Phylogenetic tree based on ITS1-5.8S-ITS2 sequences of strongly active endophytic fungi.

<p>The number of each branch point represents percentage bootstrap support from Maximum Parsimony (MP BS) and Neighbour Joining (NJ BS) with 100 replications shown on the branch. MP BS values ≥50% are shown before the slash; NJ BS values ≥50% are shown after the slash. Length; 37 steps; consistency index (CI); 0.8108; retention index (RI); 0.9391; homoplasy index (HI); 0.1892; rescaled consistency index (RC); 0.7615.</p

Scanning electron micrographs of test microorganisms with strongly active crude extracts.

<p><i>Cryptococcus neoformans</i> ATCC 90112 (A–E), <i>Cryptococcus neoformans</i> ATCC 90113 (F), <i>Candida albicans</i> NCPF 3153 (G–I) and a clinical isolate of <i>Microsporum</i><i>gypseum</i> (K–L) after incubation with 10% DMSO (A, G and J), amphotericin B (B and H), miconazole (K), hexane extract from the mycelia of <i>Penicillium</i> sp. PSU-ES43 (C), hexane extract from the mycelia of PSU-ES190 (D), ethyl acetate extract from the mycelia of <i>Fusarium</i> sp. PSU-ES73 (E and F), ethyl acetate extract from the mycelia of <i>Trichoderma</i> sp. PSU-ES38 (I), and hexane extract from the mycelia of Hypocreales sp. PSU-ES26 (L) for 16 h at 4 times their MIC values.</p

Xylariphilone: a new azaphilone derivative from the seagrass-derived fungus Xylariales sp. PSU-ES163

<div><p>One new azaphilone derivative, named xylariphilone (<b>1</b>), along with 10 known compounds was isolated from the seagrass-derived fungus Xylariales sp. PSU-ES163. Their structures were elucidated on the basis of extensive spectroscopic analysis. The absolute and relative configurations of <b>1</b> were determined by circular dichroism spectroscopy and NOEDIFF data. The antimicrobial and cytotoxic activities of the isolated compounds were evaluated.</p></div

Antimicrobial activity of endophytic fungal crude extracts against each test microorganism.

<p>SA, <i>Staphylococcus aureus</i> ATCC 25923; MRSA, methicillin-resistant <i>S. aureus</i>; EC, <i>Escherichia coli</i> ATCC 25922; PA, <i>Pseudomonas aeruginosa</i> ATCC 27853; CA1, <i>Candida albicans</i> ATCC 90028; CA2, <i>C. albicans</i> NCPF 3153; CN1, <i>Cryptococcus neoformans</i> ATCC 90112 (flucytosine-sensitive); CN2, <i>C. neoformans</i> ATCC 90113 (flucytosine-resistant); MG, <i>Microsporum</i><i>gypseum</i> clinical isolate; PM, <i>Penicillium</i><i>marneffei</i> clinical isolate.</p

Phthalide and Isocoumarin Derivatives Produced by an <i>Acremonium</i> sp. Isolated from a Mangrove <i>Rhizophora apiculata</i>

Nine new fungal metabolites, one phthalide derivative, acremonide (<b>1</b>), and eight isocoumarin derivatives, acremonones A–H (<b>2</b>–<b>9</b>), were isolated from the mangrove-derived fungus <i>Acremonium</i> sp. PSU-MA70 together with 10 known compounds. Their structures were determined by NMR analysis. The known 8-deoxytrichothecin and trichodermol exhibited moderate antifungal activity against <i>Candida albicans</i> and <i>Cryptococcus neoformanns</i>, respectively

γ‑Butyrolactone, Cytochalasin, Cyclic Carbonate, Eutypinic Acid, and Phenalenone Derivatives from the Soil Fungus <i>Aspergillus</i> sp. PSU-RSPG185

Purification of an extract from the broth of the soil fungus <i>Aspergillus</i> sp. PSU-RSPG185 resulted in the isolation of two new cyclic carbonate derivatives, aspergillusols A (<b>1</b>) and B (<b>2</b>), and one new eutypinic acid derivative, aspergillusic acid (<b>3</b>), along with six known secondary metabolites. Compounds <b>1</b> and <b>2</b> contain an unusual cyclic-carbonate functionality. In addition, the mycelial extract afforded two new phenalenones, aspergillussanones A (<b>4</b>) and B (<b>5</b>), one new cytochalasin, aspergilluchalasin (<b>6</b>), and one new γ-butyrolactone, aspergillulactone (<b>7</b>). Their structures were established by interpretation of spectroscopic evidence. Compound <b>4</b> exhibited weak activity toward KB and Vero cells with IC₅₀ values of 48.4 and 34.2 μM, respectively

Effect of statin derivatives isolated from soil fungus <i>Aspergillus sclerotiorum</i> and statin drugs on cAMP-dependent Cl^- secretion in T84 cell monolayers.

(A) Effect of α,β-dehydrolovastatin (DHLV), lovastatin, α,β-dehydrodihydromonacolin K, mevastatin, simvastatin, and pravastatin on cAMP-dependent Cl- secretion determined by ISC analysis. (B) Effect of CFTRinh-172 (CFTR inhibitor) on cAMP-dependent Cl- secretion determined by ISC analysis. All of compounds were added accumulatively in both apical and basolateral solutions at the indicated concentrations. Representative ISC tracings are shown. (C) Summary of concentration- inhibition studies. Data are fitted to Hill’s equation and expressed as means of % forskolin-stimulated ISC ± S.E.M. (n = 3–7).</p

Potential utility of α,β-dehydrolovastatin (DHLV) as an anti-diarrheal agent for secretory diarrheas.

(A) Effect of DHLV on cholera toxin (CT)- and heat-stable toxin (STa)-stimulated Cl- secretion in T84 cell monolayers determined by ISC analysis. After apical addition of CT (1 μg/ml) or STa (100 μM) to stimulate the increasing of ISC, DHLV was added in both apical and basolateral solution. Representative ISC tracings are shown. (B) Effect of DHLV on CT-induced intestinal fluid secretion determined by ileal closed-loop weight/length ratios. Ileal closed-loops were injected with PBS (control) or PBS containing CT (μg/loop) with or without intraluminal (i.l.) and intraperitoneal (i.p.) administrations of DHLV (20 μM and 2 mg/kg, respectively). Representative photographs of ileal loops are shown. Summary of data are expressed as means of ileal closed-loop weight/length ratio ± S.E.M. (n = 5–6). *** p p n = 6–7). * p < 0.05 compared with control at 1 min; NS, non-significant compared with control at the same time point (one-way ANOVA).</p

No involvement of CFTR negative regulators in CFTR inhibition by DHLV.

(A) Schematic diagrams showing the regulatory mechanism of CFTR Cl- channel activity. (B) apical Cl- current (ICl-) tracing showing the effect of α,β-dehydrolovastatin (DHLV) on forskolin-induced CFTR Cl- secretion in T84 cells pre-treatment with compound C (50 μM) or and NaF plus Na3VO4 (1 mM) for 15 min. (C) Summary of concentration-inhibition studies. Data are fitted to Hill’s equation and expressed as means of % agonist-stimulated ICl- ± S.E.M. (n = 5–9).</p

Effects of α,β-dehydrolovastatin (DHLV) on Ca²⁺-activated Cl^- channels (CaCC), the intracellular Ca²⁺ levels induced by ATP and Na⁺/K⁺ ATPases activity.

(A, left) Representative apical ICl- tracing showing the effect of ATP (100 μM) activation after 15 min pre-treatment with CFTRinh-172 (5 μM) with vehicle (control) or DHLV (20 μM). (A, right) Summary of the data of peak ICl- expressed as mean of peak ICl- ± S.E.M. (n = 5). * p 2+ levels were analyzed from the fluo-8-based fluorescence assays. Fluo-8 fluorescence intensity were measured following vehicle (control) or DHLV (20 μM) together with the addition of ATP (100 μM) and end up with EGTA (3 mM). (B) Representative values of the fractional change in fluorescence intensity relative to baseline (ΔF/F°) are shown. (C) Summary of total AUC values after baseline correction presented as the area under the curve (AUC) ± S.E.M (n = 4). NS, non-significant (Student’s t test). (D, left) Ouabain-sensitive ISC tracing showing the effect of DHLV on Na+-K+ ATPase activity. After T84 cells were permeabilized at apical membrane, ouabain (1 mM) was added. (D, right) Summary of the data are expressed as mean of ouabain-sensitive ISC ± S.E.M. (n = 4). NS, non-significant (Student’s t test).</p