Isolation,enzyme-bound structure and antibacterial activity of platencin A1 from Streptomyces platensis

Natural products continue to serve as one of the best sources for discovery of antibacterial agents as exemplified by the recent discoveries of platensimycin and platencin. Chemical modifications as well as discovery of congeners are the main sources for gaining knowledge of structure–activity relationship of natural products. Screening for congeners in the extracts of the fermentation broths of Streptomyces platensis led to the isolation of platencin A₁, a hydroxy congener of platencin. The hydroxylation of the tricyclic enone moiety negatively affected the antibacterial activity and appears to be consistent with the hydrophobic binding pocket of the FabF. Isolation, structure, enzyme-bound structure and activity of platencin A₁ and two other congeners have been described.     

Isolation, structure, and HIV-1-integrase inhibitory activity of structurally diverse fungal metabolites

HIV-1 integrase is a critical enzyme for replication of HIV, and its inhibition is one of the most promising new drug strategies for anti-retroviral therapy, with potentially significant advantages over existing therapies. In this report, a series of HIV-1 inhibitors isolated from the organic extract of fermentations from terrestrial fungi is described. These fungal species, belonging to a variety of genera, were collected from throughout the world following the strict guidelines of Rio Convention on Biodiversity. The polyketide- and terpenoid-derived inhibitors are represented by two naphthoquinones, a biphenyl and two triphenyls, a benzophenone, four aromatics with or without catechol units, a linear aliphatic terpenoid, a diterpenoid, and a sesterterpenoid. These compounds inhibited the coupled and strand-transfer reaction of HIV-1 integrase with an IC₅₀ value of 0.5–120 µM. The bioassay-directed isolation, structure elucidation, and HIV-1 inhibitory activity of these compounds are described.Abbreviations CH ₃ CN Acetonitrile - TFA Trifluoroacetic acid - CD ₃ CN Deuterio acetonitrile - CD ₂ Cl ₂ Deuterio methylene chloride - MEK Methyl ethyl ketone - ZnSe Zinc selenide - UV Ultraviolet - IR Infrared - NMR Nuclear magnetic resonance - COSY Homonuclear H-H correlation spectroscopy - HMQC Heteronuclear multiple-quantum coherence - HMBC Heteronuclear multiple-bond correlation - EIMS Electron-impact mass spectrometry - HREIMS High-resolution electron impact mass spectrometry - ESIMS Electrospray ionization mass spectrometry - HRFABMS High-resolution fast atom bombardment mass spectrometry - MES 2-(N-morpholino)-ethane sulfonic acid     

Identification of diverse microbial metabolites as potent inhibitors of HIV-1 Tat transactivation

HIV-1 Tat is one of six regulatory proteins that are required for viral replication and is an attractive target for the development of new anti-HIV agents. Screening of microbial extracts using a whole cell Tat-dependent transactivation assay, which guided the separation of the active broths, led to the identification of five structurally diverse classes (M(R) range 232-1126) of natural products. These include i) three sesquiterpenoids, namely, sporogen-AO1, petasol, and 6-dehydropetasol, ii) two resorcylic 14-membered lactones, namely monorden and monocillin IV, iii) a ten-membered lactone, iv) a quinoline and quinoxiline bicyclic octadepsipeptides, namely echinomycin and UK-63598, and v) a cyclic heptapeptide, ternatin. These compounds displayed varying degrees of potencies with IC50 values ranging from 0.0002 to 100 microM. The most active compound was the quinoxiline bicyclic octadepsipeptides, UK-63598, which inhibited Tat-dependent transactivation with an IC50 value of 0.2 nM and exhibited a 100-fold therapeutic window with respect to toxicity. In a single-cycle antiviral assay, UK-6358 inhibited viral replication with an IC50 value of 0.5 nM; however, it appeared to be equally toxic at that concentration. Monocillin IV was significantly less active (Tat transactivation inhibitory IC50 of 5 microM) but was not toxic at 100 microM in an equivalent cytotoxicity assay. The compound exhibited antiviral activity with an IC50 value of 6.2 microM in the single-cycle antiviral assay and a sixfold therapeutic window. Details of the isolation, fermentation, and biological activities of these structurally diverse natural products are described.     

A Computer Generated Model of Adenosine Receptors Rationalising Binding and Selectivity of Receptor Ligands

Abstract

Computer graphic analyses on a broad spectrum of adenosine receptor ligands has shown that both the A1 and A2 adenosine receptors have three binding sites. The spatial relationship of these three binding sites has been defined. Adenosine orientation at A1 and A2 is different.     

Isolation,structure and biological activity of phomafungin,a cyclic lipodepsipeptide from a widespread tropical Phoma sp.

We isolated a cyclic lipodepsipeptide, phomafungin, from a Phoma sp. The distinct antifungal activity of phomafungin in the crude extract was initially discovered by mechanistic profiling in the Candida albicans fitness test. The purified compound contains a 28 member ring consisting of eight amino acids and a β-hydroxy-γ-methyl-hexadecanoic acid, and displays a broad spectrum of antifungal activity against Candida spp., Aspergillus fumigatus and Trichophyton mentagrophytes with MIC of 2–8 μg/ml, and toxicity to mice at 25 mg/kg. The linear peptide derived from opening of the lactone ring was devoid of antifungal activity as well as toxicity. Phomafungin has been identified in a number of Phoma spp. collected from Africa and the Indian and Pacific Ocean islands.     

Leaf Litter Decomposition of Three Riparian Tree Species and Associated Macroinvertebrates of Eswathu Oya,a Low Order Tropical Stream in Sri Lanka

Leaf decomposition, an important component of the organic matter dynamics in streams, has been widely examined in temperate regions but much less documented in tropical regions. We report here the first study of leaf decomposition in a Sri Lankan stream. The litterbag technique was used. Coarse (8 mm) and fine (100 µm) litterbags, that included or excluded macroinvertebrates respectively, were used to enclose leaves of three dominant riparian tree species: native Ochlandra stridula (bamboo), and the introduced Alstonia macrophylla and Hevea brasiliensis (rubber). A fourth set of litterbags contained a mixture of these three species. Leaf colonization by macroinvertebrates was highest in the early stages of the decomposition process on Hevea leaves but invertebrate densities declined later. The opposite colonization effect was observed on the native Ochlandra leaves: slow colonization with continuing low densities from the beginning to the end of the process. Decomposition of all three species was significantly faster in the coarse than in the fine mesh bags. Alstonia, which has the softest leaf tissue, was most rapidly decomposed while Ochlandra, with its tough structure, was the slowest. Among the invertebrates, insects were the most important leaf colonizing animals, with Diptera, Coleoptera and Trichoptera the most dominant. The invertebrate variety in the mixed bags was higher than in the single‐species leaf bags, where Chironomidae dominated the colonizing assemblages. This study has shown that toughness, indicated by the ‘specific weight of leaf tissue’, and the quality of the leaves was more important in determining breakdown rates than their origin. (© 2011 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Reversibility of replicative senescence in Saccharomyces cerevisiae: effect of homologous recombination and cell cycle checkpoints

Primary human somatic cells grown in culture divide a finite number of times, exhibiting progressive changes in metabolism and morphology before cessation of cycling. This telomere-initiated cellular senescence occurs because cells have halted production of telomerase, a DNA polymerase required for stabilization of chromosome ends. Telomerase-deficient Saccharomyces cerevisiae cells undergo a similar process, with most cells arresting growth after approximately 60 generations. In the current study we demonstrate that senescence is largely reversible. Reactivation of telomerase (EST2) expression in the growth-arrested cells led to resumption of cycling and reversal of senescent cell characteristics. Rescue was also observed after mating of senescent haploid cells with telomerase-proficient cells to form stable diploids. Although senescence was reversible in DNA damage checkpoint response mutants (mec3 and/or rad24 cells), survival of recombination-defective rad52 mutants remained low after telomerase reactivation. Telomere lengths in rescued est2 cells were initially half those of wildtype cells, but could be restored to normal by propagation for ～70 generations in the presence of telomerase. These results place limitations on possible models for senescence and indicate that most cells, despite gross morphological changes and short, resected telomeres, do not experience lethal DNA damage and become irreversibly committed to death.     

Current status of heavy metal contamination in Asia’s rice lands

Major contribution (over 90 %) to the world’s rice production is coming from Asia, where metal contamination of agricultural lands is often reported. Thus the present paper reviews the sources and current status of heavy metal contamination of paddy lands in the region. Apart from the natural sources, agrochemicals, wastewater irrigation, sewage sludge application, livestock manures, mining and fly ash etc., could be identified as the key sources of metal contamination in Asia. Accumulation of heavy metals and metalloids (Cd, Zn, Cu, Pb, Cr, Ni, Fe, Zn, Co, Hg and As) in different parts of the rice plant (roots, straw, hull and grain) is reported at varying degrees. Rice grain accumulates the least amount of toxic metals compared to hull, straw and roots. Most importantly, a greater number of investigations confirmed that the metal contents in rice grains are within the permissible limits of Codex recommendations of joint Food and Agriculture Organization/World Health Organization Food Standards Programme and/or food regulations imposed by the respective governments. However, due to the fact that rice is a main route of human exposure to heavy metals, appropriate preventive and remedial measures should be enforced in the areas with potential risk of metal contamination.     

Discovery of a small molecule that inhibits cell division by blocking FtsZ,a novel therapeutic target of antibiotics

The emergence of bacterial resistance to antibiotics is a major health problem and, therefore, it is critical to develop new antibiotics with novel modes of action. FtsZ, a tubulin-like GTPase, plays an essential role in bacterial cell division, and its homologs are present in almost all eubacteria and archaea. During cell division, FtsZ forms polymers in the presence of GTP that recruit other division proteins to make the cell division apparatus. Therefore, inhibition of FtsZ polymerization will prevent cells from dividing, leading to cell death. Using a fluorescent FtsZ polymerization assay, the screening of >100,000 extracts of microbial fermentation broths and plants followed by fractionation led to the identification of viriditoxin, which blocked FtsZ polymerization with an IC50 of 8.2 microg/ml and concomitant GTPase inhibition with an IC50 of 7.0 microg/ml. That the mode of antibacterial action of viriditoxin is via inhibition of FtsZ was confirmed by the observation of its effects on cell morphology, macromolecular synthesis, DNA-damage response, and increased minimum inhibitory concentration as a result of an increase in the expression of the FtsZ protein. Viriditoxin exhibited broad-spectrum antibacterial activity against clinically relevant Gram-positive pathogens, including methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococci, without affecting the viability of eukaryotic cells.