Synthesis and structure of metallated macrocycle-bearing cyclophophazenes. - Part I : The (Li, Mg, Zn)/SPIR0(30203) derivatives

Syntheses and molecular structures of three metallated (Li, Zn, Mg) derivatives of the macrocycle-bearing cyclophosphazene N3P3Cl4 [HN---(CH2)3---O---(CH2)2---O---(CH2)3---NH] (coded as SPIRO 30203, 1) are reported. These three molecular structures reveal three different patterns of metal coordination. In compound 2 one of the two hydrogen atoms in SPIRO 3O2O3 is substituted by lithium generating a dimeric structure with pentacoordinated lithium centers. This structure is further stabilized by N---H hydrogen bonds. In 3 both hydrogen atoms of the macrocyclic loop are replaced by two zinc atoms through a cross-link metallation leading again to a dimeric molecule. In this compound the Zn atom is found to be in a trigonal bipyramidal environment with one very long N---Zn interaction. The origin of the dimerization of the magnesium compound 4 is analogous to 3. Magnesium is in the center of a distorted octahedron, coordinated with the O- and N-donors of the macrocyclic loop and also one nitrogen atom of the N3P3 ring. 4 is the first example of a metallic center coordinated by a neutral phosphazene ligand. Typical metal-N and metal-O distances are (in Å): Li-O, 2.05—2.07; Li-N, 2.07—2.36; Zn-O, 2.08—2.14; Zn---N, 1.94—1.95 (2.49); Mg---O, 2.09—2.14; Mg---N, 2.07—2.31

Synthesis and structure of metallated macrocycle-bearing cyclophophazenes. - Part II : The (Al) /SPIRO (30203) derivatives

Synthesis and molecular structures of two metallated (aluminium) derivatives, [C9H19Cl4AlN5O2P3]· 1.5C7H8 and [C8H16Cl5AiN5O2P3] · 1.5C7H8, of the macrocycle-bearingcyclophosphazene N3P3Cl4 [HN--- (CH2)3---O---(CH2)2---O---(CH2)3-NH] (coded as SPIRO 30203) are reported. These two molecular structures reveal the same pattern of metal coordination where the two hydrogen atoms in SPIRO 30203 are substituted by aluminium generating monomeric structures with pentacoordinated aluminium centres in the inner cavities. In the first compound the exocyclic ligand at aluminium is a methyl group, in the second a chlorine atom. Typical Al---N, Al---O and Al-X distances are (Å):1.84-1.89, 1.97-2.04 and 1.93 in the former, 1.81-1.86, 1.93-2.01 and 2.12 in the latter, respectively

Ultra-structural cell distribution of the melanoma marker iodobenzamide: improved potentiality of SIMS imaging in life sciences

BACKGROUND: Analytical imaging by secondary ion mass spectrometry (SIMS) provides images representative of the distribution of a specific ion within a sample surface. For the last fifteen years, concerted collaborative research to design a new ion microprobe with high technical standards in both mass and lateral resolution as well as in sensitivity has led to the CAMECA NanoSims 50, recently introduced onto the market. This instrument has decisive capabilities, which allow biological applications of SIMS microscopy at a level previously inaccessible. Its potential is illustrated here by the demonstration of the specific affinity of a melanoma marker for melanin. This finding is of great importance for the diagnosis and/or treatment of malignant melanoma, a tumour whose worldwide incidence is continuously growing. METHODS: The characteristics of the instrument are briefly described and an example of application is given. This example deals with the intracellular localization of an iodo-benzamide used as a diagnostic tool for the scintigraphic detection of melanic cells (e.g. metastasis of malignant melanoma). B16 melanoma cells were injected intravenously to C(57)BL(6)/J(1)/co mice. Multiple B16 melanoma colonies developed in the lungs of treated animals within three weeks. Iodobenzamide was injected intravenously in tumour bearing mice six hours before sacrifice. Small pieces of lung were prepared for SIMS analysis. RESULTS: Mouse lung B16 melanoma colonies were observed with high lateral resolution. Cyanide ions gave "histological" images of the cell, representative of the distribution of C and N containing molecules (e.g. proteins, nucleic acids, melanin, etc.) while phosphorus ions are mainly produced by nucleic acids. Iodine was detected only in melanosomes, confirming the specific affinity of the drug for melanin. No drug was found in normal lung tissue. CONCLUSION: This study demonstrates the potential of SIMS microscopy, which allows the study of ultra structural distribution of a drug within a cell. On the basis of our observations, drug internalization via membrane sigma receptors can be excluded

Promising pre-clinical validation of targeted radionuclide therapy using a [131I] labelled iodoquinoxaline derivative for an effective melanoma treatment

Targeted internal radionuclide therapy (TRT) would be an effective alternative to current therapies for dissemi- nated melanoma treatment. At our institution, a class of iodobenzamides has been developed as potent melanoma- seeking agents. This review described the preclinical vali- dations of a quinoxaline derivative molecule (ICF01012) as tracer for TRT application. It was selected for its high, specific and long-lasting uptake in tumour with rapid clear- ance from non-target organs providing suitable dosimetry parameters for TRT. Extended in vivo study of metabolic profiles confirmed durable tumoural concentration of the unchanged molecule form. Moreover melanin specificity of ICF01012 was determined by binding assay with syn- thetic melanin and in vivo by SIMS imaging. Then, we showed in vivo that [131I] ICF01012 treatment drastically inhibited growth of B16F0, B16Bl6 and M4Beu tumours whereas [131I] NaI or unlabelled ICF01012 treatment was without significant effect. Histological analysis showed that residual tumour cells exhibit a significant loss of aggres- siveness after treatment. This anti-tumoural effect was associated with a lengthening of the treated-mice survival time and an inhibition of lung dissemination for B16Bl6 model. Results presented here support the concept of TRT using a [131I] labelled iodoquinoxaline derivative for an effective melanoma treatment.<br /

Feasibility study for a microwave-powered ozone sniffer aircraft

The preliminary design of a high-altitude, remotely-piloted, atmospheric-sampling aircraft powered by microwave energy beamed from ground-based antenna was completed. The vehicle has a gross weight of 6720 pounds and is sized to carry a 1000 pound payload at an altitude of 100,000 feet. The underside of the wing serves as the surface of a rectenna designed to receive microwave energy at a power density of 700 watts per square meter and the wing has a planform area of 3634 square feet to absorb the required power at an optimum Mach number M = 0.44. The aircraft utilizes a horizontal tail and a canard for longitudinal control and to enhance the structural rigidity of the twin fuselage configuration. The wing structure is designed to withstand a gust-induced load factor n = 3 at cruise altitude but the low-wing loading of the aircraft makes it very sensitive to gusts at low altitudes, which may induce load factors in excess of 20. A structural load alleviation system is therefore proposed to limit actual loads to the designed structural limit. Losses will require transmitted power on the order of megawatts to be radiated to the aircraft from the ground station, presenting environmental problems. Since the transmitting antenna would have a diameter of several hundred feet, it would not be readily transportable, so we propose that a single antenna be constructed at a site from which the aircraft is flown. The aircraft would be towed aloft to an initial altitude at which the microwave power would be utilized. The aircraft would climb to cruise altitude in a spiral flight path and orbit the transmitter in a gentle turn

N-(4-iodophenyl)-N′-(2-chloroethyl)urea as a microtubule disrupter: in vitro and in vivo profiling of antitumoral activity on CT-26 murine colon carcinoma cell line cultured and grafted to mice

The antitumoral profile of the microtubule disrupter N-(4-iodophenyl)-N′-(2-chloroethyl)urea (ICEU) was characterised in vitro and in vivo using the CT-26 colon carcinoma cell line, on the basis of the drug uptake by the cells, the modifications of cell cycle, and β-tubulin and lipid membrane profiles. N-(4-iodophenyl)-N′-(2-chloroethyl)urea exhibited a rapid and dose-dependent uptake by CT-26 cells suggesting its passive diffusion through the membranes. Intraperitoneally injected ICEU biodistributed into the grafted CT-26 tumour, resulting thus in a significant tumour growth inhibition (TGI). N-(4-iodophenyl)-N′-(2-chloroethyl)urea was also observed to accumulate within colon tissue. Tumour growth inhibition was associated with a slight increase in the number of G2 tetraploid tumour cells in vivo, whereas G2 blockage was more obvious in vitro. The phenotype of β-tubulin alkylation that was clearly demonstrated in vitro was undetectable in vivo. Nuclear magnetic resonance analysis showed that cells blocked in G2 phase underwent apoptosis, as confirmed by an increase in the methylene group resonance of mobile lipids, parallel to sub-G1 accumulation of the cells. In vivo, a decrease of the signals of both the phospholipid precursors and the products of membrane degradation occurred concomitantly with TGI. This multi-analysis established, at least partly, the ICEU activity profile, in vitro and in vivo, providing additional data in favour of ICEU as a tubulin-interacting drug accumulating within the intestinal tract. This may provide a starting point for researches for future efficacious tubulin-interacting drugs for the treatment of colorectal cancers

Massive multiplication of genome and ribosomes in dormant cells (akinetes) of Aphanizomenon ovalisporum (Cyanobacteria)

Author Posting. © The Author(s), 2011. This is the author's version of the work. It is posted here by permission of Nature Publishing Group for personal use, not for redistribution. The definitive version was published in The ISME Journal 6 (2012): 670–679, doi:10.1038/ismej.2011.128.Akinetes are dormancy cells commonly found among filamentous cyanobacteria, many of which are toxic and/or nuisance, bloom-forming species. Development of akinetes from vegetative cells is a process that involves morphological and biochemical modifications. Here we applied a single cell approach to quantify genome and ribosome content of akinetes and vegetative cells in Aphanizomenon ovalisporum (Cyanobacteria). Vegetative cells of A. ovalisporum were naturally polyploid and contained on average 8 genome copies per cell. However, the chromosomal content of akinetes increased up to 450 copies, with an average value of 119 genome copies per akinete, 15 fold higher that in vegetative cells. Based on fluorescence in situ hybridization with a probe targeting 16S rRNA and detection with confocal laser scanning microscopy we conclude that ribosomes accumulated in akinetes to a higher level than that found in vegetative cells. We further present evidence that this massive accumulation of nucleic acids in akinetes is likely supported by phosphate supplied from inorganic polyphosphate bodies that were abundantly present in vegetative cells, but notably absent from akinetes. These results are interpreted in the context of cellular investments for proliferation following long term dormancy, as the high nucleic acid content would provide the basis for extended survival, rapid resumption of metabolic activity and cell division upon germination.Supported by the Gruss Lipper Foundation research award (AS). This study was part of the Joint German-Israeli-Project (FKZ 02WT0985, WR803) funded by the German Ministry of Research and Technology (BMBF) and Israel Ministry of Science and Technology (MOST)