Pascal Gerbaux

Qualitative and Quantitative Saponin Contents in Five Sea Cucumbers from the Indian Ocean

To avoid predation, holothuroids produce feeding-deterrent molecules in their body wall and viscera, the so-called saponins. Five tropical sea cucumber species of the family Holothuriidae were investigated in order to study their saponin content in two different organs, the body wall and the Cuvierian tubules. Mass spectrometry techniques (MALDI- and ESI-MS) were used to detect and analyze saponins. The smallest number of saponins was observed in Holothuria atra, which contained a total of four congeners, followed by Holothuria leucospilota, Pearsonothuria graeffei and Actinopyga echinites with six, eight and ten congeners, respectively. Bohadschia subrubra revealed the highest saponin diversity (19 congeners). Saponin mixtures also varied between the two body compartments within a given animal. A semi-quantitative approach completed these results and showed that a high diversity of saponins is not particularly correlated to a high saponin concentration. Although the complexity of the saponin mixtures described makes the elucidation of their respective biological roles difficult, the comparisons between species and between body compartments give some clues about how these molecules may act as predator repellents.

MALDI-ToF analysis of polythiophene: use of trans-2-[3-(4-t-butyl-phenyl)-2-methyl- 2-propenylidene]malononitrile-DCTB-as matrix.

Nowadays, numerous experimental and theoretical studies are devoted to the research field of polythiophenes and other electroconjugated polymers due to the huge potentialities of those conducting polymers. Synthetic procedures are now developed to reach the highest control over both polymerization and analytical methodologies allowing an in-depth and straightforward characterization of the polymer samples without any required doubt. Mass spectrometry methodologies and in particular MALDI-ToF measurements are definitively suitable to meet the characterization requirements. In the present study, trans-2-[3-(4-t-butyl-phenyl)-2-methyl-2-propenylidene]malononitrile (DCTB) was shown to afford better results than the reported terthiophene and dithranol matrices as far as sensitivity and signal-to-noise ratio are concerned. We tentatively proposed that the ionization of the P3HT molecules is performed by charge exchange in the condensed phase (clusters) with matrix molecule radical cations and subsequent neutral matrix molecule evaporation from the clusters. The putative key parameters to account for the really high efficiency of DCTB for the MALDI analysis of P3HT are (1) the highest ionization energy of DCTB amongst the three matrices, (2) the really high absorptivity of the matrix molecule at the laser wavelength and (3) the presence of the tertiobutyl group on the matrix molecule. The presence of this substituent is likely to decrease the intermolecular interactions in the condensed phase rendering the evaporation of the neutral matrix molecules less energy demanding. We also demonstrated for polymer samples presenting an average number molecular weight (M(n) ) below 10 000 g mol(-1) that the systematic overestimation of the low mass oligomers upon MALDI measurements ends up with wrong M(n) and polydispersity index (PDI) values. A systematic Soxhlet extraction against heptane was shown to allow the recording of absolute M(n) and PDI.

A Photoreactive Ruthenium(II) Complex Tethered to a Guanine‐Containing Oligonucleotide: A Biomolecular Tool that Behaves as a “Seppuku Molecule”

The design of specific DNA or RNA damaging agents may be achieved by anchoring a reactive species to an oligonucleotide (ODN) probe which, by duplex or triplex formation, should direct the irreversible chemical modification towards the targeted sequence. This approach is certainly interesting in the context of gene silencing, particularly for the development of anticancer agents. However, the achievement of a high level of selectivity remains a challenge because many side reactions such as interaction with proteins can occur in biological systems. In this context, the tethering of a transition metal complex to an ODN strand and subsequent activation by light is an attractive strategy. Indeed, the reactivity of the metallic compound towards the genetic material can be 1) triggered by light to act on specific tissues and 2) directed towards a DNA or RNA target sequence to cause damage. We have prepared oligonucleotides labeled with ruthenium(II) complexes (Ru-ODNs) that are able to photo-cross-link with their complementary strand under visible irradiation (Figure 1 a). The different parameters that govern the photo-cross-linking efficiency have been extensively studied, 6] these conjugates could be promising as anticancer agents based on gene silencing. Indeed, this sequence-specific photo-cross-linking process has been shown to inhibit in vitro DNA polymerase with a high efficiency. The prerequisites for this photochemical reaction are 1) the presence of at least two p-deficient polyazaaromatic ligands such as 1,4,5,8-tetraazaphenanthrene (tap) in the tethered metallic complex and 2) the presence of at least one guanine base (G) in the target sequence, in the vicinity of the tethered complex after hybridization. The mechanism involves a photoinduced electron transfer (PET) from the G base towards the oxidizing excited Ru compound with a recombination of the produced radicals, which covalently links one of the tap ligands to the G unit [Figure 1b, Eqs. (1)–(3)]. The structure of the photoadduct (Figure 1b) has been determined by ESI mass spectrometry and NMR spectroscopy. This strategy was thought to be restricted to target sequences without cytosine in order to avoid the presence of a guanine in the Ru-ODN probe, which would be unfavorable for the photo-cross-linking. In spite of these limitations, we decided to investigate the effect of the presence of a G base in the Ru-ODN probe sequence on the photoreactivity of such systems. Herein, we report the unique photochemical behavior of a new generation of photoreactive G-containing Ru-ODN probes that can selfinhibit in the absence of their specific target strands. We have named these molecules “seppuku molecules” because their photochemical behavior reminds us of the suicide of someone who has not accomplished his attributed duty in antic Japanese society. The Ru complexes contain a phenanthroline ligand derivatized by a linker, namely phen’’ (phen’’= N-(2-(1,10phenanthrolin-5-ylamino)-2-oxoethyl)-2-(aminooxy)acetaFigure 1. a) Photo-cross-linking processes that occur between an ODN probe labeled with a suitable Ru complex and the complementary G-containing ODN strand and b) formation and structure of the photoadduct produced upon illumination of [Ru(tap)3] 2+ in the presence of a guanine residue in GMP (guanosine-5’-monophosphate) or DNA after acid hydrolysis.

Elucidation of molecular diversity and body distribution of saponins in the sea cucumber Holothuria forskali (Echinodermata) by mass spectrometry

Sea cucumbers contain triterpene glycosides called saponins. We investigated the complex saponin mixture extracted from the common Mediterranean species Holothuria forskali. Two different body components were analyzed separately: the body wall (which protects the animal and is moreover the most important organ in terms of surface and weight) and the Cuvierian tubules (a defensive organ that can be expelled on predators in response to an attack). MALDI/MS and MALDI/MS/MS were used to detect saponins and describe their molecular structures. As isomers have been found in the Cuvierian tubules, LC/MS and LC/MS/MS were performed to identify each saponin separately. Twelve saponins have been detected in the body wall and 26 in the Cuvierian tubules. All the saponins from the body wall are also present in the Cuvierian tubules but the latter also contain 14 specific saponins. The presence of isomeric saponins complicated structure elucidation for the whole set but 16 saponins have been described tentatively. Among these, 3 had already been reported in the literature as holothurinosides A and C, and desholothurin A. Molecular structures have been proposed for the 13 others which, in the present work, have been provisionally named holothurinosides E, F, G, H, I, A(1), C(1), E(1), F(1), G(1), H(1) and I(1) and desholothurin A(1). The diversity and organ specificity of the saponins described here are much higher than what had been reported to date in any sea cucumber species.

Poly(ethylene imine) impurities induce n-doping reaction in organic (semi)conductors

Volatile impurities contained in polyethyleneimine (PEI), and identified as ethyleneimine dimers and trimers, are reported. These N-based molecules show a strong reducing character, as demonstrated by the change in electrical conductivity of organic (semi)conductors exposed to the PEI vapor. The results prove that electron transfer rather than a dipole effect at the electrode interface is the origin of the work-function modification by the PEI-based layers.

The triterpene glycosides of Holothuria forskali: usefulness and efficiency as a chemical defense mechanism against predatory fish

SUMMARY More than 100 triterpene glycosides (saponins) have been characterized in holothuroids in the past several decades. In particular, Holothuria forskali contains 26 saponins in its Cuvierian tubules and 12 in its body wall. This high diversity could be linked to a chemical defense mechanism, the most commonly accepted biological role for these secondary metabolites. We performed an integrated study of the body-wall saponins of H. forskali. The saponins are mainly localized in the epidermis and in the mesothelium of the body wall and appear to be released when the holothuroid is stressed. Among the saponins present in the epidermis, one (holothurinoside G) was detected in the seawater surrounding non-stressed holothuroids and three others (holohurinosides C and F, and desholothurin A) were secreted when the animals were stressed. In addition, two new congeners (detected at m/z 1301 and 1317) were also present in the immediate surroundings of stressed holothuroids. These new saponins do not originate from the epidermis and could come from an internal organ. Quantities of secreted saponins were very low compared with the body wall and Cuvierian tubules concentrations. At natural concentrations, saponins do not represent a threat to the health of predatory fish. The deterrent effect of saponins seems therefore to act as an aposematic signal, warning potential predators of the unpalatability of the holothuroid tissues.

Cobalt‐Mediated Radical Coupling (CMRC): An Unusual Route to Midchain‐Functionalized Symmetrical Macromolecules

Cobalt-mediated radical coupling (CMRC) is a straightforward approach to the synthesis of symmetrical macromolecules that relies on the addition of 1,3-diene compounds onto polymer precursors preformed by cobalt-mediated radical polymerization (CMRP). Mechanistic features that make this process so efficient for radical polymer coupling are reported here. The mechanism was established on the basis of NMR spectroscopy and MALDI-MS analyses of the coupling product and corroborated by DFT calculations. A key feature of CMRC is the preferential insertion of two diene units in the middle of the chain of the coupling product mainly according to a trans-1,4-addition pathway. The large tolerance of CMRC towards the diene structure is demonstrated and the impact of this new coupling method on macromolecular engineering is discussed, especially for midchain functionalization of polymers. It is worth noting that the interest in CMRC goes beyond the field of polymer chemistry, since it constitutes a novel carbon-carbon bond formation method that could be applied to small organic molecules.

A Rigid Dinuclear Ruthenium(II) Complex as an Efficient Photoactive Agent for Bridging Two Guanine Bases of a Duplex or Quadruplex Oligonucleotide

The rigid dinuclear [(tap)(2)Ru(tpac)Ru(tap)(2)](4+) complex (1) (TAP=1,4,5,8-tetraazaphenanthrene, TPAC=tetrapyridoacridine) is shown to be much more efficient than the mononuclear bis-TAP complexes at photodamaging oligodeoxyribonucleotides (ODNs) containing guanine (G). This is particularly striking with the G-rich telomeric sequence d(T(2)AG(3))(4). Complex 1, which interacts strongly with the ODNs as determined by surface plasmon resonance (SPR) and emission anisotropy experiments, gives rise under illumination to the formation of covalent adducts with the G units of the ODNs. The yield of photocrosslinking of the two strands of duplexes by 1 is the highest when the G bases of each strand are separated by three to four base pairs. This corresponds with each Ru(tap)(2) moiety of complex 1 forming an adduct with the G base. This separation distance of the G units of a duplex could be determined thanks to the rigidity of complex 1. On the basis of results of gel electrophoresis, mass spectrometry, and molecular modelling, it is suggested that such photocrosslinking can also occur intramolecularly in the human telomeric quadruplex d(T(2)AG(3))(4).

Cyclic Polymers by Ring-Closure Strategies

The preparation of cyclic macromolecules has always represented a challenging task for polymer science, mainly because of difficulties in connecting chain extremities together. Initiated by the pioneering studies of Jacobson and Stockmayer, preparative pathways to cyclic polymers have been considerably improved within the last two decades thanks to the advent of both controlled polymerizations and efficient coupling reactions in organic chemistry. This Review aims to provide a critical up-to-date overview and illustrate the considerable efforts that have been made in the past few years to improve the availability of macrocycles for industrial and academic investigations through the use of the ring-closure approach. Particular attention is paid to methods for the preparation of monocycles over more complex architectures, since the latter are usually derived from the former.

Characterization of ionized carbenes in the gas phase

The generation of carbene radical cations (R™C™R z1 ) and related ions in the gas phase using various mass spectrometric techniques is critically reviewed. Their reactions as well as their thermochemical properties obtained either experimentally or from ab initio quantum chemical methods are also discussed. The review covers the literature published during the last 20 years and includes more than 120 papers. (Int J Mass Spectrom 202 (2000) A8 ‐A25)