Photophysical comparative study of amylose and polyvinyle pyrrolidone/single walled carbon nanotubes complex.

Progressive addition of hydroxypropylated amylose (AmH), from 0.05 wt% to 4.5 wt%, to single-walled carbon nanotubes (SWNTs) in aqueous surfactant suspensions quenches the intrinsic near Infra-Red fluorescence of semiconducting SWNTs while dispersions obtained with a same amount of polyvinylpyrrolidone (PVP) remain luminescent. Near Infra-Red emission spectroscopy (fluorescence and Raman scattering) of the samples is used to characterize the supramolecular organization of these polymer/SWNT complexes. The SWNTs are found to be wrapped by the PVP chains and not by the AmH chains which rather form AmH/surfactant/SWNTs complexes. In PVP/SWNTs dispersion, the fluorescence line position and intensity are affected by dielectric screening. In the case of AmH/surfactant/SWNTs complex, dielectric screening plays also a role but quenching occurs above about 3 wt% of AmH. We attribute the quenching to the formation of a "composite like" microstructure by opposition to stabilized dispersion.

[A new therapy with bortezomib, an oncologic medicinal product of the year 2004]. / Une nouvelle thérapie ciblée avec le bortézomib, médicament oncologique de l'année 2004.

Proteasome-mediated proteolysis is a mechanism for mediating important regulatory proteins within the cell. Proteins that have been targeted for degradation by the proteasome are convalently tagged with a poly-ubiquitin protein chain prior to be recognized by the 19S subunit of proteasome. This degradation system controls the expression of a wide variety of cellular targets including tumor suppressors such as p53, inhibitor of nuclear factor NFkappaB, cyclin-dependent kinase inhibitors such as p21 and p27. Because of these functions, the proteasome has become a new target for cancer treatment. The potent and selective proteasome inhibitor, PS-341 or Velcade was approved in the United States and launched in may 2003 for the treatment of multiple myeloma patients who have received at least two prior therapies. On April 2004, the European commission granted marketing authorization for Velcade with the same indication. The same year 2004, the Nobel Prize in chemistry was awarded to three researchers "for the discovery of ubitiquin-mediated protein degradation", a regulated process by which proteins are cleaved into peptides inside cells.

Photobiological activities of 1,6-dioxapyrene in pro- and eukaryotic cells.

The photobiological effect of a new pyrene derivative, 1,6-dioxapyrene (1,6-DP), was studied in Salmonella typhimurium (strain TA100) and in the diploid strain D7 of the yeast Saccharomyces cerevisiae. In Salmonella, 1,6-DP shows little mutagenicity in the dark in comparison to benzo[a]pyrene (B[a]P). This mutagenic activity decreases in the presence of liver S9 homogenates from Aroclor induced XVIInc/Z mice. However, in combination with 365 nm (UVA) radiation and in the absence of S9 mix, 1,6-DP behaves as an effective photodynamic compound inducing lethal and mutagenic effects in both organisms. In yeast, its activity, like that of B[a]P, is highly dependent on the presence of oxygen. For the same incident dose of UVA, 1,6-DP is, however, at least 6 times more effective than B[a]P in inducing cytotoxic and mutagenic effects. At equitoxic doses, 1,6-DP is as photomutagenic as B[a]P, suggesting that in both cases mutagenicity is due to similar mechanisms. Spectrophotometric measurements indicate physical interaction of 1,6-DP with DNA in the dark. Laser flash photolysis experiments show that 1,6-DP generates singlet oxygen with a quantum yield of 0.17. In vitro 1,6-DP produces oxidative damage to guanine bases specific for singlet oxygen mediated reactions. Alkaline step elution analysis of 1,6-DP plus UVA treated yeast cells indicates a decrease in average molecular weights in DNA and an induction of single strand breaks (ssb) originating from alkali labile sites. This effect is enhanced by D2O and is thus likely to be due to the production of singlet oxygen. The strand breaks appear to differ from those induced by gamma-rays because little, if any, repair of these ssb occurs during 30 min of post-treatment incubation in complete growth medium. These results suggest that the photobiological effects of 1,6-DP are due to oxidative damage in DNA mostly induced by singlet oxygen.

Free radical production during photo-assisted NADPH reduction of four alpha-nitroarenofurans.

Generation of radical anions during NADPH reduction of four mutagenic and genotoxic alpha-nitroarenofurans was examined. ESR showed that free radicals were generated during reduction solely in the presence of light. Computer simulations of ESR spectra were in good agreement with the experimental ones.

Genotoxic effectivity--comparison of 36 nitrated furan and arenofuran derivatives on a quantitative scale. Statistical comparison of T7 and other short-term tests.

Thirty-six nitrated furan and arenofuran derivatives were measured and quantitatively characterized by the T7 inactivation test. A wide range of substances previously studied allowed us to compare the collected quantitative data with those obtained by other workers using different short-term tests. Based on comparative statistical evaluation of these data a borderline was determined for the genotoxic effect: compounds having in our short-term test mutagenicity index (MI) values smaller than 8.0 are positive while the higher values represent negative genotoxicity. Classification of 36 nitrofuran/nitroarenofuran derivatives is given both on the basis of the quantitative genotoxicity scale and in terms of +/- on the qualitative scale. All but one compound were found to be genotoxic and the genotoxic activities of these compounds were compared with the results of other carcinogenicity or mutagenicity tests.

Characterization of the nonenzymatic chloramphenicol resistance (cmlA) gene of the In4 integron of Tn1696: similarity of the product to transmembrane transport proteins.

Integrons constitute a novel family of DNA elements which evolved by site-specific integration of discrete units between two conserved segments. On the In4 integron of Tn1696, a precisely inserted gene cassette of 1,549 bp conferring nonenzymatic chloramphenicol resistance (cmlA) is present between the streptomycin-spectinomycin resistance (aadA2) gene cassette and the 3'-conserved segment of the integron. In this study, we present the nucleotide sequence of the cmlA gene cassette of Tn1696, show its similarity to bacterial efflux systems and other transport proteins, and present evidence for alterations that its expression exerts on bacterial membranes. The cmlA gene cassette apparently carries its own promoter(s), a situation that has not heretofore been observed in the integrons of multiresistance plasmids and transposons of gram-negative bacteria. One or more of these promoters were shown to be functionally active in expressing a cat marker gene from promoter-probe vectors. The putative CmlA polypeptide appears to provoke a reduction of the content of the major porins OmpA and OmpC.

Genotoxicity testing: phage T7 inactivation test of various furan and arenofuran derivatives.

The genotoxic activities of 28 furan and arenofuran derivatives were tested by the phage T7-inactivation test. The genotoxic activity of the compounds was characterized quantitatively. All the compounds studied have pronounced genotoxic activities in our system. Empirical rules relating structure to genotoxic activity were found. Data obtained with our system were compared with the results of other biological systems (Salmonella assay, SOS Chromotest, CHO/HGPRT, gene amplification) in the case of some compounds included as references.

DNA adduct formation by 7-methoxy-2-nitro-naphtho[2,1-b]furan (R7000), an extremely potent mutagen.

The effects on DNA, in bacteria, of 7-methoxy-2-nitro-naphtho[2,1-b]furan (R7000), a very potent genotoxic product from the 2-nitronaphthofuran series, were investigated with two different approaches: (i) measurement of the binding of the radiolabelled mutagen to DNA and (ii) detection by the '32P-postlabelling' method of DNA adducts following treatment with unlabelled mutagens. The covalent binding of R7000 to DNA in Escherichia coli was demonstrated by both methods, and in the latter case it was found to involve the formation of nine different adducts. Formation of adducts by R7000 was shown to require metabolic activation of the compound.

In-vivo carcinogenicity of 2-nitro-oxaphenalenes.

2-Nitro-oxaphenalenes are synthetic chemicals which were synthesized in the authors' laboratory. They are the most efficient mutagenic compounds on mammalian cells in culture. They are chemically related to the nitro-naphthofuran family by the displacement of the heterocycle on the naphthalene ring. Since nitro-naphthofurans have a strong mutagenic activity in bacterial tests without metabolic activation and are active in-vivo carcinogens, the purpose of this study was to demonstrate the carcinogenic activity of two 2-nitro-oxaphenalenes. The two compounds were injected s.c. into Wistar rats initially 6-weeks-old. They were dissolved in dimethylsulfoxide (DMSO) at a concentration of 1 mg/ml. A s.c. injection of 0.5 ml containing 0.5 mg of carcinogen was given once a week in the neck of each animal tested. Five control animals were not injected and five animals received 0.5-ml injection of DMSO every week to serve as a control. The animals developed tumors only at the site of injection. The tumors were classified as high grade fibrosarcomas. This experiment demonstrates that: (i) 2-nitro-oxaphenalenes are very active in-vivo carcinogens in rats; (ii) there is a good correlation between the high mutagenic activity especially in mammalian tests and the strong carcinogenicity of the compounds; and (iii) the presence of a 6-methoxy group increases by two-fold the carcinogenic potential.

Oxidative metabolism of 7-methoxy-2-nitro-naphtho[2,1-b]furan (R 7000) by the microsomal system isolated from 3-methylcholanthrene-induced rat liver.

The metabolism of 7-methoxy-2-nitro-naphtho[2,1-b]furan and the subsequent binding to DNA, under aerobic conditions, were investigated using liver microsomes of both untreated rats and rats pre-treated with 3-methylcholanthrene [3-MC]. The metabolites were analyzed by HPLC. The following compounds: 7-hydroxy-2-nitro-naphtho[2,1-b]-furan-6,9-dione; 6,7-dihydro-2-nitro-naphtho[2,1-b]furan; 7-hydroxy-2-nitro-naphtho[2,1-b]furan and 6-hydroxy-7-methoxy-2-nitro-naphtho[2,1-b]furan have been identified by their UV-visible, mass spectra, NMR spectra and by comparison to an authentic reference sample. Qualitative and quantitative metabolic charts involving only ring oxidation have been established.

DNA ploidy in 1,2-dimethylhydrazine-induced rat tumors.

The DNA content of 1,2-dimethylhydrazine (DMH)-induced intestinal tumors of male Wistar rats was analyzed by using flow cytometry. All adenomas and carcinomas were DNA diploid. S-phase fractions of tumors showed no significant difference from those of normal mucosae. Histological evidence of adenoma-carcinoma sequence was observed in our series. The lack of DNA aneuploidy is one of the principal differences between the DMH model and human colorectal cancers.

Comparison of the carcinogenic effects of two 2-nitro-naphthofurans injected sub-cutaneously in rats.

7-Methoxy-2-nitro-naphtho[2,1-b]furan (R 7000) and its methylated homolog in position 1 (R 7372) are among the most mutagenic agents presently known, as shown by the results obtained both in the Ames test and in the SOS Chromotest. Their carcinogenic effects were tested in rats. We were able to confirm the carcinogenic effects of these nitro-naphthofurans, the presence of a methyl group--while increasing the mutagenic effect of R 7000 10 times--induces a significant decrease of the carcinogenic effects in R 7372. The discrepancy between the mutagenic effects in bacterial assays and the carcinogenic effects of these two 2-nitro-naphthofurans remains to be explained.

Mode of action of nitro-heterocyclic compounds on Escherichia coli.

The inhibitory and bactericidal activities of several different nitro-heterocyclic compounds, such as nitrofuran, nitronaphthofuran, nitrobenzofuran, nitroimidazole and nitrothiazole, were assessed in vitro. All these substances except nitroimidazole were active against Escherichia coli, though to different degrees. Under anaerobic test conditions the antibacterial activity increased slightly. Nitroreductase-deficient mutants, however, were highly resistant to all nitro-compounds, indicating that only when the nitro-group is reduced to these agents get into an active antibacterial form. SOS repair-deficient strains were much more susceptible to the nitro-containing substances than repair-proficient counterparts, indicating that damage to bacterial DNA is the essential mechanism of antibacterial activity of all nitro-heterocyclic compounds.

Effect of different nitroheterocyclic compounds on aerobic, microaerophilic, and anaerobic bacteria.

The antibacterial activities of different nitroheterocyclic compounds were assessed by an agar dilution method against aerobic, microaerophilic, and anaerobic bacteria. Nitronaphthofurans inhibited the multiplication of aerobic bacteria at low concentrations (MIC for 50% of strains tested [MIC50], 1 mg/liter). Under anaerobic growth conditions the MICs were found to be even lower. The rough, DNA repair-deficient mutants of Salmonella typhimurium were more susceptible, whereas nitroreductase-deficient strains were resistant. Microaerophilic campylobacter isolates could be divided into two groups, one of which was as susceptible as aerobic bacteria (MIC50, 1 mg/liter) and the other of which was more highly susceptible (MIC50, 0.015 mg/liter). All anaerobic bacteria tested were susceptible to nitronaphthofurans (MIC50, 0.125 mg/liter). Nitrothiazole exerted antibacterial activities similar to those of the nitronaphthofurans. Metronidazole, a nitroimidazole derivative, and nitrofurans were definitely less active. Nitrobenzofurans showed relatively high MICs.

Carcinogenic effect of 7-methoxy-2-nitro-naphtho[2,1-b] furan (R 7000) in the forestomach of rats.

7-methoxy-2-nitro-naphtho[2,1-b] furan (R 7000), known as a very potent mutagen and a very active in vivo carcinogen was employed here to develop squamous cell carcinoma in the rat. Seventy male Wistar rats received R 7000 p.o. dissolved in deionized water with 5% ethanol for periods from 1 to 21 months, while 20 served as controls receiving either water or 5% ethanol. All the animals were killed 21 months after the beginning of the experiment. Microscopic lesions were noticed in the forestomach after only 1 month of R 7000 administration. Histologic features varied from slight dysplasia to invasive carcinoma. Their sizes and invasive character increased significantly with the amount of R 7000 administered (P less than 0.05). R 7000 can be considered as a locally acting carcinogen since its carcinogenic effects appear at a place where the compound collects after swallowing. R 7000 is a weak carcinogen at the concentration employed here, when compared to some nitrosamines used in the development of forestomach cancer in rodents. The exact mechanism of the carcinogenic effects remains to be explained.

Genotoxic activities of 2-nitronaphthofurans and related molecules.

The genotoxic activities of 63, 2-nitronaphthofurans and related molecules were examined using two bacterial short-term tests, the Salmonella mammalian microsome assay test or Mutatest, a mutagenesis assay, and/or the SOS Chromotest, an assay for induction of an SOS function in Escherichia coli. Seven compounds were also investigated in the Chinese hamster ovary cells/hypoxanthine-guanine phosphoribosyl transferase (CHO/HGPRT) test, a mammalian gene mutation assay. Our main conclusions are the following: (a) Simple empirical rules relating structure to mutagenic activity in the Mutatest can be derived for some of the compounds. In particular, they account for the extremely high Mutagenic Potency of 7-methoxy-1-methyl-2-nitronaphtho[2,1-b]furan (R7372), approximately 2 X 10(6) mutants/nmol on strain TA100. (b) There is a good quantitative correlation between the Mutagenic Potency in the Salmonella/mammalian microsomes assay and the SOS-inducing potency in the SOS Chromotest. This, and previous evidence, suggests strongly that the 2-nitronaphthofurans derivatives are essentially recA and thus probably umuDC-dependent mutagens. (c) Four out of seven compounds tested in the CHO/HGPRT assay gave responses correlated with the bacterial responses. One of them, 7-methoxy-2-nitronaphtho[2,1-b]furan (R7000), is among, or is, the strongest mutagen described for mammalian cells. We briefly discuss the practical and theoretical implications of these results.

[Epidermoid cancer of the rumen induced in the C3H mouse by 7-methoxy-2-nitronaphto (2,1-b)furan (R7000)]. / Cancer épidermoïde du rumen induit chez la Souris C3H par le méthoxy-7 nitro-2 naphto [2,1-b] furanne (R 7000).

Nitro naphtofurans are powerful mutagenic agents. Subcutaneous injections of 7-methoxy 2-nitro naphtho [2,1-b] furan (R 7000) in C3H mice induce subcutaneous fibrosarcomas at the site of injection and squamous cell carcinomas of the stomach.

Naphthofurans induced chromosomal aberrations detected in metaphase, anaphase and telophase V79 Chinese hamster cells.

The mutagenic activities of 5 newly synthesized naphthofurans were analysed in two in vitro cytogenetic assays: the metaphase chromosomal aberration assay and the anaphase telophase bridge-fragment assay. Both assays were conducted using V79 Chinese hamster cells. The compounds included: 2-nitro-7-methoxynaphtho[2,1-b]furan (A), 2-nitro-8-methoxynaphtho[2,1-b]furan (B), 2-nitro-naphtho[2,1-b]furan (C), 2-nitro-7-bromonaphtho[2,1-b]furan (D) and 7-methoxynaphtho[2,1-b]furan (E). The cells were treated with 3 concentrations (0.1, 0.2 and 0.4 microgram/ml) of each compound, in the dose range already tested in studies on the mutagenic properties of the same compounds realised with other systems. The highest concentration, only, was used in the anaphase-telophase assay. In the first approach, compounds A, B and C were active while compounds D and E did not increase significantly the aberration frequency above that of the DMSO controls. The results were confirmed in the second approach. They demonstrated that the two studies were complementary. Based on their genotoxic activities, the 5 compounds were ranked in the following decreasing order of potency: A congruent to B much greater than C greater than D congruent to E congruent to DMSO; which is comparable to the ranking order obtained in different in vitro mutagenic and carcinogenic assays. All these activities are closely related to the highly specific molecular structure of each compound, particularly to the nature and position of the different substituents introduced on the skeleton.

Disposition in rats and mice of 7-methoxy-2-nitronaphtho[2,1-b]furan.

The disposition of 7-methoxy-2-nitronaphtho[2,1-b]furan (MNNF), labelled with 14C in the furan ring (label 1) and in the methoxy group (label 2) has been studied in rats and mice. After i.p. administration to rat (5 mg/kg), both labelled species were absorbed by the lymphatics; and after oral administration, through the intestinal lumen. Excretion of the furan ring (label 1) is mainly urinary (44% dose in 24 h); label 2 was mostly expired as 14CO2 (48% dose in 24 h), indicating considerable demethylation. No target organ was found for MNNF, except liver and kidney. For both labelled species given orally, radioactivity was bound to the intestinal wall. Preliminary metabolic studies, using t.l.c. and h.p.l.c., have shown the presence of an urinary metabolite, namely, the glucuronide of 7-hydroxy-2-nitronaphtho[2,1-b]furan (15-20% of the urinary radioactivity). The remaining radioactivity comprises basic compounds, that bind to a cationic resin, which might be formed by enzymic reduction of the nitro group.