RNA transcriptions of <i>egl4</i>, <i>cbh1</i> and <i>bgl1</i> genes at 24 h are up to 500-fold higher in populations F1 (circle) and F4 (square) than in the ancestor (triangle).

<p>Culture of CMC media (40 mL) were inoculated by conidia (~ 5 × 10⁶) and RNA was extracted from hyphae collected at 16, 20, 24, 28, 36, 48 and 72 h and measured by qPCR. The error bar means standard error of the mean.</p

Extracellular protein amounts (panel a) in ancestral and selected <i>T</i>. <i>citrinoviride</i> populations are similar (range 0.9–1.5-fold of the ancestor), but the extracellular specific activity (panel b) is at least two fold higher in four of the ten selected populations.

<p>The symbol indicates the ancestor and the numbers indicate the selected populations that named as “F” and the number. Conditions: 50 mL media containing 10 g/L CMC inoculated with ~ 5 × 10⁶ conidia, 5 d growth.</p

The relative amounts of secreted proteins in the major molecular weight bands as determined by SDS-PAGE.^a

<p>The relative amounts of secreted proteins in the major molecular weight bands as determined by SDS-PAGE.<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0147024#t002fn001" target="_blank">^a</a></p

The selected (45^th, 85^th and 125^th transfer) populations of <i>T</i>. <i>citrinoviride</i> deconstructed more filter paper than the ancestral population.

<p>The horizontal lines indicate the average of each cluster and the error bars indicate the standard deviation. Filter paper deconstruction increased greatly during the first 45 transfers; then more slowly during additional transfers.</p

The transcription levels of positive and negative regulators of expression of cellulase in <i>T</i>. <i>citrinoviride</i>.

<p>The transcription levels of positive and negative regulators of expression of cellulase in <i>T</i>. <i>citrinoviride</i>.</p

Amounts of secreted protein and specific activity in culture for several populations.

<p>a) The amount of extracellular protein during days 2–5 in population F1 (closed square, 125 transfers) was about twice the amount in population F4 (closed circle, 125 transfers) and in the ancestral (open square) <i>T</i>. <i>citrinoviride</i>. During selection, the populations were transferred at 2–3 days (gray shading), so changes after this time do not influence adaptation. b) The specific activity of the secreted proteins remained constant over days 2–7 for both populations F1 (closed square, 125 transfers), F4 (closed circle, 125 transfers) and the ancestral population (open square): CMC activities (dashed line, estimates endocelluase activity), Avicel activities (solid line, estimates cellobiohydrolase activity) and β-glucosidase activities (dot-dash line). The populations F1 and F4 showed similar CMC activity as the ancestor, but approximately two-fold higher of Avicel activity and three-fold higher β-glucosidase activities. Conditions: CMC-media was inoculated with the same number of conidia from each strain and cultured at 28°C. The secreted proteins were harvested at days 2, 3, 5 and 7, and assayed for activity. See Supporting Information for details.</p

Oligonucleotides for amplifying the internal transcribed spacer (ITS) region, amplifying the cellulase genes <i>cbh1</i>, <i>cbh2</i> and <i>bgl1</i>, and for qPCR of various <i>T</i>. <i>citrinoviride</i> genes.

<p>Oligonucleotides for amplifying the internal transcribed spacer (ITS) region, amplifying the cellulase genes <i>cbh1</i>, <i>cbh2</i> and <i>bgl1</i>, and for qPCR of various <i>T</i>. <i>citrinoviride</i> genes.</p

The Fungus <i>Trichoderma</i> Regulates Submerged Conidiation Using the Steroid Pregnenolone

In previous work, we evolved a population of <i>Trichoderma citrinoviride</i> in liquid cultures to speed up its asexual development cycle. The evolved population, called T-6, formed conidia 3 times sooner and in >1000-fold greater numbers. Here, we identify the steroid pregnenolone as a molecular signal for this different behavior. Media in which the ancestral <i>T. citrinoviride</i> population was grown (called ancestral spent media) contained a submerged conidiation inhibitor. Growing the evolved population T-6 in ancestral spent media eliminated the abundant formation of conidia. This inhibition depended on the amount and age of the ancestral spent medium and the time that the ancestral spent medium was added to the T-6 culture. Fractionation of the ancestral spent medium identified a hydrophobic inhibiting compound with a molecular weight less than 2000 g/mol. A combination of GC-MS, ELISA, and reaction with cholesterol oxidase identified it as pregnenolone. The addition of pregnenolone to cultures of T-6 inhibited submerged conidiation by inhibiting formation of conidiophores, while 10 other analogous steroids did not. Pregnenolone also inhibited submerged conidiation of <i>Fusarium graminearum</i> PH-1, a plant pathogen that causes head blight in wheat and barley. This identification of steroids as signal molecules in fungi creates opportunities to disrupt this signaling to control fungal behavior

DataSheet_1_Somatostatin-SSTR3-GSK3 modulates human T-cell responses by inhibiting OXPHOS.pdf

IntroductionSomatostatin (SST) is a peptide hormone primarily synthesized in the digestive and nervous systems. While its impact on the endocrine system is well-established, accumulating evidence suggests a crucial role for SST and its analogues in modulating immune responses. Despite this, the precise mechanism through which SST regulates T cells has remained largely unknown.MethodsTo elucidate the impact of SST on human T cells, we conducted a series of experiments involving cell culture assays, molecular analyses, and metabolic profiling. Human T cells were treated with SST, and various parameters including proliferation, cytokine production, and metabolic activities were assessed. Additionally, we employed pharmacological inhibitors and genetic manipulations to dissect the signaling pathways mediating SST's effects on T cells.ResultsWe showed that SST diminishes T-cell proliferation by influencing IL-2 production and T-cell mitochondrial respiration, while having no discernible impact on TCR-induced glycolysis. Our findings also identified that the regulatory influence of SST on T-cell responses and metabolism is contingent on its receptor, SSTR3. Moreover, we demonstrated that SST governs T-cell responses and metabolism by acting through the T-cell metabolic checkpoint GSK3.DiscussionOur study provides novel insights into the immunoregulatory function of SST in human T cells, highlighting the complex interplay between hormonal signaling and immune regulation. Understanding the molecular mechanisms underlying SST's effects on T cells may offer therapeutic opportunities for manipulating immune responses in various pathological conditions.</p

Theoretical and Experimental Insights into the Electrochemical Mineralization Mechanism of Perfluorooctanoic Acid

The electrochemical mineralization mechanism of environmentally persistent perfluorooctanoic acid (PFOA) at a Ce-doped modified porous nanocrystalline PbO₂ film anode was investigated using density functional theory (DFT) simulation and further validated experimentally. The potential energy surface was mapped out for all possible reactions during electrochemical mineralization reaction of PFOA. The hydroxyl radical (·OH), O₂ and H₂O took part in the mineralization process and played different roles. The ·OH-initiated process was found to be the main degradation pathway, and the existence of O₂ obviously accelerated the degradation process of PFOA in aqueous solution. On the basis of the DFT calculations, an optimal electrochemical mineralization mechanism of PFOA was proposed, which involved the electronic migration, decarboxylation, radical reaction, hydrogen abstraction reaction, and radical fragmentation reaction. The proposed mechanism was verified by the dynamics and intermediate determination experiments. Furthermore, the observed ·OH concentration showed that the electrolysis system could produce enough ·OH for PFOA mineralization process, indicating that the proposed ·OH-initiated process derived from DFT calculations was feasible. These insightful findings are instrumental for a comprehensive understanding of the mineralization of PFOA in the electrolysis system