Low MW Compounds Research Articles

Soluble microbial products (SMPs) in the effluent from a submerged anaerobic membrane bioreactor (SAMBR) under different HRTs and transient loading conditions

This study investigated the performance of a submerged anaerobic membrane bioreactor (SAMBR) fed with synthetic wastewater (544±22mgCOD/L) operating at different hydraulic retention times (HRTs-12h, 8h, 6h, 4h, 2h, and 1h) at both steady state, and under transient load conditions (2 and 1h), and the SMPs produced under these conditions. COD removal at decreasing HRTs (12h, 8h, 6h, 4h, and 2h) was high (>94%), but decreased to 80% when operating at 1h HRT. VFAs accumulated when the HRT was decreased to 2h and 1h, accounting for 69% and 89% of the effluent COD, respectively. Effluent SMPs accounted for an average of 14±2mgCOD/L at steady state, but this fluctuated more during transient conditions (12±6mgCOD/L). The COD equivalent of dissolved methane in the effluent was 17% at 4h HRT, exceeding the saturation value of methane. Low MW compounds were identified using gas chromatography–mass spectrometry (GC–MS), with solid phase extraction (SPE) as the pre-treatment. 120 compounds were identified in the effluent at steady state, and were alkanes (39), alkenes (3), esters (11), alcohols (7), nitrogenated compounds (11), phenols (11), and others (9). Increases in cyclooctasulfur, N-butyl-benzenesulfonamide, alkanes, 1-naphthalenol, camphor, 2-methylphenol, and (Z)-9-octadecenamide were also found during transient conditions, and these compounds were not found in the feed; hence it is possible that these compounds were produced by microorganism as by-products from substrate utilization.

Abstract A17: Identification and characterization of small molecules targeting MYC-cofactor interactions

Abstract Our aim is to develop specific inhibitors targeting the interaction between the oncoprotein/transcription factor MYC and its cofactors. MYC-family genes (MYC, MYCN, MYCL) are involved in over half of all cancers, including many childhood cancers such as neuroblastoma, medulloblastoma, leukemia and lymphoma, and is often strongly associated with poor prognosis. This emphasizes the need to identify clinically relevant MYC inhibitors. It regulates numerous genes involved in cell growth, metabolism, apoptosis, senescence and other processes. MYC is functionally dependent on interaction with the dimerization partner MAX and with several cofactors involved in different aspects of transcription regulation. Targeting such interactions with low Mw compounds is a plausible way to inhibit the transcriptional function of MYC. We have established several techniques to visualize MYC-cofactor interactions in living (BiFC, split Gaussia Luc) or fixed (isPLA) cells to screen small Mw compound libraries for MYC/cofactor-interaction inhibitors. In MYC/MAX BiFC and Gaussia Luc screens, 15 compounds were identified that directly or indirectly inhibit MYC/MAX interactions in cells. Several of these compounds including analogues are potent inhibitors of MYC/MYCN-dependent proliferation and transformation in neuroblastoma, leukemia/lymphoma and other cancer cells by inducing apoptosis or senescence. One of these, #2, and analogues completely inhibit MYC/MAX interactions in vitro by binding MYC. Another compound, 2XU, inhibit MYC/MAX interactions in sub-nanomolar concentration. We are evaluating the biological activities of these molecules further in different MYC-driven cell culture systems, and are elucidating their mechanisms of action by X-ray crystallography, FRET and SPR. In vivo studies with some of these molecules in mouse tumor models is ongoing. Furthermore, extended screens for finding molecules targeting MYC and other cofactor interactions, such as E3-ligases, are ongoing. The MYC inhibitors identified here have the potential to become important tools in the studies of MYC function in cells and in vivo as well as lead molecules for drug development for treatment of MYC-driven tumors. Citation Format: Alina Castell, Qinzi Yan, Karin Ridderstråle, Lars-Gunnar Larsson. Identification and characterization of small molecules targeting MYC-cofactor interactions. [abstract]. In: Proceedings of the AACR Special Conference on Myc: From Biology to Therapy; Jan 7-10, 2015; La Jolla, CA. Philadelphia (PA): AACR; Mol Cancer Res 2015;13(10 Suppl):Abstract nr A17.

Prediction of Gestational Diabetes through NMR Metabolomics of Maternal Blood

Metabolic biomarkers of pre- and postdiagnosis gestational diabetes mellitus (GDM) were sought, using nuclear magnetic resonance (NMR) metabolomics of maternal plasma and corresponding lipid extracts. Metabolite differences between controls and disease were identified through multivariate analysis of variable selected (1)H NMR spectra. For postdiagnosis GDM, partial least squares regression identified metabolites with higher dependence on normal gestational age evolution. Variable selection of NMR spectra produced good classification models for both pre- and postdiagnostic GDM. Prediagnosis GDM was accompanied by cholesterol increase and minor increases in lipoproteins (plasma), fatty acids, and triglycerides (extracts). Small metabolite changes comprised variations in glucose (up regulated), amino acids, betaine, urea, creatine, and metabolites related to gut microflora. Most changes were enhanced upon GDM diagnosis, in addition to newly observed changes in low-Mw compounds. GDM prediction seems possible exploiting multivariate profile changes rather than a set of univariate changes. Postdiagnosis GDM is successfully classified using a 26-resonance plasma biomarker. Plasma and extracts display comparable classification performance, the former enabling direct and more rapid analysis. Results and putative biochemical hypotheses require further confirmation in larger cohorts of distinct ethnicities.

Low MW Peptides and Carcinogenesis

Problem: Low MW peptides many of them N-substituted, have growth inhibitory effects. Are the peptide levels different in malignant cells from normal cells? Method: Deprimerones dissociated from DNA at pH 9.5 was measured in normal and malignant cells. The level of Low MW compounds was also compared to large MW compounds [ratios] after gel filtration. Result: Chalones and some other low MW compounds were not as in normal tissues and cells found in these. However, these peptides could be found in the incubation fluid or ascites. DNA bound peptides were decreased in malignant cells.

Targeting the cis-dimerization of LINGO-1 with low MW compounds affects its downstream signalling.

The transmembrane protein LINGO-1 is a negative regulator in the nervous system mainly affecting axonal regeneration, neuronal survival, oligodendrocyte differentiation and myelination. However, the molecular mechanisms regulating its functions are poorly understood. In the present study, we investigated the formation and the role of LINGO-1 cis-dimers in the regulation of its biological activity. LINGO-1 homodimers were identified in both HEK293 and SH-SY5Y cells using co-immunoprecipitation experiments and BRET saturation analysis. We performed a hypothesis-driven screen for identification of small-molecule protein-protein interaction modulators of LINGO-1 using a BRET-based assay, adapted for screening. The compound identified was further assessed for effects on LINGO-1 downstream signalling pathways using Western blotting analysis and AlphaScreen technology. LINGO-1 was present as homodimers in primary neuronal cultures. LINGO-1 interacted homotypically in cis-orientation and LINGO-1 cis-dimers were formed early during LINGO-1 biosynthesis. A BRET-based assay allowed us to identify phenoxybenzamine as the first conformational modulator of LINGO-1 dimers. In HEK-293 cells, phenoxybenzamine was a positive modulator of LINGO-1 function, increasing the LINGO-1-mediated inhibition of EGF receptor signalling and Erk phosphorylation. Our data suggest that LINGO-1 forms constitutive cis-dimers at the plasma membrane and that low MW compounds affecting the conformational state of these dimers can regulate LINGO-1 downstream signalling pathways. We propose that targeting the LINGO-1 dimerization interface opens a new pharmacological approach to the modulation of its function and provides a new strategy for drug discovery.

Neuroprotective effects of the anti-cancer drug sunitinib in models of HIV neurotoxicity suggests potential for the treatment of neurodegenerative disorders.

Anti-retrovirals have improved and extended the life expectancy of patients with HIV. However, as this population ages, the prevalence of cognitive changes is increasing. Aberrant activation of kinases, such as receptor tyrosine kinases (RTKs) and cyclin-dependent kinase 5 (CDK5), play a role in the mechanisms of HIV neurotoxicity. Inhibitors of CDK5, such as roscovitine, have neuroprotective effects; however, CNS penetration is low. Interestingly, tyrosine kinase inhibitors (TKIs) display some CDK inhibitory activity and ability to cross the blood-brain barrier. We screened a small group of known TKIs for a candidate with additional CDK5 inhibitory activity and tested the efficacy of the candidate in in vitro and in vivo models of HIV-gp120 neurotoxicity. Among 12 different compounds, sunitinib inhibited CDK5 with an IC50 of 4.2 μM. In silico analysis revealed that, similarly to roscovitine, sunitinib fitted 6 of 10 features of the CDK5 pharmacophore. In a cell-based model, sunitinib reduced CDK5 phosphorylation (pCDK5), calpain-dependent p35/p25 conversion and protected neuronal cells from the toxic effects of gp120. In glial fibrillary acidic protein-gp120 transgenic (tg) mice, sunitinib reduced levels of pCDK5, p35/p25 and phosphorylated tau protein, along with amelioration of the neurodegenerative pathology. Compounds such as sunitinib with dual kinase inhibitory activity could ameliorate the cognitive impairment associated with chronic HIV infection of the CNS. Moreover, repositioning existing low MW compounds holds promise for the treatment of patients with neurodegenerative disorders.

1E7‐03, a low MW compound targeting host protein phosphatase‐1, inhibits HIV‐1 transcription

HIV-1 transcription is activated by the Tat protein which recruits the cyclin-dependent kinase CDK9/cyclin T1 to TAR RNA. Tat binds to protein phosphatase-1 (PP1) through the Q(35) VCF(38) sequence and translocates PP1 to the nucleus. PP1 dephosphorylates CDK9 and activates HIV-1 transcription. We have synthesized a low MW compound 1H4, that targets PP1 and prevents HIV-1 Tat interaction with PP1 and inhibits HIV-1 gene transcription. Here, we report our further work with the 1H4-derived compounds and analysis of their mechanism of action. Using the 1H4-PP1 complex as a model, we iteratively designed and synthesized follow-up libraries that were analysed for the inhibition of HIV-1 transcription and toxicity. We also confirmed the mechanism of action of the PP1-targeting molecules by determining the affinity of binding of these molecules to PP1, by analysing their effects on PP1 activity, disruption of PP1 binding to Tat and shuttling of PP1 to the nucleus. We identified a tetrahydroquinoline derivative, compound 7, which disrupted the interaction of Tat with PP1. We further optimized compound 7 and obtained compound 7c, renamed 1E7-03, which inhibited HIV-1 with low IC50 (fivefold lower than the previously reported compound, 1H4), showed no cytotoxicity and displayed a plasma half-life greater than 8 h in mice. 1E7-03 bound to PP1 in vitro and prevented shuttling of PP1 into the nucleus. Our study shows that low MW compounds that functionally mimic the PP1-binding RVxF peptide can inhibit HIV-1 transcription by deregulating PP1.

Novel TAZ modulators enhance myogenic differentiation and muscle regeneration.

The transcriptional co-activator with PDZ-binding motif (TAZ) is a key controller of mesenchymal stem cell differentiation through its nuclear localization and subsequent interaction with master transcription factors. In particular, TAZ directly associates with myoblast determining protein D (MyoD) and activates MyoD-induced myogenic gene expression, thereby enhancing myogenic differentiation. Here, we have synthesized and characterized low MW compounds modulating myogenic differentiation via induction of TAZ nuclear localization. COS7 cells stably transfected with GFP-TAZ were used in a high content imaging screen for compounds specifically enhancing nuclear localization of TAZ. We then studied the effects of such TAZ modulators on myocyte differentiation of C2C12 cells and myogenic transdifferentiation of mouse embryonic fibroblast cells in vitro and muscle regeneration in vivo. We identified two TAZ modulators, TM-53, and its structural isomer, TM-54. Each compound strongly enhanced nuclear localization of TAZ by reducing S89-phosphorylation and dose-dependently augmented myogenic differentiation and MyoD-mediated myogenic transdifferentiation through an activation of MyoD-TAZ interaction. The myogenic stimulatory effects of TM-53 and TM-54 were impaired in the absence of TAZ, but retrieved by the restoration of TAZ. In addition, administration of TM-53 and TM-54 enhanced injury-induced muscle regeneration in vivo and attenuated myofiber injury in vitro. The novel TAZ modulators TM-53 and TM-54 accelerated myogenic differentiation and improved muscle regeneration and function after injury, demonstrating that low MW compounds targeting the nuclear localization of TAZ have beneficial effects in skeletal muscle regeneration and in recovery from muscle degenerative diseases.

Impact of e-beam irradiation of municipal secondary effluent on MF and RO membranes performances

Due to water scarcity, development of innovative processes for wastewater reuse has become a priority. Hence, hybrid processes coupling membrane separation with advanced oxidation processes could play a decisive role in the production of safe water. However, membrane fouling is still one of the main limitations to process widespread application. Thus, the present study investigates the impact of e-beam irradiation on the fouling ability of a secondary effluent (SE) filtered on MF and subsequently on RO membranes. For this, two electron beam radiation doses (1 and 5kGy) were investigated. Results demonstrate that whatever the radiation dose investigated, no change in terms of TOC in SE was observed (prior to filtration). However, a strong decrease in UV254 and fluorescence signal was observed for organic matter (OM) exhibiting a MW ranging from 135 to 10kDa. Pre-oxidation was found to efficiently optimize MF filtration permeate flux whereas no improvement could be observed regarding RO separation. Moreover it was demonstrated that MF efficiently removed the high-MW compounds and was barely efficient in low-MW compounds removal. Energetic balance based on experimental results demonstrates that 1kGy was a sufficient dose to operate the hybrid e-beam irradiation and MF process.

A comparison of the character of algal extracellular versus cellular organic matter produced by cyanobacterium, diatom and green alga

This study investigated characteristics of algal organic matter (AOM) derived from three species (cyanobacterium Microcystis aeruginosa, diatom Fragilaria crotonensis and green alga Chlamydomonas geitleri) which dominate phytoplanktonic populations in reservoirs supplying drinking water treatment plants. Algal growth was monitored by cell counting, optical density and dissolved organic carbon concentration measurements. Extracellular organic matter (EOM) released at exponential and stationary growth phases and cellular organic matter (COM) were characterised in terms of specific UV absorbance (SUVA), peptide/protein and non-peptide content, hydrophobicity and molecular weight (MW). It was found that both EOM and COM were predominantly hydrophilic with low SUVA. COM was richer in peptides/proteins, more hydrophilic (with about 89% of hydrophilic fraction for all three species) and had lower SUVA than EOM. MW fractionation showed that both EOM and COM of all three species contain large portions of low-MW (<1 kDa) compounds and high-MW (>100 kDa) polysaccharides. Peptides/proteins exhibited narrower MW distribution than non-peptide fraction and it widened as the cultures grew. The highest amount of peptides/proteins with a significant portion of high-MW ones (22%) was observed in COM of M. aeruginosa. The results imply that the knowledge of AOM composition and characteristics predetermine which processes would be effective in the treatment of AOM laden water.

Use of Potentiometric Sensors To Study (Bio)molecular Interactions

Potentiometric sensors were used to study molecular interactions in liquid environments with sensorgram methodology. This is demonstrated with a lipophilic rubber-based and a collagen-based hydrogel sensor coating. The investigated molecules were promazine and tartaric acid, respectively. The sensors were placed in a hydrodynamic wall-jet system for the recording of sensorgrams. Millivolt sensor responses were first converted to a signal, expressing the concentration of adsorbed organic ions. Using a linearization method, a pseudo-first order-kinetic model of adsorption was shown to fit the experimental results perfectly. K(assoc), k(on), and k(off) values were calculated. The technique can be used over 4 decades of concentration, and it is very sensitive to low-MW compounds as well as to multiply charged large biomolecules. This study is the first to demonstrate the application of potentiometric sensors as an alternative and complement to surface plasmon resonance methods.

21 Concomitant activation of G-protein coupled receptors (GPCR) and sigma1 receptor (Sig1R): a novel pharmacological concept in treatment of Alzheimer's disease (AD)

The M1 muscarinic agonists AF102B (Evoxac: prescribed in Sjogren’s syndrome) and AF267B {successful in Phases I and IIa (Sjogren’s syndrome)} are effective cognitive enhancers and disease modifiers with a wide safety margin. Notably, i) AF102B decreased CSF Abeta in Alzheimer’s disease (AD) patients (Nitsch et al, Ann Neurol 2000); ii) AF267B rescued cognitive deficits and decreased Abeta42 and tau pathologies in 3xTg-AD mice (Caccamo et al, Neuron, 2006); and iii) in transgenic mice overexpressing human alpha-synuclein, AF102B and AF267B decreased brain alpha-synuclein aggregates (A. Fisher, E. Mazliah, B. Hutter-Paier, D. Havas, M. Windisch, ADPD 2011). Thus the M1 muscarinic GPCR appears to be a pivotal target for treatment of AD, Parkinson’s disease (PD) and Lewy body dementia (LBD). We have also hypothesized that concomitant activation of such a GPCR and the molecular chaperone Sig1R may have broader therapeutic implications in AD and related diseases. To achieve such combined activity we designed a low MW (357.5) compound, AF710B. This compound shows a novel mechanism of enhancing neuroprotection and cognition via Sig1R activation and M1 muscarinic modulation, but not resembling sigma1, M1 muscarinic (allosteric or orthosteric) and dual sigma1/M1 agonists, respectively. The effects of AF710B at low concentrations in vitro against neurodegeneration, oxidative stress, Abeta, Tau-phosphorylation and GSK-3beta activation translate into down-regulation of the apoptotic protein Bax and mitochondrial dysfunction, up-regulation of antiapoptotic Bcl2 and possible modulations of downstream kinases (e.g. PKC, PKB, GSK-3beta). AF710B has an exceptional pharmacology being an excellent cognitive enhancer in rats (at 1-30 and 10-100 mcg/kg, po in trihexyphenidyland in MK801-induced passive avoidance impairments, respectively). Furthermore, AF710B is devoid of side effects, having an unprecedented safety margin 50000 (po). In summary, while M1 agonists can alter pathologies in AD, PD and LBD, the unique profile of AF710B indicates widespread therapeutic advantage, inter alia, also on mitochondrial dysfunctions in these and other protein-aggregation related diseases. 22 NPT002: A NOVEL APPROACH FOR TARGETING -AMYLOID AND TAU AGGREGATES IN ALZHEIMER’S DISEASE

Preventing Pseudomonas aeruginosa and Chromobacterium violaceum infections by anti-adhesion-active components of edible seeds

BackgroundPseudomonas aeruginosa adhesion to animal/human cells for infection establishment involves adhesive proteins, including its galactose- and fucose-binding lectins PA-IL (LecA) and PA-IIL (LecB). The lectin binding to the target-cell receptors may be blocked by compatible glycans that compete with those of the receptors, functioning as anti-adhesion glycodecoys. The anti-adhesion treatment is of the utmost importance for abrogating devastating antibiotic-resistant P. aeruginosa infections in immunodeficient and cystic fibrosis (CF) patients. This strategy functions in nature in protecting embryos and neonates. We have shown that PA-IL, PA-IIL, and also CV-IIL (a PA-IIL homolog produced in the related pathogen Chromobacterium violaceum) are highly useful for revealing natural glycodecoys that surround embryos in diverse avian eggs and are supplied to neonates in milks and royal jelly. In the present study, these lectins were used as probes to search for seed embryo-protecting glycodecoys.MethodsThe lectin-blocking glycodecoy activities were shown by the hemagglutination-inhibition test. Lectin-binding glycoproteins were detected by Western blotting with peroxidase-labeled lectins.ResultsThe present work reports the finding - by using PA-IL, PA-IIL, and CV-IIL - of rich glycodecoy activities of low (< 10 KDa) and high MW (> 10 kDa) compounds (including glycoproteins) in extracts of cashew, cocoa, coffee, pumpkin, and tomato seeds, resembling those of avian egg whites, mammal milks, and royal jelly.ConclusionsEdible seed extracts possess lectin-blocking glycodecoys that might protect their embryos from infections and also might be useful for hampering human and animal infections.

TM‐25659 enhances osteogenic differentiation and suppresses adipogenic differentiation by modulating the transcriptional co‐activator TAZ

The transcriptional co-activator with PDZ-binding motif (TAZ) is characterized as a transcriptional modulator of mesenchymal stem cell differentiation into osteoblasts and adipocytes. Moreover, increased TAZ activity in the nucleus enhances osteoblast differentiation and suppresses adipocyte development by interacting with runt-related transcription factor 2 (RUNX2) and PPARγ, respectively. Therefore, it would be of interest to identify low MW compounds that modulate nuclear TAZ activity. High-throughput screening was performed using a library of low MW compounds in order to identify TAZ modulators that enhance nuclear TAZ localization. The effects and molecular mechanisms of a TAZ modulator have been characterized in osteoblast and adipocyte differentiation. We identified 2-butyl-5-methyl-6-(pyridine-3-yl)-3-[2'-(1H-tetrazole-5-yl)-biphenyl-4-ylmethyl]-3H-imidazo[4,5-b]pyridine] (TM-25659) as a TAZ modulator. TM-25659 enhanced nuclear TAZ localization in a dose-dependent manner and attenuated PPARγ-mediated adipocyte differentiation by facilitating PPARγ suppression activity of TAZ. In addition, TAZ-induced RUNX2 activity activation was further increased in osteoblasts, causing increased osteoblast differentiation. Accordingly, TM-25659 suppressed bone loss in vivo and decreased weight gain in an obesity model. After oral administration, TM-25659 had a favourable pharmacokinetic profile. TM-25659 stimulated nuclear TAZ localization and thus caused TAZ to suppress PPARγ-dependent adipogenesis and enhance RUNX2-induced osteoblast differentiation in vitro and in vivo. Our data suggest that TM-25659 could be beneficial in the control of obesity and bone loss.

Recent Advances in Metabolomics in Oncology

The burden of cancer is growing worldwide and with it a more desperate need for better tools to detect, diagnose and monitor the disease is required. It is well recognized that cancer cells are characterized by distinct metabolic perturbations. The metabolomics approach involves the comprehensive profiling of the full complement of low MW compounds in a biological system. By applying advanced analytical and statistical tools, the 'metabolome' is mined for biomarkers that are associated with the state of cancer. This review presents an introduction to the main analytical platforms used in metabolomics analyses, such as NMR spectroscopy and MS, as well as the statistical tools used to mine these datasets. The discussion focuses on 'state-of-the-art' investigations on the four cancer types that have received the most study by metabolomics, namely breast, prostate, colorectal and liver cancer.

Post-treatment of the permeate of a submerged anaerobic membrane bioreactor (SAMBR) treating landfill leachate

In this study, various methods were compared to reduce the Chemical Oxygen Demand (COD) content of stabilised leachate from a Submerged Anaerobic Membrane Bioreactor (SAMBR). It was found that Powdered Activated Carbon (PAC) resulted in greater COD removals (84 %) than Granular Activated Carbon (GAC-80 %), an ultrafiltration membrane of 1kDa (75 %), coagulation-flocculation with FeCl3 and polyelectrolyte (45 %), FeCl3 alone (32 %), and polymeric adsorbents such as XAD7HP (46 %) and XAD4 (32 %). Results obtained on the <1 kDa fraction showed that PAC and GAC had a similar adsorption efficiency of about 60 % COD removal, followed by XAD7HP (48 %), XAD4 (27 %) and then FeCl3 (23 %). The post-treatment sequence UF+GAC would result in a final effluent with less than 100 mg COD/L. Size Exclusion Chromatography (SEC) revealed that the extent of adsorption of low MW compounds onto PAC was limited due to low MW hydrophilic compounds, whereas the kinetics of PAC adsorption depended mainly on the adsorption of high MW aromatics.

ChemInform Abstract: Styrylpyrone‐Class Compounds from Medicinal Fungi Phellinus and Inonotus spp., and Their Medicinal Importance

AbstractReview: 76 refs.

New path in the thermal cracking of triacylglycerols (canola and soybean oil)

Triacylglycerols (TGs) are naturally occurring oils abundant in many crops. A series of batch uncatalyzed thermal decomposition experiments were performed using canola and soybean oils to explore pathways of TG cracking. A detailed gas chromatographic protocol based on mass spectrometric identification and flame ionization quantification was applied to the organic liquid product generated upon cracking. Reaction conditions were identified that resulted in a novel organic liquid product (OLP) composition compared to previously reported work. Under these conditions (temperatures within a 420–440°C range) a new route for TG thermolysis was discovered in which cracking reactions of original TG-bound fatty acids were nearly complete and led to the formation of 15–25wt.% C2–C10 linear saturated monocarboxylic acids and ca. 30% linear alkanes. Less than 2wt.% C16–C18 fatty acids which were originally present in the feedstocks as glycerol triesters were found in the OLP. These reactions appear to be kinetically controlled due to abundant hydrogen formation. This route provides a significant enrichment of low-MW compounds in the OLP (65–70wt.% being <C11) and thus may be considered as a new option for the production of replacement products for petroleum-based fuels and chemicals.

Mechanisms of Humic Acids Degradation by White Rot Fungi Explored Using 1H NMR Spectroscopy and FTICR Mass Spectrometry

Enzymatic activities involved in decay processes of natural aromatic macromolecules, such as humic acids (HA) and lignin by white rot fungi, have been widely investigated. However, the physical and chemical analysis of degradation products of these materials has not been intensively explored. Fourier transform cyclotron resonance mass spectrometry (FTICR MS) and 1H NMR as well as CHNOS and size exclusion chromatography were employed to study the mechanisms of HA degradation by Trametes sp. M23 and Phanerochaete sp. Y6. Size exclusion chromatography analyses demonstrate and provide evidence for HA breakdown into low MW compounds. The 1H NMR analysis revealed oxidation, a decrease in the aromatic content, and an indication of demethylation of the HA during biodegradation. Evidence for oxidation was also obtained using CHNOS. Analysis of FTICR MS results using a new software program developed by our group (David Mass Sort) revealed consecutive series of masses suggesting biochemical degradation trends such as oxidation, aromatic cleavage, and demethylation. These results are in agreement with the 1H NMR analysis and with the suggested role of the ligninolytic system leading to HA degradation.

Styrylpyrone-class compounds from medicinal fungi Phellinus and Inonotus spp., and their medicinal importance

Members of the genera Phellinus and Inonotus, including P. linteus, P. igniarius, P. ribis, I. obliquus and I. xeranticus are well-known medicinal fungi (mushrooms) and have been used in treatment of cancer, diabetes, bacterial and viral infections and ulcer. Adverse effects of these medicinal mushrooms have not yet been reported, indicating the safe nature of these mushrooms. Polysaccharides, particularly β-glucan, are considered the compounds responsible for the biological activity of medicinal mushrooms. However, there is only a limited amount of evidence to indicate that polysaccharides are in fact responsible for the biological effects of these medicinal mushrooms. Recently, many research groups have begun identification of active low-MW compounds in medicinal mushrooms, with a focus on the yellow polyphenol pigments, which are composed of a styrylpyrone class of compounds. Interestingly, a representative group of medicinal fungi, including P. linteus, P. igniarius, P. ribis, I. obliquus and I. xeranticus were shown to produce a large and diverse range of styrylpyrone-type polyphenol pigments that exhibited various biological activities, including anti-oxidative, anti-inflammatory, cytotoxic, anti-platelet aggregation, anti-diabetic, anti-dementia and anti-viral effects. Styrylpyrone pigments in mushrooms are thought to have a role similar to that of flavonoids in plants. The unique and unprecedented carbon skeleton of fused styrylpyrone might be an attractive molecular scaffold for pharmacological applications. In this review, the structural diversity, biological effects and biogenesis of styrylpyrone-class polyphenols from medicinal fungi are described.