecido Adiposo/citologia , Tecido Adiposo/metabolismo , Criança , DNA Mitocondrial/metabolismo , Humanos , Masculino , Proteínas Mitocondriais/análise , Proteínas Mitocondriais/química , Proteínas Mitocondriais/metabolismo , Estresse Oxidativo/fisiologia , Obesidade Infantil/metabolismo
15.
Metab Eng ; 47: 10-20, 2018 05.
Artigo em Inglês | MEDLINE | ID: mdl-29477855

RESUMO

Strain engineering for industrial production requires a targeted improvement of multiple complex traits, which range from pathway flux to tolerance to mixed sugar utilization. Here, we report the use of an iterative CRISPR EnAbled Trackable genome Engineering (iCREATE) method to engineer rapid glucose and xylose co-consumption and tolerance to hydrolysate inhibitors in E. coli. Deep mutagenesis libraries were rationally designed, constructed, and screened to target ~40,000 mutations across 30 genes. These libraries included global and high-level regulators that regulate global gene expression, transcription factors that play important roles in genome-level transcription, enzymes that function in the sugar transport system, NAD(P)H metabolism, and the aldehyde reduction system. Specific mutants that conferred increased growth in mixed sugars and hydrolysate tolerance conditions were isolated, confirmed, and evaluated for changes in genome-wide expression levels. We tested the strain with positive combinatorial mutations for 3-hydroxypropionic acid (3HP) production under high furfural and high acetate hydrolysate fermentation, which demonstrated a 7- and 8-fold increase in 3HP productivity relative to the parent strain, respectively.


Assuntos
Escherichia coli/genética , Edição de Genes/métodos , Engenharia Metabólica/métodos , Mutagênese , Escherichia coli/metabolismo
16.
Metab Eng ; 48: 52-62, 2018 07.
Artigo em Inglês | MEDLINE | ID: mdl-29852272

RESUMO

Medium- and long-chain methyl ketones are fatty acid-derived compounds that can be used as biofuel blending agents, flavors and fragrances. However, their large-scale production from sustainable feedstocks is currently limited due to the lack of robust microbial biocatalysts. The oleaginous yeast Yarrowia lipolytica is a promising biorefinery platform strain for the production of methyl ketones from renewable lignocellulosic biomass due to its natively high flux towards fatty acid biosynthesis. In this study, we report the metabolic engineering of Y. lipolytica to produce long- and very long-chain methyl ketones. Truncation of peroxisomal ß-oxidation by chromosomal deletion of pot1 resulted in the biosynthesis of saturated, mono-, and diunsaturated methyl ketones in the C13-C23 range. Additional overexpression and peroxisomal targeting of a heterologous bacterial methyl ketone biosynthesis pathway yielded an initial titer of 151.5 mg/L of saturated methyl ketones. Dissolved oxygen concentrations in the cultures were found to substantially impact cell morphology and methyl ketone biosynthesis. Bioreactor cultivation under optimized conditions resulted in a titer of 314.8 mg/L of total methyl ketones, representing more than a 6000-fold increase over the parental strain. This work highlights the potential of Y. lipolytica to serve as chassis organism for the biosynthesis of acyl-thioester derived long- and very long-chain methyl ketones.


Assuntos
Deleção de Genes , Genes Fúngicos , Cetonas/metabolismo , Engenharia Metabólica , Peroxissomos , Yarrowia , Oxirredução , Peroxissomos/genética , Peroxissomos/metabolismo , Yarrowia/genética , Yarrowia/crescimento & desenvolvimento
17.
Clin Genet ; 93(1): 3-14, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-28369730

RESUMO

Hypertrophic cardiomyopathy (HCM) is the most common inherited cardiovascular disease. HCM is a highly complex and heterogeneous disease regarding not only the number of associated mutations but also the severity of phenotype, symptom burden, and the risk of complications, such as heart failure and sudden death. The penetrance is incomplete and it is age and gender dependent. It is accepted as a disease of the sarcomere. Sixty percent of HCM cases carry mutations in 1 of 8 sarcomere protein genes, mainly non-sense MYBPC3 and missense MYH7 variants. Young patients with severe phenotype and other clinical features are included in proposed scores for prediction of high positive genetic result. The number of genes reported as disease-causing has increased in the last few years, in some cases without robust evidence. Currently available in silico tools are not always useful for differentiation between benign and deleterious variants. There is enough information on genotype-phenotype correlations to start understanding the mechanisms of the disease. Genetic and environmental modifiers have been explored with some interesting insights from miRNA studies with potential as biomarkers and therapeutic agents. There is an additional value of genetic testing in HCM for prognosis. Knowledge about genetics and functional studies are the basis of near future therapies.


Assuntos
Cardiomiopatia Hipertrófica/genética , Predisposição Genética para Doença/genética , Mutação , Sarcômeros/genética , Cardiomiopatia Hipertrófica/complicações , Cardiomiopatia Hipertrófica/diagnóstico , Morte Súbita Cardíaca/etiologia , Estudos de Associação Genética , Testes Genéticos , Humanos
18.
BMC Vet Res ; 14(1): 141, 2018 Apr 27.
Artigo em Inglês | MEDLINE | ID: mdl-29699565

RESUMO

BACKGROUND: During the storage of packed red blood cells (pRBC), packed cell volume (PCV), bacterial contamination and percentage of haemolysis [percentage of free haemoglobin (HGB) in relation to the total HGB] are important quality parameters. Both PCV and haemolysis are indicators of the cellular integrity of stored units. There are no published experimental studies that evaluated these parameters during storage of feline pRBC using SAGM (adenine, dextrose, mannitol and sodium chloride) as the additive solution. The present study aims to (1) evaluate the quality of feline pRBCs stored in SAGM; (2) test for the semi-closed system's suitability for use and risk of bacterial contamination; (3) establish the maximum storage time that may be appropriate to meet the criteria established by the United States Food and Drug Administration (US-FDA) guidelines for human blood banking; and (4) evaluate the need to calculate the percentage of haemolysis prior to the administration of units stored for more than 4 weeks. Four hundred eighty nine feline pRBC units were analyzed. Bacterial culture, PCV and percentage of haemolysis were determined within 6 h after processing (t0). One hundred and eighty units were re-tested for haemolysis and PCV after 29-35 days of storage (t1) and 118 units after 36-42 days (t2). RESULTS: Bacterial contamination was not detected in any pRBC unit. Mean PCV at t0 was 52.25% (SD: ±5.27) and decreased significantly (p < 0.001) during storage to 48.15% (SD: ±3.79) at t1 and to 49.34% (SD: ±4.45) at t2. Mean percentage of haemolysis at t0 was 0.07% (SD: ±0.06) and increased significantly (p < 0.001) to 0.69% (SD: ±0.40) at t1 and to 0.81% (SD: ±0.47) at t2. In addition, 13.88% and 19.49% of pRBC units exceeded 1% haemolysis at t1 and t2, respectively. CONCLUSIONS: According to the US-FDA guidelines for human blood banking that recommend a maximum of 1% haemolysis, the results of this study show that all feline pRBC units with less than 24 h of shelf life have low levels of haemolysis. However, units preserved up to 28 days can only be administered if tested for haemolysis before use, since 13.88% units exceeded the 1% limit. The semi-closed system was considered safe for use as bacterial contamination was not detected in any pRBC unit.


Assuntos
Armazenamento de Sangue , Bancos de Sangue , Coleta de Amostras Sanguíneas/veterinária , Gatos/sangue , Eritrócitos , Animais , Bancos de Sangue/normas , Coleta de Amostras Sanguíneas/normas , Hematócrito/veterinária , Hemoglobinas/análise , Hemólise , Técnicas In Vitro , Controle de Qualidade , Fatores de Tempo , Armazenamento de Sangue/métodos
19.
J Investig Allergol Clin Immunol ; 28(6): 379-391, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30004024

RESUMO

Atopic dermatitis (AD) is a recurrent and chronic skin disease characterized by dysfunction of the epithelial barrier, skin inflammation, and immune dysregulation, with changes in the skin microbiota and colonization by Staphylococcus aureus being common. For this reason, the therapeutic approach to AD is complex and should be directed at restoring skin barrier function, reducing dehydration, maintaining acidic pH, and avoiding superinfection and exposure to possible allergens. There is no curative treatment for AD. However, a series of measures are recommended to alleviate the disease and enable patients to improve their quality of life. These include adequate skin hydration and restoration of the skin barrier with the use of emollients, antibacterial measures, specific approaches to reduce pruritus and scratching, wet wrap applications, avoidance of typical AD triggers, and topical anti-inflammatory drugs. Anti-inflammatory treatment is generally recommended during acute flares or, more recently, for preventive management. Nevertheless, the selection of the pharmacologic agent, as well as its potency, duration, and frequency of application must be in accordance with the severity of the disease and the distribution and type of the lesion. The objectives of this review are to emphasize the importance of basic skin care and to describe current and novel topical therapies for AD.


Assuntos
Dermatite Atópica/tratamento farmacológico , Pele/efeitos dos fármacos , Animais , Humanos , Qualidade de Vida , Higiene da Pele/métodos
20.
Water Sci Technol ; 78(1-2): 49-56, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30101788

RESUMO

In two pilot-scale experiments, fingerlings and juvenile of tilapia were reared in high rate algal pond (HRAP) effluent. The combination of three different total ammonia nitrogen (TAN) surface loading rates (SLR1 = 0.6, SLR2 = 1.2; SLR3 = 2.4 kg TAN·ha-1·d-1) and two fish stocking densities (D1 = 4 and D2 = 8 fish per tank) was evaluated during two 12-week experiments. Fingerlings total weight gain varied from 4.9 to 18.9 g, with the highest value (equivalent to 0.225 g·d-1) being recorded in SLR2-D1 treatment; however, high mortality (up to 67%) was recorded, probably due to sensitivity to ammonia and wide daily temperature variations. At lower water temperatures, juvenile tilapia showed no mortality, but very low weight gain. The fish rearing tanks worked as wastewater polishing units, adding the following approximate average removal figures on top of those achieved at the HRAP: 63% of total Kjeldahl nitrogen; 54% of ammonia nitrogen; 42% of total phosphorus; 37% of chemical oxygen demand; 1.1 log units of Escherichia coli.


Assuntos
Amônia/metabolismo , Lagoas/química , Tilápia/fisiologia , Animais , Aquicultura , Microalgas , Nitrogênio/metabolismo , Tilápia/metabolismo
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA