The effect of viscosity, friction, and sonication on the morphology and metabolite production from Aspergillus terreus ATCC 20542

This study investigates the effects of viscosity, friction, and sonication on the morphology and the production of lovastatin, (+)-geodin, and sulochrin by Aspergillus terreus ATCC 20542. Sodium alginate and gelatine were used to protect the fungal pellet from mechanical force by increasing the media viscosity. Sodium alginate stimulated the production of lovastatin by up to 329.0% and sulochrin by 128.7%, with inhibitory effect on (+)-geodin production at all concentrations used. However, the use of gelatine to increase viscosity significantly suppressed lovastatin, (+)-geodin, and sulochrin’s production (maximum reduction at day 9 of 42.7, 60.8, and 68.3%, respectively), which indicated that the types of chemical play a major role in metabolite production. Higher viscosity increased both pellet biomass and size in all conditions. Friction significantly increased (+)-geodin’s titre by 1527.5%, lovastatin by 511.1%, and sulochrin by 784.4% while reducing pellet biomass and size. Conversely, sonication produced disperse filamentous morphology with significantly lower metabolites. Sodium alginate-induced lovastatin and sulochrin production suggest that these metabolites are not affected by viscosity; rather, their production is affected by the specific action of certain chemicals. In contrast, low viscosity adversely affected (+)-geodin’s production, while pellet disintegration can cause a significant production of (+)-geodin

Performance of mycelial biomass and exopolysaccharide from Malaysian Ganoderma lucidum for the fungivore red hybrid Tilapia (Oreochromis sp.) in Zebrafish embryo

Natural mycelial biomass (MB) and extracted exopolysaccharide (EPS) from the pre-grown Malaysian Ganoderma lucidum mushroom are both considered as high-end materials due to their high commercial value in the aquaculture industry. To evaluate their potential toxicity as a fish-feed supplement for the fungivore red hybrid Tilapia (Oreochromis sp.), both MB (250−5000 μg/mL) and EPS (62.5−3000 μg/mL) were subjected to zebrafish embryo toxicity (ZFET) assay, and the effects on zebrafish embryos (ZE) early development were analyzed between 24−120 hours of post-exposure (HPE). MB and EPS showed no toxic effect towards the ZE with LC50 of 1650 μg/mL and 2648.38 μg/mL, respectively. MB at concentrations between 250−5000 μg/mL and EPS at 3000 μg/mL showed no significant changes in ZE hatching. No significant changes in the ZE heart rate were detected following treatment with both tested compounds (MB: 250−2000 μg/mL and EPS: 62.5−3000 μg/mL) as compared to untreated embryos (135.5 beats/min). Furthermore, teratogenic effects of both MB and EPS (<3000 μg/mL) on zebrafish embryonic development were not observed. Together, both natural compounds MB and EPS can be considered non-toxic, suggesting that these can be safely applied as feed substances in the fish-feed aquaculture industry. © 2020 The Author

Calcium-induced calcification is blocked by TRAIL in VSMCs.

<p>(<b>A</b>) Human VSMCs were treated with TRAIL and calcium (3.3 mM) every 2–3 d. Cells were serum-starved for 24 h prior to treatment for 10 d. Representative photograph of each treatment group. Alizarin red staining indicates calcification. Cells were fixed and calcification assessed using an alizarin red-based assay. (<b>B</b>) WT and TRAIL^−/− VSMCs were treated with increasing concentrations of calcium (2.45–3.3 mM). Cells were serum-starved for 24 h prior to treatment for 2 d. Representative photograph of each treatment group. Alizarin red staining indicates calcification. Cells were fixed and calcification assessed using an alizarin red-based assay as described in the methods. NT, no treatment. ANOVA (n = 3 experiments); ****p<0.0001.</p

Aortas from 20 w TRAIL^−/−ApoE^−/− mice display increased inflammation.

<p>ANOVA (n = 5/genotype); *p<0.05.</p

Aortas from 12 w HFD-fed TRAIL^−/−ApoE^−/− mice display altered expression of bone markers and inflammation.

<p>ANOVA (n = 4-6/genotype); *p<0.05, **p<0.01.</p

Calcium differentially regulates RANKL, OPG and TRAIL expression in VSMCs.

<p>Relative changes in mRNA levels of (<b>A</b>) RANKL, (<b>B</b>) OPG and (<b>C</b>) TRAIL in WT VSMCs treated with calcium using the 2ΔΔCt method. WT cells were serum-starved for 24 h prior to treatment with total 3.3 mM calcium. RNA was extracted 24 h later and real-time PCR was performed in triplicate. Expression was normalised to 18S RNA. Changes in mRNA expression were compared with untreated WT expression. Mann-Whitney t-test (n = 3 experiments); *p<0.05; **p<0.01.</p

Murine primer sequences.

<p>Murine primer sequences.</p

Model for the protective action of TRAIL in vascular calcification.

<p>Calcium differentially regulates RANKL, OPG and TRAIL expression in VSMCs. As a consequence, a deficiency in TRAIL with the addition of exogenous calcium leads to vascular calcification <i>in vitro</i>. <i>In vivo</i>, TRAIL-deficiency in ApoE^−/− mice on a HFD increases the expression of cellular RANKL and inflammatory cytokines within the vessel wall, simultaneously leading to an acceleration of cartilage development and subsequent calcification.</p