Effect of Nigella sativa on Selected Gastrointestinal Diseases.

Nigella sativa L. (family Ranunculaceae), also known as black cumin, has been used in cuisine around the world for many years. Due to its health-promoting properties, it can be used not only in the food industry but also in medicine. The main bioactive compound contained in the black cumin extract is thymoquinone (TQ), which has a special therapeutic role. The results of research in recent years confirmed its hypoglycemic, hypolipemic, and hepatoprotective effects, among others. In addition, the results of laboratory tests also indicate its immunomodulatory and anticancer effects, although there is still a lack of data on the mechanisms of how they are involved in the fight against cancer. Including this plant material in one's diet can be both an element of prophylaxis and therapy supporting the treatment process, including pharmacological treatment. However, attention should be paid to its potential interactions with drugs used in the treatment of chronic diseases.

Perturbation of astroglial Slc38 glutamine transporters by NH4+ contributes to neurophysiologic manifestations in acute liver failure.

Ammonia is considered the main pathogenic toxin in hepatic encephalopathy (HE). However, the molecular mechanisms involved have been disputed. As altered glutamatergic and GABAergic neurotransmission has been reported in HE, we investigated whether four members of the solute carrier 38 (Slc38) family of amino acid transporters-involved in the replenishment of glutamate and GABA-contribute to ammonia neurotoxicity in HE. We show that ammonium ion exerts multiple actions on the Slc38 transporters: It competes with glutamine for the binding to the system N transporters Slc38a3 and Slc38a5, consequently inhibiting bidirectional astroglial glutamine transport. It also competes with H+ , Na+ , and K+ for uncoupled permeation through the same transporters, which may perturb astroglial intracellular pH, membrane potential, and K+ -buffering. Knockdown of Slc38a3 in mice results in cerebral cortical edema and disrupted neurotransmitter synthesis mimicking events contributing to HE development. Finally, in a mouse model of acute liver failure (ALF), we demonstrate the downregulation of Slc38a3 protein, impeded astroglial glutamine release, and cytotoxic edema. Altogether, we demonstrate contribution of Slc38 transporters to the ammonia-induced impairment of glutamine recycling between astrocytes and neurons, a phenomenon underlying acute ammonia neurotoxicity in the setting of ALF.

Mitochondrial Changes in Rat Brain Endothelial Cells Associated with Hepatic Encephalopathy: Relation to the Blood-Brain Barrier Dysfunction.

The mechanisms underlying cerebral vascular dysfunction and edema during hepatic encephalopathy (HE) are unclear. Blood-brain barrier (BBB) impairment, resulting from increased vascular permeability, has been reported in acute and chronic HE. Mitochondrial dysfunction is a well-documented result of HE mainly affecting astrocytes, but much less so in the BBB-forming endothelial cells. Here we review literature reports and own experimental data obtained in HE models emphasizing alterations in mitochondrial dynamics and function as a possible contributor to the status of brain endothelial cell mitochondria in HE. Own studies on the expression of the mitochondrial fusion-fission controlling genes rendered HE animal model-dependent effects: increase of mitochondrial fusion controlling genes opa1, mfn1 in cerebral vessels in ammonium acetate-induced hyperammonemia, but a decrease of the two former genes and increase of fis1 in vessels in thioacetamide-induced HE. In endothelial cell line (RBE4) after 24 h ammonia and/or TNFα treatment, conditions mimicking crucial aspects of HE in vivo, we observed altered expression of mitochondrial fission/fusion genes: a decrease of opa1, mfn1, and, increase of the fission related fis1 gene. The effect in vitro was paralleled by the generation of reactive oxygen species, decreased total antioxidant capacity, decreased mitochondrial membrane potential, as well as increased permeability of RBE4 cell monolayer to fluorescein isothiocyanate dextran. Electron microscopy documented enlarged mitochondria in the brain endothelial cells of rats in both in vivo models. Collectively, the here observed alterations of cerebral endothelial mitochondria are indicative of their fission, and decreased potential of endothelial mitochondria are likely to contribute to BBB dysfunction in HE.

The Effect of TGF-ß1 Reduced Functionality on the Expression of Selected Synaptic Proteins and Electrophysiological Parameters: Implications of Changes Observed in Acute Hepatic Encephalopathy.

Decreased platelet count represents a feature of acute liver failure (ALF) pathogenesis. Platelets are the reservoir of transforming growth factor 1 (TGF-ß1), a multipotent cytokine involved in the maintenance of, i.a., central nervous system homeostasis. Here, we analyzed the effect of a decrease in TGF-ß1 active form on synaptic proteins levels, and brain electrophysiology, in mice after intraperitoneal (ip) administration of TGF-ß1 antibody (anti-TGF-ß1; 1 mg/mL). Next, we correlated it with a thrombocytopenia-induced TGF-ß1 decrease, documented in an azoxymethane-induced (AOM; 100 mM ip) model of ALF, and clarified the impact of TGF-ß1 decrease on blood-brain barrier functionality. The increase of both synaptophysin and synaptotagmin in the cytosolic fraction, and its reduction in a membrane fraction, were confirmed in the AOM mice brains. Both proteins' decrease in analyzed fractions occurred in anti-TGF-ß1 mice. In turn, an increase in postsynaptic (NR1 subunit of N-methyl-D-aspartate receptor, postsynaptic density protein 95, gephyrin) proteins in the AOM brain cortex, but a selective compensatory increase of NR1 subunit in anti-TGF-ß mice, was observed. The alterations of synaptic proteins levels were not translated on electrophysiological parameters in the anti-TGF-ß1 model. The results suggest the impairment of synaptic vesicles docking to the postsynaptic membrane in the AOM model. Nevertheless, changes in synaptic protein level in the anti-TGF-ß1 mice do not affect neurotransmission and may not contribute to neurologic deficits in AOM mice.

Subcritical water extraction of bioactive phenolic compounds from distillery stillage.

Distilleries generate huge amounts of by-products that have a negative impact on the environment, so the management of wastes generated by this sector should be improved. Because distillery by-products are a source of bioactive compounds, the recovery of these compounds not only reduces issues with environmental protection but also provides the basis for a waste-to-profit solution. Following the latest trends in the search for so-called green extraction techniques for recovering valuable products, this study investigated the effect of subcritical water extraction (SWE) conditions (temperature (25-260 °C), time (5-90 min), and solid-to-solvent ratio (1:5-1:50, w:v)) on the efficiency of recovery of bioactive compounds, i.e., polyphenols from distillery stillage, and on the antioxidant activity of the extracts. The highest extraction yield was obtained with 30-min SWE with a solid-to-solvent ratio of 1:15 at either 140 °C (for total polyphenol content and phenolic acid content) or 200 °C (for total flavonoid content), as indicated by the Response Surface Methodology analysis. Phenolic acids in the extracts were present mainly in free forms (up to 88% of the total content). The antioxidant activity, which was measured using several assays, correlated positively with the content of phenolic acids, which confirmed their significant contribution to the bioactive properties of the extracts. The antioxidant effects of the extracts were mostly due to hydroxycinnamic acids (especially ferulic and p-coumaric acids). Principal component analysis showed that the temperature and time of SWE were the factors that can explain the greatest amount of variation in the extraction yield, composition, and bioactive properties of the polyphenols. These results will influence the design of further processes, such as purification and concentration, which are necessary before using the extracted compounds as substrates that are applicable in various industries. Based on the analysis of the elemental composition, the biomass remaining after SWE was evaluated to consider the possibilities of its further utilization.

Valorization of distillery stillage by polyphenol recovery using microwave-assisted, ultrasound-assisted and conventional extractions.

An increasing understanding of the negative environmental effects of waste discharges has made valorization of distillery by-products to recover added-value compounds a sound option for distillery stillage management. In this study, the recovery of bioactive compounds, i.e. polyphenols, from distillery stillage was performed by microwave-assisted extraction (MAE), ultrasound-assisted extraction (UAE) and conventional solid-liquid extraction (CSLE) to investigate the effects of extraction time, the concentration of ethyl acetate (EA) in a solvent mixture with ethanol and water, and solid-to-solvent ratio on the recovery yield and antioxidant activity of the extracts. The highest yields of total polyphenol content (TPC) (3.73 mg gallic acid equivalent/g) and phenolic acid content (2.51 µg/g) were obtained with 8-min MAE with 70% EA. MAE provided 1.2- and 1.4-times higher yield of phenolic acids and 1.2- and 1.6-times higher antioxidant activity than UAE and CSLE, respectively. Due to the approximately 3-times higher rate of extraction, the ratio between energy consumption and extraction yield was better in MAE than in UAE. Principal component analysis (PCA) showed that the antioxidant activity of the extracts was positively correlated with TPC and phenolic acid content. Six phenolic acids that were identified were present mainly in their free forms (up to 95% of the total), with a predominance of ferulic (up to 0.80 µg/g) and p-coumaric (up to 0.72 µg/g) acids.

The effect of high humidity hot air impingement blanching on the changes in cell wall polysaccharides and phytochemicals of okra pods.

BACKGROUND: Okra pods contain heat-sensitive substances, such as phenolic compounds and other phytochemicals that can be degraded when okra pods are subjected to heat treatment. The understanding of the impact of high humidity hot air impingement blanching (HHAIB) on the changes in physicochemical properties of polysaccharides and phytochemicals of okra pods is of great importance because over-blanching may result in cell membrane disruption and changes in biologically active compounds under prolonged exposure to the thermal treatment. Therefore, the present study aimed to investigate the effect of HHAIB on the changes in physicochemical properties of pectins and phytochemicals extracted from okra pods. RESULTS: Both the HHAIB time and method of extraction influenced their physicochemical characteristics and biological activity. Pectin fractions subjected to HHAIB were composed of polygalacturonic acid, rhamnogalacturonan, glucomannan, galactan, mannose, arabinose, rhamnose, calcium pectate and arabinogalactan. The contents of total phenolics, total flavonoids and antioxidant activity of extracts mostly increased during HHAIB (i.e. up to 19.0%, 13.2% and 35.3%, respectively). However, HHAIB reduced the chlorophyll-a (up to 55.7%) and lycopene (up to 52.6%) contents of okra pods. CONCLUSION: The acquired knowledge may be useful for better understanding and optimization of technologies based on HHAIB treatment. The HHAIB treated okra can be a promising natural alternative in different applications, including its use as a replacement of some ingredients in food or non-food systems as a result of richness in polysaccharides and polyphenols, as well as high antioxidant properties. © 2022 Society of Chemical Industry.

Insight into microRNAs-Mediated Communication between Liver and Brain: A Possible Approach for Understanding Acute Liver Failure?

Acute liver failure (ALF) is a life-threatening consequence of hepatic function rapid loss without preexisting liver disease. ALF may result in a spectrum of neuropsychiatric symptoms that encompasses cognitive impairment, coma, and often death, collectively defined as acute hepatic encephalopathy. Micro RNAs are small non-coding RNAs that modulate gene expression and are extensively verified as biomarker candidates in various diseases. Our systematic literature review based on the last decade's reports involving a total of 852 ALF patients, determined 205 altered circulating miRNAs, of which 25 miRNAs were altered in the blood, regardless of study design and methodology. Selected 25 miRNAs, emerging predominantly from the analyses of samples obtained from acetaminophen overdosed patients, represent the most promising biomarker candidates for a diagnostic panel for symptomatic ALF. We discussed the role of selected miRNAs in the context of tissue-specific origin and its possible regulatory role for molecular pathways involved in blood-brain barrier function. The defined several common pathways for 15 differently altered miRNAs were relevant to cellular community processes, indicating loss of intercellular, structural, and functional components, which may result in blood-brain barrier impairment and brain dysfunction. However, a causational relationship between circulating miRNAs differential expression, and particular clinical features of ALF, has to be demonstrated in a further study.

The Role of Nrf2 Transcription Factor and Sp1-Nrf2 Protein Complex in Glutamine Transporter SN1 Regulation in Mouse Cortical Astrocytes Exposed to Ammonia.

Ammonia toxicity in the brain primarily affects astrocytes via a mechanism in which oxidative stress (OS), is coupled to the imbalance between glutamatergic and GABAergic transmission. Ammonia also downregulates the astrocytic N system transporter SN1 that controls glutamine supply from astrocytes to neurons for the replenishment of both neurotransmitters. Here, we tested the hypothesis that activation of Nrf2 is the process that links ammonia-induced OS formation in astrocytes to downregulation and inactivation of SN1 and that it may involve the formation of a complex between Nrf2 and Sp1. Treatment of cultured cortical mouse astrocytes with ammonia (5 mM NH4Cl for 24 h) evoked Nrf2 nuclear translocation, increased its activity in a p38 MAPK pathway-dependent manner, and enhanced Nrf2 binding to Slc38a3 promoter. Nrf2 silencing increased SN1 mRNA and protein level without influencing astrocytic [3H]glutamine transport. Ammonia decreased SN1 expression in Nrf2 siRNA treated astrocytes and reduced [3H]glutamine uptake. In addition, while Nrf2 formed a complex with Sp1 in ammonia-treated astrocytes less efficiently than in control cells, treatment of astrocytes with hybrid-mode inactivated Sp1-Nrf2 complex (Nrf2 silencing + pharmacological inhibition of Sp1) did not affect SN1 protein level in ammonia-treated astrocytes. In summary, the results document that SN1 transporter dysregulation by ammonia in astrocytes involves activation of Nrf2 but does not require the formation of the Sp1-Nrf2 complex.

Decreased Expression and Uncoupling of Endothelial Nitric Oxide Synthase in the Cerebral Cortex of Rats with Thioacetamide-Induced Acute Liver Failure.

Acute liver failure (ALF) is associated with deregulated nitric oxide (NO) signaling in the brain, which is one of the key molecular abnormalities leading to the neuropsychiatric disorder called hepatic encephalopathy (HE). This study focuses on the effect of ALF on the relatively unexplored endothelial NOS isoform (eNOS). The cerebral prefrontal cortices of rats with thioacetamide (TAA)-induced ALF showed decreased eNOS expression, which resulted in an overall reduction of NOS activity. ALF also decreased the content of the NOS cofactor, tetrahydro-L-biopterin (BH4), and evoked eNOS uncoupling (reduction of the eNOS dimer/monomer ratio). The addition of the NO precursor L-arginine in the absence of BH4 potentiated ROS accumulation, whereas nonspecific NOS inhibitor L-NAME or EDTA attenuated ROS increase. The ALF-induced decrease of eNOS content and its uncoupling concurred with, and was likely causally related to, both increased brain content of reactive oxidative species (ROS) and decreased cerebral cortical blood flow (CBF) in the same model.

Review of recent applications and research progress in hybrid and combined microwave-assisted drying of food products: Quality properties.

The growing concerns over product quality have increased demand for high quality dried food products and encouraged researchers to explore and producers of such products to implement novel microwave (MW)-assisted drying methods. This paper presents a critical review of the key principles and drawbacks of MW-assisted drying as well as needs for future research. In this article, recent research into application of microwaves as an alternative heat source, applications and progress in hybrid MW-assisted drying that rely on various drying media and combined two or three stages of MW-assisted drying for the preservation of food products is reviewed critically. The effect of different MW-assisted drying methods, conditions and initial pretreatments on the thermophysical properties, color, nutritional value and rehydration potential of dried food products is discussed in detail along with the discussion on how the material properties evolve and change in structure, color, and composition during MW-assisted drying and recent attempts at mathematical modeling of these changes made for different fruits and vegetables. It should be noted that most of the published results were obtained in laboratory-scale dryers. Pilot-scale testing is needed to bridge the gap between laboratory research and industrial applications to fulfill the potential for novel hybrid and combined MW-assisted drying methods and to expand their role in food processing.

Emerging chemical and physical disinfection technologies of fruits and vegetables: a comprehensive review.

With a growing demand for safe, nutritious, and fresh-like produce, a number of disinfection technologies have been developed. This review comprehensively examines the working principles and applications of several emerging disinfection technologies. The chemical treatments, including chlorine dioxide, ozone, electrolyzed water, essential oils, high-pressure carbon dioxide, and organic acids, have been improved as alternatives to traditional disinfection methods to meet current safety standards. Non-thermal physical treatments, such as UV-light, pulsed light, ionizing radiation, high hydrostatic pressure, cold plasma, and high-intensity ultrasound, have shown significant advantages in improving microbial safety and maintaining the desirable quality of produce. However, using these disinfection technologies alone may not meet the requirement of food safety and high product quality. Several hurdle technologies have been developed, which achieved synergistic effects to maximize lethality against microorganisms and minimize deterioration of produce quality. The review also identifies further research opportunities for the cost-effective commercialization of these technologies.

The Status of Bile Acids and Farnesoid X Receptor in Brain and Liver of Rats with Thioacetamide-Induced Acute Liver Failure.

Acute liver failure (ALF) leads to neurological symptoms defined as hepatic encephalopathy (HE). Although accumulation of ammonia and neuroinflammation are generally accepted as main contributors to HE pathomechanism, a buildup of bile acids (BA) in the blood is a frequent component of liver injury in HE patients. Recent studies have identified the nuclear farnesoid X receptor (FXR) acting via small heterodimer partner (SHP) as a mediator of BA-induced effects in the brain of ALF animals. The present study investigated the status of the BA-FXR axis in the brain and the liver, including selective changes in pertinent genes in thioacetamide (TAA)-induced ALF in Sprague-Dawley rats. FXR was found in rat neurons, confirming earlier reports for mouse and human brain. BA accumulated in blood but not in the brain tissue. Expression of mRNAs coding for Fxr and Shp was reduced in the hippocampus and of Fxr mRNA also in the cerebellum. Changes in Fxr mRNA levels were not followed by changes in FXR protein. The results leave open the possibility that mobilization of the BA-FXR axis in the brain may not be necessarily pathognomonic to HE but may depend upon ALF-related confounding factors.

TNFα increases STAT3-mediated expression of glutaminase isoform KGA in cultured rat astrocytes.

Glutamate related excitotoxicity and excess of cerebral levels of tumor necrosis factor alpha (TNFα) are interrelated and well documented abnormalities noticed in many central nervous system diseases. Contribution of kidney type glutaminase (KGA) and shorter alternative splicing form (GAC) to glutamine degradation in astrocytes has been recently a matter of dispute and extensive study but the regulation of the GLS isoforms by inflammatory factors is still not well known. Here we show that treatment of cultured rat cortical astrocytes with pathophysiologically relevant (50 ng/ml) concentration of TNFα specifically increases the expression of KGA but not GAC and increases activity of GLS. No changes in the expression of either of two GLS isoforms were observed following treatment with other tested cytokines IL-1ß and IL-6. The TNFα mediated KGA expression was associated with increased phosphorylation of signal transducer and activator of transcription 3 (STAT3). Stimulatory effect of TNF-α on KGA expression was reduced by selective inhibition of (STAT3) but not by inhibition of STAT1 nor nuclear transcription factor kappa. Additionally, the role of miRNA in TNFα-induced expression of KGA in astrocytes was excluded, since the expression of miR-23a/b and miR-200c, potential regulators of KGA expression, was unchanged. This study documents increased KGA expression in the astrocytes under inflammatory stimulation, identifying TNFα as a cytokine mediating this response, and demonstrates the specific and selective involvement of STAT3.

Silencing of Transcription Factor Sp1 Promotes SN1 Transporter Regulation by Ammonia in Mouse Cortical Astrocytes.

The involvement of the astrocytic SN1 (SNAT3) transporter in ammonia-induced l-glutamine retention was recently documented in mouse-cultured astrocytes. Here we investigated the involvement of specificity protein 1 (Sp1) transcription factor in SN1 regulation in ammonium chloride ("ammonia")-treated astrocytes. Sp1 expression and its cellular localization were determined using real-time qPCR, Western blot, and confocal microscopy. Sp1 binding to Snat3 promoter was analyzed by chromatin immunoprecipitation. The role of Sp1 in SN1 expression and SN1-mediated [³H]glutamine uptake in ammonia-treated astrocytes was verified using siRNA and mithramycin A. The involvement of protein kinase C (PKC) isoforms in Sp1 level/phosphorylation status was verified using siRNA technology. Sp1 translocation to the nuclei and its enhanced binding to the Snat3 promoter, along with Sp1 dependence of system N-mediated [³H]glutamine uptake, were observed in astrocytes upon ammonia exposure. Ammonia decreased the level of phosphorylated Sp1, and the effect was reinforced by long-term incubation with PKC modulator, phorbol 12-myristate 13-acetate, which is a treatment likely to dephosphorylate Sp1. Furthermore, silencing of the PKCδ isoform appears to enhance the ammonia effect on the Sp1 level. Collectively, the results demonstrate the regulatory role of Sp1 in regulation of SN1 expression and activity in ammonia-treated astrocytes and implicate altered Sp1 phosphorylation status in this capacity.

NMDA Receptors in Astrocytes: In Search for Roles in Neurotransmission and Astrocytic Homeostasis.

Studies of the last two decades have demonstrated the presence in astrocytic cell membranes of N-methyl-d-aspartate (NMDA) receptors (NMDARs), albeit their apparently low abundance makes demonstration of their presence and function more difficult than of other glutamate (Glu) receptor classes residing in astrocytes. Activation of astrocytic NMDARs directly in brain slices and in acutely isolated or cultured astrocytes evokes intracellular calcium increase, by mutually unexclusive ionotropic and metabotropic mechanisms. However, other than one report on the contribution of astrocyte-located NMDARs to astrocyte-dependent modulation of presynaptic strength in the hippocampus, there is no sound evidence for the significant role of astrocytic NMDARs in astrocytic-neuronal interaction in neurotransmission, as yet. Durable exposure of astrocytic and neuronal co-cultures to NMDA has been reported to upregulate astrocytic synthesis of glutathione, and in this way to increase the antioxidative capacity of neurons. On the other hand, overexposure to NMDA decreases, by an as yet unknown mechanism, the ability of cultured astrocytes to express glutamine synthetase (GS), aquaporin-4 (AQP4), and the inward rectifying potassium channel Kir4.1, the three astroglia-specific proteins critical for homeostatic function of astrocytes. The beneficial or detrimental effects of astrocytic NMDAR stimulation revealed in the in vitro studies remain to be proven in the in vivo setting.

Cerebrovascular reactivity and cerebral perfusion of rats with acute liver failure: role of L-glutamine and asymmetric dimethylarginine in L-arginine-induced response.

Cerebral blood flow (CBF) is impaired in acute liver failure (ALF), however, the complexity of the underlying mechanisms has often led to inconclusive interpretations. Regulation of CBF depends at least partially on variations in the local brain L-arginine concentration and/or its metabolic rate. In ALF, other factors, like an increased concentration of asymmetric dimethylarginine (ADMA), an endogenous nitric oxide synthase inhibitor and elevated level of L-glutamine, may contribute to CBF alteration. This study demonstrated strong differences in the reactivity of the middle cerebral arteries and their response to extravascular L-arginine application between vessels isolated from rats with thioacetamide (TAA)-induced ALF and control animals. Our results also showed the decrease in the cerebral perfusion in TAA rats measured by arterial spin labeling perfusion magnetic resonance. Subsequently, we aimed to investigate the importance of balance between the concentration of ADMA and L-arginine in the CBF regulation. In vivo, intraperitoneal L-arginine administration in TAA rats corrected: (i) decrease in cerebral perfusion, (ii) decrease in brain extracellular L-arginine/ADMA ratio and (iii) increase in brain L-glutamine concentration. Our study implicates that impaired vascular tone of cerebral arteries is most likely associated with exposure to high ADMA and L-glutamine levels resulting in limited availability of L-arginine and might be responsible for reduced cerebral perfusion observed in ALF.

Microbial composition of biofilm treating wastewater rich in bisphenol A.

Although microbial degradation plays a major role in the removal of bisphenol A (BPA) from water environments, there is little information on the effect of BPA on microorganisms in wastewater treatment systems. The aim of this study was to determine the dynamics of the microbial communities in biofilm growing on porous ceramic supports in a column up-flow reactor during exposure to BPA at increasing concentrations from 0 to 10 mg L-1. Independent of BPA load, the efficiency of BPA removal was about 90%. Groups of microorganisms that differ in their sensitivity to the presence of BPA in wastewater were identified. The core microbial genera in the biofilm were Acidovorax, Pseudoxanthomonas and Acinetobacter. Arenimonas sp., Thauera sp. and Acidobacterium sp. were the main components of the biofilm in the absence of BPA in wastewater. Increased abundances of Pseudomonas sp., Acidovorax sp. and Luteimonas sp. in BPA-exposed biofilm indicate that these genera may have played important roles in BPA biodegradation. A correlation between Pseudomonas sp. abundance and BPA removal efficiency indicates that BPA was used directly as a source of carbon and energy for growth. This study indicates that the use of the biofilm reactor enables effective BPA removal from wastewater and expands knowledge about the microbial structure of communities responsible for BPA degradation.

Mechanisms of Excessive Extracellular Glutamate Accumulation in Temporal Lobe Epilepsy.

There is compelling evidence that initiation and maintenance of epileptic seizures in temporal lobe epilepsy (TLE) is facilitated by excessive accumulation in the extracellular (perisynaptic) space of the excitatory neurotransmitter glutamate (Glu). This review discusses the mechanisms underlying this phenomenon. Glu released from neurons is taken up by astrocytes and activated there by glutamine synthetase (GS) to form glutamine (Gln) which upon entry to neurons is degraded back to Glu by phosphate-activated glutaminase (PAG): this chain of reactions has been defined as the glutamine/glutamate/cycle (GGC). In the initial phase of epileptogenesis, increased Glu supply is a consequence of activation of its turnover in GGC by Glu released by a primary chemical or physical stimulus. In chronic TLE, profound astrogliosis and demise of neurons which culminate in hippocampal sclerosis, are associated with changes in GGC which act in concert towards increasing the extracellular Glu concentration. Deficiency of GS and of the astrocytic Glu transporter, GLT-1, impede Glu inactivation, whereas Glu release from neurons appears facilitated by activation of PAG and increased activity of the neuronal Glu transporter EAAC1. Conclusions derived from measurements of activities/expression patterns of the GGC enzymes and transporter moieties find support in metabolic studies employing 13C labeled Glu precursors. Glu reuptake by astrocytes is additionally impeded by unfavorable ion gradients resulting from ion and water dyshomeostasis, and extracellular Glu concentration is further increased by reduction of extracellular space due to edema and altered cytoarchitecture of the hippocampus. Missing links in the scenario are discussed in concluding comments.