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The serotonin transporter (SERT) is a member of the SLC6 neurotransmitter transporter family that mediates serotonin reuptake at presynaptic nerve terminals. SERT is the target of both therapeutic antidepressant drugs and psychostimulant substances such as cocaine and methamphetamines, which are small molecules that perturb normal serotonergic transmission by interfering with serotonin transport. Despite decades of studies, important functional aspects of SERT such as the oligomerization state of native SERT and its interactions with potential proteins remain unresolved. Here, we develop methods to isolate SERT from porcine brain (pSERT) using a mild, nonionic detergent, utilize fluorescence-detection size-exclusion chromatography to investigate its oligomerization state and interactions with other proteins, and employ single-particle cryo-electron microscopy to elucidate the structures of pSERT in complexes with methamphetamine or cocaine, providing structural insights into psychostimulant recognition and accompanying pSERT conformations. Methamphetamine and cocaine both bind to the central site, stabilizing the transporter in an outward open conformation. We also identify densities attributable to multiple cholesterol or cholesteryl hemisuccinate (CHS) molecules, as well as to a detergent molecule bound to the pSERT allosteric site. Under our conditions of isolation, we find that pSERT is best described as a monomeric entity, isolated without interacting proteins, and is ensconced by multiple cholesterol or CHS molecules.
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Estimulantes del Sistema Nervioso Central , Cocaína , Metanfetamina , Animales , Porcinos , Proteínas de Transporte de Serotonina en la Membrana Plasmática , Microscopía por Crioelectrón , Detergentes , Serotonina , Cocaína/farmacología , Metanfetamina/farmacologíaRESUMEN
Bacteria possess various receptors that sense different signals and transmit information to enable an optimal adaptation to the environment. A major limitation in microbiology is the lack of information on the signal molecules that activate receptors. Signals recognized by sensor domains are poorly reflected in overall sequence identity, and therefore, the identification of signals from the amino acid sequence of the sensor alone presents a challenge. Biogenic amines are of great physiological importance for microorganisms and humans. They serve as substrates for aerobic and anaerobic growth and play a role of neurotransmitters and osmoprotectants. Here, we report the identification of a sequence motif that is specific for amine-sensing sensor domains that belong to the Cache superfamily of the most abundant extracellular sensors in prokaryotes. We identified approximately 13,000 sensor histidine kinases, chemoreceptors, receptors involved in second messenger homeostasis and Ser/Thr phosphatases from 8,000 bacterial and archaeal species that contain the amine-recognizing motif. The screening of compound libraries and microcalorimetric titrations of selected sensor domains confirmed their ability to specifically bind biogenic amines. Mutants in the amine-binding motif or domains that contain a single mismatch in the binding motif had either no or a largely reduced affinity for amines. We demonstrate that the amine-recognizing domain originated from the universal amino acid-sensing Cache domain, thus providing insight into receptor evolution. Our approach enables precise "wet"-lab experiments to define the function of regulatory systems and therefore holds a strong promise to enable the identification of signals stimulating numerous receptors.
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Aminoácidos , Archaea , Humanos , Archaea/genética , Archaea/metabolismo , Aminoácidos/metabolismo , Proteínas Bacterianas/metabolismo , Bacterias/genética , Bacterias/metabolismo , Aminas Biogénicas/metabolismoRESUMEN
BACKGROUND: This study investigates the safety evaluation of enterocin-producing 11 E. mundtii and two E. faecium strains previously isolated from small livestock colostrums. Enterococcus species do not possess Generally Recognized as Safe (GRAS) status. Hence, it is critical to scrutinize enterococci's antibiotic resistance, virulence characteristics, and biogenic amine production capabilities in order to assess their safety before using them as starter or adjunct cultures. RESULTS: Enterococcus strains showed susceptibility to medically significant antibiotics. Multiple-drug resistance (MDR) was found in only E. faecium HC121.4, and its multiple antibiotic resistance (MAR) index was detected to be 0.22. The tetL and aph(3')-IIIa were the most commonly found antibiotic resistance genes in the strains. However, E. mundtii strains HC56.3, HC73.1, HC147.1, and E. faecium strain HC121.4 were detected to lack any of the antibiotic resistance genes examined in this study. Only E. mundtii HC166.3 showed hemolytic activity, while none of the strains engage in gelatinase activity. The strains were identified to have virulence factor genes with a low rate. None of the virulence factor genes could be detected in E. mundtii HC26.1, HC56.3, HC73.1, HC165.3, HC166.8, and E. faecium HC121.4. The E. mundtii HC73.2 strain displayed the highest presence of virulence factor genes, namely gelE, efaAfs, cpd, and ccf. Similarly, the E. mundtii HC112.1 strain showed a significant presence of genes efaAfm, ccf, and acm. There was no decarboxylation of histidine, ornithine, or lysine seen in any of the strains. Nevertheless, E. faecium HC121.4 and HC161.1 strains could decarboxylate tyrosine, but E. mundtii HC26.1, HC56.3, HC73.1, HC73.2, HC112.1, HC147.1, HC155.2, HC165.3, HC166.3, HC166.5, and HC166.8 strains only showed a limited capacity for tyrosine decarboxylation. None of the strains possessed the hdc, odc, or ldc genes, but all of them had the tdc gene. CONCLUSION: The E. mundtii HC56.3 and HC73.1 strains were deemed appropriate for utilization in food production. Using the remaining 11 strains as live cultures in food production activities could pose a possible risk to consumer health.
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Antibacterianos , Calostro , Enterococcus , Cabras , Pruebas de Sensibilidad Microbiana , Animales , Ovinos , Enterococcus/genética , Enterococcus/aislamiento & purificación , Enterococcus/metabolismo , Enterococcus/patogenicidad , Enterococcus/clasificación , Enterococcus/efectos de los fármacos , Antibacterianos/farmacología , Calostro/microbiología , Hidrocarburos Aromáticos con Puentes/metabolismo , Farmacorresistencia Bacteriana Múltiple/genética , Factores de Virulencia/genética , Enterococcus faecium/genética , Enterococcus faecium/aislamiento & purificación , Enterococcus faecium/metabolismo , Enterococcus faecium/patogenicidad , Enterococcus faecium/efectos de los fármacos , Virulencia/genéticaRESUMEN
We have developed and validated a novel LC-MS/MS method for simultaneously analyzing amino acids, biogenic amines, and their acetylated and methylated derivatives in plants. This method involves a one-step extraction of 2-5 mg of lyophilized plant material followed by fractionation of different biogenic amine forms, and exploits an efficient combination of hydrophilic interaction liquid chromatography (HILIC), reversed phase (RP) chromatography with pre-column derivatization, and tandem mass spectrometry (MS). This approach enables high-throughput processing of plant samples, significantly reducing the time needed for analysis and its cost. We also present a new synthetic route for deuterium-labeled polyamines. The LC-MS/MS method was rigorously validated by quantifying levels of nitrogen-related metabolites in seedlings of seven plant species, including Arabidopsis, maize, and barley, all of which are commonly used model organisms in plant science research. Our results revealed substantial variations in the abundance of these metabolites between species, developmental stages, and growth conditions, particularly for the acetylated and methylated derivatives and the various polyamine fractions. However, the biological relevance of these plant metabolites is currently unclear. Overall, this work contributes significantly to plant science by providing a powerful analytical tool and setting the stage for future investigations into the functions of these nitrogen-related metabolites in plants.
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Nitrógeno , Espectrometría de Masas en Tándem , Espectrometría de Masas en Tándem/métodos , Cromatografía Liquida , Nitrógeno/metabolismo , Arabidopsis/metabolismo , Arabidopsis/crecimiento & desarrollo , Zea mays/metabolismo , Zea mays/crecimiento & desarrollo , Hordeum/metabolismo , Hordeum/crecimiento & desarrollo , Poliaminas/metabolismo , Poliaminas/análisis , Plantas/metabolismo , Cromatografía Líquida con Espectrometría de MasasRESUMEN
Biogenic amines, produced by bacterial enzymatic reactions in food storage or processing, serve as indicators in food processing industries to assess food quality and freshness. Biogenic amines also often associated with various health problems, including abnormal immune responses and gastrointestinal disease. Previously, salphen base complexes have been reported but still exhibited low fluorescence enhancement upon biogenic amines. This research focused on synthesizing and characterizing new Zn(II) Schiff base complex with indole sidechain to enhance the fluorescence property and exploring their binding behaviour with the biogenic amines, which were phenylethylamine and cadaverine. The Zn(II) indole Schiff base complex's structure was verified by diverse spectroscopic techniques. Then, the binding behaviours between the Zn(II) indole Schiff base complex with the biogenic amines were analyzed using UV-Vis, fluorescence spectroscopy, and Job's plot analysis. UV-Vis binding study results indicated that the synthesized complexes could bind stronger with phenylethylamine than cadaverine, with binding constant, Kb= (8.21 ± 0.58) × 104 M- 1 and (2.506 ± 0.004) × 104 M- 1 respectively. Moreover, Zn(II) indole Schiff base complex-phenylethylamine binding also generated higher fluorescence enhancement than cadaverine, which were 54% and 51% respectively. Based on Job's plot analysis, the complex and biogenic amines were bound in the ratio of 1:1. To conclude, the synthesized complex has promising potential as a sensing material for biogenic amines detection in food. The complex is recommended to be deployed in the development of solid-state fluorescence sensor for biogenic amines detection for monitoring the food spoilage in the food industry in the future.
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INTRODUCTION: Social experience early in life appears to be necessary for the development of species-typical behavior. Although isolation during critical periods of maturation has been shown to impact behavior by altering gene expression and brain development in invertebrates and vertebrates, workers of some ant species appear resilient to social deprivation and other neurobiological challenges that occur during senescence or due to loss of sensory input. It is unclear if and to what degree neuroanatomy, neurochemistry, and behavior will show deficiencies if social experience in the early adult life of worker ants is compromised. METHODS: We reared newly eclosed adult workers of Camponotus floridanus under conditions of social isolation for 2-53 days, quantified brain compartment volumes, recorded biogenic amine levels in individual brains, and evaluated movement and behavioral performance to compare the neuroanatomy, neurochemistry, brood-care behavior, and foraging (predatory behavior) of isolated workers with that of workers experiencing natural social contact after adult eclosion. RESULTS: We found that the volume of the antennal lobe, which processes olfactory inputs, was significantly reduced in workers isolated for an average of 40 days, whereas the size of the mushroom bodies, centers of higher-order sensory processing, increased after eclosion and was not significantly different from controls. Titers of the neuromodulators serotonin, dopamine, and octopamine remained stable and were not significantly different in isolation treatments and controls. Brood care, predation, and overall movement were reduced in workers lacking social contact early in life. CONCLUSION: These results suggest that the behavioral development of isolated workers of C. floridanus is specifically impacted by a reduction in the size of the antennal lobe. Task performance and locomotor ability therefore appear to be sensitive to a loss of social contact through a reduction of olfactory processing ability rather than change in the size of the mushroom bodies, which serve important functions in learning and memory, or the central complex, which controls movement.
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Hormigas , Encéfalo , Aislamiento Social , Animales , Hormigas/fisiología , Hormigas/anatomía & histología , Encéfalo/metabolismo , Encéfalo/anatomía & histología , Conducta Social , Conducta Animal/fisiología , Cuerpos Pedunculados/metabolismo , Cuerpos Pedunculados/anatomía & histología , Aminas Biogénicas/metabolismo , Dopamina/metabolismoRESUMEN
The dynamic properties of neural systems throughout life can be hijacked by so-called manipulative parasites. This study investigated changes in the brain chemistry of the amphipod Gammarus fossarum in response to infection with two trophically-transmitted helminth parasites known to induce distinct behavioral alterations: the bird acanthocephalan Polymorphus minutus and the fish acanthocephalan Pomphorhynchus tereticollis. We quantified brain antioxidant capacity as a common marker of homeostasis and neuroprotection, and brain total protein, on 72 pools of six brains. We analyzed the concentration of serotonin (5HT), dopamine (DA) and tyramine in 52 pools of six brains, by using ultrafast high performance liquid chromatography with electrochemical detection (UHPLC-ECD). Brain total protein concentration scaled hypo-allometrically to dry body weight, and was increased in infected gammarids compared to uninfected ones. The brain of gammarids infected with P. minutus had significantly lower total antioxidant capacity relative to total proteins. Infection with P. tereticollis impacted DA level compared to uninfected ones, and in opposite direction between spring and summer. Brain 5HT level was higher in summer compared to spring independently of infection status, and was decreased by infection after correcting for brain total protein concentration estimated from dry whole-body weight. The potential implication of 5HT/DA balance in parasite manipulation, as a major modulator of the reward-punishment axis, is discussed. Taken together, these findings highlight the need to consider both brain homeostatic and/or structural changes (antioxidant and total protein content) together with neurotransmission balance and flexibility, in studies investigating the impact of parasites on brain and behavior.
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Acantocéfalos , Anfípodos , Química Encefálica , Encéfalo , Serotonina , Animales , Acantocéfalos/fisiología , Serotonina/análisis , Serotonina/metabolismo , Encéfalo/parasitología , Encéfalo/metabolismo , Anfípodos/parasitología , Anfípodos/fisiología , Estaciones del Año , Dopamina/análisis , Dopamina/metabolismo , Cromatografía Líquida de Alta Presión , Antioxidantes/análisis , Antioxidantes/metabolismoRESUMEN
Oxidative stress from generation of increased reactive oxygen species or has been reported to play an important role in dementia. Oxidative stress due to free radicals of oxygen or reactive oxygen species could be precipitating factors in the etiology of dementia. Apomorphine has been reported to have neuroprotective effects. To monitor memory enhancing and neuroprotective effects of apomorphine, we determined the antioxidant enzymes activities, lipid peroxidation, acetylcholine esterase (AChE) activity in brain and plasma, following repetitive administration of apomorphine in rat model of dementia. Biogenic amine levels were also monitored in hippocampus. Repeated administration of scopolamine was taken as an animal model of dementia. Decreased glutathione peroxidase, superoxide dismutase and catalase activities were observed in these animal models of dementia. While increased lipid peroxidation was also observed in the brain and plasma samples. The results showed significant effects of apomorphine. The activities of antioxidant enzymes displayed increased activities in both brain and plasma. Glutathione peroxidase and catalase activities were found to be significantly higher in brain and plasma of apomorphine treated rats. Superoxide dismutase (SOD) was significantly decreased in plasma of scopolamine injected rats; and a decreased tendency (non-significant) of SOD in brain was also observed. AChE activity in brain and plasma was significantly decreased in scopolamine treated rats. Learning and memory of rats in the present study was assessed by Morris Water Maze (MWM). Short-term memory and long-term memory was impaired significantly in scopolamine treated rats, which was prevented by apomorphine. Moreover, a marked decrease in biogenic amines was also found in the brain of scopolamine treated rats and was reverted in apomorphine treated rats. Results showed that scopolamine-treatment induced memory impairment and induced oxidative stress in rats as compared to saline-treated controls. These impairments were significantly restored by apomorphine administration. In conclusion, our data suggests that apomorphine at the dose of 1 mg/kg could be a potential therapeutic agent to treat dementia and related disorders.
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Apomorfina , Demencia , Modelos Animales de Enfermedad , Memoria , Fármacos Neuroprotectores , Ratas Wistar , Escopolamina , Animales , Apomorfina/farmacología , Ratas , Fármacos Neuroprotectores/farmacología , Fármacos Neuroprotectores/uso terapéutico , Masculino , Demencia/tratamiento farmacológico , Demencia/metabolismo , Demencia/prevención & control , Memoria/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Encéfalo/metabolismo , Encéfalo/efectos de los fármacos , Catalasa/metabolismo , Superóxido Dismutasa/metabolismo , Peroxidación de Lípido/efectos de los fármacos , Acetilcolinesterasa/metabolismo , Glutatión Peroxidasa/metabolismo , Antioxidantes/farmacología , Antioxidantes/uso terapéuticoRESUMEN
There is growing interest in using autochthonous lactic acid bacteria (LAB) that provide unique sensory characteristics to dairy products without affecting their safety and quality. This work studied the capacity of three Brazilian indigenous nonstarter LABs (NSLAB) to produce biogenic amines (BAs) and evaluated their effect on the volatile organic compounds (VOCs), microbial LAB communities, and physicochemical profile of short-aged cheese. Initially, the strain's potential for biosynthesis of BAs was assessed by PCR and in vitro assays. Then, a pilot-scale cheese was produced, including the NSLAB, and the microbial and VOC profiles were analyzed after 25 and 45 days of ripening. As a results, the strains did not present genes related to relevant BAs and did not produce them in vitro. During cheese ripening, the Lactococci counts were reduced, probably in the production of alcohols and acid compounds by the NSLAB. Each strain produces a unique VOC profile that changes over the ripening time without the main VOCs related to rancid or old cheese. Particularly, the use of the strain Lacticaseibacillus. paracasei ItalPN16 resulted in production of ester compounds with fruity notes. Thus, indigenous NSLAB could be a valuable tool for the enhancement and diversification of flavor in short-aged cheese.
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Queso , Lactobacillales , Compuestos Orgánicos Volátiles , Lactobacillales/genética , Queso/microbiología , Compuestos Orgánicos Volátiles/análisis , Brasil , LactobacillusRESUMEN
Fino Sherry wine undergoes biological aging carried out by a velum of flor yeast within a traditional dynamic system known as "criaderas and solera". The complex microbiota of biofilm-forming Saccharomyces cerevisiae strains play a crucial role in shaping the distinctive organoleptic profile of these types of wines. For this reason, the aim of this study is to analyze the changes produced by different flor yeast strains in the volatilome and the aminogram of different wines from the criaderas and solera system during biological aging in the laboratory, simulating a flor yeast velum condition at different stages of the system. Results suggest that each strain metabolizes wine differently, finding that depending on the wine, some strains are better suited for the process than others. In addition, it is found that the content of biogenic amines in Fino Sherry wines, previously attributed to malolactic bacteria, varies according to the yeast strain metabolizing the wine, suggesting that flor yeast could be used to modify biogenic amines content during biological aging. Results indicate that the use of selected flor yeast starters in biological aging may be of interest to modulate some parameters during Fino Sherry wine aging.
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Fermentación , Saccharomyces cerevisiae , Compuestos Orgánicos Volátiles , Vino , Vino/análisis , Vino/microbiología , Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/crecimiento & desarrollo , Compuestos Orgánicos Volátiles/metabolismo , Compuestos Orgánicos Volátiles/análisis , Compuestos de Nitrógeno/metabolismo , Aminas Biogénicas/metabolismo , Aminas Biogénicas/análisisRESUMEN
Exploring new methodologies for simple and on-demand methods of manipulating the emission and sensing ability of fluorescence sensor devices with solid-state emission molecular systems is important for realizing on-site sensing platforms. In this regard, although conjugated polymers (CPs) are some of the best candidates for preparing molecular sensor devices owing to their luminescent and molecular recognition properties, the development of CP-based sensor devices is still in its early stages. In this study, we herein propose a novel strategy for preparing a chemical stimuli-responsive solid-state emission system based on supramacromolecular assembly-induced emission enhancement (SmAIEE). The system was spontaneously developed by mixing only the component polymers (i.e., polythiophene and a transient cross-linking polymer). The proposed strategy can be applied to the facile preparation of molecular sensor devices. The analyte-induced fluorescent response of polythiophene originated from the dynamic displacement of the transient cross-linker in the polythiophene ensemble and the generation of the polythiophene-analyte complex. Our successful demonstration of the spontaneous preparation of the fluorescence sensor system by mixing two component polymers could lead to the development of on-site molecular analyzers including the determination of multiple analytes.
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Recently, prokaryotic laccases from lactic acid bacteria (LAB), which can degrade biogenic amines, were discovered. A laccase enzyme has been cloned from Oenococcus oeni, a very important LAB in winemaking, and it has been expressed in Escherichia coli. This enzyme has similar characteristics to those previously isolated from LAB as the ability to oxidize canonical substrates such as 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 2,6-dimethoxyphenol (2,6-DMP), and potassium ferrocyanide K4[Fe(CN6)], and non-conventional substrates as biogenic amines. However, it presents some distinctiveness, the most characteristic being its psychrophilic behaviour, not seen before among these enzymes. Psychrophilic enzymes capable of efficient catalysis at low temperatures are of great interest due to their potential applications in various biotechnological processes. In this study, we report the discovery and characterization of a new psychrophilic laccase, a multicopper oxidase (MCO), from the bacterium Oenococcus oeni. The psychrophilic laccase gene, designated as LcOe 229, was identified through the genomic analysis of O. oeni, a Gram-positive bacterium commonly found in wine fermentation. The gene was successfully cloned and heterologously expressed in Escherichia coli, and the recombinant enzyme was purified to homogeneity. Biochemical characterization of the psychrophilic laccase revealed its optimal activity at low temperatures, with a peak at 10 °C. To our knowledge, this is the lowest optimum temperature described so far for laccases. Furthermore, the psychrophilic laccase demonstrated remarkable stability and activity at low pH (optimum pH 2.5 for ABTS), suggesting its potential for diverse biotechnological applications. The kinetic properties of LcOe 229 were determined, revealing a high catalytic efficiency (kcat/Km) for several substrates at low temperatures. This exceptional cold adaptation of LcOe 229 indicates its potential as a biocatalyst in cold environments or applications requiring low-temperature processes. The crystal structure of the psychrophilic laccase was determined using X-ray crystallography demonstrating structural features similar to other LAB laccases, such as an extended N-terminal and an extended C-terminal end, with the latter containing a disulphide bond. Also, the structure shows two Met residues at the entrance of the T1Cu site, common in LAB laccases, which we suggest could be involved in substrate binding, thus expanding the substrate-binding pocket for laccases. A structural comparison of LcOe 229 with Antarctic laccases has not revealed specific features assigned to cold-active laccases versus mesophilic. Thus, further investigation of this psychrophilic laccase and its engineering could lead to enhanced cold-active enzymes with improved properties for future biotechnological applications. Overall, the discovery of this novel psychrophilic laccase from O. oeni expands our understanding of cold-adapted enzymes and presents new opportunities for their industrial applications in cold environments.
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Lacasa , Oenococcus , Oenococcus/enzimología , Oenococcus/genética , Lacasa/metabolismo , Lacasa/genética , Lacasa/química , Especificidad por Sustrato , Proteínas Bacterianas/metabolismo , Proteínas Bacterianas/genética , Proteínas Bacterianas/química , Secuencia de Aminoácidos , Escherichia coli/genética , Escherichia coli/metabolismo , Clonación Molecular , Cinética , Modelos Moleculares , Cristalografía por Rayos X , Concentración de Iones de HidrógenoRESUMEN
Sideritis scardica Griseb. and Clinopodium vulgare L., belonging to the Lamiaceae family, are rich in terpenoids and phenolics and exhibit various pharmacological effects, including antioxidant, anti-inflammatory and anti-cancer activities. While the memory-enhancing impacts of S. scardica are well documented, the cognitive benefits of C. vulgare remain unexplored. This study assessed the potential effect of C. vulgare on learning and memory in healthy and scopolamine (Sco)-induced memory-impaired male Wistar rats, comparing it with the effects of S. scardica. Over a 21-day period, rats orally received extracts of cultivated S. scardica (200 mg/kg) and C. vulgare (100 mg/kg), either individually or in combination, with administration starting 10 days before and continuing 11 days simultaneously with Sco injection at a dose of 2 mg/kg intraperitoneally. The results showed that both extracts effectively mitigated Sco-induced memory impairment. Their combination significantly improved recognition memory and maintained monoaminergic function. S. scardica excelled in preserving spatial working memory, while C. vulgare exhibited comparable retention of recognition memory, robust antioxidant activity and acetylcholinesterase inhibitory activity. The extracts alleviated Sco-induced downregulation of p-CREB/BDNF signaling, suggesting neuroprotective mechanisms. The extract combination positively affected most of the Sco-induced impairments, underscoring the potential for further investigation of these extracts for therapeutic development.
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Disfunción Cognitiva , Demencia , Sideritis , Ratas , Masculino , Animales , Escopolamina/efectos adversos , Ratas Wistar , Acetilcolinesterasa , Extractos Vegetales/farmacología , Extractos Vegetales/uso terapéutico , Antioxidantes/farmacología , Antioxidantes/uso terapéutico , Trastornos de la Memoria/inducido químicamente , Trastornos de la Memoria/tratamiento farmacológico , Disfunción Cognitiva/inducido químicamente , Disfunción Cognitiva/tratamiento farmacológico , Demencia/inducido químicamente , Demencia/tratamiento farmacológico , Aprendizaje por LaberintoRESUMEN
Tyramine (TRM) is a biogenic catecholamine neurotransmitter, which can trigger migraines and hypertension. TRM accumulated in foods is reduced and detected using additive cyclodextrins (CDs) while their association characteristics remain unclear. Here, single-crystal X-ray diffraction and density functional theory (DFT) calculation have been performed, demonstrating the elusive pseudopolymorphs in ß-CD inclusion complexes with TRM base/HCl, ß-CD·0.5TRM·7.6H2O (1) and ß-CD·TRM HCl·4H2O (2) and the rare α-CD·0.5(TRM HCl)·10H2O (3) exclusion complex. Both 1 and 2 share the common inclusion mode with similar TRM structures in the round and elliptical ß-CD cavities, belong to the monoclinic space group P21, and have similar herringbone packing structures. Furthermore, 3 differs from 2, as the smaller twofold symmetry-related, round α-CD prefers an exclusion complex with the twofold disordered TRM-H+ sites. In the orthorhombic P21212 lattice, α-CDs are packed in a channel-type structure, where the column-like cavity is occupied by disordered water sites. DFT results indicate that ß-CD remains elliptical to suitably accommodate TRM, yielding an energetically favorable inclusion complex, which is significantly contributed by the ß-CD deformation, and the inclusion complex of α-CD with the TRM aminoethyl side chain is also energetically favorable compared to the exclusion mode. This study suggests the CD implications for food safety and drug/bioactive formulation and delivery.
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Tiramina , Tiramina/química , beta-Ciclodextrinas/química , Modelos Moleculares , Ciclodextrinas/química , alfa-Ciclodextrinas/química , Teoría Funcional de la Densidad , Cristalografía por Rayos X , Difracción de Rayos XRESUMEN
Ostrich meat is an interesting alternative to poultry or beef due to its nutritional value. The addition of three plant species (hot peppers, acerola, Schisandra chinesis) was suggested as a method to improve the quality, safety, and consumer acceptance of sausages prepared from ostrich meat. A series of microbiological and chemical analyses (including, inter alia, content of biogenic amines, heavy metals, and bioactive compounds) of the products as well as their sensory evaluation was performed to verify this claim. The microflora of all sausages was dominated by lactic acid bacteria. The biggest threat to consumers' health could be connected to the presence of biogenic amines formed through the enzymatic activity of lactic acid bacteria. The sausages with plant additives had better antioxidative and anti-inflammatory properties and lower fat oxidation-these features were correlated with the presence of vitamin C. Sausages with plant additives had a higher acceptability in terms of taste and smell.
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Productos de la Carne , Struthioniformes , Productos de la Carne/análisis , Productos de la Carne/microbiología , Animales , Aditivos Alimentarios/análisis , Antioxidantes/análisis , Gusto , Aminas Biogénicas/análisisRESUMEN
BACKGROUND: Biogenic amines (BAs) in high concentrations are toxic and may cause a series of health symptoms. A sensitive measurement of BA levels is essential for human health. Capillary electrophoresis (CE) has emerged for the separation of eight BAs due to simple sample preparation and highly efficient separation. However, an important drawback for CE is low sensitivity. Magnetic solid-phase extraction (MSPE) has become a technique of interest owing to its brief operation and low solvent consumption. Hence, MSPE as a pretreatment has great potential to improve CE sensitivity for the analysis of BAs in complex food. RESULTS: Results showed that the Pt-Co-MWCNTs-COOH possessed strong magnetism, good reusability, and high adsorptive ability toward eight biogenic amines based on the hydrogen bonding between the -COOH of Pt-Co-MWCNTs-COOH and -NH2 groups of BAs. Using it as an adsorbent, a magnetic solid-phase extraction coupled with capillary electrophoresis (MSPE-CE) method was developed to effectively extract and sensitively analyze eight BAs. Under optimal conditions, the MSPE-CE method has wide linearities (10.0-1000.0 µg L-1 ) and low limits of detection (1.0-6.1 µg L-1 ). The accuracy of the developed method yielded recovery values from 82.07% to 102.58%. Meanwhile, the BAs contents in two samples were analyzed using the MSPE-CE method, with the results consistent with those detected by a high-performance liquid chromatography method. CONCLUSION: Given those advantages, the established MSPE-CE method promises the practical guidance of monitoring a variety of BAs and provides a foundation for the detection of other food hazards. © 2023 Society of Chemical Industry.
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Aminas Biogénicas , Electroforesis Capilar , Humanos , Electroforesis Capilar/métodos , Cromatografía Líquida de Alta Presión/métodos , Aminas Biogénicas/análisis , Extracción en Fase Sólida/métodos , Fenómenos Magnéticos , Límite de DetecciónRESUMEN
BACKGROUND: The excessive salt intake associated with Douchi has become a topic of controversy. Addressing this concern and enhancing its market competitiveness necessitates the application of salt reduction fermentation in Douchi. Therefore, to promote the application of salt reduction fermentation in Douchi, a comprehensive study was undertaken aiming to investigate the differences in biogenic amines, volatile compounds and non-volatile compounds in Douchi with varying salt content. RESULTS: The findings unequivocally demonstrate that salt hampers the formation of metabolites in Douchi. As the salt content increased, there was a significant decrease (P < 0.05) in the levels of total acid, amino-type nitrogen and free amino acids in Douchi. Notably, when the salt content exceeded 80 g kg-1, there was a substantial reduction (P < 0.05) in putrescine, lactic acid and malic acid levels. Similarly, when the salt content surpassed 40 g kg-1, ß-phenethylamine and oxalic acid levels exhibited a significant decline (P < 0.05). Furthermore, the results of E-nose and principal component analysis based on headspace solid phase microextraction gas chromatography-mass spectrometry revealed notable discrepancies in the volatile compound content between Douchi samples with relatively low salt content (40 and 80 g kg-1) and those with relatively high salt content (120, 160 and 200 g kg-1) (P < 0.05). By employing partial least squares discriminant analysis, eight distinct volatile compounds, including o-xylene, benzaldehyde and 1-octen-one, were identified. These compounds exhibited higher concentrations in Douchi samples with relatively low salt content (40 and 80 g kg-1). The sensory results showed that Douchi samples with lower salt content exhibited higher scores in the soy sauce-like and Douchi aroma attributes. CONCLUSION: In conclusion, this study significantly enhances our understanding of the impact of salt on metabolites in Douchi and provides invaluable insights for the development of salt reduction fermentation in this context. © 2024 Society of Chemical Industry.
Asunto(s)
Aminas Biogénicas , Fermentación , Cromatografía de Gases y Espectrometría de Masas , Compuestos Orgánicos Volátiles , Aminas Biogénicas/análisis , Aminas Biogénicas/metabolismo , Compuestos Orgánicos Volátiles/metabolismo , Compuestos Orgánicos Volátiles/química , Compuestos Orgánicos Volátiles/análisis , Cloruro de Sodio/análisis , Cloruro de Sodio/metabolismoRESUMEN
Background and Objectives: Metabolic disorders cause many skin issues, including acne vulgaris. This research investigated the function of glutathione peroxidase (GTPx) and biogenic amines as a potential novel pathophysiological link between metabolic syndrome (MetS) and acne vulgaris. Materials and Methods: The patients were distributed into two groups: metabolic precondition (MPG, n = 78) and control (CG, n = 81). To determine the extent of acne and metabolic preconditioning, patients were subjected to extensive clinical/paraclinical investigations. Additionally, catecholamine levels in urine and GTPx levels in blood were measured. Results: Mild acne was more common in the CG (32.1 vs. 6.4, p < 0.001), and severe acne was more common in the MPG (61.54 vs. 25.9, p < 0.001), with the average age being substantially higher in the MPG (23.81 vs. 21.05, p = 0.002). Significant variations were observed in the paraclinical levels for catecholamines (p < 0.05). In the MPG, most severe acne patients were overweight (52.1%), insulin-resistant (48.8%), or obese (47.9%). Moderate acne was most often linked to obesity (56%), overweight (44%), and insulin resistance (20%). Patients with severe acne (48.83%) had a considerably greater incidence of insulin resistance syndrome (p = 0.039) than those with moderate or severe acne (20%). The presence of two or three metabolic disorders considerably raised the risk of severe acne. Significant differences between groups were observed only in the subgroup of patients with severe acne, with lower values in the MPG (p = 0.015). Significant differences between groups were observed regarding the subgroup of patients with severe acne, with lower DTPx values in the MPG. At the group level, only CG patients with severe acne had reduced GTPx levels. Significant differences in catecholamine values were seen between groups (p < 0.05), independent of acne severity, except for adrenaline in mild acne patients (p = 0.059). Conclusions: The complex connection between GTPx and catecholamines in MetS suggests a significant role of these factors in the pathogenesis of acne associated with this condition, opening new perspectives in the research and treatment of acne in the context of MetS.
Asunto(s)
Acné Vulgar , Resistencia a la Insulina , Síndrome Metabólico , Humanos , Síndrome Metabólico/complicaciones , Sobrepeso/complicaciones , Acné Vulgar/etiología , Aminas Biogénicas/uso terapéutico , Obesidad/complicaciones , Catecolaminas/uso terapéuticoRESUMEN
Strain specificity (within-species variation) of microorganisms occurs widely in nature. It might affect microbiome construction and function in a complex microbial environment. Tetragenococcus halophilus, a halophilic bacterium that generally is used in high salt food fermentation, consists of two histamine-producing and non-histamine-producing subgroups. It is unclear whether and how the strain specificity of histamine-producing capacity influences the microbial community function during food fermentation. Here, based on systematic bioinformatic analysis, histamine production dynamic analysis, clone library construction analysis, and cultivation-based identification, we identified that T. halophilus is the focal histamine-producing microorganism during soy sauce fermentation. Furthermore, we discovered that a larger number and ratio of histamine-producing subgroups of T. halophilus significantly contributed more histamine production. We were able to artificially decrease the ratio of histamine-producing to non-histamine-producing subgroups of T. halophilus in complex soy sauce microbiota and realized the reduction of histamine by 34%. This study emphasizes the significance of strain specificity in regulating microbiome function. This study investigated how strain specificity influenced microbial community function and developed an efficient technique for histamine control. IMPORTANCE Inhibiting the production of microbiological hazards under the assumption of stable and high-quality fermentation is a critical and time-consuming task for the food fermentation industry. For spontaneously fermented food, it can be realized theoretically by finding and controlling the focal hazard-producing microorganism in complex microbiota. This work used histamine control in soy sauce as a model and developed a system-level approach to identify and regulate the focal hazard-producing microorganism. We discovered that the strain specificity of focal hazard-producing microorganisms had an important impact on hazard accumulation. Microorganisms frequently exhibit strain specificity. Strain specificity is receiving increasing interest since it determines not only microbial robustness but also microbial community assembly and microbiome function. This study creatively explored how the strain specificity of microorganisms influenced microbiome function. In addition, we believe that this work provides an excellent model for microbiological hazard control which can promote future work in other systems.
Asunto(s)
Alimentos de Soja , Alimentos de Soja/análisis , Alimentos de Soja/microbiología , Histamina , Fermentación , EnterococcaceaeRESUMEN
Carotid body (CB) glomus cells in most mammals, including humans, contain a broad diversity of classical neurotransmitters, neuropeptides and gaseous signaling molecules as well as their cognate receptors. Among them, acetylcholine, adenosine triphosphate and dopamine have been proposed to be the main excitatory transmitters in the mammalian CB, although subsequently dopamine has been considered an inhibitory neuromodulator in almost all mammalian species except the rabbit. In addition, co-existence of biogenic amines and neuropeptides has been reported in the glomus cells, thus suggesting that they store and release more than one transmitter in response to natural stimuli. Furthermore, certain metabolic and transmitter-degrading enzymes are involved in the chemotransduction and chemotransmission in various mammals. However, the presence of the corresponding biosynthetic enzyme for some transmitter candidates has not been confirmed, and neuroactive substances like serotonin, gamma-aminobutyric acid and adenosine, neuropeptides including opioids, substance P and endothelin, and gaseous molecules such as nitric oxide have been shown to modulate the chemosensory process through direct actions on glomus cells and/or by producing tonic effects on CB blood vessels. It is likely that the fine balance between excitatory and inhibitory transmitters and their complex interactions might play a more important than suggested role in CB plasticity.