intravital-microscopy-imaging-liver-following-leishmania-infection-an

Intravital Microscopy Imaging of the Liver following <em>Leishmania</em> Infection: An Assessment of Hepatic Hemodynamics

Intravital microscopy (IVM) is a powerful optical imaging technique that has made possible the visualization, monitoring and quantification of various ...

University of Aix-Marseille

Taxol treatment of HeLa cells resulted in a transient recruitment of Caveolin-1 to the cell surface followed by internalization. Interestingly, 20min after 10-deacetylbaccatinIII (10-DAB) treatment, the caveolae displayed faster 'kiss and run' dynamics while BaccatinIII (BacIII) did not induce any change. Sustained phosphorylation of Caveolin-1 is observed upon treatment and between Taxol and 10-DAB, the former shows phosphorylated Raf-1, ERK1/2 and hyperphosphorylated Bcl-2 while the later showed much less magnitude of the same. BacIII treatment did not induce phosphorylation of Raf-1 or Bcl-2. It is possible that Taxol might act on multiple targets and the side chain may be crucial.

Taxol and 10-deacetylbaccatinIII induce distinct changes in the dynamics of caveolae.

We have isolated endophytic fungi from Indian yew tree, Taxus baccata and then screened for taxol production. Out of the forty fungal cultures screened, one fungus Gliocladium sp. was found to produce taxol and 10DAB III (10 Deacetyl baccatin III). These compounds were purified by TLC, HPLC and characterized using UV-Spectroscopy, ESI-MS, MS/MS and proton NMR. One liter of Gliocladium sp. culture yielded 10 microg of taxol and 65 microg of 10 DAB III. The purified taxol from the fungus showed cytotoxicity towards cancer lines HL-60 (leukemia), A431 (epidermal carcinoma) and MCF-7 (breast cancer).

Production, purification and characterization of taxol and 10DAB III from a new endophytic fungus Gliocladium sp. isolated from the Indian yew tree, Taxus baccata.

The title compound (AME; systematic name: 3,7-dihy-droxy-9-meth-oxy-1-methyl-6H-benzo[c]chromen-6-one), C(15)H(12)O(5), was isolated from an endophytic fungi Alternaria sp., from Catharanthus roseus (common name: Madagascar periwinkle). There is an intramolecular O-H⋯O hydrogen bond in the essentially planar mol-ecule (r.m.s. deviation 0.02 Å). In the crystal, the molecule forms an O-H⋯O hydrogen bond with its centrosymmetric counterpart with four bridging inter-actions (two O-H⋯O and two C-H⋯O). The almost planar sheets of the dimeric units thus formed are stacked along b axis via C-H⋯π and π-π contacts [with C⋯C short contacts between aromatic moieties of 3.324 (3), 3.296 (3) and 3.374 (3) Å].

Alternariol 9-O-methyl ether.

3,7-dihy-droxy-9-meth-oxy-1-methyl-6H-benzo[c]chromen-6-one dimethyl sulfoxide monosolvate), C15H12O5·C2H6OS, was isolated from an unidentified endophytic fungus (belonging to class Ascomycetes) of Taxus sp. In the crystal, both the alternariol 9-O-methyl ether (AME) and the dimethyl sulfoxide (DMSO) mol-ecules exhibit crystallographic mirror symmetry. One of the hy-droxy groups makes bifurcated hydrogen bonds, viz. an intra-molecular bond with the carbonyl group and an inter-molecular bond with the carbonyl group in an inversion-related AME mol-ecule. In the crystal, the AME mol-ecules are organized into stacks parallel with the b axis by π-π inter-actions between centrosymmetrically related mol-ecules [the distance between the centroid of the central ring and the centroid of the meth-oxy-substituted benzene ring in the next mol-ecule of the stack is 3.6184 (5) Å]. Pairs of DMSO mol-ecules, linked via centrosymmetric C-H⋯O contacts, are inserted into the voids created by the AME mol-ecules, making O-H⋯O and C-H⋯O contacts with the hosts.

Alternariol 9-O-methyl ether dimethyl sulfoxide monosolvate.

Leishmaniasis is a vector-borne disease for which only limited therapeutic options are available. The disease is ranked among the six most important tropical infectious diseases and represents the second-largest parasitic killer in the world. The development of new therapies has been hampered by the lack of technologies and methodologies that can be integrated into the complex physiological environment of a cell or organism and adapted to suitable in vitro and in vivo Leishmania models. Recent advances in microscopy imaging offer the possibility to assess the efficacy of potential drug candidates against Leishmania within host cells. This technology allows the simultaneous visualization of relevant phenotypes in parasite and host cells and the quantification of a variety of cellular events. In this review, we present the powerful cellular imaging methodologies that have been developed for drug screening in a biologically relevant context, addressing both high-content and high-throughput needs. Furthermore, we discuss the potential of intra-vital microscopy imaging in the context of the anti-leishmanial drug discovery process.

Simultaneous multi-parametric analysis of Leishmania and of its hosting mammal cells: A high content imaging-based method enabling sound drug discovery process.

Article	Year
Taxol and 10-deacetylbaccatinIII induce distinct changes in the dynamics of caveolae. FEBS letters\| PubMed ID: 18817775	2008
Production, purification and characterization of taxol and 10DAB III from a new endophytic fungus Gliocladium sp. isolated from the Indian yew tree, Taxus baccata. Journal of microbiology and biotechnology\| PubMed ID: 19996685	2009
Alternariol 9-O-methyl ether. Acta crystallographica. Section E, Structure reports online\| PubMed ID: 22590344	2012
Alternariol 9-O-methyl ether dimethyl sulfoxide monosolvate. Acta crystallographica. Section E, Structure reports online\| PubMed ID: 23795051	2013
Simultaneous multi-parametric analysis of Leishmania and of its hosting mammal cells: A high content imaging-based method enabling sound drug discovery process. Microbial pathogenesis\| PubMed ID: 25448129	2014

Sreekanth Dasari

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