The preventive effect of Gastrodia elata Blume extract on vancomycin-induced acute kidney injury in rats.

BACKGROUND: Gastrodia elata Blume (GEB), a traditional medicinal herb, has been reported to have pharmacological effect including protection against liver, neuron and kidney toxicity. However, explanation of its underlying mechanisms remains a great challenge. This study investigated the protective effects of GEB extract on vancomycin (VAN)-induced nephrotoxicity in rats and underlying mechanisms with emphasis on the anti-oxidative stress, anti-inflammation and anti-apoptosis. The male Sprague-Dawley rats were randomly divided three groups: control (CON) group, VAN group and GEB group with duration of 14 days. RESULTS: The kidney weight and the serum levels of blood urea nitrogen and creatinine in the GEB group were lower than the VAN group. Histological analysis using hematoxylin & eosin and periodic acid Schiff staining revealed pathological changes of the VAN group. Immunohistochemical analysis revealed that the expression levels of N-acetyl-D-glucosaminidase, myeloperoxidase and tumor necrosis factor-alpha in the GEB group were decreased when compared with the VAN group. The number of terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling-positive cells, phosphohistone and malondialdehyde levels were lower in the GEB group than VAN group. The levels of total glutathione in the GEB group were higher than the VAN group. CONCLUSIONS: The findings of this study suggested that GEB extract prevents VAN-induced renal tissue damage through anti-oxidation, anti-inflammation and anti-apoptosis.

Proximity Proteome Analysis Reveals Novel TREM2 Interactors in the ER-Mitochondria Interface of Human Microglia.

Triggering receptor expressed on myeloid cells 2 (TREM2) plays a central role in microglial biology and the pathogenesis of Alzheimer's disease (AD). Besides DNAX-activating protein 12 (DAP12), a communal adaptor for TREM2 and many other receptors, other cellular interactors of TREM2 remain largely elusive. We employed a 'proximity labeling' approach using a biotin ligase, TurboID, for mapping protein-protein interactions in live mammalian cells. We discovered novel TREM2-proximal proteins with diverse functions, including those localized to the Mitochondria-ER contact sites (MERCs), a dynamic subcellular 'hub' implicated in a number of crucial cell physiology such as lipid metabolism. TREM2 deficiency alters the thickness (inter-organelle distance) of MERCs, a structural parameter of metabolic state, in microglia derived from human induced pluripotent stem cells. Our TurboID-based TREM2 interactome study suggest novel roles for TREM2 in the structural plasticity of the MERCs, raising the possibility that dysregulation of MERC-related TREM2 functions contribute to AD pathobiology.

A distinct astrocyte subtype in the aging mouse brain characterized by impaired protein homeostasis.

The aging brain exhibits a region-specific reduction in synapse number and plasticity. Although astrocytes play central roles in regulating synapses, it is unclear how changes in astrocytes contribute to age-dependent cognitive decline and vulnerability to neurodegenerative diseases. Here, we identified a unique astrocyte subtype that exhibits dysregulated autophagy and morphology in aging hippocampus. In these autophagy-dysregulated astrocytes (APDAs), autophagosomes abnormally accumulate in swollen processes, impairing protein trafficking and secretion. We found that reduced mammalian target of rapamycin (mTOR) and proteasome activities with lysosomal dysfunction generate APDAs in an age-dependent manner. Secretion of synaptogenic molecules and astrocytic synapse elimination were significantly impaired in APDAs, suggesting that APDAs have lost their ability to control synapse number and homeostasis. Indeed, excitatory synapses and dendritic spines associated with APDAs were significantly reduced. Finally, we found that mouse brains with Alzheimer's disease showed a significantly accelerated increase in APDAs, suggesting potential roles for APDAs in age- and Alzheimer's disease-related cognitive decline and synaptic pathology.

Enhancement of catabolite regulatory genes in Saccharomyces cerevisiae to increase ethanol production using hydrolysate from red seaweed Gloiopeltis furcata.

Glucose and galactose are monosaccharides obtained from Gloiopeltis furcata (Red algae). A total monosaccharide yield of 62.3 g/L was obtained from G. furcata using thermal acid hydrolysis and enzymatic saccharification. Activated carbon was used to eliminate hydroxymethylfurfural (HMF) from the hydrolysate. Previously obtained monosaccharides are used for ethanol production by Saccharomyces cerevisiae. S. cerevisiae consumes glucose first, then galactose. The methods for reducing fermentation time and increasing the ethanol yield coefficient using the simultaneous consumption of glucose and galactose have been evaluated. Gal3p and Gal80p of S. cerevisiae act as signal transducers that govern the galactose inducer Gal4p mediated transcriptional activation of the Gal gene family. Gal80p binds to Gal4p for transcription deactivation. Therefore, Gal80p was deleted for Gal4p expression without interruption.

Enhancement of Galactose Uptake from Kappaphycus alvarezii Using Saccharomyces cerevisiae through Deletion of Negative Regulators of GAL Genes.

This study was aimed at enhancing galactose consumption from the red seaweed Kappaphycus alvarezii. The optimal pretreatment condition of thermal acid hydrolysis was treated with 350 mM HNO3 for 60 min at 121 °C. The enzymatic saccharification with a 1:1 mixture of Celluclast 1.5 L and Viscozyme L showed the maximum yield of glucose; 42-g/L monosaccharide concentration was obtained with the highest yield of pretreatment and enzymatic saccharification (EPS) and the lowest inhibitory compound concentration. The deletion of the GAL80, MIG1, CYC8, or TUP1 gene was performed to improve the galactose consumption rate. The strains with the deletion of the MIG1 gene (mig1Δ) showed higher galactose consumption rate and ethanol yield than other strains. High transcription levels of regulatory genes revealed that the mig1Δ relieved glucose repression. These results show that the mig1Δ enhances galactose consumption rate from K. alvarezii.

Enhancement of Galactose Uptake from Kappaphycus alvarezii Hydrolysate Using Saccharomyces cerevisiae Through Overexpression of Leloir Pathway Genes.

A total 42.68 g/L monosaccharide with 0.10 g/L HMF was obtained from 10% (w/v) Kappaphycus alvarezii with thermal acid hydrolysis using 350 mM HNO3 at 121 °C for 60 min and enzymatic saccharification with a 1:1 mixture of Viscozyme L and Celluclast 1.5 L for 72 h. To enhance the galactose utilization rate, fermentation was performed with overexpression of GAL1 (galactokinase), GAL7 (galactose-1-phosphate uridyltransferase), GAL10 (UDP-glucose-4-epimerase), and PGM2 (phosphoglucomutase 2) in Saccharomyces cerevisiae CEN.PK2 using CCW12 as a strong promoter. Among the strains, the overexpression of PGM2 showed twofold high galactose utilization rate (URgal) and produced ethanol 1.4-fold more than that of the control. Transcriptional analysis revealed the increase of PGM2 transcription level leading to enhance glucose-6-phosphate and fructose-6-phosphate and plays a key role in ensuring a higher glycolytic flux in the PGM2 strain. This finding shows particular importance in biofuel production from seaweed because galactose is one of the major monosaccharides in seaweeds such as K. alvarezii.

Bioethanol Production from Azolla filiculoides by Saccharomyces cerevisiae, Pichia stipitis, Candida lusitaniae, and Kluyveromyces marxianus.

Ethanol was produced by separate hydrolysis and fermentation using Azolla filiculoides as a biomass. Thermal acid hydrolysis and enzymatic saccharification were used as pretreatment methods to produce monosaccharides from Azolla. The optimal content for thermal acid hydrolysis of 14% (w/v) Azolla weed slurry produced 16.7-g/L monosaccharides by using 200 mM H2SO4 at 121 °C for 60 min. Enzymatic saccharification using 16 U/mL Viscozyme produced 61.6 g/L monosaccharide at 48 h. Ethanol productions with ethanol yield coefficients from Azolla weed hydrolysate using Kluyveromyces marxianus, Candida lusitaniae Saccharomyces cerevisiae, and Pichia stipitis were 26.8 g/L (YEtOH = 0.43), 23.2 g/L (YEtOH = 0.37), 18.2 g/L (YEtOH = 0.29), and 13.7 g/L (YEtOH = 0.22), respectively. Saccharomyces cerevisiae produces the lowest yield as it utilized only glucose. Bioethanol from Azolla weed hydrolysate can be successfully produced by using Kluyveromyces marxianus because it consumed the mixture of glucose and xylose completely within 60 h.

Enhanced Bioethanol Fermentation by Sonication Using Three Yeasts Species and Kariba Weed (Salvinia molesta) as Biomass Collected from Lake Victoria, Uganda.

Kariba weed (Salvinia molesta) was used as biomass feedstock for ethanol production by separate hydrolysis and fermentation (SHF). Monosaccharides from Kariba weed hydrolysate were produced using thermal acid hydrolysis, sonication, and enzymatic saccharification. The optimal conditions for thermal acid hydrolysis of 12% (w/v) Kariba weed slurry were evaluated as 200 mM HNO3 at 121 °C for 60 min yielding 10.2 g/L monosaccharides. Sonication for 45 min before enzymatic saccharification yielded more monosaccharides to 18.7 g/L. Enzymatic saccharification with 16 U/mL Cellic CTec2 produced 35.4 g/L monosaccharides. Fermentation was performed using Saccharomyces cerevisiae, Kluyveromyces marxianus, or Pichia stipitis with sonicated Kariba weed hydrolysate. The control fermentations were carried out using Kariba weed hydrolysate without sonication. The improvement of ethanol production from sonicated Kariba weed hydrolysate using P. stipitis produced 15.9 g/L ethanol with ethanol yield coefficient YEtOH = 0.45, K. marxianus produced 14.7 g/L ethanol with YEtOH = 0.41. S. cerevisiae produced the lowest yield of 13.2 g/L ethanol with YEtOH = 0.37 as it utilized only glucose not xylose. Sonication of Kariba weed was essential in the ethanol production to enhance the productivity of monosaccharides. P. stipitis was determined as the best yeast species using hydrolysates with the mixture of glucose and xylose to produce ethanol.