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1.
Sensors (Basel) ; 22(6)2022 Mar 18.
Artigo em Inglês | MEDLINE | ID: mdl-35336530

RESUMO

In this paper, we aim to open up new perspectives in the field of autonomous aerial surveillance and target tracking systems, by exploring an alternative that, surprisingly, and to the best of the authors' knowledge, has not been addressed in that context by the research community thus far. It can be summarized by the following two questions. Under the scope of such applications, what are the implications and possibilities offered by mounting several steerable cameras onboard of each aerial agent? Second, how can optimization algorithms benefit from this new framework, in their attempt to provide more efficient and cost-effective solutions on these areas? The paper presents the idea as an additional degree of freedom to be exploited, which can enable more efficient alternatives in the deployment of such applications. As an initial approach, the problem of the optimal positioning with respect to a set of targets of one single agent, equipped with several onboard tracking cameras with different or variable focal lengths, is addressed. As a consequence of this allowed heterogeneity in focal lengths, the notion of distance needs to be adapted into a notion of optical range, as the agent can trade longer Euclidean distances for correspondingly longer focal lengths. Moreover, the proposed optimization indices try to balance, in an optimal way, the verticality of the viewpoints along with the optical range to the targets. Under these premises, several positioning strategies are proposed and comparatively evaluated.


Assuntos
Esportes , Dispositivos Aéreos não Tripulados , Algoritmos
2.
Nature ; 517(7535): 509-12, 2015 Jan 22.
Artigo em Inglês | MEDLINE | ID: mdl-25363770

RESUMO

Lantibiotics are a class of peptide antibiotics that contain one or more thioether bonds. The lantibiotic nisin is an antimicrobial peptide that is widely used as a food preservative to combat food-borne pathogens. Nisin contains dehydroalanine and dehydrobutyrine residues that are formed by the dehydration of Ser/Thr by the lantibiotic dehydratase NisB (ref. 2). Recent biochemical studies revealed that NisB glutamylates Ser/Thr side chains as part of the dehydration process. However, the molecular mechanism by which NisB uses glutamate to catalyse dehydration remains unresolved. Here we show that this process involves glutamyl-tRNA(Glu) to activate Ser/Thr residues. In addition, the 2.9-Å crystal structure of NisB in complex with its substrate peptide NisA reveals the presence of two separate domains that catalyse the Ser/Thr glutamylation and glutamate elimination steps. The co-crystal structure also provides insights into substrate recognition by lantibiotic dehydratases. Our findings demonstrate an unexpected role for aminoacyl-tRNA in the formation of dehydroamino acids in lantibiotics, and serve as a basis for the functional characterization of the many lantibiotic-like dehydratases involved in the biosynthesis of other classes of natural products.


Assuntos
Proteínas de Bactérias/química , Proteínas de Bactérias/metabolismo , Bacteriocinas/metabolismo , Hidroliases/química , Hidroliases/metabolismo , Lactococcus lactis/enzimologia , Proteínas de Membrana/química , Proteínas de Membrana/metabolismo , RNA de Transferência de Ácido Glutâmico/metabolismo , Proteínas de Bactérias/classificação , Bacteriocinas/biossíntese , Cristalografia por Raios X , Escherichia coli/genética , Ácido Glutâmico/metabolismo , Hidroliases/classificação , Lactococcus lactis/genética , Proteínas de Membrana/classificação , Modelos Moleculares , Nisina/biossíntese , Nisina/metabolismo , Filogenia , Estrutura Terciária de Proteína , RNA de Transferência de Ácido Glutâmico/genética , Serina/metabolismo , Treonina/metabolismo
3.
Molecules ; 26(19)2021 Sep 30.
Artigo em Inglês | MEDLINE | ID: mdl-34641471

RESUMO

Orange peel by-products generated in the food industry are an important source of value-added compounds that can be potentially reused. In the current research, the effect of oven-drying (50-70 °C) and freeze-drying on the bioactive compounds and antioxidant potential from Navelina, Salustriana, and Sanguina peel waste was investigated using pressurized extraction (ASE). Sixty volatile components were identified by ASE-GC-MS. The levels of terpene derivatives (sesquitenenes, alcohols, aldehydes, hydrocarbons, and esters) remained practically unaffected among fresh and freeze-dried orange peels, whereas drying at 70 °C caused significative decreases in Navelina, Salustriana, and Sanguina peels. Hesperidin and narirutin were the main flavonoids quantified by HPLC-MS. Freeze-dried Sanguina peels showed the highest levels of total-polyphenols (113.3 mg GAE·g-1), total flavonoids (39.0 mg QE·g-1), outstanding values of hesperedin (187.6 µg·g-1), phenol acids (16.54 mg·g-1 DW), and the greatest antioxidant values (DPPH•, FRAP, and ABTS•+ assays) in comparison with oven-dried samples and the other varieties. Nanotechnology approaches allowed the formulation of antioxidant-loaded nanoemulsions, stabilized with lecithin, starting from orange peel extracts. Those provided 70-80% of protection against oxidative UV-radiation, also decreasing the ROS levels into the Caco-2 cells. Overall, pressurized extracts from freeze-drying orange peel can be considered a good source of natural antioxidants that could be exploited in food applications for the development of new products of commercial interest.


Assuntos
Antioxidantes/isolamento & purificação , Citrus sinensis/química , Flavonoides/análise , Estresse Oxidativo/efeitos dos fármacos , Extratos Vegetais/farmacologia , Polifenóis/análise , Antioxidantes/farmacologia , Células CACO-2 , Sobrevivência Celular , Emulsões , Temperatura Alta , Humanos , Extratos Vegetais/isolamento & purificação , Pressão
4.
J Food Sci Technol ; 58(10): 4034-4044, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34471327

RESUMO

The objective of this study was to use accelerated-solvent-extraction to achieve antioxidant extracts from chia seeds oils, enriched in tocopherols and tocotrienols, namely tocochromanols. Nanotechnology applications have been also incorporated to develop an innovative formulation of chia seeds oil nanoemulsion that preserve its antioxidant potential after conditions of oxidative stress. Chia seeds oils proved to be a valuable source of tocochromanols, from 568.84 to 855.98 µg g-1, depending on the geographical provenance. Quantitative data obtained by LC-DAD-ESI-MS/MS showed outstanding levels of γ-Tocopherol, over 83%, followed far behind by Tocopherols-(α, ß, δ) and Tocotrienols-(α, ß, δ, γ)-tocotrienols. The characteristic tocochromanols fingerprint of chia seeds oils was positively correlated with the FRAP and DPPH antioxidant activity of the extracts (between 18.81 and 138.48 mg Trolox/g). Formulation of the Chia seeds oils as nanoemulsions did not compromised the antioxidant properties of fresh extracts. Interestingly, nanoemulsions retained about the 80% of the initial antioxidant capacity after UV-induced stress, where the non-emulsified oils displayed a remarkable reduction (50-60%) on its antioxidant capacity under the same conditions. These antioxidant chia seeds formulations can constitute a promising strategy to vectorizing vitamin E isomers, in order to be used for food fortification, natural additives and to increase the self-life of food products during packing.

5.
Microb Pathog ; 128: 112-118, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30583020

RESUMO

Sigma factor B (SigB) controls the expression of Staphylococcus aureus genes including virulence factors and plays a role in the bacterial secretion system through membrane vesicle production. Inhibition of SigB could attenuate SigB dependent virulence and secretion system. The objective of this study was to determine the effects of rhodomyrtone on SigB and virulence factors related to SigB. Minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) values of rhodomyrtone against 67 clinical methicillin-resistant S. aureus isolates were 0.25-8 µg/ml, which were similar to those of vancomycin. Using luciferase gene fused to SigB dependent promoters of asp23, five time reduction in SigB activity was observed when the bacteria were treated with rhodomyrtone for 3 h. Rhodomyrtone significantly reduced SigB activity in a concentration dependent manner in exponentially growing cells (P < 0.05). In addition, sigB mutant was more sensitive towards increasing concentrations of rhodomyrtone than the wild type and yabJ-spoVG mutant. Rhodomyrtone at 0.625 µg/ml reduced the growth of sigB mutant by approximately 99%, compared with the yabJ-spoVG mutant and the wild type. Membrane vesicles were significantly reduced in the bacterial cells when treated with 0.5 × MIC rhodomyrtone (P < 0.05). Decreased haemolytic activity was detected within rhodomyrtone-treated membrane vesicles. The results indicated that rhodomyrtone inhibited S. aureus SigB activity during exponentially growing phase and inhibited haemolytic activity within membrane vesicles.


Assuntos
Antibacterianos/farmacologia , Proteínas de Bactérias/efeitos dos fármacos , Proteínas de Bactérias/metabolismo , Membrana Celular/efeitos dos fármacos , Fator sigma/efeitos dos fármacos , Fator sigma/metabolismo , Staphylococcus aureus/efeitos dos fármacos , Xantonas/farmacologia , Proteínas de Bactérias/genética , Membrana Celular/metabolismo , Proteínas de Ligação a DNA/genética , Humanos , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Staphylococcus aureus Resistente à Meticilina/crescimento & desenvolvimento , Testes de Sensibilidade Microbiana , Mutação , Fator sigma/genética , Infecções Estafilocócicas , Staphylococcus aureus/crescimento & desenvolvimento , Vancomicina/farmacologia , Virulência/efeitos dos fármacos , Fatores de Virulência/metabolismo
6.
Mol Cell Proteomics ; 14(10): 2591-608, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26183717

RESUMO

Pneumonia is one of the most common and severe diseases associated with Streptococcus pneumoniae infections in children and adults. Etiological diagnosis of pneumococcal pneumonia in children is generally challenging because of limitations of diagnostic tests and interference with nasopharyngeal colonizing strains. Serological assays have recently gained interest to overcome some problems found with current diagnostic tests in pediatric pneumococcal pneumonia. To provide insight into this field, we have developed a protein array to screen the antibody response to many antigens simultaneously. Proteins were selected by experimental identification from a collection of 24 highly prevalent pediatric clinical isolates in Spain, using a proteomics approach consisting of "shaving" the cell surface with proteases and further LC/MS/MS analysis. Ninety-five proteins were recombinantly produced and printed on an array. We probed it with a collection of sera from children with pneumococcal pneumonia. From the set of the most seroprevalent antigens, we obtained a clear discriminant response for a group of three proteins (PblB, PulA, and PrtA) in children under 4 years old. We validated the results by ELISA and an immunostrip assay showed the translation to easy-to-use, affordable tests. Thus, the protein array here developed presents a tool for broad use in serodiagnostics.


Assuntos
Anticorpos Antibacterianos , Antígenos de Bactérias , Proteínas de Bactérias , Infecções Pneumocócicas , Análise Serial de Proteínas , Streptococcus pneumoniae/imunologia , Anticorpos Antibacterianos/sangue , Anticorpos Antibacterianos/imunologia , Antígenos de Bactérias/sangue , Antígenos de Bactérias/imunologia , Proteínas de Bactérias/sangue , Proteínas de Bactérias/imunologia , Criança , Pré-Escolar , Humanos , Testes Imunológicos , Lactente , Infecções Pneumocócicas/sangue , Infecções Pneumocócicas/diagnóstico , Infecções Pneumocócicas/imunologia , Proteômica , Reprodutibilidade dos Testes , Testes Sorológicos
7.
Proc Natl Acad Sci U S A ; 111(33): 12031-6, 2014 Aug 19.
Artigo em Inglês | MEDLINE | ID: mdl-25092299

RESUMO

Ribosomally synthesized and posttranslationally modified peptides (RiPPs) are a growing class of natural products that are found in all domains of life. These compounds possess vast structural diversity and have a wide range of biological activities, promising a fertile ground for exploring novel natural products. One challenging aspect of RiPP research is the difficulty of structure determination due to their architectural complexity. We here describe a method for automated structural characterization of RiPPs by tandem mass spectrometry. This method is based on the combined analysis of multiple mass spectra and evaluation of a collection of hypothetical structures predicted based on the biosynthetic gene cluster and molecular weight. We show that this method is effective in structural characterization of complex RiPPs, including lanthipeptides, glycopeptides, and azole-containing peptides. Using this method, we have determined the structure of a previously structurally uncharacterized lanthipeptide, prochlorosin 1.2, and investigated the order of the posttranslational modifications in three biosynthetic systems.


Assuntos
Produtos Biológicos/química , Peptídeos/química , Ribossomos/metabolismo , Espectrometria de Massas em Tandem/métodos , Sequência de Aminoácidos , Produtos Biológicos/metabolismo , Dados de Sequência Molecular , Estrutura Molecular , Peptídeos/metabolismo
8.
Proteomics ; 15(5-6): 1089-112, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-25487722

RESUMO

In this article, the topic of plant proteomics is reviewed based on related papers published in the journal Proteomics since publication of the first issue in 2001. In total, around 300 original papers and 41 reviews published in Proteomics between 2000 and 2014 have been surveyed. Our main objective for this review is to help bridge the gap between plant biologists and proteomics technologists, two often very separate groups. Over the past years a number of reviews on plant proteomics have been published . To avoid repetition we have focused on more recent literature published after 2010, and have chosen to rather make continuous reference to older publications. The use of the latest proteomics techniques and their integration with other approaches in the "systems biology" direction are discussed more in detail. Finally we comment on the recent history, state of the art, and future directions of plant proteomics, using publications in Proteomics to illustrate the progress in the field. The review is organized into two major blocks, the first devoted to provide an overview of experimental systems (plants, plant organs, biological processes) and the second one to the methodology.


Assuntos
Proteínas de Plantas , Estruturas Vegetais , Proteômica/métodos , Eletroforese em Gel Bidimensional , Espectrometria de Massas , Modelos Biológicos , Mapeamento de Peptídeos , Proteínas de Plantas/análise , Proteínas de Plantas/química , Proteínas de Plantas/isolamento & purificação , Estruturas Vegetais/química , Estruturas Vegetais/metabolismo
9.
Nanomedicine ; 11(2): 369-77, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25267700

RESUMO

Gemcitabine is currently the standard therapy for pancreatic cancer. However, growing concerns over gemcitabine resistance mean that new combinatory therapies are required to prevent loss of efficacy with prolonged treatment. Here, we suggest that this could be achieved through co-administration of RNA interference agents targeting the ubiquitin ligase ITCH. Stable anti-ITCH siRNA and shRNA dendriplexes with a desirable safety profile were prepared using generation 3 poly(propylenimine) dendrimers (DAB-Am16). The complexes were efficiently taken up by human pancreatic cancer cells and produced a 40-60% decrease in ITCH RNA and protein expression in vitro (si/shRNA) and in a xenograft model of pancreatic cancer (shRNA). When co-administered with gemcitabine (100 mg/kg/week) at a subtherapeutic dose, treatment with ITCH-shRNA (3x 50 mg/week) was able to fully suppress tumour growth for 17 days, suggesting that downregulation of ITCH mediated by DAB-Am16/shRNA sensitizes pancreatic cancer to gemcitabine in an efficient and specific manner. FROM THE CLINICAL EDITOR: Gemcitabine delivery to pancreatic cancer often results in the common problem of drug resistance. This team overcame the problem through co-administration of siRNA and shRNA dendriplexes targeting the ubiquitin ligase ITCH.


Assuntos
Proliferação de Células/efeitos dos fármacos , Desoxicitidina/análogos & derivados , Neoplasias Pancreáticas/tratamento farmacológico , Proteínas Repressoras/antagonistas & inibidores , Ubiquitina-Proteína Ligases/antagonistas & inibidores , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Desoxicitidina/administração & dosagem , Desoxicitidina/química , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Neoplasias Pancreáticas/genética , Neoplasias Pancreáticas/patologia , Polipropilenos/administração & dosagem , Polipropilenos/química , Interferência de RNA , Proteínas Repressoras/biossíntese , Ubiquitina-Proteína Ligases/biossíntese , Gencitabina
10.
Acta Cardiol ; 70(6): 728-34, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26717223

RESUMO

INTRODUCTION: Diabetic patients with an acute coronary syndrome undergoing percutaneous coronary intervention frequently exhibit high platelet reactivity while on clopidogrel. We hypothesized that in diabetic patients undergoing percutaneous coronary intervention, who exhibit high-platelet-reactivity after standard treatment with clopidogrel, a 60-mg prasugrel loading dose is superior to standard treatment with clopidogrel for optimal P2Y12 inhibition within the first 24-36 h post-angioplasty. METHODS: VERDI was a prospective, randomized, single-centre, single-blind, parallel-design study (NCT01684813). Consecutive diabetic patients with an non-ST-segment elevation acute coronary syndrome undergoing percutaneous coronary intervention and loaded with clopidogrel were considered for platelet reactivity assessment immediately before angioplasty with the VerifyNow assay measured in P2Y12 reaction units (PRU). Fifty of 63 screened patients (79.4%) had high platelet reactivity (PRU ≥ 208) and were randomized to receive a 60-mg prasugrel loading dose (n = 25) versus clopidogrel standard dose (n = 25). Platelet function was assessed again 24 hours post-angioplasty. RESULTS: Prasugrel achieved greater platelet inhibition than clopidogrel 24 hours post-angioplasty (median [interquartile range], 38 [9-72] vs 285 [240-337], respectively; P < 0.001). The non-high-platelet-reactivity rate (PRU < 208) at 24 h post-angioplasty (primary end point) was higher with prasugrel; 25 patients (100%) in the prasugrel group achieved optimal antiaggregation vs 4 patients (16%) in the clopidogrel group (P < 0.001). No significant acute bleeding was documented in either group. CONCLUSION: Among type 2 diabetic patients suffering an acute coronary syndrome with high-platelet-reactivity undergoing percutaneous coronary intervention, switching from clopidogrel to prasugrel was superior to standard treatment with clopidogrel for the achievement of optimal antiaggregation within the first 24 hours post-angioplasty.


Assuntos
Síndrome Coronariana Aguda/terapia , Plaquetas/efeitos dos fármacos , Diabetes Mellitus Tipo 2/complicações , Cloridrato de Prasugrel/farmacocinética , Ticlopidina/análogos & derivados , Síndrome Coronariana Aguda/sangue , Síndrome Coronariana Aguda/complicações , Idoso , Clopidogrel , Diabetes Mellitus Tipo 2/sangue , Diabetes Mellitus Tipo 2/tratamento farmacológico , Relação Dose-Resposta a Droga , Feminino , Seguimentos , Humanos , Masculino , Intervenção Coronária Percutânea , Inibidores da Agregação Plaquetária/administração & dosagem , Inibidores da Agregação Plaquetária/farmacocinética , Testes de Função Plaquetária , Cloridrato de Prasugrel/administração & dosagem , Estudos Prospectivos , Método Simples-Cego , Ticlopidina/administração & dosagem , Ticlopidina/farmacocinética , Resultado do Tratamento
11.
Langmuir ; 30(20): 5911-20, 2014 May 27.
Artigo em Inglês | MEDLINE | ID: mdl-24801765

RESUMO

The colloidal and chemical stability of nanoparticles of the nontoxic and biodegradable iron(III) trimesate MIL-100(Fe) nanocarrier have been evaluated in the presence of a series of simulated physiological fluids for intravenous and oral administration. MIL-100(Fe) nanoparticles exhibit an appropriate colloidal stability and biodegradability, mainly dependent on both the nature of their physicochemical surface and the media composition, being a priori compatible with their biomedical use.


Assuntos
Portadores de Fármacos/química , Ferro/química , Nanopartículas Metálicas/química , Ácidos Tricarboxílicos/química , Coloides , Estabilidade de Medicamentos
12.
Eur Heart J Case Rep ; 8(2): ytae045, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38332924

RESUMO

Background: Ventricular septal defect (VSD) is an unusual complication of transcatheter aortic valve implantation (TAVI). The risk factors are not well understood but may include oversizing, calcification amount and location, left-ventricular chamber morphology, and valve-in-valve (ViV) procedures. Percutaneous treatment is challenging but is usually the preferred option. Case summary: An 80-year-old woman with two previous surgical aortic valve replacements was admitted to our Cardiology Department for decompensated heart failure. New bioprosthesis degeneration (19 mm Mitroflow™, Sorin Group, Canada) was observed with severe intraprosthetic aortic regurgitation. After evaluation, the heart team chose to perform ViV TAVI. Because of the high risk of coronary obstruction, chimney stenting of both coronary arteries was performed. A 23 mm self-expandable Navitor™ valve (Abbott, IL, USA) was implanted, but the Mitroflow™ valve had to be cracked to minimize the persistent high gradient. During valve fracture, the non-compliant balloon broke and a small iatrogenic VSD appeared. However, the patient remained stable, so conservative management was selected. During follow-up, she developed severe haemolytic anaemia and heart failure; therefore, percutaneous closure of the iatrogenic VSD was performed twice, which was a difficult challenge. Discussion: A viable alternative to redo surgery is ViV TAVI. Risks include higher rates of prosthesis-patient mismatch and coronary obstruction. Occasionally, bioprosthetic valve fracture is required, particularly in small bioprostheses, to achieve low gradients. Anecdotally, fracture has led to annular rupture and VSD. Most VSDs are small and without clinical or haemodynamic repercussions; however, in symptomatic cases, percutaneous closure is a viable alternative to surgery.

13.
Artigo em Inglês | MEDLINE | ID: mdl-38856952

RESUMO

The success of colon-targeted oral hybrid systems relies in the proper control over the release of the entrapped nanostructures at the colon. This work describes the design of hybrid systems for their colonic enzyme-triggered release. The hybrid systems were constituted by nanoemulsions, with adequate characteristics for the treatment of ulcerative colitis, included in a pectin hydrogel-like matrix. For that purpose, pectins with similar degrees of methylation (< 50%) and increasing degree of amidation, i.e. 0, 13 and 20%, were selected. Hybrid systems were formulated by a novel aggregation induced gelation method, using Ca2+, Ba2+ or Zn2+ as aggregating agents, as well as by a polyelectrolyte condensation approach, obtaining structures in the micrometric range (< 10 µm). Despite the resistance of pectins to the upper gastrointestinal tract stimuli, the analysis of the behaviour of the different prototypes showed that the non-covalent crosslinks that allow the formation of the hybrid structure may play a relevant role on the performance of the formulation.Our results indicated that the partial disassembling of the hybrid system's microstructure due to the intestinal conditions may facilitate the stimuli-triggered release of the nanoemulsions at the colon. More interestingly, the particle tracking experiments showed that the condensation process that occurs during the formation of the system may affect to the enzymatic degradation of pectin. In this sense, the effect of the high degree of amidation of pectin may be more prevalent as structural feature rather than as a promoter of the enzyme-triggered release.

14.
Biomolecules ; 14(7)2024 Jul 09.
Artigo em Inglês | MEDLINE | ID: mdl-39062530

RESUMO

Dairy products are an important source of protein and other nutrients in the Mediterranean diet. In these countries, the most common sources of milk for producing dairy products are cow, goat, sheep, and buffalo. Andalusia is traditionally the largest producer of goat milk in Spain. Kefir is a fermented product made from bacteria and yeasts and has health benefits beyond its nutritional properties. There is a lack of knowledge about the molecular mechanisms and metabolites that bring about these benefits. In this work, the combination of analytical techniques (GC-FID, UHPLC-MS-QToF, GC-QqQ-MS, and GC-ToF-MS) resulted in the detection of 105 metabolites in kefir produced with goat milk from two different thermal treatments (raw and pasteurized) fermented at four time points (12, 24, 36, and 48 h, using 0 h as the control). Of these, 27 metabolites differed between kefir produced with raw and pasteurized milk. These changes could possibly be caused by the effect of pasteurization on the microbial population in the starting milk. Some interesting molecules were identified, such as shikimic acid, dehydroabietic acid, GABA, and tyramine, which could be related to antibacterial properties, strengthening of the immune system, and arterial pressure. Moreover, a viability assay of the NIRS technique was performed to evaluate its use in monitoring the fermentation and classification of samples, which resulted in a 90% accuracy in comparison to correctly classified samples according to their fermentation time. This study represents the most comprehensive metabolomic analysis of goat milk kefir so far, revealing the intricate changes in metabolites during fermentation and the impact of milk treatment.


Assuntos
Fermentação , Cabras , Kefir , Metabolômica , Leite , Animais , Kefir/microbiologia , Metabolômica/métodos , Leite/metabolismo , Leite/química , Leite/microbiologia , Temperatura Alta , Cromatografia Líquida de Alta Pressão
15.
Vaccines (Basel) ; 12(4)2024 Apr 09.
Artigo em Inglês | MEDLINE | ID: mdl-38675780

RESUMO

Burkholderia cepacia complex infections remain life-threatening to cystic fibrosis patients, and due to the limited eradication efficiency of current treatments, novel antimicrobial therapies are urgently needed. Surface proteins are among the best targets to develop new therapeutic strategies since they are exposed to the host's immune system. A surface-shaving approach was performed using Burkholderia cenocepacia J2315 to quantitatively compare the relative abundance of surface-exposed proteins (SEPs) expressed by the bacterium when grown under aerobic and microaerophilic conditions. After trypsin incubation of live bacteria and identification of resulting peptides by liquid chromatography coupled with mass spectrometry, a total of 461 proteins with ≥2 unique peptides were identified. Bioinformatics analyses revealed a total of 53 proteins predicted as localized at the outer membrane (OM) or extracellularly (E). Additionally, 37 proteins were predicted as moonlight proteins with OM or E secondary localization. B-cell linear epitope bioinformatics analysis of the proteins predicted to be OM and E-localized revealed 71 SEP moieties with predicted immunogenic epitopes. The protegenicity higher scores of proteins BCAM2761, BCAS0104, BCAL0151, and BCAL0849 point out these proteins as the best antigens for vaccine development. Additionally, 10 of the OM proteins also presented a high probability of playing important roles in adhesion to host cells, making them potential targets for passive immunotherapeutic approaches. The immunoreactivity of three of the OM proteins identified was experimentally demonstrated using serum samples from cystic fibrosis patients, validating our strategy for identifying immunoreactive moieties from surface-exposed proteins of potential interest for future immunotherapies development.

16.
Int J Clin Health Psychol ; 24(2): 100465, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38737628

RESUMO

Background: Although significant progress has been made in the rights of the LGBTQ+ community, even today this population still faces stigma and discrimination that impacts their mental health. In the case of men who have sex with men, it has been demonstrated that the use of drugs in a sexual context (chemsex) is one of the coping mechanisms and means of escape to deal with these situations. Method: We assessed 284 native Spanish speakers' participants, 45,4 % were not engaged in sexualised drug use (n = 129) while 54,6 % were chemsex users (n = 155) using 18,7 % of them the injected via. The participants completed six questionnaires about life and sexual satisfaction, depression, anxiety, internalised homophobia and personality. Bivariate and multivariable logistic regression were performed to assess the associations between sexual behaviour-related and psychological variables. Kruskal-Wallis H test was used to analysed the impact on mental health of the administration via. Results: Aged, unprotected sexual relationships, positive serostatus, homonegativity and conscientiousness predicted the chemsex engagement. Furthermore, we found differences regarding the administration via. Conclusions: We conclude that mental health significantly correlates with the practice of chemsex, highlighting the importance of integrating mental health considerations into the prevention of risky sexual behaviors.

17.
Foods ; 12(15)2023 Aug 07.
Artigo em Inglês | MEDLINE | ID: mdl-37569243

RESUMO

Sheep farming is an important socioeconomic activity in most Mediterranean countries, particularly Spain, where it contributes added value to rural areas. Sheep milk is used in Spain mainly for making cheese, but it can be used also for making other dairy products, such as the lactic-alcoholic fermentation product known as kefir. Dairy products have health benefits because, among other reasons, they contain molecules with biological activity. In this work, we performed a proteomics strategy to identify the peptidome, i.e., the set of peptides contained in sheep milk kefir fermented for four different periods of time, aiming to understand changes in the pattern of digestion of milk proteins, as well as to identify potential bioactive peptides. In total, we identified 1942 peptides coming from 11 different proteins, and found that the unique peptides differed qualitatively among samples and their numbers increased along the fermentation time. These changes were supported by the increase in ethanol, lactic acid, and D-galactose concentrations, as well as proteolytic activity, as the fermentation progressed. By searching in databases, we found that 78 of the identified peptides, all belonging to caseins, had potential biological activity. Of these, 62 were not previously found in any milk kefir from other animal species. This is the first peptidomic study of sheep milk kefir comprising time-course comparison.

18.
Nat Biotechnol ; 41(5): 686-697, 2023 05.
Artigo em Inglês | MEDLINE | ID: mdl-36624149

RESUMO

Cytosine base editors (CBEs) enable programmable genomic C·G-to-T·A transition mutations and typically comprise a modified CRISPR-Cas enzyme, a naturally occurring cytidine deaminase, and an inhibitor of uracil repair. Previous studies have shown that CBEs utilizing naturally occurring cytidine deaminases may cause unguided, genome-wide cytosine deamination. While improved CBEs that decrease stochastic genome-wide off-targets have subsequently been reported, these editors can suffer from suboptimal on-target performance. Here, we report the generation and characterization of CBEs that use engineered variants of TadA (CBE-T) that enable high on-target C·G to T·A across a sequence-diverse set of genomic loci, demonstrate robust activity in primary cells and cause no detectable elevation in genome-wide mutation. Additionally, we report cytosine and adenine base editors (CABEs) catalyzing both A-to-I and C-to-U editing (CABE-Ts). Together with ABEs, CBE-Ts and CABE-Ts enable the programmable installation of all transition mutations using laboratory-evolved TadA variants with improved properties relative to previously reported CBEs.


Assuntos
Citosina , Edição de Genes , Mutação/genética , Citidina Desaminase/genética , Genoma , Sistemas CRISPR-Cas/genética
19.
Urol Res ; 40(5): 587-92, 2012 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-22388467

RESUMO

Our objective was to analyze the advantages of the percutaneous nephrolithotomy in oblique supine decubitus compared to the prone and dorsal supine position. In 87 patients diagnosed with urolithiasis (495.5-530.8 mm(2)), percutaneous nephrolithotomy (PNL) was performed from 2000 to 2011. The patients were divided into three groups: Group A, 32 patients, PNL in the prone decubitus position; Group B, 24 patients, PNL in the dorsal supine position; Group C, 31 patients, PNL in the oblique supine position. We analyzed intraoperative parameters, complications, and results among the three groups. The three procedures were performed with a single access, 24-30 Ch. No statistically significant differences were found among the three groups regarding the patients' characteristics, or the morphology or size of the kidney stone treated. The operation time was shorter in the cases of PNL in dorsal supine and oblique supine compared to the prone position. The complication rate was very similar in the three groups. The main advantage of the PNL in oblique supine compared to the dorsal supine was that the puncture could in all cases be directed by ultrasonography, with greater precision, more safety, and more control of the percutaneous renal access. The oblique supine decubitus position is a safe position for the percutaneous treatment of urolithiasis and it becomes easier when the puncture is guided by ultrasound.


Assuntos
Nefrostomia Percutânea/métodos , Posicionamento do Paciente , Urolitíase/cirurgia , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Decúbito Ventral , Estudos Retrospectivos , Decúbito Dorsal
20.
Med Clin (Barc) ; 159(3): 109-115, 2022 08 12.
Artigo em Inglês, Espanhol | MEDLINE | ID: mdl-34972550

RESUMO

INTRODUCTION AND OBJECTIVE: Excess weight can cause structural and functional cardiac disorders. The presence of left ventricular hypertrophy in the obese patient is an independent predictor of cardiovascular morbidity and mortality. The major aim of the present study is to know the prevalence of cardiac morphofunctional disorders in obese patients, before and after weight loss due to bariatric surgery (BS). PATIENTS AND METHODS: Prospective cohort study of 75 patients with obesity without known heart disease referred to gastric bypass. Anthropometric, analytical and echocardiographic parameters were measured before and after 6 and 12 months after BS. RESULTS: The study included 75 patients (66.6% women, mean age 39.3 [9.7] years and BMI 47.8 [7.1] kg/m2). At 6 and 12 months after BS there was a significant reduction in body weight and an improvement in metabolic, inflammatory and prothrombotic parameters and in cardiovascular risk factors associated with obesity (hypertension, type 2 diabetes, dyslipidemia and obstructive sleep apnea-hypopnea syndrome). Before surgery, cardiac remodeling was present in 62.7%, most frequently in the form of concentric remodeling (38.7%). Diastolic dysfunction occurred in 50.7% of the patients. One year after surgery, the ventricular pattern was normal in 92% of cases and the diastolic function improved significantly. CONCLUSIONS: Our results support the negative effect of obesity on cardiac geometry and function and the potential reversibility of these cardiac alterations after marked weight loss due to BS.


Assuntos
Cirurgia Bariátrica , Diabetes Mellitus Tipo 2 , Cardiopatias , Obesidade Mórbida , Apneia Obstrutiva do Sono , Adulto , Cirurgia Bariátrica/efeitos adversos , Diabetes Mellitus Tipo 2/complicações , Feminino , Cardiopatias/complicações , Humanos , Masculino , Obesidade/complicações , Obesidade/cirurgia , Obesidade Mórbida/complicações , Estudos Prospectivos , Apneia Obstrutiva do Sono/complicações , Redução de Peso
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