Monalbidins A–E,Decalins with Potential Cytotoxic Activities from Marine Derived Fungus Monascus albidus BB3

Five new decalins, monalbidins A–E ( 1 , 2 and 7–9 ), together with 16 known compounds ( 3 – 6 and 10–21 ), were isolated from the AcOEt extract of marine derived fungus Monascus albidus BB3 cultured in GPY medium. Among the known compounds, 1-hydroxymonacolin L ( 11 ), dehydromonacolin J ( 15 ), 8-O-acetylmonacolin J ( 19 ) and O-acetylmonacolin K ( 21 ) were separated from natural sources for the first time. Their structures were determined by comprehensive analysis on the 1D and 2D NMR, HR-ESI-MS, UV and IR data, and their absolute configurations were assigned by experimental and calculated ECD data, and X-ray single-crystal diffraction analysis. Monalbidins C and D ( 7 and 8 ), monacolin K methyl ester ( 13 ), dehydromonacolin L ( 14 ), dehydromonacolin K ( 16 ), monacolin K ( 20 ) and O-acetylmonacolin K ( 21 ) showed moderate cytotoxicity against human cancer cell lines SUNE1, HepG2, QGY7701, HCT116 and MDA-MB-231.     

Hyposterolactone A,a 3α-Hydroxy Steroidal Lactone from the Deep-Sea-Derived Fungus Hypocrea sp. ZEN14

Chemical investigation of the deep-sea-derived fungus Hypocrea sp. ZEN14 afforded a new 3α-hydroxy steroidal lactone, hyposterolactone A ( 1 ) and 25 known secondary metabolites ( 2 – 26 ). The structure of the new compound was established by detailed spectroscopic analysis, electronic circular dichroism (ECD) calculation as well as a J-based configuration analysis. Compound 10 showed potent cytotoxicity against Huh7 and Jurkat cells with IC₅₀ values of 1.4 μM and 6.7 μM, respectively.     

Structural Studies on “Isosclerotan”, a New Glucan Isolated from Sclerotinia Fungus,by Physical,Chemical and Enzymatic Methods

“Isosclerotan”, a polysaccharide constituent extracted with a sodium hydroxide solution from sclerotia of Sclerotinia libertiana, could be purified by the successive precipitation with the followings; a mixture of copper sulfate and sodium hydroxide, ammonium sulfate, and ethyl alcohol. The preparation proved homogeneous by ultracentrifugal analysis. From sedimentation and viscosity measurements, the molecular weight of isosclerotan was calculated as 6.13 × 10⁶, andas 1.60 × 10⁵ after treatment with a dilute oxalic acid solution. Isosclerotan was scarecely soluble in cold water but soluble in hot water, yielding a highly viscous solution. It exhibited a low positive optical rotation, + 23.0° (in water), and infrared spectrum had a sharp absorption at 890~898 cm^?1, which indicated the prevalence of the β-glycosidic linkage in isosclerotan. Through degradation by acids and enzymes of isosclerotan, there were obtained various oligosaccharides containing β-1.3, β-1.4, and β-1.6 linkages. From results obtained by periodate oxidation and methylation, it is assumed that the polysaccharide involves the 1.3, 1.4, and 1.6 linkages in 47.7%, 16.6% and 35.7%, respectively, and a branching structure about 12.5%.     

Trichocarotins A–H and trichocadinin A,nine sesquiterpenes from the marine-alga-epiphytic fungus Trichoderma virens

In addition to CAF-603, 14-hydroxy CAF-603 (trichocarane B), 7-β-hydroxy CAF-603, and trichocarane A, eight new carotane sesquiterpenes, trichocarotins A–H, and one new cadinane sesquiterpene, trichocadinin A, were isolated from the culture of Trichoderma virens Y13-3, obtained from the surface of a marine red alga. Their structures and relative configurations were unambiguously assigned by interpretation of 1D/2D NMR and MS data, and their absolute configurations were established by X-ray diffraction or ECD spectra aided by quantum chemical calculations. These compounds represent two rarely occurring sesquiterpene types from filamentous fungi, and six of them feature potent inhibition against some marine plankton species.     

Gynostemosides A–E,megastigmane glycosides from Gynostemma pentaphyllum

Megastigmane glycosides (1–5) together with seven (6–12) related known compounds were isolated from the whole plants of Gynostemma pentaphyllum. The structures were elucidated by means of spectroscopic methods, including 2D NMR, HR-ESIMS, and circular dichroism (CD), as well as chemical transformations to be (3R, 4R, 5S, 6S, 7E)-3,4,6-trihydroxymegastigmane-7-en-9-one-3-O-β-d-glucopyranoside (gynostemoside A, 1), (3S, 4S, 5R, 6R, 7E, 9R)-3,4,6,9-tetrahydroxymegastigmane-7-en-3-O-β-d-glucopyranoside (gynostemoside B, 2), (3S, 4S, 5S, 6S, 7E, 9R)-3,4,9-trihydroxymegastigmane-7-en-9-O-β-d-glucopyranoside (gynostemoside C, 3), (3S, 4S, 5S, 6S, 7E, 9R)-3,4,9-trihydroxymegastigmane-7-en-3-O-β-d-glucopyranoside (gynostemoside D, 4), and (3S, 4S, 5S, 6S, 7E, 9R)-3,4,9-trihydroxymegastigmane-7-en-4-O-β-d-glucopyranoside (gynostemoside E, 5), respectively.     

9-Phenanthrol,a TRPM4 Inhibitor,Protects Isolated Rat Hearts from Ischemia–Reperfusion Injury

Despite efforts to elucidate its pathophysiology, ischemia–reperfusion injury lacks an effective preventative intervention. Because transient receptor potential cation channel subfamily M member 4 (TRPM4) is functionally expressed by many cell types in the cardiovascular system and is involved in the pathogenesis of various cardiovascular diseases, we decided to assess its suitability as a target of therapy. Thus, the aim of this study was to examine the possible cardioprotective effect of 9-phenanthrol, a specific inhibitor of TRPM4. Isolated Langendorff-perfused rat hearts were pretreated with Krebs–Henseleit (K–H) solution (control), 9-phenanthrol, or 5-hydroxydecanoate (5-HD, a blocker of the ATP-sensitive potassium channel) and then subjected to global ischemia followed by reperfusion with the K–H solution. To evaluate the extent of heart damage, lactate dehydrogenase (LDH) activity in the effluent solution was measured, and the size of infarcted area of the heart was measured by 2,3,5-triphenyltetrazolium chloride staining. In controls, cardiac contractility decreased, and LDH activity and the infarcted area size increased. In contrast, in hearts pretreated with 9-phenanthrol, contractile function recovered dramatically, and the infarcted area size significantly decreased. The cardioprotective effects of 9-phenanthrol was not completely blocked by 5-HD. These findings show that 9-phenanthrol exerts a cardioprotective effect against ischemia in the isolated rat heart and suggest that its mechanism of action is largely independent of ATP-sensitive potassium channels.     

Tumstatin,a Matrikine Derived from Collagen Type IVα3, is Elevated in Serum from Patients with Non–Small Cell Lung Cancer

OBJECTIVES: Fibrosis and cancer are characterized by extracellular matrix (ECM) remodeling. The basement membrane is mainly composed by collagen type IV and laminin. Tumstatin is a matrix metalloproteinase-9 (MMP-9) generated matrikine of collagen type IV α3 chain. We evaluated the potential of tumstatin as a diagnostic biomarker of lung disorders. METHODS: A monoclonal antibody was raised against the neo-epitope tumstatin. A novel competitive enzyme-linked immunosorbent assay for detection of tumstatin (TUM), was developed and technically characterized. Levels of TUM were measured in serum of patients with idiopathic pulmonary fibrosis (IPF), chronic obstructive pulmonary disease (COPD), and non–small cell lung cancer (NSCLC) belonging to two cohorts. RESULTS: The developed TUM enzyme-linked immunosorbent assay (ELISA) was technically robust. In cohort 1, levels of TUM were significantly higher in NSCLC compared to healthy controls, IPF, and COPD (P = 0.007, P = 0.03 and P = 0.001, respectively). The area under the receiver operating characteristics (AUROC) for separation of patients with NSCLC from healthy controls was 0.97, for separation of NSCLC and IPF patients was 0.98, and for separation of NSCLC and COPD patients was 1.0. In cohort 2, levels of TUM were also significantly higher in patients with NSCLC compared to healthy controls (P = 0.002), and the AUROC for separation of NSCLC and healthy controls was 0.73. CONCLUSIONS: We developed a technically robust competitive ELISA targeting the fragment tumstatin. The level of TUM in circulation was significantly higher in patients with NSCLC compared to patients with IPF, COPD and healthy controls. The assay provided high diagnostic accuracy in separating NSCLC patients from other lung disorders and from healthy controls.     

Amenyunnaosides A–C,Three New Neolignans Isolated from Amentotaxus yunnanensis and Their Anti-inflammatory Activities

Phytochemical study on the leaves of Amentotaxus yunnanensis led to the isolation of seventeen phenolic compounds including sixteen neolignans and lignans, and one flavone glycoside. Three among the isolates were previously unreported neolignans and named as amenyunnaosides A–C, respectively. Their structures were elucidated by extensive analyses of HR-ESI-MS, 1D and 2D NMR, and ECD spectra. The isolated neolignans potentially inhibited NO production in LPS-activated RAW264.7 cells with their IC₅₀ values ranging from 11.05 to 44.07 μM, compared to that of the positive control compound, dexamethasone, IC₅₀ value of 16.93 μM. Additionally, amenyunnaoside A dose-dependently reduced production of IL-6 and COX-2 but did not effect to that of TNF-α at concentrations of 0.8, 4, and 20 μM.     

Boseongazepines A–C,pyrrolobenzodiazepine derivatives from a Streptomyces sp. 11A057

Three new pyrrolobenzodiazepine derivatives, boseongazepines A–C (1–3), were isolated from a culture broth of Streptomyces sp. 11A057, together with the known compound usabamycin B (4). The structures of 1–4 were determined through the analysis of spectroscopic data including extensive 1D-, 2D-NMR, and MS techniques. Cell growth inhibition effects of these compounds were evaluated against Jurkat, K-562, HL-60, and HepG2 cell lines.     

Thermostable α-1,4- and α-1,6-glucosidase enzymes from Bacillus sp. Isolated from a marine environment

Penaeus vannamei (the shrimp) is an omnivorous species and it can be assumed that a high level of carbohydrates is necessary for its growth. -1,4- and 1,6-glucosidases are important enzymes necessary for the ultimate liberation of glucose residues from various carbohydrates, principally starch. However, the shrimp's hepatopancreas produces only -1,4-glucosidases, which limits the growth rate in different sources of starch. In order to identify strains with -1,4- and 1,6-glucosidase enzymes with potential uses in shrimp feed production, Bacillus strains were isolated from marine environments. One strain produced large amounts of an extracellular thermostable -glucosidase that permitted good growth on starch. The organism was identified by polymorphism (restriction-fragment-length polymorphism, RFLP), sequenced, and named B. subtilis LMM-12.     

Hexahydrohippuric Acid,a Newly Isolated Compound from the Cattle’s Urine

A cosmid library and physical maps of mitochondrial DNA (mtDNA) from a liverwort, Marchantia polymorpha, were constructed using the cosmid clones. Electrophoresis profile and the physical maps indicated that the liverwort mtDNA was approximately 183 kb long, the smallest among plant mtDNAs, and that it consisted of a single circular molecule. Southern hybridization analysis showed that genes typical to the mitochondrial genome existed in a single copy, and also that there was no incorporation of chloroplast DNA fragments into the mitochondrial genome.     

A Rare Fungal Species,Quambalaria cyanescens,Isolated from a Patient after Augmentation Mammoplasty – Environmental Contaminant or Pathogen?

Some emerging but less common human fungal pathogens are known environmental species and could be of low virulence. Meanwhile, some species have natural antifungal drug resistance, which may pose significant clinical diagnosis and treatment challenges. Implant breast augmentation is one of the most frequently performed surgical procedures in China, and fungal infection of breast implants is considered rare. Here we report the isolation of a rare human fungal species, Quambalaria cyanescens, from a female patient in China. The patient had undergone bilateral augmentation mammoplasty 11 years ago and was admitted to Peking Union Medical College Hospital on 15 September 2011 with primary diagnosis of breast infection. She underwent surgery to remove the implant and fully recovered thereafter. During surgery, implants and surrounding tissues were removed and sent for histopathology and microbiology examination. Our careful review showed that there was no solid histopathologic evidence of infection apart from inflammation. However, a fungal strain, which was initially misidentified as “Candida tropicalis” because of the similar appearance on CHROMagar Candida, was recovered. The organism was later on re-identified as Q. cyanescens, based on sequencing of the rDNA internal transcribed spacer region rather than the D1/D2 domain of 26S rDNA. It exhibited high MICs to 5-flucytosine and all echinocandins, but appeared more susceptible to amphotericin B and azoles tested. The possible pathogenic role of Q. cyanescens in breast implants is discussed in this case, and the increased potential for misidentification of the isolate is a cause for concern as it may lead to inappropriate antifungal treatment.     

Miospore assemblages from the Silurian–Devonian boundary,in borehole A1-61, Ghadamis Basin,Libya

Well preserved and diversified miospore assemblages have been recorded from a relatively continuous sequence in borehole A1-61 which spans the Silurian–Devonian boundary in the northwestern part of the Ghadamis Basin, Libya. The sequence is represented by early Devonian Lochkovian beds of the Tadrart Formation that transgress onto the Silurian Ludlow-Pridoli beds of the upper part of the ‘Alternances Argilo-gréseuses’ Formation. The present work demonstrates a succession of miospore assemblages from closely sampled layers that have been stratigraphically dated as Ludlow–middle Pridoli and early Lochkovian by chitinozoans and acritarchs. Over 80 species of cryptospores and trilete spores have been identified. Modified detailed morphological interpretations are given. The miospore assemblages are correlated with miospore zonation schemes established for the type sequences of the Welsh Borderland, and those previously described from Libya. Early occurrences of some species as Streelispora newportensis on the western Gondwana plate, are put forward by comparison with the Old Red Sandstone continent. Phytogeographic and palaeobotanic implications based on these observations are discussed.     

Pestalofones A–E,bioactive cyclohexanone derivatives from the plant endophytic fungus Pestalotiopsis fici

Pestalofones A–E (1–5), five new cyclohexanone derivatives, have been isolated from cultures of the plant endophytic fungus Pestalotiopsis fici, along with the known compounds, isosulochrin (6), isosulochrin dehydrate (7), and iso-A82775C (8). The structures of 1–5 were determined by NMR spectroscopy, and the absolute configuration of 1 was assigned using the modified Mosher method. Compounds 1, 2, and 5 displayed inhibitory effects on HIV-1 replication in C8166 cells, whereas 3 and 5 showed significant antifungal activity against Aspergillus fumigatus.     

Synthesis of β-1, 3-Glucan and β-1, 3 and β-1, 4-Mixed Glucan from UDP-α-d-Glucose

A particulate enzyme preparation from Phaseolus aureus (mung bean) seedlings catalyzed the synthesis of a water insoluble β-1,3-glucan from UDP-α-d-glucose (UDPG) at high concentrations (0.4~20 mm) and an alkaline insoluble β-1,3 and β-1,4-mixed glucan from UDPG at a low concentration (8.5 µm).

Furthermore, the two kinds of β-glucan synthetases which were investigated with two reaction systems at high and low concentrations of UDPG had different properties in optimal pH, stability of enzyme activity, and metallic ion requirement.     

Antibacterial Activity and Mechanism of a Bacteriocin Derived from the Valine-Cecropin A(1–8)-Plantaricin ZJ5(1–18) Hybrid Peptide Against Escherichia coli O104

Food Biophysics - Peptide fragment hybridization is an effective method to obtain novel hybrid antimicrobial peptides with higher antibacterial activities. The novel peptide, valine-cecropin...     

Production, Purification and Characterization of Alkaline β-Glucosidase Isolated from Agrobacterium sp. PJD-1-1

In order to screen novel β-glucosidase producing strains from environment, one targeted novel strain PJD-1-1 producing β-glucosidase were isolated from putrefied sugarcane leaves with screening and spreading plate. 16S rDNA analysis revealed it was a novel Agrobacterium sp. When the strain was incubated at initial pH 7.0, 20 ℃ with lactose as carbon and NaNO₃ as nitrogen sources, the maximum enzyme activity was 3.92 U/mg. β-glucosidase from this strain was purified using (NH₄)₂SO₄ precipitation followed by dextran gel filtration chromatography and ion exchange chromatography. A purifying fold of 4.85 with gaining rate of 8.0% was obtained. SDA-PAGE analysis of the purified enzyme showed that it was a clear and pure band with molecular mass of ca. 40 kDa. The most optimum activity of the enzyme was at 50 ℃ and pH at 8.0. The enzyme could maintain stability under the conditions below 50 ℃. Hg²⁺ and Ag⁺ heavily inhibited the enzyme activity suggesting that the active catalytic sites of the enzymes might possess thiol radical. Ba²⁺, Ca²⁺, Pb²⁺, Co²⁺, Zn²⁺, Mn²⁺, Na⁺, K⁺, EDTA, and urea had no obvious effects on the enzyme activity. It is concluded that the novel strain Agrobacterium sp. PJD-1-1 producing β-glucosidase was successfully screened from putrefied sugar cane leaves. The produced enzyme had thermal stability, alkaline feature and metal ions tolerance made it useful in the food and broad potential applications in other fields.