Critical Role of Astrocytic Polyamine and GABA Metabolism in Epileptogenesis

Accumulating evidence indicate that astrocytes are essential players of the excitatory and inhibitory signaling during normal and epileptiform activity via uptake and release of gliotransmitters, ions, and other substances. Polyamines can be regarded as gliotransmitters since they are almost exclusively stored in astrocytes and can be released by various mechanisms. The polyamine putrescine (PUT) is utilized to synthesize GABA, which can also be released from astrocytes and provide tonic inhibition on neurons. The polyamine spermine (SPM), synthesized form PUT through spermidine (SPD), is known to unblock astrocytic Cx43 gap junction channels and therefore facilitate astrocytic synchronization. In addition, SPM released from astrocytes may also modulate neuronal NMDA, AMPA, and kainate receptors. As a consequence, astrocytic polyamines possess the capability to significantly modulate epileptiform activity. In this study, we investigated different steps in polyamine metabolism and coupled GABA release to assess their potential to control seizure generation and maintenance in two different epilepsy models: the low-[Mg2+] model of temporal lobe epilepsy in vitro and in the WAG/Rij rat model of absence epilepsy in vivo. We show that SPM is a gliotransmitter that is released from astrocytes and significantly contributes to network excitation. Importantly, we found that inhibition of SPD synthesis completely prevented seizure generation in WAG/Rij rats. We hypothesize that this antiepileptic effect is attributed to the subsequent enhancement of PUT to GABA conversion in astrocytes, leading to GABA release through GAT-2/3 transporters. This interpretation is supported by the observation that antiepileptic potential of the Food and Drug Administration (FDA)-approved drug levetiracetam can be diminished by specifically blocking astrocytic GAT-2/3 with SNAP-5114, suggesting that levetiracetam exerts its effect by increasing surface expression of GAT-2/3. Our findings conclusively suggest that the major pathway through which astrocytic polyamines contribute to epileptiform activity is the production of GABA. Modulation of astrocytic polyamine levels, therefore, may serve for a more effective antiepileptic drug development in the future.

Microwave Soil Treatment along with Biochar Application Alleviates Arsenic Phytotoxicity and Reduces Rice Grain Arsenic Concentration

Rice grain arsenic (As) is a major pathway of human dietary As exposure. This study was conducted to reduce rice grain As concentration through microwave (MW) and biochar soil treatment. Collected soils were spiked to five levels of As concentration (As-0, As-20, As-40, As-60, and As-80 mg kg−1) prior to applying three levels of biochar (BC-0, BC-10, and BC-20 t ha−1) and three levels of MW treatment (MW-0, MW-3, and MW-6 min). The results revealed that MW soil treatment alleviates As phytotoxicity as rice plant growth and grain yield increase significantly and facilitate less grain As concentration compared with the control. For instance, the highest grain As concentration (912.90 µg kg−1) was recorded in the control while it was significantly lower (442.40 µg kg−1) in the MW-6 treatment at As-80. Although the BC-10 treatment had some positive effects, unexpectedly, BC-20 had a negative effect on plant growth, grain yield, and grain As concentration. The combination of BC-10 and MW-6 treatment was found to reduce grain As concentration (498.00 µg kg−1) compared with the control (913.7 µg kg−1). Thus, either MW-6 soil treatment alone or in combination with the BC-10 treatment can be used to reduce dietary As exposure through rice consumption. Nevertheless, further study is needed to explore the effectiveness and economic feasibility of this novel technique in field conditions.

A Kalirin missense mutation enhances dendritic RhoA signaling and leads to regression of cortical dendritic arbors across development

Normally, dendritic size is established prior to adolescence and then remains relatively constant into adulthood due to a homeostatic balance between growth and retraction pathways. However, schizophrenia is characterized by accelerated reductions of cerebral cortex gray matter volume and onset of clinical symptoms during adolescence, with reductions in layer 3 pyramidal neuron dendritic length, complexity, and spine density identified in multiple cortical regions postmortem. Nogo receptor 1 (NGR1) activation of the GTPase RhoA is a major pathway restricting dendritic growth in the cerebral cortex. We show that the NGR1 pathway is stimulated by OMGp and requires the Rho guanine nucleotide exchange factor Kalirin-9 (KAL9). Using a genetically encoded RhoA sensor, we demonstrate that a naturally occurring missense mutation in Kalrn, KAL-PT, that was identified in a schizophrenia cohort, confers enhanced RhoA activitation in neuronal dendrites compared to wild-type KAL. In mice containing this missense mutation at the endogenous locus, there is an adolescent-onset reduction in dendritic length and complexity of layer 3 pyramidal neurons in the primary auditory cortex. Spine density per unit length of dendrite is unaffected. Early adult mice with these structural deficits exhibited impaired detection of short gap durations. These findings provide a neuropsychiatric model of disease capturing how a mild genetic vulnerability may interact with normal developmental processes such that pathology only emerges around adolescence. This interplay between genetic susceptibility and normal adolescent development, both of which possess inherent individual variability, may contribute to heterogeneity seen in phenotypes in human neuropsychiatric disease.

Crosstalk Between Autophagy and the cGAS–STING Signaling Pathway in Type I Interferon Production

Innate immunity is the front-line defense against infectious microorganisms, including viruses and bacteria. Type I interferons are pleiotropic cytokines that perform antiviral, antiproliferative, and immunomodulatory functions in cells. The cGAS–STING pathway, comprising the main DNA sensor cyclic guanosine monophosphate/adenosine monophosphate synthase (cGAS) and stimulator of IFN genes (STING), is a major pathway that mediates immune reactions and is involved in the strong induction of type I IFN production, which can fight against microbial infections. Autophagy is an evolutionarily conserved degradation process that is required to maintain host health and facilitate capture and elimination of invading pathogens by the immune system. Mounting evidence indicates that autophagy plays an important role in cGAS–STING signaling pathway-mediated type I IFN production. This review briefly summarizes the research progress on how autophagy regulates the cGAS–STING pathway, regulating type I IFN production, with a particular focus on the crosstalk between autophagy and cGAS–STING signaling during infection by pathogenic microorganisms.

Transcriptional regulation of sphingolipid metabolism in budding yeast

Global control for the synthesis of lipids constituting a bilayer of cell membranes is known to be with a small number of transcription factors called master transcriptional regulators, which target a wide range of genes encoding lipid metabolism enzymes and/or their regulators. Although master transcriptional regulators of glycerophospholipids and sterols have been identified in both yeast and mammals, this aspect of sphingolipid metabolism is not yet understood. In the present study, we identified the C2H2-type zinc finger transcription factor, Com2, as a master transcriptional regulator of sphingolipid metabolism in the budding yeast, Saccharomyces cerevisiae. The target of rapamycin complex 2 (TORC2)-activated protein kinase Ypk1 is known to regulate sphingolipid metabolism. Activated Ypk1 stimulates the activity of serine palmitoyl transferase (SPT), the first-step enzyme in sphingolipid biosynthesis, by phosphorylating and inhibiting Orm1/2, a negative regulator of SPT. This regulation of SPT activity is thought to be a major pathway in the regulation of sphingolipid metabolism. In the present study, we found that inhibition of sphingolipid synthesis upregulates the expression of Com2, which in turn leads to the concomitant expression of Ypk1. The upregulation of Ypk1 expression was found to be dependent on a putative Com2-binding site in the YPK1 promoter. Our results also suggested that Com2 senses intracellular sphingolipid levels through a pathway independent of TORC2-Ypk1-mediated sensing of sphingolipids. Our results revealed an additional layer of mechanistic regulation that allows cells to maintain appropriate levels of sphingolipid biosynthesis and to rapidly induce this process in response to environmental stresses.

Proteolytic Transformation and Stimulation of SARSCov-2 Spike Protein with Human ACE-2 Receptor

Severe acute respiratory syndrome coronavirus has a great role in causing respiratory illness in humans and has the most important relationship of its spike proteins with host ACE-2 receptors. After entry into the human body, the viral S protein receptor-binding domain binds to human ACE-2 receptor. Two modes explained in this paper of an ACE-2 shedding. The shedding induces the process of viral entry to host cells by binding SARS-CoV-2 proteins. The residues of arginine and lysine in the ACE-2 receptor from 652 to 659 amino acid cleavage by ADAM17 but in TMPRSS2 the residues can be seen on amino acid from 697 to 716. Corona virus genome shows some structural proteins that are responsible for the cellular entry and facilitate the attachment of a virus to the host cell. Virus recognizes the attachment site and binds with it and enter into the cell. Spike protein is split from the cleavage site along its two subunits S1 and S2 then during this process. S2 subunit release RBD (Receptor- Binding Domain) of S1 mediated to the ACE-2. The RBD of S1 consists of 200 amino acid domains. The unknown protein B6ATI which is a neutral amino acid transporter located in ileum is the basic cause for formation of ACE-2 homodimer. In this way S1 domain provides site for another S2 domain. This leads to concealing of the ACE-2 ectodomain cleavage-sites, shedding. It prevents endocytosis of the receptor blocking a major pathway in the viral entry.

Caveolae respond to acute oxidative stress through membrane lipid peroxidation, cytosolic release of CAVIN1, and downstream regulation of NRF2

Caveolae have been linked to many biological functions, but their precise roles are unclear. Using quantitative whole cell proteomics of genome-edited cells, we show that the oxidative stress response is the major pathway dysregulated in cells lacking the key caveola structural protein, CAVIN1. CAVIN1 deletion compromised sensitivity to oxidative stress in cultured cells and in animals. Wound-induced accumulation of reactive oxygen species and apoptosis were suppressed in Cavin1-null zebrafish, negatively affecting regeneration. Oxidative stress triggered lipid peroxidation (LPO) and induced caveolar disassembly. The resulting release of CAVIN1 from caveolae allowed direct interaction between CAVIN1 and NRF2, a key regulator of the antioxidant response, facilitating NRF2 degradation. CAVIN1-null cells with impaired negative regulation of NRF2 showed resistance to LPO-induced ferroptosis. Thus, caveolae, via LPO and CAVIN1 release, maintain cellular susceptibility to oxidative stress-induced cell death demonstrating a crucial role for this enigmatic organelle in cellular homeostasis and wound response.

Mitotic inactivation of the cGAS‒MITA/STING pathways

The cyclic GMP‒AMP synthase (cGAS)‒mediator of interferon response factor 3 activation/stimulator of interferon genes (MITA/STING) axis has emerged as a major pathway, which senses microbial or mis-located cellular DNA in the cytosol to trigger innate immune responses. cGAS senses cytosolic DNA without a preference of self or non-self DNA. How the cGAS‒MITA/STING axis is inactivated upon nuclear membrane breakdown (NEBD) at mitotic entry in vertebrate cells to avoid self DNA sensing remains unclear until very recently. In this review, we summarize the recent advances on how cGAS responds to chromosomes upon NEBD and the mechanisms involved in the inactivation of the cGAS‒MITA/STING pathways in mitosis.

Riverine antibacterial resistance gradient determined by environmental factors

Wastewater emission to surface waters is a major pathway for antibacterials and antibacterial-resistant bacteria. Polluted waterbodies such as rivers provide a reservoir for bacterial resistance. We studied water quality and bacterial antibacterial resistance along the subtropical Qishan River in Taiwan as a case study of environmental resistance spread in a pristine to rural area. Human settlement densities increased generally from pristine mountain sites to the more polluted lowlands generally. Accordingly, as a working hypothesis, we expected antibacterial resistance level to increase towards downstream. We collected sediment samples from 8 stations along the Qishan river and where the Qishan river reaches the Kaoping river. The samples were processed in the lab for bacteriological and physicochemical analysis. Antibacterial resistance was tested by disk diffusion and micro-dilution with ten common antibacterials. A comparison was made among the sites where isolates began to occur at the upstream (site 1–6) with the downstream, including site 7 (Qishan town), site 8 (wastewater treatment plant) and site 9 (Kaoping river). The results of multivariate analysis for bacteriological and physicochemical parameters showed increasing water pollution levels downstream of the Qishan river. Ten bacteria including Escherichia coli, Klebsiella pneumoniae, Serratia marcescens, Enterobacter sp., Acinetobacter sp., Staphylococcus spp. and Bacillus spp. were analyzed and tested in the study. Their percentage of occurrence varied at each site. The resistance level was determined from the growth inhibition zone diameter (disk diffusion) and the minimum inhibitory concentration (micro-dilution). The results indicated that antibacterial resistance was related to certain environmental factors. Besides, the usage pattern of different classes of antibacterials in different places could alter trends of their resistance. Bacteria were found with increased resistance to antibacterials used in agriculture through the downstream sites according to the results. The WWTP emitting wastewater was demonstrated to be a hotspot of resistance in aquatic environments. In conclusion, bacterial resistance against antibacterials from the Qishan river has become a potential public health threat. This study could assist authorities by providing a reference for water quality risk assessment and management in Kaohsiung city and southern Taiwan.

Riverine antibacterial resistance gradient determined by environmental factors

Wastewater emission to surface waters is a major pathway for antibacterials and antibacterial-resistant bacteria. Polluted waterbodies such as rivers provide a reservoir for bacterial resistance. We studied water quality and bacterial antibacterial resistance along the subtropical Qishan River in Taiwan as a case study of environmental resistance spread in a pristine to rural area. Human settlement densities increased generally from pristine mountain sites to the more polluted lowlands generally. Accordingly, as a working hypothesis, we expected antibacterial resistance level to increase towards downstream. We collected sediment samples from 8 stations along the Qishan river and where the Qishan river reaches the Kaoping river. The samples were processed in the lab for bacteriological and physicochemical analysis. Antibacterial resistance was tested by disk diffusion and micro-dilution with ten common antibacterials. A comparison was made among the sites where isolates began to occur at the upstream (site 1-6) with the downstream, including site 7 (Qishan town), site 8 (wastewater treatment plant) and site 9 (Kaoping river). The results of multivariate analysis for bacteriological and physicochemical parameters showed increasing water pollution levels downstream of the Qishan river. Ten bacteria including Escherichia coli, Klebsiella pneumoniae, Serratia marcescens, Enterobacter sp., Acinetobacter sp., Staphylococcus spp. and Bacillus spp. were analyzed and tested in the study. Their percentage of occurrence varied at each site. The resistance level was determined from the growth inhibition zone diameter (disk diffusion) and the minimum inhibitory concentration (micro-dilution). The results indicated that antibacterial resistance was related to certain environmental factors. Besides, the usage pattern of different classes of antibacterials in different places could alter trends of their resistance. Bacteria were found with increased resistance to antibacterials used in agriculture through the downstream sites according to the results. The WWTP emitting wastewater was demonstrated to be a hotspot of resistance in aquatic environments. In conclusion, bacterial resistance against antibacterials from the Qishan river has become a potential public health threat. This study could assist authorities by providing a reference for water quality risk assessment and management in Kaohsiung city and southern Taiwan.