[Holmium laser enucleation of the prostate as a day case surgery: prospective evaluation of the first 30 patients]. / Énucléation de la prostate au laser Holmium en chirurgie ambulatoire : évaluation prospective des 30 premiers patients.

OBJECTIVES: To evaluate the feasibility of holmium laser enucleation of the prostate (HoLEP) as a day case surgery. MATERIAL AND METHODS: Observational prospective study including 30 consecutive patients after exclusion of unstable diseases and anticoagulant therapy. Patients were discharged before 8PM and the urinary catheter was removed at home the next morning. The monitoring included a phone call after 24hours and clinical evaluations after 1 and 3month follow-up. Clinical data were prospectively collected and complications were classified according to the Clavien-Dindo classification. RESULTS: The mean age of the study population was 63.8, prostate volume was 75.3cc, maximum urinary flow rate was 9.5mL/s, and IPSS was 22.9. The conversion rate to conventional hospitalization was 3.3%. After 3months follow-up, readmission and reoperation rates were respectively 16.6% and 3.3%. The overall complication rate was 66% (Clavien I=57.7%, II=38.5%, III=3.8%). The satisfaction rate was 100% (score=9.2/10). The mean prostate volume at 3months follow-up was 23.3cc, maximum urinary flow was 25.6mL/s, and IPSS was 4.7. CONCLUSION: This study confirmed the feasibility of HoLEP as a day case surgery for selected patients. Conversion rate to conventional hospitalization and complications of grade >2 were less than 5% while the satisfaction rate was high. LEVEL OF EVIDENCE: 3.

Purification and characterization of a NADPH-dependent aldehyde reductase from mung bean that detoxifies eutypine, a toxin from eutypa lata1

Eutypine (4-hydroxy-3-[3-methyl-3-butene-1-ynyl] benzaldehyde) is a toxin produced by Eutypa lata, the causal agent of eutypa dieback in the grapevine (Vitis vinifera). Eutypine is enzymatically converted by numerous plant tissues into eutypinol (4-hydroxy-3-[3-methyl-3-butene-1-ynyl] benzyl alcohol), a metabolite that is nontoxic to grapevine. We report a four-step procedure for the purification to apparent electrophoretic homogeneity of a eutypine-reducing enzyme (ERE) from etiolated mung bean (Vigna radiata) hypocotyls. The purified protein is a monomer of 36 kD, uses NADPH as a cofactor, and exhibits a Km value of 6.3 &mgr;M for eutypine and a high affinity for 3- and 4-nitro-benzaldehyde. The enzyme failed to catalyze the reverse reaction using eutypinol as a substrate. ERE detoxifies eutypine efficiently over a pH range from 6.2 to 7.5. These data strongly suggest that ERE is an aldehyde reductase that could probably be classified into the aldo-keto reductase superfamily. We discuss the possible role of this enzyme in eutypine detoxification.

An evolutionary analytical model of a complementary circular code.

The subset X0=[AAC,AAT,ACC,ATC,ATT,CAG,CTC,CTG, GAA,GAC,GAG,GAT,GCC,GGC,GGT,GTA,GTC,GTT,TAC,TTC] of 20 trinucleotides has a preferential occurrence in the frame 0 (reading frame established by the ATG start trinucleotide) of protein (coding) genes of both prokaryotes and eukaryotes. This subset X0 is a complementary maximal circular code with two permutated maximal circular codes X1 and X2 in the frames 1 and 2 respectively (frame 0 shifted by one and two nucleotides respectively in the 5'-3' direction). X0 is called a C3 code (Arquès and Michel, 1997, J. Biosyst 44, 107-134). A quantitative study of these three subsets X0, X1 and X2 in the three frames 0, 1 and 2 of eukaryotic protein genes shows that their occurrence frequencies are constant functions of the trinucleotide positions in the sequences. The frequencies of X0, X1 and X2 in the frame 0 of eukaryotic protein genes are 48.5%, 29% and 22.5% respectively. These properties are not observed in the 5' and 3' regions of eukaryotes where X0, X1 and X2 occur with variable frequencies around the random value (1/3). Several frequency asymmetries unexpectedly observed, e.g. the frequency difference between X1 and X2 in the frame 0, are related to a new property of the C3 code X0 involving substitutions. An evolutionary analytical model at three parameters (p, q, t) based on an independent mixing of the 20 codons (trinucleotides in the frame 0) of X0 with equiprobability (1/20) followed by t approximately 4 substitutions per codon according to the proportions p approximately 0.1, q approximately 0.1 and r = 1 - p - q approximately 0.8 in the three codon sites respectively, retrieves the frequencies of X0, X1 and X2 observed in the three frames of protein genes and explains these asymmetries. The complex behaviour of these analytical curves is totally unexpected and a priori difficult to imagine. Finally, the evolutionary analytical method developed could be applied to the phylogenetic tree reconstruction and the DNA sequence alignment.

Identification of several types of periodicities in the collagens and their simulation.

The collagens constitute an important population of proteins providing the structural support in vertebrate tissues A collagen is mainly based on a series of tripeptides of the type GX1X2 (G = Glycine, X1 and X2 being any residues). The nine amino acids occurring with significant frequencies in the X1 and X2 residue sites and G form the reduced protein alphabet Q = [A,D,E,G,K,L,P,Q,R,S] (A = Alanine, D = Aspartic acid, E = Glutamic acid, K = Lysine, L = Leucine, P = Proline, Q = Glutamine, R = Arginine, S = Serine). Surprisingly, the method based on the autocorrelation function w(X)iw' analysing the probability that an amino acid w' in Q occurs any i residues X after an amino acid w in Q (called i-motif w(X)iw'), identifies six types of modulo 3 periodicities in collagens: three basic types 0, 1 and 2 modulo 3 and three combined types 0,1, 0,2 and 1,2 modulo 3. Furthermore, the classification of these 100 i-motifs according to the types of periodicities shows several strong relations between four sub-sets of Q [G], [A,D,P,S], [E,L] and [K,Q,R]. Then, these relations allow the construction of a simple automaton for the generation of model collagen sequences. Indeed, this automaton can simulate the six types of periodicities and it retrieves the types of periodicities for almost all i-motifs. Finally, the autocorrelation function based on the sub-set [K,Q,R] identifies segments of 18 amino acids in collagens which may correspond to the exons (segments of genes of 54 nucleotides) coding for those collagens.

Early psychological care of the French victims of the Costa Concordia shipwreck.

Most of the French passengers who survived the shipwreck of the cruise ship Costa Concordia were repatriatedfrom Italy to Marseille, one of the stopovers of the cruise. The shipwreck happened during the nightof 13th-14th January 2012 and entailed the forced evacuation of 4195 passengers and crewmembers.Thirty-two persons died and 2 others are still reported missing. The massive and unexpected inflow of402 French citizens in the port of Marseille required the quick setting up of welcome facilities, not only tosolve logistical problems, but also to address psychological and sometimes even medical problems. ThePrehospital Psychological Emergency Service (CUMP) and the Prehospital Emergency Medical Service(SAMU) of Marseille examined 196 persons in total, and were able to avoid a great number of emergencyadmissions deemed necessary because of difficult psychological situations (death, missing or lost persons,acute stress). The objective of this report is to rapidly present the emergency committee as a whole andto describe in more detail the work that the CUMP accomplished during the 36 hours necessary to takecharge of the majority of the French passengers of the Costa Concordia.

Biogenesis of monoterpenes : Bioconversion of citral by a cell suspension culture of muscat grapes.

Cell suspension cultures of "Muscat de Frontignan" grapes Vitis vinifera L. are able to convert citral (a mixture of neral and geranial) into the corresponding monoterpenic alcohols, nerol and geraniol. The geraniol formed is esterified into geranyl acetate. Bioconversion of nerol or geraniol added alone to the cell suspension was also studied. Interconversions between these different monoterpenic compounds are described and discussed.

Stimulation of somatic embryogenesis and plant regeneration from anther culture of Vitis vinifera cv. Cabernet-Sauvignon.

Somatic embryogenesis and subsequent diploid plants have been obtained from anthers of Vitis vinifera Cabernet-Sauvignon, a cultivar so far considered as recalcitrant to in vitro regeneration. Anthers enclosing microspores near the first pollen mitosis were found to be the most responsive. However, from a practical point of view anther length proved to be an easier criterium for determining the optimal physiological anther stage. Calli derived from the anther somatic tissues produced embryoids only when cultured on a medium supplemented with casein hydrolysate. Glutamine and adenine were found to stimulate this embryoid production. Evidence is presented that early removal of cotyledons increases the frequency of normal development of embryoids into plantlets.

Genetic analysis of organogenesis in the cotyledons of zygotic embryos of sunflower (Helianthus annuus L.).

Crosses were made between five cytoplasmic male-sterile and five restorer sunflower inbred lines. Twenty-five F1 hybrids and their parents were studied for their organogenesis ability in a randomized block design with four replications. Each replication per genotype consisted of ten petri dishes with four expiants. Regeneration medium consisted of full MS medium modified by the addition of hormones and solidified with 6 g/l agar. Statistical analysis showed that both general and specific combining abilities were significant for all of the organogenesis parameters studied, and both showed several significant positive or negative values. General combining ability values were usually higher than those of specific combining ability, indicating the importance of additive genetic control for organogenesis parameters in sunflower. Narrow-sense heritability for the number of explants producing shoots and roots was 65.8%, which suggests that organogenesis of currently inferior inbred lines in sunflower should be improved in a crossing program.

Some Physiological Changes Occurring during the Senescence of Auxin-Deprived Pear Cells in Culture.

Part of the changes in the hormonal balance involved in plant senescence is due to an auxin limitation. Some of its physiological consequences are studied using pear (Pyrus communis L.) cells cultured in a continuously renewed medium in which 2,4-dichlorophenoxyacetic acid (2,4-D) was absent. In these conditions, an assessment was made of the absence of nutrient deficiency.In the period preceding cell death, the rate of respiration and ethylene production remain low, and no major changes were observed in the total protein and RNA content of the cells. Beginning around day 9, an important efflux of three amino acids (serine, threonine, and aspartic acid) occurs among which serine represents more than 52%. However, exogenous serine supplied to the medium fails to show any senescence promoting effect. At the same time, leucine uptake and incorporation sharply and simultaneously increased. The presence of 2,4-D inhibits both these phenomena and prevents cell death. It is proposed that auxin deprivation is responsible for unmasking a program of synthesis of new proteins involved in cell death.

An evolutionary analytical model of a complementary circular code simulating the protein coding genes, the 5' and 3' regions.

The self-complementary subset T0 = X0 [symbol: see text] ¿AAA, TTT¿ with X0 = ¿AAC, AAT, ACC, ATC, ATT, CAG, CTC, CTG, GAA, GAC, GAG, GAT, GCC, GGC, GGT, GTA, GTC, GTT, TAC, TTC¿ of 22 trinucleotides has a preferential occurrence in the frame 0 (reading frame established by the ATG start trinucleotide) of protein (coding) genes of both prokaryotes and eukaryotes. The subsets T1 = X1 [symbol: see text] ¿CCC¿ and T2 = X2 [symbol: see text] ¿GGG¿ of 21 trinucleotides have a preferential occurrence in the shifted frames 1 and 2 respectively (frame 0 shifted by one and two nucleotides respectively in the 5'-3' direction). T1 and T2 are complementary to each other. The subset T0 contains the subset X0 which has the rarity property (6 x 10(-8) to be a complementary maximal circular code with two permutated maximal circular codes X1 and X2 in the frames 1 and 2 respectively. X0 is called a C3 code. A quantitative study of these three subsets T0, T1, T2 in the three frames 0, 1, 2 of protein genes, and the 5' and 3' regions of eukaryotes, shows that their occurrence frequencies are constant functions of the trinucleotide positions in the sequences. The frequencies of T0, T1, T2 in the frame 0 of protein genes are 49, 28.5 and 22.5% respectively. In contrast, the frequencies of T0, T1, T2 in the 5' and 3' regions of eukaryotes, are independent of the frame. Indeed, the frequency of T0 in the three frames of 5' (respectively 3') regions is equal to 35.5% (respectively 38%) and is greater than the frequencies T1 and T2, both equal to 32.25% (respectively 31%) in the three frames. Several frequency asymmetries unexpectedly observed (e.g. the frequency difference between T1 and T2 in the frame 0), are related to a new property of the subset T0 involving substitutions. An evolutionary analytical model at three parameters (p, q, t) based on an independent mixing of the 22 codons (trinucleotides in frame 0) of T0 with equiprobability (1/22) followed by t approximately 4 substitutions per codon according to the proportions p approximately 0.1, q approximately 0.1 and r = 1 - p - q approximately 0.8 in the three codon sites respectively, retrieves the frequencies of T0, T1, T2 observed in the three frames of protein genes and explains these asymmetries. Furthermore, the same model (0.1, 0.1, t) after t approximately 22 substitutions per codon, retrieves the statistical properties observed in the three frames of the 5' and 3' regions. The complex behaviour of these analytical curves is totally unexpected and a priori difficult to imagine.

Stimulation ofDaucus carota somatic embryogenesis by inhibitors of ethylene synthesis: cobalt and nickel.

The effects of Co(2+) and Ni(2+) on ethylene production and somatic embryogenesis by carrot (Daucus carota L.) cell cultures were studied. At concentrations of 10 µM to 50 µM, CoCl2 effectively inhibited ethylene production by embryogenic cultures and significantly stimulated somatic embryogenesis. The observed increase of embryo number was proportional to the inhibition level of ethylene production. However, CoCl2 had no effect when Ethephon was supplied. Nickel also reduced ethylene production, but to a slightly lesser extent than CoCl2, bringing about a lower increase in the number of somatic embryos. The role of ethylene on somatic embryogenesis is discussed.

An evolutionary model of a complementary circular code.

The subset X0 = [sequence: see text] of 20 trinucleotides has a preferential occurrence in frame 0 (a reading frame established by the ATG start trinucleotide) of protein (coding) genes of both prokaryotes and eukaryotes. This subset X0++ has the rarity property (6 x 10(-8)) to be a complementary maximal circular code with two permutated maximal circular codes X1 and X2 in frames 1 and 2 respectively (frame 0 shifted by one and two nucleotides respectively in the 5'-3' direction). X0 is called a C3 code. A quantitative study of these three subsets X0, X1 and X2 in the three frames 0, 1 and 2 of eukaryotic protein genes shows that their occurrence frequencies are constant functions of the trinucleotide positions in the sequences. The frequencies of X0, X1 and X2 in frame 0 of the eukaryotic protein genes are 48.5%, 29% and 22.5% respectively. These properties are not observed in the 5' and 3' regions of eukaryotes where X0, X1 and X2 occur with variable frequencies around the random value (1/3). Several frequency asymmetries unexpectedly observed, e.g. the frequency difference between X1 and X2 in the frame 0, are related to a new property of the C3 code X0 involving substitutions. An evolutionary model at three parameters (p, q, k) based on an independent mixing of the 20 codons (trinucleotides in frame 0) of X0 with equiprobability (1/20) followed by k approximately 5 substitutions per codon in the three codon sites in proportions p approximately 0.1, q approximately 0.1 and r = 1-p-q approximately 0.8 respectively, retrieves the frequencies of X0, X1 and X2 observed in the three frames of protein genes and explains these asymmetries.

Stimulation of shoot regeneration from cotyledons of Helianthus annuus by the ethylene inhibitors, silver and cobalt.

The effects of CoCl2, AgNO3 and ethylene released by exogenous 2-chloroethylphosphonic acid (Ethephon), were studied on shoot regeneration from cotyledons of Helianthus annuus cv. E8206R, a poorly regenerative cultivar. Inhibition of ethylene biosynthesis by CoCl2, at concentrations of 20 µK, provoked a substantial enhancement of shoot regeneration (30 %): the control was poorly regenerative. However, CoCl2 had no effect when Ethephon was supplied. Inhibition of ethylene action by AgNO3, at concentrations of 10-25 µM, caused a significant increase in plant regeneration: 25 % instead of 1.2 % in the control. Furthermore, addition of Ethephon to AgNO3-treated tissues failed to reduce the stimulation of shoot regeneration caused by AgNO3. On the basis of these findings, it is suggested that ethylene inhibits the regeneration process from cotyledons of sunflower.

Enhancement of shoot regeneration potential by liquid medium culture from mature cotyledons of sunflower (Helianthus annuus L.).

We describe here a liquid culture system for the regeneration of shoots at high frequencies from mature cotyledon tissues of three genotypes of sunflower (Helianthus annuus L.) one of which had previously been found to be recalcitrant to regeneration when cotyledons were cultured on solid medium. Cotyledons were excised from 2-day-old seedlings and incubated in liquid Murashige and Skoog's modified medium supplemented with 5.4 µM naphthaleneacetic acid (NAA) and 4.4 µM benzylaminopurine (BAP). After two weeks in culture, the whole upper surface of regenerating explants was covered with green shootlets. The percentages of regenerating explants of three genotypes varied between 60 and 70%, and the number of shoots per regenerating explant was highly increased. The shootlets were transferred to solid Murashige and Skoog's medium allowing shoot development, then to rooting medium. Rooted plantlets were successfully acclimatized and gave fertile plants. The role of liquid medium culture in the induction of sunflower regeneration is discussed.

Protonophoric activity of eutypine, a toxin from Eutypa lata, in plant mitochondria.

Eutypine is a toxin produced by Eutypa lata, the causal agent of the dying-arm disease of grapevine. We have previously shown that this toxin behaves as a lipophylic weak acid (pK = 6.2) and induces drastic changes in both the respiration and energy balance of grapevine cells. In the present study, the molecular mode of action of eutypine at the mitochondrial level, using methyl-eutypine, the unprotonable derivative of the toxin, has been investigated. The effects of these molecules on mitochondrial respiration and membrane potential were compared using isolated mitochondria from grapevine cells in suspension cultures or potato tuber mitochondria. Eutypine induces marked stimulation of oxygen consumption and a depolarizing effect, while methyl-eutypine exhibits a very small effect on both the rate of oxygen uptake and membrane potential. For high eutypine concentrations, a mixed effect corresponding to a direct inhibition of electron transport and uncoupling can be observed. In addition, below 200 microM, eutypine displays a linear relationship between oxidation rate and membrane potential similar to that of the classical protonophore carbonyl cyanide-m-chlorophenylhydrazone (CCCP). However, unlike CCCP, eutypine induces a potential-dependent proton conductance that can be due to the potential-dependent migration of the dissociated form of the toxin across the membrane. It is concluded that eutypine uncouples mitochondrial oxidative phosphorylation and decreases the ADP/O ratio in grapevine cells by increasing the proton leaks via a cyclic protonophore mechanism. The physiological aspects of these results are discussed.

A novel NADPH-dependent aldehyde reductase gene from Vigna radiata confers resistance to the grapevine fungal toxin eutypine.

Eutypine, 4-hydroxy-3-(3-methyl-3-butene-1-ynyl) benzyl aldehyde, is a toxin produced by Eutypa lata, the causal agent of eutypa dieback of grapevines. It has previously been demonstrated that tolerance of some cultivars to this disease was correlated with their capacity to convert eutypine to the corresponding alcohol, eutypinol, which lacks phytotoxicity. We have thus purified to homogeneity a protein from Vigna radiata that exhibited eutypine-reducing activity and have isolated the corresponding cDNA. This encodes an NADPH-dependent reductase of 36 kDa that we have named Vigna radiata eutypine-reducing enzyme (VR-ERE), based on the capacity of a recombinant form of the protein to reduce eutypine into eutypinol. The strongest homologies (86.8%) of VR-ERE at the amino acid level were found with CPRD14, a drought-inducible gene of unknown function, isolated from Vigna unguiculata and with an aromatic alcohol dehydrogenase (71.7%) from Eucalyptus gunnii. Biochemical characterization of VR-ERE revealed that a variety of compounds containing an aldehyde group can act as substrates. However, the highest affinity was observed with 3-substituted benzaldehydes. Expression of a VR-ERE transgene in Vitis vinifera cells cultured in vitro conferred resistance to the toxin. This discovery opens up new biotechnological approaches for the generation of grapevines resistant to eutypa dieback.