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1.
Molecules ; 29(14)2024 Jul 17.
Artículo en Inglés | MEDLINE | ID: mdl-39064928

RESUMEN

Angomonas deanei belongs to Trypanosomatidae family, a family of parasites that only infect insects. It hosts a bacterial endosymbiont in a mutualistic relationship, constituting an excellent model for studying organelle origin and cellular evolution. A lipidomic approach, which allows for a comprehensive analysis of all lipids in a biological system (lipidome), is a useful tool for identifying and measuring different expression patterns of lipid classes. The present study applied GC-MS and NMR techniques, coupled with principal component analysis (PCA), in order to perform a comparative lipidomic study of wild and aposymbiotic A. deanei grown in the presence or absence of FBS. Unusual contents of branched-chain iso C17:0 and C19:0-cis-9,10 and-11,12 fatty acids were identified in A. deanei cultures, and it was interesting to note that their content slightly decreased at the log phase culture, indicating that in the latter growth stages the cell must promote the remodeling of lipid synthesis in order to maintain the fluidity of the membrane. The combination of analytical techniques used in this work allowed for the detection and characterization of lipids and relevant contributors in a variety of A. deanei growth conditions.


Asunto(s)
Ácidos Grasos , Lipidómica , Trypanosomatina , Lipidómica/métodos , Ácidos Grasos/metabolismo , Ácidos Grasos/análisis , Trypanosomatina/metabolismo , Cromatografía de Gases y Espectrometría de Masas , Análisis de Componente Principal , Espectroscopía de Resonancia Magnética/métodos
2.
Mar Drugs ; 21(7)2023 Jun 29.
Artículo en Inglés | MEDLINE | ID: mdl-37504919

RESUMEN

A blend refers to the combination of two or more components to achieve properties that are superior to those found in the individual products used for their production. Gracilaria birdiae agaran (SPGb) and chromium picolinate (ChrPic) are both antioxidant agents. However, there is no documentation of blends that incorporate agarans and ChrPic. Hence, the objective of this study was to generate blends containing SPGb and ChrPic that exhibit enhanced antioxidant activity compared to SPGb or ChrPic alone. ChrPic was commercially acquired, while SPGb was extracted from the seaweed. Five blends (B1; B2; B3; B4; B5) were produced, and tests indicated B5 as the best antioxidant blend. B5 was not cytotoxic or genotoxic. H2O2 (0.6 mM) induced toxicity in fibroblasts (3T3), and this effect was abolished by B5 (0.05 mg·mL-1); neither ChrPic nor SPGb showed this effect. The cells also showed no signs of toxicity when exposed to H2O2 after being incubated with B5 and ChrPic for 24 h. In another experiment, cells were incubated with H2O2 and later exposed to SPGb, ChrPic, or B5. Again, SPGb was not effective, while cells exposed to ChrPic and B5 reduced MTT by 100%. The data demonstrated that B5 has activity superior to SPGb and ChrPic and points to B5 as a product to be used in future in vivo tests to confirm its antioxidant action. It may also be indicated as a possible nutraceutical agent.


Asunto(s)
Gracilaria , Rhodophyta , Algas Marinas , Antioxidantes/farmacología , Peróxido de Hidrógeno/farmacología , Verduras
3.
Molecules ; 28(7)2023 Mar 30.
Artículo en Inglés | MEDLINE | ID: mdl-37049845

RESUMEN

Oreochromis niloticus (tilapia) is one of the most cultivated fish species worldwide. Tilapia farming generates organic waste from fish removal processes in nurseries. Visceral waste can damage natural ecosystems. Therefore, the use of this material as a source of biomolecules helps reduce environmental impacts and improve pharmacological studies. Tilapia viscera were subjected to proteolysis and complexation with an ion-exchange resin. The obtained glycosaminoglycans were purified using ion exchange chromatography (DEAE-Sephacel). The electrophoretic profile and analysis of 1H/13C nuclear magnetic resonance (NMR) spectra allowed for the characterization of the compound as chondroitin sulfate and its sulfation position. This chondroitin was named CST. We tested the ability of CST to reduce leukocyte influx in acute peritonitis models induced by sodium thioglycolate and found a significant reduction in leukocyte migration to the peritoneal cavity, similar to the polymorphonuclear population of the three tested doses of CST. This study shows, for the first time, the potential of CST obtained from O. niloticus waste as an anti-inflammatory drug, thereby contributing to the expansion of the study of molecules with pharmacological functions.


Asunto(s)
Cíclidos , Peritonitis , Tilapia , Animales , Sulfatos de Condroitina , Ecosistema , Peritonitis/inducido químicamente , Peritonitis/tratamiento farmacológico
4.
Appl Environ Microbiol ; 86(17)2020 08 18.
Artículo en Inglés | MEDLINE | ID: mdl-32631857

RESUMEN

Under conditions of carbon starvation or thermal, osmotic, or oxidative shock, mutants affected in the synthesis or mobilization of poly-3-hydroxybutyrate (PHB) are known to survive less well. It is still unclear if the synthesis and accumulation of PHB are sufficient to protect bacteria against stress conditions or if the stored PHB has to be mobilized. Here, we demonstrated that mobilization of PHB in Herbaspirillum seropedicae SmR1 was heat-shock activated at 45°C. In situ proton (1H) nuclear magnetic resonance spectroscopy (i.e., 1H-nuclear magnetic resonance) showed that heat shock increased amounts of 3-hydroxybutyrate (3HB) only in H. seropedicae strains able to synthesize and mobilize PHB. H. seropedicae SmR1 mutants unable to synthesize or mobilize PHB were more susceptible to heat shock and survived less well than the parental strain. When 100 mM 3-hydroxybutyrate was added to the medium, the ΔphaC1 strain (an H. seropedicae mutant unable to synthesize PHB) and the double mutant with deletion of both phaZ1 and phaZ2 (i.e., ΔphaZ1.2) (unable to mobilize PHB) showed partial rescue of heat adaptability (from 0% survival without 3HB to 40% of the initial viable population). Addition of 200 mM 3HB before the imposition of heat shock reduced protein aggregation to 15% in the ΔphaC1 mutant and 12% in the ΔphaZ1.2 mutant. We conclude that H. seropedicae SmR1 is naturally protected by 3HB released by PHB mobilization, while mutants unable to generate large amounts of 3HB under heat shock conditions are less able to cope with heat damage.IMPORTANCE Bacteria are subject to abrupt changes in environmental conditions affecting their growth, requiring rapid adaptation. Increasing the concentration of some metabolites can protect bacteria from hostile conditions that lead to protein denaturation and precipitation, as well as damage to plasma membranes. In this work, we demonstrated that under thermal shock, the bacterium Herbaspirillum seropedicae depolymerized its intracellular stock polymer known as poly-3-hydroxybutyrate (PHB), rapidly increasing the concentration of 3-hydroxybutyrate (3HB) and decreasing protein precipitation by thermal denaturation. Mutant H. seropedicae strains unable to produce or depolymerize PHB suffered irreparable damage during thermal shock, resulting in fast death when incubated at 45°C. Our results will contribute to the development of bacteria better adapted to high temperatures found either in natural conditions or in industrial processes. In the case of H. seropedicae and other bacteria that interact beneficially with plants, the understanding of PHB metabolism can be decisive for the development of more-competitive strains and their application as biofertilizers in agriculture.


Asunto(s)
Ácido 3-Hidroxibutírico/metabolismo , Respuesta al Choque Térmico , Herbaspirillum/fisiología , Hidroxibutiratos/metabolismo , Poliésteres/metabolismo , Agregado de Proteínas
5.
Mar Drugs ; 18(5)2020 Apr 29.
Artículo en Inglés | MEDLINE | ID: mdl-32365741

RESUMEN

Sulfated polysaccharides (SPs) obtained from green seaweeds are structurally heterogeneous molecules with multifunctional bioactivities. In this work, two sulfated and pyruvated galactans were purified from Caulerpa cupressoides var. flabellata (named SP1 and SP2), and their immunostimulatory effect was evaluated using cultured murine macrophage cells. Both SPs equally increased the production of nitric oxide, reactive oxygen species, and the proinflammatory cytokines TNF-α and IL-6. NMR spectroscopy revealed that both galactans were composed primarily of 3)-ß-d-Galp-(1→3) units. Pyruvate groups were also found, forming five-membered cyclic ketals as 4,6-O-(1'carboxy)-ethylidene-ß-d-Galp residues. Some galactoses are sulfated at C-2. In addition, only SP2 showed some galactose units sulfated at C-4, indicating that sulfation at this position is not essential for the immunomodulatory activity of these galactans. Overall, the data showed that the galactans of C. cupressoides exhibited immunostimulating activity with potential therapeutic applications, which can be used in the development of new biomedical products.


Asunto(s)
Adyuvantes Inmunológicos/metabolismo , Caulerpa/metabolismo , Galactanos/metabolismo , Macrófagos/efectos de los fármacos , Algas Marinas , Adyuvantes Inmunológicos/farmacología , Animales , Supervivencia Celular/efectos de los fármacos , Citocinas/metabolismo , Galactanos/farmacología , Macrófagos/metabolismo
6.
Chem Biodivers ; 17(9): e2000369, 2020 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-32644295

RESUMEN

The current study was carried out by a bioguided fractionation of a hexane extract of the latex of Euphorbia umbellata against leukemic cells. Samples were analyzed by NMR, GC/MS, triterpenes quantification, and MTT reduction assay. Morphological, cell cycle, mitochondrial membrane potential and caspases 3/7 analyses were performed for the dichloromethane and ethanol fractions, and selectivity index for the dichloromethane fraction. NMR analysis presented characteristic signals of terpenes and steroids, data were confirmed by the quantification of triterpenes and GC/MS analysis. MTT reduction assay demonstrated that HL-60 was the most sensitive cell lineage against dichloromethane and ethanol fractions. Compounds of these matrices caused morphological changes compatible with apoptosis induction, altered cell cycle, increment of depolarized population cells and activation of caspases 3/7. Selectivity indices were higher than 22.44. Bioguided-fractionation study showed that samples of the latex of E. umbellata raised the activity of the phytocomplex against leukemic cells, and the cytotoxicity can be associated with an apoptosis pathway.


Asunto(s)
Antineoplásicos Fitogénicos/farmacología , Apoptosis/efectos de los fármacos , Euphorbia/química , Látex/química , Terpenos/farmacología , Antineoplásicos Fitogénicos/química , Antineoplásicos Fitogénicos/aislamiento & purificación , Ciclo Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Ensayos de Selección de Medicamentos Antitumorales , Humanos , Terpenos/química , Terpenos/aislamiento & purificación , Células Tumorales Cultivadas
7.
Arch Microbiol ; 201(2): 209-214, 2019 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-30506165

RESUMEN

The committed and rate-limiting step in fatty acid biosynthesis is catalyzed by acetyl-CoA carboxylase (ACC). In previous studies we showed that ACC activity is inhibited through interactions with the PII signaling proteins in vitro. Here we provide in vivo support for that model; we noted that PII proteins are able to reduce malonyl-CoA levels in vivo in Escherichia coli. Furthermore, we show that fatty acid biosynthesis is strongly enhanced in E. coli strains carrying deletions in PII coding genes. Given that PII proteins act as conserved negative regulators of ACC in Bacteria, our findings may be explored to engineer other prokaryotes to improve fatty acid yields, thereby turning microbial biofuel production economically competitive in the future.


Asunto(s)
Proteínas de Escherichia coli/genética , Escherichia coli/metabolismo , Ácidos Grasos/biosíntesis , Acetil-CoA Carboxilasa/metabolismo , Biocombustibles , Escherichia coli/genética , Eliminación de Gen , Transducción de Señal
8.
Molecules ; 24(11)2019 May 31.
Artículo en Inglés | MEDLINE | ID: mdl-31151328

RESUMEN

It has recently been shown that chitosan (Chit) induces the formation of calcium oxalate (CaOx) crystals, which are mainly responsible for the appearance of kidney stones, and this might limit the use of Chit in vivo. Here, Chit was conjugated with gallic acid (Chit-Gal) to decrease the formation of CaOx crystal. This conjugation was confirmed by FTIR and NMR analyses. Chit-Gal contains 10.2 ± 1.5 mg GA per g of Chit. Compared to the control group, Chit increased the number of crystals by six-fold, mainly in the number of monohydrated CaOx crystals, which are the most harmful CaOx crystals. In addition, Chit increased the zeta potential (ζ) of CaOx crystals by three-fold, indicating that Chit was associated with the crystals. These alterations were abolished when Chit-gal was used in these tests. As oxidative stress is related to renal calculus formation, Chit and Chit-Gal were also evaluated as antioxidants using total antioxidant Capacity (TAC), reducing power, ferrous chelation, and copper chelation tests. Chit-gal was more efficient antioxidant agent in TAC (2 times), in ferrous chelation (90 times), and in reducing Power (5 times) than Chit. Overall, Chit-gal has higher antioxidant activity than Chit, does not induce the formation of CaOx crystals. Thus, Chit-Gal has potential to be used as a chit substitute.


Asunto(s)
Oxalato de Calcio/química , Quitosano/química , Ácido Gálico/química , Antioxidantes/química , Cristalización , Quelantes del Hierro/química , Cálculos Renales/química , Espectroscopía de Resonancia Magnética , Peso Molecular , Espectroscopía Infrarroja por Transformada de Fourier
9.
Biol Chem ; 399(8): 859-867, 2018 07 26.
Artículo en Inglés | MEDLINE | ID: mdl-29664729

RESUMEN

Exocyclic DNA adducts are considered as potential tools for the study of oxidative stress-related diseases, but an important aspect is their chemical reactivity towards oxidant species. We report here the oxidation of 1-N2-etheno-2'-deoxyguanosine (1,N2-εdGuo) by singlet molecular oxygen (1O2) generated by a non-ionic water-soluble endoperoxide [N,N'-di(2,3-dihydroxypropyl)-1,4-naphthalenedipropanamide endoperoxide (DHPNO2)] and its corresponding oxygen isotopically labeled [18O]-[N,N'-di(2,3-dihydroxypropyl)-1,4- naphthalenedipropanamide endoperoxide (DHPN18O2)], and by photosensitization with two different photosensitizers [methylene blue (MB) and Rose Bengal (RB)]. Products detection and characterization were achieved using high performance liquid chromatography (HPLC) coupled to ultraviolet and electrospray ionization (ESI) tandem mass spectrometry, and nuclear magnetic resonance (NMR) analyses. We found that dGuo is regenerated via reaction of 1O2 with the ε-linkage, and we propose a dioxetane as an intermediate, which cleaves and loses the aldehyde groups as formate residues, or alternatively, it generates a 1,2-ethanediol adduct. We also report herein the quenching rate constants of 1O2 by 1,N2-εdGuo and other etheno modified nucleosides. The rate constant (kt) values obtained for etheno nucleosides are comparable to the kt of dGuo. From these results, we suggest a possible role of 1O2 in the cleanup of etheno adducts by regenerating the normal base.


Asunto(s)
Daño del ADN , Desoxiguanosina/química , Oxígeno Singlete/química , Desoxiguanosina/análogos & derivados , Desoxiguanosina/aislamiento & purificación , Espectroscopía de Resonancia Magnética , Espectrometría de Masas , Estructura Molecular , Oxidación-Reducción
10.
Biochemistry ; 56(5): 779-792, 2017 02 07.
Artículo en Inglés | MEDLINE | ID: mdl-28125217

RESUMEN

Xylella fastidiosa is a plant-infecting bacillus, responsible for many important crop diseases, such as Pierce's disease of vineyards, citrus variegated chlorosis, and coffee leaf scorch (CLS), among others. Recent genomic comparisons involving two CLS-related strains, belonging to X. fastidiosa subsp. pauca, revealed that one of them carries a frameshift mutation that inactivates a gene encoding an oxidoreductase of the short-chain dehydrogenase/reductase (SDR) superfamily, which may play important roles in determining structural variations in bacterial glycans and glycoconjugates. However, the exact nature of this SDR has been a matter of controversy, as different annotations of X. fastidiosa genomes have implicated it in distinct reactions. To confirm the nature of this mutated SDR, a comparative analysis was initially performed, suggesting that it belongs to a subgroup of SDR decarboxylases, representing a UDP-xylose synthase (Uxs). Functional assays, using a recombinant derivative of this enzyme, confirmed its nature as XfUxs, and carbohydrate composition analyses, performed with lipopolysaccharide (LPS) molecules obtained from different strains, indicate that inactivation of the X. fastidiosa uxs gene affects the LPS structure among CLS-related X. fastidiosa strains. Finally, a comparative sequence analysis suggests that this mutation is likely to result in a morphological and evolutionary hallmark that differentiates two subgroups of CLS-related strains, which may influence interactions between these bacteria and their plant and/or insect hosts.


Asunto(s)
Carboxiliasas/química , Evolución Molecular , Lipopolisacáridos/química , Filogenia , Proteínas de Plantas/química , Xylella/genética , Secuencia de Aminoácidos , Secuencia de Bases , Carboxiliasas/genética , Carboxiliasas/metabolismo , Clonación Molecular , Coffea/microbiología , Escherichia coli/genética , Escherichia coli/metabolismo , Mutación del Sistema de Lectura , Expresión Génica , Hidrólisis , Lipopolisacáridos/biosíntesis , Monosacáridos/análisis , Enfermedades de las Plantas/microbiología , Hojas de la Planta/microbiología , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Alineación de Secuencia , Xylella/clasificación , Xylella/enzimología , Xylella/aislamiento & purificación
11.
Bull Environ Contam Toxicol ; 96(1): 49-54, 2016 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-26589900

RESUMEN

Juveniles Rhamdia quelen fish species were exposed to diclofenac for 96 h at concentrations of 0.2, 2, and 20 µg/L. Biochemical, genetic, and hematological biomarkers were assessed in the liver, kidney, and blood in order to evaluate the toxic effects. No oxidative stress was observed in liver. In kidney the superoxide dismutase activity increased in all concentrations, suggesting an alteration in the hydrogen peroxide production, but DNA damage and lipid peroxidation were not detected. Diclofenac exposure increased the red blood cells number at concentrations of 0.2 and 2 µg/L, and monocytes and neutrophils at 2 and 20 µg/L, respectively. These results suggest that acute exposure to diclofenac, even at low concentrations, caused hematologic and renal enzymatic alterations in R. quelen.


Asunto(s)
Bagres/sangre , Diclofenaco/toxicidad , Riñón/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Contaminantes Químicos del Agua/toxicidad , Animales , Acuicultura , Biomarcadores/sangre , Daño del ADN , Riñón/metabolismo , Peroxidación de Lípido/efectos de los fármacos , Hígado/efectos de los fármacos , Hígado/metabolismo , Alimentos Marinos , Pruebas de Toxicidad Aguda
12.
Mar Drugs ; 13(1): 141-58, 2014 Dec 29.
Artículo en Inglés | MEDLINE | ID: mdl-25551781

RESUMEN

Chitosan is widely used in the biomedical field due its chemical and pharmacological properties. However, intake of chitosan results in renal tissue accumulation of chitosan and promotes an increase in calcium excretion. On the other hand, the effect of chitosan on the formation of calcium oxalate crystals (CaOx) has not been described. In this work, we evaluated the antioxidant capacity of chitosan and its interference in the formation of CaOx crystals in vitro. Here, the chitosan obtained commercially had its identity confirmed by nuclear magnetic resonance and infrared spectroscopy. In several tests, this chitosan showed low or no antioxidant activity. However, it also showed excellent copper-chelating activity. In vitro, chitosan acted as an inducer mainly of monohydrate CaOx crystal formation, which is more prevalent in patients with urolithiasis. We also observed that chitosan modifies the morphology and size of these crystals, as well as changes the surface charge of the crystals, making them even more positive, which can facilitate the interaction of these crystals with renal cells. Chitosan greatly influences the formation of crystals in vitro, and in vivo analyses should be conducted to assess the risk of using chitosan.


Asunto(s)
Quitosano/efectos adversos , Cálculos Renales/etiología , Antioxidantes/farmacología , Oxalato de Calcio/metabolismo , Quitosano/farmacología , Cristalización , Humanos , Riñón/efectos de los fármacos , Espectroscopía de Resonancia Magnética , Espectroscopía Infrarroja por Transformada de Fourier
13.
Int J Biol Macromol ; 262(Pt 2): 130121, 2024 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-38350588

RESUMEN

This study identified a rhamnose-containing cell wall polysaccharide (RhaCWP) in an alkaline extract prepared to analyze intracellular polysaccharides (IPS) from Streptococcus mutans biofilm. IPS was an 1,4-α-D-glucan with branchpoints introduced by 1,6-α-glucan while RhaCWP presented 1,2-α-L-and 1,3-α-L rhamnose backbone and side chains connected by 1,2-α-D-glucans, as identified by nuclear magnetic resonance (NMR) spectroscopy and methylation analyses. The MW of IPS and RhaCWP was 11,298 Da, as determined by diffusion-ordered NMR spectroscopy. Therefore, this study analyzed the chemical structure of RhaCWP and IPS from biofilm in a single fraction prepared via a convenient hot-alkali extraction method. This method could be a feasible approach to obtain such molecules and improve the comprehension of the structure-function relationships in polymers from S. mutans in future studies.


Asunto(s)
Ramnosa , Streptococcus mutans , Ramnosa/análisis , Polisacáridos/análisis , Glucanos/química , Pared Celular/química
14.
Int J Biol Macromol ; 257(Pt 1): 128612, 2024 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-38070366

RESUMEN

Eucalypt kraft lignin isolated in a LignoBoost™ pilot plant was characterized by GC-MS, ICP-OES, DSC, HPSEC, 31P NMR, and HSQC 2D-NMR to be used without any further processing to produce lignin nanoparticles (LNPs) by nanoprecipitation. Tetrahydrofuran (THF) was used as a solvent, and water as a non-solvent. Microscopic analysis (TEM) showed that LNPs were regularly spherical with some hollow particles dispersed in-between, and sizes were tunable by changing the solvent dripping rate onto the non-solvent. LNP particle sizes had a bimodal distribution, with the largest population having an average apparent hydrodynamic diameter ranging from 105.6 to 75.6 nm. Colloidal dispersions of LNPs in water presented good stability in different dilutions without significant size changes upon storage at pH close to neutral for as long as 45 days. Zeta potentials around -40 mV were obtained for LNP suspensions at pH ranging from 7 to 9. The high carbohydrate content (circa 10 % on a dry basis, mostly xylans) of the lignin precursor did not interfere in LNP formation, whose antioxidant activity was expressive as demonstrated by the ABTS assay at pH 7.4, with an EC50 of 4.04 µg mL-1. Also, the Trolox® equivalent antioxidant capacity (TEAC) of LNPs reached 1.90 after 40 min reaction time.


Asunto(s)
Antioxidantes , Nanopartículas , Antioxidantes/química , Lignina/química , Nanopartículas/química , Solventes , Agua
15.
Food Chem ; 451: 139531, 2024 Sep 01.
Artículo en Inglés | MEDLINE | ID: mdl-38704992

RESUMEN

Winemaking production is old knowledge of the combination of saccharification and fermentation processes. During the fermentation process, ethanol concentration is one of the main key parameters that provides the quality of wine and is linked to the consumption of carbohydrates present in wine. In this work was determined the better fermentation time, where the wine retains its highest concentration of ethanol and a higher concentration of the polysaccharides of Bordo wine of Vitis labrusca by 1D and 2D NMR measurements. The study provides information on the polysaccharide content for improving features and quality control of winemaking. Moreover, following previous studies by our group (de Lacerda Bezerra et al., 2018, de Lacerda Bezerra, Caillot, de Oliveira, Santana-Filho, & Sassaki, 2019; Stipp et al., 2023) showed that the soluble polysaccharides also inhibited the production of inflammatory cytokines (TNF-α and IL-1ß) and mediator (NO) in macrophage cells stimulated with LPS, bringing some important health benefits of wine.


Asunto(s)
Etanol , Fermentación , Espectroscopía de Resonancia Magnética , Polisacáridos , Vitis , Vino , Vino/análisis , Polisacáridos/química , Polisacáridos/metabolismo , Polisacáridos/farmacología , Etanol/metabolismo , Etanol/análisis , Animales , Vitis/química , Vitis/metabolismo , Vitis/microbiología , Ratones , Células RAW 264.7 , Factor de Necrosis Tumoral alfa/metabolismo , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Interleucina-1beta/metabolismo
16.
Talanta ; 270: 125501, 2024 Apr 01.
Artículo en Inglés | MEDLINE | ID: mdl-38091749

RESUMEN

Biocatalytic processes play a crucial role in the valorization of lignin; therefore, methods enabling the monitoring of enzymes such as ß-etherases, capable of breaking ß-O-4 aryl-ether bonds, are of significant biotechnological interest. A novel method for quantifying ß-etherase activity was developed based on the ß-ester bond formation between a chromophore and acetovainillone. The chromogenic substrate ß-(ρ-nitrophenoxy)-α-acetovanillone (PNPAV), was chemically synthesized. Kintetic monitoring of ρ-nitrophenolate release at 410 nm over 10 min, using recombinant LigF from Sphingobium sp SYK-6, LigF-AB and LigE-AB from Althererytrobacter sp B11, yielded enzimatic activities of 404. 3 mU/mg, 72 mU/mg, and 50 mU/mg, respectively. This method is applicable in a pH range of 7.0-9.0, with a sensitivity of up to 50 ng of enzyme, exhibiting no interference with lipolytic, glycolytic, proteolytic, and oxidoreductase enzymes.


Asunto(s)
Compuestos Cromogénicos , Sphingomonadaceae , Oxidorreductasas/química , Proteínas Bacterianas/química , Lignina/química
17.
Int J Biol Macromol ; 259(Pt 1): 129108, 2024 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-38158055

RESUMEN

ß-D-glucan has significant implications in regulating lipid metabolism and preventing diseases associated with lipid accumulation. Schizophyllan (SPG) from Schizophyllum commune fungus is a commercially important ß-glucan with applications in the health food industry, pharmacy, and cosmetics. However, SPG was obtained by submerged culture of the wood-rotting and filamentous fungus S. commune BRM 060008, which may have been isolated from the Cerrado Biome of Brazil. In this study, to confirm that the polysaccharide produced by BRM 060008 strain fermentation was indeed (1→3)(1→6)-ß-D-glucan, it was purified and characterized using Fourier transform infrared spectroscopy, thermogravimetric analysis, high-performance size exclusion chromatography, nuclear magnetic resonance, and methylation analysis. The polysaccharide produced was identified as the ß-D-glucan expected with a high molecular weight (1.093 × 106 g/mol) and the thermogravimetric analysis indicated a maximum degradation temperature of ~324 °C and a 60 % residual weight, lower than commercial SPG. The molecular structure and thermal properties of the ß-D-glucan were similar to the commercial sample. Additionally, the in vitro pancreatic lipase inhibitory activity was evaluated, investigating anti-obesity and anti-lipidemic properties. The results showed unprecedented lipase inhibition activity to SPG prepared using the S. commune strain BRM 060008, making it promising for food and pharmaceutical applications.


Asunto(s)
Schizophyllum , Sizofirano , Sizofirano/farmacología , Sizofirano/química , Schizophyllum/metabolismo , Glucanos/metabolismo , Lipasa/metabolismo , Polisacáridos/metabolismo
18.
Lipids Health Dis ; 12: 146, 2013 Oct 16.
Artículo en Inglés | MEDLINE | ID: mdl-24131597

RESUMEN

BACKGROUND: Shark liver oil (SLOil) and fish oil (FOil), which are respectively rich in alkylglycerols (AKGs) and n-3 polyunsaturated fatty acids (PUFAs), are able to reduce the growth of some tumors and the burden of cachexia. It is known that FOil is able to reduce proliferation rate and increase apoptotic cells and lipid peroxidation of tumor cells efficiently. However, there are few reports revealing the influence of SLOil on these parameters. In the current study, effects of FOil chronic supplementation on tumor growth and cachexia were taken as reference to compare the results obtained with SLOil supplementation. Also, we evaluated if the association of SLOil and FOil was able to promote additive effects. METHODS: Weanling male Wistar rats were divided into 4 groups: fed regular chow (C), supplemented (1 g/kg body weight) with SLOil (CSLO), FOil (CFO) and both (CSLO + FO). After 8 weeks half of each group was inoculated with Walker 256 cells originating new groups (W, WSLO, WFO and WSLO + FO). Biochemical parameters of cachexia, tumor weight, hydroperoxide content, proliferation rate and percentage of apoptotic tumor cells were analysed. Fatty acids and AKG composition of tumor and oils were obtained by high performance liquid chromatography and gas chromatography - mass spectrometry, respectively. Statistical analysis was performed by unpaired t-test and one-way ANOVA followed by a post hoc Tukey test. RESULTS: Fourteen days after inoculation, SLOil was able to restore cachexia parameters to control levels, similarly to FOil. WSLO rats presented significantly lower tumor weight (40%), greater tumor cell apoptosis (~3-fold), decreased tumor cell proliferation (35%), and higher tumor content of lipid hydroperoxides (40%) than observed in W rats, but FOil showed more potent effects. Supplementation with SLOil + FOil did not promote additive effects. Additionally, chromatographic results suggested a potential incorporation competition between the n-3 fatty acids and the AKGs in the tumor cells' membranes. CONCLUSIONS: SLOil is another marine source of lipids with similar FOil anti-cachectic capacity. Furthermore, despite being less potent than FOil, SLOil presented significant in vivo antitumor effects. These results suggest that the chronic supplementation with SLOil may be adjuvant of the anti-cancer therapy.


Asunto(s)
Antineoplásicos/farmacología , Caquexia/dietoterapia , Carcinoma 256 de Walker/dietoterapia , Suplementos Dietéticos , Aceites de Pescado/farmacología , Hígado/química , Animales , Antineoplásicos/aislamiento & purificación , Apoptosis/efectos de los fármacos , Caquexia/complicaciones , Caquexia/metabolismo , Caquexia/patología , Carcinoma 256 de Walker/complicaciones , Carcinoma 256 de Walker/metabolismo , Carcinoma 256 de Walker/patología , Proliferación Celular/efectos de los fármacos , Cromatografía Líquida de Alta Presión , Ácidos Grasos/metabolismo , Aceites de Pescado/aislamiento & purificación , Cromatografía de Gases y Espectrometría de Masas , Peróxido de Hidrógeno/agonistas , Peróxido de Hidrógeno/metabolismo , Masculino , Ratas , Ratas Wistar , Tiburones/metabolismo , Carga Tumoral/efectos de los fármacos , Destete
19.
Carbohydr Polym ; 321: 121333, 2023 Dec 01.
Artículo en Inglés | MEDLINE | ID: mdl-37739546

RESUMEN

Halophilic archaea are capable of producing fructans, which are fructose-based polysaccharides. However, their biochemical characterization and biological and technological properties have been scarcely studied. The aim of this study was to evaluate the production, chemical characterization, biological and technological properties of a fructan inulin-type biosynthesized by a halophilic archaeon. Fructan extraction was performed through ethanol precipitation and purification by diafiltration. The chemical structure was elucidated using Fourier Transform-Infrared Spectroscopy and Nuclear Magnetic Resonance (NMR). Haloarcula sp. M1 biosynthesizes inulin with an average molecular weight of 8.37 × 106 Da. The maximal production reached 3.9 g of inulin per liter of culture within seven days. The glass transition temperature of inulin was measured at 138.85 °C, and it exhibited an emulsifying index of 36.47 %, which is higher than that of inulin derived from chicory. Inulin from Haloarcula sp. M1 (InuH) demonstrates prebiotic capacity. This study represents the first report on the biological and technological properties of inulin derived from halophilic archaea.


Asunto(s)
Cichorium intybus , Haloarcula , Inulina , Fructanos , Etanol
20.
Int J Biol Macromol ; 240: 124385, 2023 Jun 15.
Artículo en Inglés | MEDLINE | ID: mdl-37060983

RESUMEN

The soluble fraction of polysaccharides from cabernet franc red wine (SFP) previously showed antitumoral effects by modulating the immune system. The present study tested the hypothesis that the SFP can regulate CYPs in vitro in HepG2 cells and in vivo in Walker-256 tumor-bearing rats. The SFP was used in the following protocols: (i) solid tumor, (ii) liquid tumor, and (iii) chemopreventive solid tumor. The SFP reduced solid tumor growth in both solid tumor protocols but did not inhibit liquid tumor development. The SFP reduced total CYP levels in the solid and liquid tumor protocols and reduced the gene expression of Cyp1a1 and Cyp2e1 in rats and CYP1A2 in HepG2 cells. An increase of N-acetylglucosaminidase activity was observed in all SFP-treated rats, and TNF-α levels increased in the solid tumor protocol in the vehicle, SFP, and vincristine (positive control) groups. The chemopreventive solid tumor protocol did not modify CYP levels in the liver or intestine or N-acetylglucosaminidase and myeloperoxidase activity in the liver. The in vitro digestion and nuclear magnetic resonance analyses suggested that SFP was minimally modified in the gastrointestinal system. In conclusion, SFP inhibited CYPs both in vivo and in vitro, likely as a result of its immunoinflammatory actions.


Asunto(s)
Vino , Ratas , Animales , Acetilglucosaminidasa , Sistema Enzimático del Citocromo P-450/metabolismo , Citocromo P-450 CYP1A1/genética , Citocromo P-450 CYP1A1/metabolismo , Polisacáridos/farmacología
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