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1.
Microorganisms ; 8(4)2020 Mar 25.
Artigo em Inglês | MEDLINE | ID: mdl-32218311

RESUMO

Within this work, the microbial synthesis of (2R,3S)-isocitric acid (ICA), a metabolite of the nonconventional yeast Yarrowia lipolytica, from biodiesel waste, has been studied. The selected strain Y. lipolytica VKM Y-2373 synthesized ICA with citric acid (CA) as a byproduct. This process can be regulated by changing cultivation conditions. The maximal production of ICA with the minimal formation of the byproduct was provided by the use of a concentration of (NH4)2SO4 (6 g/L); the addition of biodiesel waste to cultivation medium in 20-60 g/L portions; maintaining the pH of the cultivation medium at 6, and degree of aeration between 25% and 60% of saturation. Itaconic acid at a concentration of 15 mM favorably influenced the production of ICA by the selected strain. The optimization of cultivation conditions allowed us to increase the concentration of ICA in the culture liquid from 58.32 to 90.2 g/L, the product yield (Y) by 40%, and the ICA/CA ratio from 1.1:1 to 3:1. Research on laboratory animals indicated that ICA counteracted the negative effect of ammonium molybdate (10-5 М) and lead diacetate (10-7 М) on the learning and spatial memory of rats, including those exposed to emotional stress.

2.
Learn Behav ; 48(3): 364-372, 2020 09.
Artigo em Inglês | MEDLINE | ID: mdl-32212100

RESUMO

The main features of the Shuttle Box Active Avoidance paradigm (e.g., the use of simple locomotor response as an operant and electrical current as a primary reinforcer) make this task easily automated. However, learning in this paradigm cannot be easily separated from the specificity of fear motivation. Punishment and negative reinforcement highly affect behavior in this task and complicate learning. In the present study, we describe a novel computer-controlled appetitive task in a shuttle box and compare it with active avoidance. The appetitive task was performed in the same shuttle box apparatus, additionally equipped with food dispensers in each compartment, and using a similar protocol. The reinforced reaction included the transition to the feeder in the opposite compartment in response to a stimulus. Animals mastered the appetitive task faster than the active avoidance task in the shuttle box. Other major differences between the models were the number and dynamics of intertrial responses (ITRs). Whereas in active avoidance the number of ITRs was low during learning, in the appetitive task rates were higher and they persisted throughout learning. Overall, the findings demonstrate some benefits of the appetitive task as a control condition to active avoidance: the use of a similar reaction and apparatus, no prior habituation, and fast acquisition.


Assuntos
Aprendizagem da Esquiva , Condicionamento Operante , Animais , Comportamento Apetitivo , Medo , Punição , Reforço Psicológico
3.
Acta Neurobiol Exp (Wars) ; 79(3): 232-237, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31587015

RESUMO

Emotional stress is considered a serious pathogenetic factor of depression. In this study an ultrasound model of emotional stress developed in our laboratory was applied. It is characterized by the use of ultrasound as the stressor agent. Animals are triggered not by any organic or physical disturbances but by the perception of adverse information. This type of stress can induce depressive-like behavioral changes in rodents, manifested by decreased sucrose preference and increased time of immobility in a forced swim test. Ultrasound stress also increased the levels of oxidative stress markers. This is important, as stress has an established association with increased oxidative processes in the central nervous system. Total glutathione and carbonyl protein content were selected as relevant brain markers, as glutathione plays a critical role in cellular defensive mechanisms during oxidative stress and the level of protein carbonyls can be a measure of global protein oxidation. We demonstrated that two weeks of chronic exposure to ultrasound was enough to cause depressive-like behavioral changes in rats. Increased levels of oxidative stress markers in the hippocampus and prefrontal cortex were also observed after two weeks of such stress. The current study has two goals: the first is to study the relationship of depression and oxidative stress; the second is an additional validation of our approach to modeling stress­induced depressive-like states in rats. The present data further support the validity of the ultrasound model by expanding information related to the influence of ultrasound stress on behavioral and physiological parameters, which are of great importance in the development of stress-induced depression. A time correlation between the onset of symptoms and a change in the level of oxidative stress markers in the brain is also demonstrated.


Assuntos
Comportamento Animal/fisiologia , Depressão/fisiopatologia , Estresse Oxidativo/fisiologia , Estresse Psicológico/fisiopatologia , Animais , Depressão/metabolismo , Transtorno Depressivo/fisiopatologia , Modelos Animais de Doenças , Glutationa/metabolismo , Hipocampo/metabolismo , Hipocampo/fisiopatologia , Masculino , Córtex Pré-Frontal/fisiopatologia , Ratos
4.
Appl Microbiol Biotechnol ; 103(8): 3549-3558, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30852660

RESUMO

Biosynthesis of Ds(+)-threo-isocitric acid from ethanol in the Yarrowia lipolytica batch and repeated-batch cultures was studied. Repeated-batch cultivation was found to provide for a good biosynthetic efficiency of the producer for as long as 748 h, probably due to maintenance of high activities of enzymes involved in the biosynthesis of isocitric acid. Under optimal repeated-batch cultivation conditions, the producer accumulated 109.6 g/L Ds(+)-threo-isocitric acid with a production rate of 1.346 g/L h. The monopotassium salt of isocitric acid isolated from the culture liquid and purified to 99.9% was found to remove neurointoxication, to restore memory, and to improve the learning of laboratory rats intoxicated with lead and molybdenum salts. Taking into account the fact that the neurotoxic effect of heavy metals is mainly determined by oxidative stress, the aforementioned favorable action of isocitric acid on the intoxicated rats can be explained by its antioxidant activity among other pharmacological effects.


Assuntos
Antioxidantes/metabolismo , Antioxidantes/farmacologia , Técnicas de Cultura Celular por Lotes/métodos , Isocitratos/metabolismo , Isocitratos/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Animais , Antioxidantes/isolamento & purificação , Reatores Biológicos/microbiologia , Etanol/metabolismo , Isocitratos/isolamento & purificação , Masculino , Metais Pesados/toxicidade , Ratos , Yarrowia/metabolismo
5.
Artigo em Inglês | MEDLINE | ID: mdl-30472146

RESUMO

Emotional stress is a form of stress evoked by processing negative mental experience rather than an organic or physical disturbance and is a frequent cause of neuropsychiatric pathologies, including depression. Susceptibility to emotional stress is commonly regarded as a human-specific trait that is challenging to model in other species. Recently, we showed that a 3-week-long exposure to ultrasound of unpredictable alternating frequencies within the ranges of 20-25 kHz and 25-45 kHz can induce depression-like characteristics in laboratory mice and rats. In an anti-depressant sensitive manner, exposure decreases sucrose preference, elevates behavioural despair, increases aggression, and alters serotonin-related gene expression. To further investigate this paradigm, we studied depression/distress-associated markers of neuroinflammation, neuroplasticity, oxidative stress and the activity of glycogen synthase kinase-3 (GSK-3) isoforms in the hippocampus of male mice. Stressed mice exhibited a decreased density of Ki67-positive and DCX-positive cells in the subgranular zone of hippocampus, and altered expression of brain-derived neurotrophic factor (BDNF), its receptor TrkB, and anti-apoptotic protein kinase B phosphorylated at serine 473 (AktpSer473). The mice also exhibited increased densities of Iba-1-positive cells, increased oxidative stress, increased levels of interleukin-1ß (IL-1ß), interleukin-6 (IL-6) in the hippocampus and plasma, and elevated activity of GSK-3 isoforms. Together, the results of our investigation have revealed that unpredictable alternating ultrasound evokes behavioural and molecular changes that are characteristic of the depressive syndrome and validates this new and simple method of modeling emotional stress in rodents.


Assuntos
Transtorno Depressivo/fisiopatologia , Hipocampo/fisiopatologia , Inflamação/fisiopatologia , Plasticidade Neuronal/fisiologia , Estresse Psicológico/fisiopatologia , Estimulação Acústica/métodos , Animais , Fator Neurotrófico Derivado do Encéfalo/metabolismo , Modelos Animais de Doenças , Proteína Duplacortina , Quinase 3 da Glicogênio Sintase/metabolismo , Interleucina-1beta/metabolismo , Interleucina-6/metabolismo , Isoenzimas/metabolismo , Masculino , Camundongos Endogâmicos BALB C , Estresse Oxidativo/fisiologia , Receptor trkB/metabolismo , Ultrassom
6.
Prep Biochem Biotechnol ; 48(1): 1-5, 2018 Jan 02.
Artigo em Inglês | MEDLINE | ID: mdl-28976247

RESUMO

The effect of biologically active form (threo-Ds-) of isocitric acid (ICA) on oxidative stress was studied using the infusorian Paramecium caudatum stressed by hydrogen peroxide and salts of some heavy metals (Cu, Pb, Zn, and Cd). ICA at concentrations between 0.5 and 10 mM favorably influenced the infusorian cells with oxidative stress induced by the toxicants studied. The maximal antioxidant effect of ICA was observed at its concentration 10 mM irrespective of the toxicant used (either H2O2 or heavy metal ions). ICA was found to be a more active antioxidant than ascorbic acid. Biologically active pharmaceutically pure threo-Ds-ICA was produced through cultivation of the yeast Yarrowia lipolytica and isolated from the culture liquid in the form of crystalline monopotassium salt with a purity of 99.9%.


Assuntos
Isocitratos/metabolismo , Paramecium caudatum/metabolismo , Antioxidantes/metabolismo , Antioxidantes/farmacologia , Meios de Cultura/metabolismo , Peróxido de Hidrogênio/metabolismo , Isocitratos/farmacologia , Metais Pesados/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Paramecium caudatum/efeitos dos fármacos , Yarrowia/metabolismo
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