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PURPOSE: To assess the potential adjunctive role of a 3D electromagnetic (EM) navigational system for use in above-knee vessels afflicted with peripheral artery disease (PAD). Peripheral artery disease can be challenging to operators encountering significant vessel tortuosity, calcium, and stenoses, which may require prolonged procedure times and excessive use of nephrotoxic iodinated contrast when performed with conventional fluoroscopy. MATERIALS AND METHODS: Following appropriate ethical oversight, five 3D-printed bench phantoms modeling tortuous calcified PAD were created based on source CTA (computed tomography angiography) data sets from real patients. Investigational software was developed based on a commercially available aortic EM navigation platform (Intraoperative Positioning System [IOPS]; Centerline Biomedical, Inc., Cleveland, Ohio), with patient-specific structural maps of vessel lumens and calcification. Using a sensorized prototype 6 French (Fr) catheter and 0.035" guidewire, 15 interventionalists traversed each phantom using the EM platform as well as 2D simulated fluoroscopy-like image guidance and the times were recorded. Participants completed a 10-item standard system usability scale (SUS) questionnaire (score 1-5, 5=strongly agree) evaluating system usability and user satisfaction. Navigation times and SUS scores were compared with a 1-tailed statistical t test. RESULTS: Participants demonstrated a statistically significant reduction in navigation times using EM guidance, performing 0.7 minutes (42 seconds) faster on average (P < .001), corresponding to a 25% average relative reduction. Participants reported sufficiently high levels of usability satisfaction, with a mean SUS score of 4.29 (P < .001), exceeding the acceptance criterion (score ≥3.5). CONCLUSION: This preclinical phantom study highlights the future potential of Centerline Biomedical's EM navigation technology as a possible adjunct to fluoroscopy for highly precise visualization and navigation of PAD-afflicted vasculature. CLINICAL IMPACT: This preclinical proof-of-concept study highlights the feasibility of EM navigation not only for branch vessel cannulation, but also for inline navigation of peripheral vessels afflicted with calcified plaques via benchtop iliofemoral phantom simulations. The navigation platform studied addresses the need for improvements in EM technology through modelling algorithms that facilitate 3D visualization of calcified plaque in any projection in real time, in addition to sensorization of both catheter and guidewire in a compact 6Fr system.
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Nitrite is a ubiquitous pollutant in modern society. Developing new strategies for its determination is very important, and electroanalytical methods present outstanding performance on this task. However, the use of bare electrodes is not recommended because of their predisposition to poisoning and passivation. We herein report a procedure to overcome these limitations on carbon fiber microelectrodes through pulsed amperometry. A three-pulse amperometry approach was used to reduce the current decay from 47% (after 20 min under constant potential) to virtually 0%. Repeatability and reproducibility were found to have an RSD lower than 0.5% and 7%, respectively. Tap water and synthetic inorganic saliva samples were fortified with nitrite, and the results obtained with the proposed sensor were in good agreement with the amount added.
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Incrustaciones Biológicas , Nitritos , Microelectrodos , Fibra de Carbono , Nitritos/análisis , Reproducibilidad de los Resultados , Incrustaciones Biológicas/prevención & control , Electrodos , Carbono , Técnicas Electroquímicas/métodosRESUMEN
PURPOSE: The aim of the study was to evaluate the reliability and validity of the Visual Prostate Symptom Score (VPSS), its correlation with the International Prostate Symptom Score (IPSS), and the time required and need for help when completing the instrument. We also aimed to identify variables associated with difficulty completing these instruments. DESIGN: Longitudinal, prospective correlational study. SUBJECTS AND SETTING: The target population was Brazilian men from different cities within the state of São Paulo with lower urinary tract symptoms followed in urological consultation at a university hospital during October 2017. The sample comprised 59 patients with a mean age of 66.53 years. METHODS: Reliability of the VPSS was evaluated using a test-retest approach and its convergent validity with the IPSS. Cronbach α coefficient was also calculated. Both questionnaires were self-administered and, in case of difficulty of comprehension, assistance was provided. RESULTS: Mean completion time of the IPSS was 6.56 minutes and for VPSS was 5.02 minutes. The variables skin color and educational level were associated with the difficulty in completing the IPSS and for comprehending meaning of the VPSS pictograms. Internal consistency evaluated using Cronbach α coefficient was 0.74 for IPSS and 0.15 for VPSS, respectively. Test-retest reliability testing revealed that both instruments had a high intraclass correlation index (>0.75). There was a significant correlation between the health-related quality of life (QoL) scores of the 2 instruments (0.71, P = .0001) and between the total score of each instrument with its corresponding QoL score. CONCLUSION: Although time for response of the VPSS was shorter and it demonstrated good test-retest reliability, it more frequently required help to answer. The VPSS showed low internal consistency and low correlation with the IPSS (except for the QoL item).
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Próstata , Calidad de Vida , Masculino , Humanos , Anciano , Estudios Prospectivos , Reproducibilidad de los Resultados , Brasil , Encuestas y CuestionariosRESUMEN
Coenzyme F420 is involved in bioprocesses such as biosynthesis of antibiotics by streptomycetes, prodrug activation in Mycobacterium tuberculosis, and methanogenesis in archaea. F420-dependent enzymes also attract interest as biocatalysts in organic chemistry. However, as only low F420 levels are produced in microorganisms, F420 availability is a serious bottleneck for research and application. Recent advances in our understanding of the F420 biosynthesis enabled heterologous overproduction of F420 in Escherichia coli, but the yields remained moderate. To address this issue, we rationally designed a synthetic operon for F420 biosynthesis in E. coli. However, it still led to the production of low amounts of F420 and undesired side-products. In order to strongly improve yield and purity, a screening approach was chosen to interrogate the gene expression-space of a combinatorial library based on diversified promotors and ribosome binding sites. The whole pathway was encoded by a two-operon construct. The first module ("core") addressed parts of the riboflavin biosynthesis pathway and FO synthase for the conversion of GTP to the stable F420 intermediate FO. The enzymes of the second module ("decoration") were chosen to turn FO into F420. The final construct included variations of T7 promoter strengths and ribosome binding site activity to vary the expression ratio for the eight genes involved in the pathway. Fluorescence-activated cell sorting was used to isolate clones of this library displaying strong F420-derived fluorescence. This approach yielded the highest titer of coenzyme F420 produced in the widely used organism E. coli so far. Production in standard LB medium offers a highly effective and simple production process that will facilitate basic research into unexplored F420-dependent bioprocesses as well as applications of F420-dependent enzymes in biocatalysis.
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Escherichia coli , Riboflavina , Escherichia coli/genética , Escherichia coli/metabolismo , Fluorescencia , Expresión Génica , Riboflavina/análogos & derivados , Riboflavina/genéticaRESUMEN
Coenzyme F420 is a redox cofactor involved in hydride transfer reactions in archaea and bacteria. Since F420-dependent enzymes are attracting increasing interest as tools in biocatalysis, F420 biosynthesis is being revisited. While it was commonly accepted for a long time that the 2-phospho-l-lactate (2-PL) moiety of F420 is formed from free 2-PL, it was recently shown that phosphoenolpyruvate is incorporated in Actinobacteria and that the C-terminal domain of the FbiB protein, a member of the nitroreductase (NTR) superfamily, converts dehydro-F420 into saturated F420 Outside the Actinobacteria, however, the situation is still unclear because FbiB is missing in these organisms and enzymes of the NTR family are highly diversified. Here, we show by heterologous expression and in vitro assays that stand-alone NTR enzymes from Thermomicrobia exhibit dehydro-F420 reductase activity. Metabolome analysis and proteomics studies confirmed the proposed biosynthetic pathway in Thermomicrobium roseum These results clarify the biosynthetic route of coenzyme F420 in a class of Gram-negative bacteria, redefine functional subgroups of the NTR superfamily, and offer an alternative for large-scale production of F420 in Escherichia coli in the future.IMPORTANCE Coenzyme F420 is a redox cofactor of Archaea and Actinobacteria, as well as some Gram-negative bacteria. Its involvement in processes such as the biosynthesis of antibiotics, the degradation of xenobiotics, and asymmetric enzymatic reductions renders F420 of great relevance for biotechnology. Recently, a new biosynthetic step during the formation of F420 in Actinobacteria was discovered, involving an enzyme domain belonging to the versatile nitroreductase (NTR) superfamily, while this process remained blurred in Gram-negative bacteria. Here, we show that a similar biosynthetic route exists in Thermomicrobia, although key biosynthetic enzymes show different domain architectures and are only distantly related. Our results shed light on the biosynthesis of F420 in Gram-negative bacteria and refine the knowledge about sequence-function relationships within the NTR superfamily of enzymes. Appreciably, these results offer an alternative route to produce F420 in Gram-negative model organisms and unveil yet another biochemical facet of this pathway to be explored by synthetic microbiologists.
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Chloroflexi/metabolismo , Nitrorreductasas/metabolismo , Riboflavina/análogos & derivados , Vías Biosintéticas , Chloroflexi/enzimología , Oxidación-Reducción , Riboflavina/biosíntesisRESUMEN
Upon injury, psychotropic psilocybin-producing mushrooms instantly develop an intense blue color, the chemical basis and mode of formation of which has remained elusive. We report two enzymes from Psilocybe cubensis that carry out a two-step cascade to prepare psilocybin for oxidative oligomerization that leads to blue products. The phosphatase PsiP removes the 4-O-phosphate group to yield psilocin, while PsiL oxidizes its 4-hydroxy group. The PsiL reaction was monitored by inâ situ 13 Câ NMR spectroscopy, which indicated that oxidative coupling of psilocyl residues occurs primarily via C-5. MS and IR spectroscopy indicated the formation of a heterogeneous mixture of preferentially psilocyl 3- to 13-mers and suggest multiple oligomerization routes, depending on oxidative power and substrate concentration. The results also imply that phosphate ester of psilocybin serves a reversible protective function.
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Agaricales/química , Productos Biológicos/química , Alucinógenos/efectos adversos , Psilocybe/enzimologíaRESUMEN
Among the invaluable benefits of basidiomycete genomics is the dramatically enhanced insight into the potential capacity to biosynthesize natural products. This study focuses on adenylate-forming reductases, which is a group of natural product biosynthesis enzymes that resembles non-ribosomal peptide synthetases, yet serves to modify one substrate, rather than to condense two or more building blocks. Phylogenetically, these reductases fall in four classes. The phylogeny of Heterobasidion annosum (Russulales) and Serpula lacrymans (Boletales) adenylate-forming reductases was investigated. We identified a previously unrecognized phylogenetic branch within class III adenylate-forming reductases. Three representatives were heterologously produced and their substrate preferences determined in vitro: NPS9 and NPS11 of S. lacrymans preferred l-threonine and benzoic acid, respectively, while NPS10 of H. annosum accepted phenylpyruvic acid best. We also investigated two class IV adenylate-forming reductases of Coprinopsis cinerea, which each were active with l-alanine, l-valine, and l-serine as substrates. Our results show that adenylate-forming reductases are functionally more diverse than previously recognized. As none of the natural products known from the species investigated in this study includes the identified substrates of their respective reductases, our findings may help further explore the diversity of these basidiomycete secondary metabolomes.
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Agaricales/enzimología , Variación Genética , Genoma Fúngico , Oxidorreductasas/clasificación , Oxidorreductasas/genética , Filogenia , Agaricales/genética , Oxidorreductasas/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/aislamiento & purificación , Proteínas Recombinantes/metabolismo , Especificidad por SustratoRESUMEN
Cyclic peptides containing non-proteinogenic amino acids often exhibit a broad bioactivity spectrum and many have entered clinical trials with good prospects for drug development. We recently reported the discovery of six cyclic tetrapeptides, pseudoxylallemycins A-F (1-6), from a termite-associated Pseudoxylaria sp. X802. These compounds contain a rare O-homoallenyl-l-tyrosine moiety and show promising antimicrobial activity against the Gram-negative pathogenic bacterium Pseudomonas aeruginosa. To perform more detailed structure-activity studies, we pursued a precursor-directed diversification strategy. Herein, we report the purification, identification, and testing of 21 new pseudoxylallemycin derivatives.
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Antibacterianos/química , Antibacterianos/farmacología , Ascomicetos/química , Péptidos Cíclicos/química , Péptidos Cíclicos/farmacología , Pseudomonas aeruginosa/efectos de los fármacos , Animales , Isópteros/microbiología , Pruebas de Sensibilidad Microbiana , Infecciones por Pseudomonas/tratamiento farmacológico , Infecciones por Pseudomonas/microbiología , Relación Estructura-ActividadAsunto(s)
Fístula Arteriovenosa , Endofuga , Fístula Arteriovenosa/diagnóstico por imagen , Fístula Arteriovenosa/etiología , Fístula Arteriovenosa/cirugía , Endofuga/diagnóstico por imagen , Endofuga/etiología , Endofuga/cirugía , Humanos , Arteria Ilíaca/diagnóstico por imagen , Arteria Ilíaca/cirugía , Vena Ilíaca/diagnóstico por imagen , Vena Ilíaca/cirugíaRESUMEN
Auriculamide is the first natural product known from the predatory bacterium Herpetosiphon aurantiacus. It is composed of three unusual building blocks, including the non-proteinogenic amino acid 3-chloro-L-tyrosine, the α-hydroxy acid L-isoleucic acid, and a methylmalonyl-CoA-derived ethane unit. A candidate genetic locus for auriculamide biosynthesis was identified and encodes four enzymes. Among them, the non-canonical 199 kDa four-domain nonribosomal peptide synthetase, AulA, is extraordinary in that it features two consecutive adenylation domains. Here, we describe the functional characterization of the recombinantly produced AulA. The observed activation of 3-methyl-2-oxovaleric acid by the enzyme supports the hypothesis that it participates in the biosynthesis of auriculamide. An artificially truncated version of AulA that lacks the first adenylation domain activated this substrate like the full-length enzyme which shows that the first adenylation domain is dispensable. Additionally, we provide evidence that the enzyme tolerates structural variation of the substrate. α-Carbon substituents significantly affected the substrate turnover. While all tested aliphatic α-keto acids were accepted by the enzyme and minor differences in chain size and branches did not interfere with the enzymatic activity, molecules with methylene α-carbons led to low turnover. Such enzymatic plasticity is an important attribute to help in the perpetual search for novel molecules and to access a greater structural diversity by mutasynthesis.
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With the increasing demand for lighter, more environmentally friendly, and affordable solutions in the mobility sector, designers and engineers are actively promoting the use of innovative integral dissimilar structures. In this field, friction stir-based technologies offer unique advantages compared with conventional joining technologies, such as mechanical fastening and adhesive bonding, which recently demonstrated promising results. In this study, an aluminum alloy and a glass fiber-reinforced polymer were friction stir joined in an overlap configuration. To assess the main effects, interactions, and influence of processing parameters on the mechanical strength and processing temperature of the fabricated joints, a full factorial design study with three factors and two levels was carried out. The design of experiments resulted in statistical models with excellent fit to the experimental data, enabling a thorough understanding of the influence of rotational speed, travel speed, and tool tilt angle on dissimilar metal-to-polymer friction stir composite joints. The mechanical strength of the composite joints ranged from 1708.1 ± 45.5 N to 3414.2 ± 317.1, while the processing temperature was between 203.6 ± 10.7 °C and 251.5 ± 9.7.
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Ultraviolet irradiation C (UVC) has emerged as an effective strategy for microbial control in indoor public spaces. UVC is commonly applied for air, surface, and water disinfection. Unlike common 254 nm UVC, far-UVC at 222 nm is considered non-harmful to human health, being safe for occupied spaces, and still effective for disinfection purposes. Therefore, and allied to the urgency to mitigate the current pandemic of SARS-CoV-2, an increase in UVC-based technology devices appeared in the market with levels of pathogens reduction higher than 99.9 %. This environmentally friendly technology has the potential to overcome many of the limitations of traditional chemical-based disinfection approaches. The novel UVC-based devices were thought to be used in public indoor spaces such as hospitals, schools, and public transport to minimize the risk of pathogens contamination and propagation, saving costs by reducing manual cleaning and equipment maintenance provided by manpower. However, a lack of information about UVC-based parameters and protocols for disinfection, and controversies regarding health and environmental risks still exist. In this review, fundamentals on UVC disinfection are presented. Furthermore, a deep analysis of UVC-based technologies available in the market for the disinfection of public spaces is addressed, as well as their advantages and limitations. This comprehensive analysis provides valuable inputs and strategies for the development of effective, reliable, and safe UVC disinfection systems.
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COVID-19 , Desinfección , Humanos , Desinfección/métodos , SARS-CoV-2 , COVID-19/prevención & control , Rayos UltravioletaRESUMEN
In this work we present a methodology for the non-destructive elemental determination of formalin-fixed paraffin-embedded (FFPE) human tissue samples based on the Fundamental Parameters method for the quantification of micro Energy Dispersive X Ray Fluorescence (micro-EDXRF) area scans. This methodology intended to overcome two major constraints in the analysis of paraffin embedded tissue samples - retrieval of optimal region of analysis of the tissue within the paraffin block and the determination of the dark matrix composition of the biopsied sample. This way, an image treatment algorithm, based on R® tool to select the regions of the micro-EDXRF area scans was developed. Also, different dark matrix compositions were evaluated using varying combinations of H, C, N and O until the most accurate matrix was found: 8% H, 15% C, 1% N and 60% O for breast FFPE samples and 8% H, 23% C, 2% N and 55% O for colon. The developed methodology was applied to paired normal-tumour samples of breast and colon biopsied tissues in order to gauge potential elemental biomarkers for carcinogenesis in these tissues. The obtained results showed distinctive biomarkers for breast and for colon: there was a significant increase of P, S, K and Fe in both tissues, while a significant increase of Ca an Zn concentrations was also determined for breast tumour samples.
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Neoplasias del Colon , Formaldehído , Humanos , Fijación del Tejido/métodos , Rayos X , Adhesión en Parafina , BiomarcadoresRESUMEN
Coenzyme F420 is a microbial redox cofactor that mediates diverse physiological functions and is increasingly used for biocatalytic applications. Recently, diversified biosynthetic routes to F420 and the discovery of a derivative, 3PG-F420, were reported. 3PG-F420 is formed via activation of 3-phospho-d-glycerate (3-PG) by CofC, but the structural basis of substrate binding, its evolution, as well as the role of CofD in substrate selection remained elusive. Here, we present a crystal structure of the 3-PG-activating CofC from Mycetohabitans sp. B3 and define amino acids governing substrate specificity. Site-directed mutagenesis enabled bidirectional switching of specificity and thereby revealed the short evolutionary trajectory to 3PG-F420 formation. Furthermore, CofC stabilized its product, thus confirming the structure of the unstable molecule and revealing its binding mode. The CofD enzyme was shown to significantly contribute to the selection of related intermediates to control the specificity of the combined biosynthetic CofC/D step. These results imply the need to change the design of combined CofC/D activity assays. Taken together, this work presents novel mechanistic and structural insights into 3PG-F420 biosynthesis and evolution and opens perspectives for the discovery and enhanced biotechnological production of coenzyme F420 derivatives in the future. IMPORTANCE The microbial cofactor F420 is crucial for processes like methanogenesis, antibiotics biosynthesis, drug resistance, and biocatalysis. Recently, a novel derivative of F420 (3PG-F420) was discovered, enabling the production and use of F420 in heterologous hosts. By analyzing the crystal structure of a CofC homolog whose substrate choice leads to formation of 3PG-F420, we defined amino acid residues governing the special substrate selectivity. A diagnostic residue enabled reprogramming of the substrate specificity, thus mimicking the evolution of the novel cofactor derivative. Furthermore, a labile reaction product of CofC was revealed that has not been directly detected so far. CofD was shown to provide another layer of specificity of the combined CofC/D reaction, thus controlling the initial substrate choice of CofC. The latter finding resolves a current debate in the literature about the starting point of F420 biosynthesis in various organisms.
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Riboflavina , Riboflavina/metabolismo , Oxidación-Reducción , BiocatálisisRESUMEN
The urgent need to treat type 2 diabetes mellitus (T2DM), which is currently reaching epidemic proportions, has been a major focus of healthcare systems and policy makers worldwide. Pharmacological treatment and lifestyle interventions together with the control of cardiovascular risk factors are the main strategies to prevent or delay the onset of T2DM. The present review discusses the state of the art knowledge of effective therapeutic approaches (metformin, thiazolidinediones, nateglinides, α-glucosidase inhibitors, incretin-based and angiotensin-based therapies, weight reducers, statins, fibric acid derivatives), including surgery, and identifies the major lifestyle changes for specific target groups.
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Diabetes Mellitus Tipo 2/prevención & control , Humanos , Hipoglucemiantes/uso terapéuticoRESUMEN
Morphotype switches frequently occur in Actinobacteria and are often associated with disparate natural product production. Here, we report on differences in the secondary metabolomes of two morphotypes of a Streptomyces species, including the discovery of a novel antimicrobial glycosylated macrolide, which we named termidomycin A. While exhibiting an unusual 46-member polyene backbone, termidomycin A (1) shares structural features with the clinically important antifungal agents amphotericin B and nystatin A1. Genomic analyses revealed a biosynthetic gene cluster encoding for a putative giant type I polyketide synthase (PKS), whose domain structure allowed us to propose the relative configuration of the 46-member macrolide. The architecture of the biosynthetic gene cluster was different in both morphotypes, thus leading to diversification of the product spectrum. Given the high frequency of genomic rearrangements in Streptomycetes, the metabolic analysis of distinct morphotypes as exemplified in this study is a promising approach for the discovery of bioactive natural products and pathways of diversification.
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Antibacterianos/farmacología , Antifúngicos/farmacología , Macrólidos/farmacología , Streptomyces/química , Antibacterianos/química , Antibacterianos/aislamiento & purificación , Antifúngicos/química , Antifúngicos/aislamiento & purificación , Bacterias/efectos de los fármacos , Hongos/efectos de los fármacos , Genómica , Macrólidos/química , Macrólidos/aislamiento & purificación , Metabolómica , Pruebas de Sensibilidad Microbiana , Familia de Multigenes , Sintasas Poliquetidas/genética , Sintasas Poliquetidas/metabolismo , Streptomyces/genética , Streptomyces/metabolismoRESUMEN
The zebrafish (Danio rerio) is an emerging model system in several research areas worldwide, especially in the Global South. In this context, the present study revised the historical use and trends of zebrafish as experimental models in Brazil. The data concerning the bibliometric parameters, research areas, geographic distribution, experimental design, zebrafish strain, and reporter lines, as well as recent advances were revised. In addition, the comparative trends of Brazilian and global research were discussed. Revised data showed the rapid growth of Brazilian scientific production using zebrafish as a model, especially in three main research areas (Neuroscience &and Behavior, Pharmacology and Toxicology, and Environment/Ecology). Studies were conducted in 19 Brazilian states (70.37%), confirming the wide geographic distribution and importance of zebrafish research. Results indicated that research related to toxicological approaches are widespread in Global South countries such as Brazil. Studies were performed mainly using in vivo tests (89.58%) with adult fish (59.75%) and embryos (30.67%). Moreover, significant research gaps and recommendations for future research are presented. The present study shows that the zebrafish is a suitable vertebrate model system in the Global South.
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Modelos Animales , Pez Cebra/fisiología , Animales , BrasilRESUMEN
Mycofactocin (MFT) is a redox cofactor belonging to the family of ribosomally synthesized and post-translationally modified peptides (RiPPs) and is involved in alcohol metabolism of mycobacteria including Mycobacterium tuberculosis. A preliminary biosynthetic model had been established by bioinformatics and in vitro studies, while the structure of natural MFT and key biosynthetic steps remained elusive. Here, we report the discovery of glycosylated MFT by 13C-labeling metabolomics and establish a model of its biosynthesis in Mycolicibacterium smegmatis. Extensive structure elucidation including NMR revealed that MFT is decorated with up to nine ß-1,4-linked glucose residues including 2-O-methylglucose. Dissection of biosynthetic genes demonstrated that the oligoglycosylation is catalyzed by the glycosyltransferase MftF. Furthermore, we confirm the redox cofactor function of glycosylated MFTs by activity-based metabolic profiling using the carveol dehydrogenase LimC and show that the MFT pool expands during cultivation on ethanol. Our results will guide future studies into the biochemical functions and physiological roles of MFT in bacteria.
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PURPOSE: To evaluate and compare the effects of homologous and heterologous PRP (Platelet-Rich Plasma) on the quality and speed of skin wound healing, compared to Poor Platelet Plasma (PPP). METHODS: Twenty-one male adult rabbits were used; two for preparing homologous PRP, with the rest of them separated randomly in three groups, according to the treatment received: PPP - control (n=5), homologous PRP (n=7), heterologous (n=7). Excisional skin wounds were made on the back of the animals, for the application of homologous and heterologous PPP and PRP. At the 14th post-operative day (POD), the animals were subjected to a new wound, and the treatments were inverted. The wounds were evaluated macroscopically and histologically. RESULTS: A larger percentage of scar retraction was observed on the group treated with heterologous PRP, compared to homologous PRP, at the third POD, an increase of 25.03% (p=0.01). No other statistically significant differences among treatments were observed. Among every group, skin healing was efficient, without local adverse effects. CONCLUSIONS: Heterologous PRP contributed with more tissue retraction at the beginning of the wound healing process. After this, there were no differences on the wound healing skin process treated with PRP or PPP. However, our findings suggest the presence of others plasmatic factors, besides platelets, which could also contribute to the wound healing process, and thus, should be further investigated.
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Plaquetas , Plasma Rico en Plaquetas , Animales , Masculino , Conejos , Piel , Cicatrización de HeridasRESUMEN
Combinatorial biosynthesis has great potential for designing synthetic circuits and amplifying the production of new active compounds. Studies on multienzyme cascades are extremely useful for improving our knowledge on enzymatic catalysis. In particular, the elucidation of enzyme substrate promiscuity can be potentially used for bioretrosynthetic approaches, leading to the design of alternative and more convenient routes to produce relevant molecules. In this perspective, plant-derived polyketides are extremely adaptable to those synthetic biological applications. Here, we present a combination of an in vitro CoA ligase activity assay coupled with a bacterial multigene expression system that leads to precursor-directed biosynthesis of 21 flavonoid derivatives. When the vast knowledge from protein databases is exploited, the herein presented procedure can be easily repeated with additional plant-derived polyketides. Lastly, we report an efficient in vivo expression system that can be further exploited to heterologously express pathways not necessarily related to plant polyketide synthases.