Characterization of Biological Properties of Individual Phenolamides and Phenolamide-Enriched Leaf Tomato Extracts.

Resistance to conventional treatments renders urgent the discovery of new therapeutic molecules. Plant specialized metabolites such as phenolamides, a subclass of phenolic compounds, whose accumulation in tomato plants is mediated by the biotic and abiotic environment, constitute a source of natural molecules endowed with potential antioxidant, antimicrobial as well as anti-inflammatory properties. The aim of our study was to investigate whether three major phenolamides found in Tuta absoluta-infested tomato leaves exhibit antimicrobial, cytotoxic and/or anti-inflammatory properties. One of them, N1,N5,N14-tris(dihydrocaffeoyl)spermine, was specifically synthesized for this study. The three phenolamides showed low to moderate antibacterial activities but were able to counteract the LPS pro-inflammatory effect on THP-1 cells differentiated into macrophages. Extracts made from healthy but not T. absoluta-infested tomato leaf extracts were also able to reduce inflammation using the same cellular approach. Taken together, these results show that phenolamides from tomato leaves could be interesting alternatives to conventional drugs.

Comparison of outcomes in patients with or without ARC-HBR criteria undergoing PCI with polymer-free biolimus coated stents: The BioFreedom France study.

BACKGROUND: The polymer-free biolimus coated stent (BioFreedom) was shown to be superior to bare metal stents in the LEADERS FREE randomized trial in high bleeding risk (HBR) patients treated with 1-month dual antiplatelet therapy (DAPT). However, there is limited outcome data with this device in an all-comers' population. METHODS: We conducted a prospective single-arm study of patients undergoing percutaneous coronary intervention with the polymer-free biolimus coated stent in 25 centers in France with wide inclusion criteria including multivessel disease, complex lesions, and acute coronary syndromes. The primary endpoint was the incidence of target lesion failure (TLF), a composite of cardiac death or target-vessel myocardial infarction (MI) or clinically indicated target lesion revascularization (ci-TLR) at 1-year. The patient population was classified according to the presence (or not) of HBR criteria according to the recent ARC-HBR definition. RESULTS: Between April 2019 and April 2020, 1497 patients were enrolled. TLF occurred in 101 (6.9%) patients, including cardiac death in 35 (2.4%), target vessel MI in 20 (1.4%) and ci-TLR in 65 (4.5%) of them. There were 491 HBR patients (32.8%) and 1006 non-HBR patients. The median duration of DAPT was 74 days in the HBR group versus 348 days in the non-HBR group (p < 0.0001). TLF occurred in 44 (9.2%) of the HBR group and in 57 (5.8%) of the non-HBR group (relative risk 1.62 [95% confidence interval: 1.10-2.41], p = 0015). Compared to the non-HBR group, HBR patients had higher rates of cardiac death (4.4% vs. 1.4%, p = 0.0005) and target vessel MI (2.9% vs. 0.6%, p = 0.0003), but similar rates of ci-TLR. BARC 3-5 bleeding occurred in 6.2% of the HBR group versus 1.4% of the non-HBR group (p < 0.0001). CONCLUSION: In this multicenter all-comers study, HBR patients treated with a polymer-free biolimus coated stent had, compared to non-HBR patients, an increased risk of cardiac death and MI, and despite a shorter duration of DAPT, continued to have higher rates of BARC 3-5 bleeding.

Transcriptomics and Metabolomics Analyses Reveal High Induction of the Phenolamide Pathway in Tomato Plants Attacked by the Leafminer Tuta absoluta.

Tomato plants are attacked by a variety of herbivore pests and among them, the leafminer Tuta absoluta, which is currently a major threat to global tomato production. Although the commercial tomato is susceptible to T. absoluta attacks, a better understanding of the defensive plant responses to this pest will help in defining plant resistance traits and broaden the range of agronomic levers that can be used for an effective integrated pest management strategy over the crop cycle. In this study, we developed an integrative approach combining untargeted metabolomic and transcriptomic analyses to characterize the local and systemic metabolic responses of young tomato plants to T. absoluta larvae herbivory. From metabolomic analyses, the tomato response appeared to be both local and systemic, with a local response in infested leaves being much more intense than in other parts of the plant. The main response was a massive accumulation of phenolamides with great structural diversity, including rare derivatives composed of spermine and dihydrocinnamic acids. The accumulation of this family of specialized metabolites was supported by transcriptomic data, which showed induction of both phenylpropanoid and polyamine precursor pathways. Moreover, our transcriptomic data identified two genes strongly induced by T. absoluta herbivory, that we functionally characterized as putrescine hydroxycinnamoyl transferases. They catalyze the biosynthesis of several phenolamides, among which is caffeoylputrescine. Overall, this study provided new mechanistic clues of the tomato/T. absoluta interaction.

Post-Traumatic Arteriovenous Fistula of Left Renal Vessels: Case Report of a Silent Complication for 23 Years.

Abdominal trauma leads rarely to severe renal injury such as acquired arterioveinous fistula. Here, we present the case of a 46-year-old man with a history of suicide attempt by a gunshot in the abdomen. At that time, explorative laparotomy was unremarkable. He consulted 23 years later for chronic left lumbar pain and was diagnosed with an arterioveinous fistula of left renal vessels with a-10-cm aneurysm of the left renal artery. We performed a left nephrectomy and endovascular clamping was the best option to manage this giant aneurysm in a hostile abdomen.

Phenolamides in plants: an update on their function, regulation, and origin of their biosynthetic enzymes.

Phenolamides represent a family of specialized metabolites, consisting of the association of hydroxycinnamic acid derivatives with aliphatic or aromatic amines. Since the discovery of the first phenolamide in the late 1940s, decades of phytochemical analyses have revealed a high structural diversity for this family and a wide distribution in the plant kingdom. The occurrence of structurally diverse phenolamides in almost all plant organs has led to early hypotheses on their involvement in floral initiation and fertility, as well as plant defense against biotic and abiotic stress. In the present work, we critically review the literature ascribing functional hypotheses to phenolamides and recent evidence on the control of their biosynthesis in response to biotic stress. We additionally provide a phylogenetic analysis of the numerous N-hydroxycinnamoyltransferases involved in the synthesis of phenolamides and discuss the potential role of other enzyme families in their diversification. The data presented suggest multiple evolutionary events that contributed to the extension of the taxonomic distribution and diversity of phenolamides.

Phenolamides: Plant specialized metabolites with a wide range of promising pharmacological and health-promoting interests.

Phenolamides constitute a family of metabolites, widely represented in the plant kingdom, that can be found in all plant organs with a predominance in flowers and pollen grains. They represent a large and structurally diverse family, resulting from the association of phenolic acids with aliphatic or aromatic amines. Initially revealed as active compounds in several medicinal plant extracts, phenolamides have been extensively studied for their health-promoting and pharmacological properties. Indeed, phenolamides have been shown to exhibit antioxidant, anti-inflammatory, anti-cancer and antimicrobial properties, but also protective effects against metabolic syndrome and neurodegenerative diseases. The purpose of this review is to summarise this large body of literature, including in vitro and in vivo studies, by describing the diversity of their biological properties and our actual knowledge of the molecular mechanisms behind them. With regard to their considerable pharmacological interest, the question of industrial production is also tackled through chemical and biological syntheses in engineered microorganisms. The diversity of biological activities already described, together with the active discovery of the broad structural diversity of this metabolite family, make phenolamides a promising source of new active compounds on which future studies should be focused.

Datura innoxia plants hydroponically-inoculated with Agrobacterium rhizogenes display an enhanced growth and alkaloid metabolism.

BACKGROUND: The production of secondary metabolites through the culture of entire plants is of great interest. Soilless culture, such as hydroponics, enables the control of plant growth and metabolism. Specific environmental conditions must be developed to maximize the productivity of medicinal plants used as efficient natural bioreactors. METHODS: The nutrient solution of newly established hydroponic cultures ofDatura innoxia Mill. were inoculated with Agrobacterium rhizogenes (A.r.) wild strains (TR7, TR107, 11325 or 15834). Growth and the alkaloid contents of roots and aerial parts were analyzed. Axenic cultures were also performed with modified TR7 strains containing the egfp or gus reporter gene. In vitro isolated root cultures enabled the phenological and molecular demonstration of gene transfer. RESULTS: A.r.TR 7 led to a greater improvement in plant secondary metabolism and growth. Positive expression of the reporter genes occurred. Isolation and subculture of some of the roots of these plants showed a hairy root phenotype; molecular tests proved the transfer of bacterial genes into the roots isolated from the plants. CONCLUSIONS: Hyoscyamine and scopolamine productivity is enhanced after A.r. inoculation in the nutrient solution of hydroponic plants. Transformation events occur in the original roots of the plants. This leads to chimeric plants with a part of their roots harboring a hairy root phenotype. Such semi-composite plants could be used for successful specialized metabolite bioproduction in greenhouses.

Five years investigation of female and male genotypes in périgord black truffle (Tuber melanosporum Vittad.) revealed contrasted reproduction strategies.

The Périgord black truffle (Tuber melanosporum Vittad.) is a heterothallic ascomycete that establishes ectomycorrhizal symbiosis with trees and shrubs. Small-scale genetic structures of female genotypes in truffle orchards are known, but it has not yet been studied in male genotypes. In this study, our aim was to characterize the small-scale genetic structure of both male and female genotypes over five years in an orchard to better understand the T. melanosporum sexual reproduction strategy, male genotype dynamics, and origins. Two-hundred forty-one ascocarps, 475 ectomycorrhizas, and 20 soil cores were harvested and genotyped using microsatellites and mating type genes. Isolation by distance analysis revealed pronounced small-scale genetic structures for both female and male genotypes. The genotypic diversity was higher for male than female genotypes with numerous small size genotypes suggesting an important turnover due to ascospore recruitment. Larger and perennial female and male genotypes were also detected. Only three genotypes (1.5%) were found as both female and male genotypes (hermaphrodites) while most were detected only as female or male genotype (dioecy). Our results suggest that germinating ascospores act as male genotypes, but we also proposed that soil mycelium could be a reservoir of male genotypes.

Nitrogen nutrition of tomato plant alters leafminer dietary intake dynamics.

The leafminer Tuta absoluta (Meyrick) is a major pest of the tomato crop and its development rate is known to decline when nitrogen availability for crop growth is limited. Because N limitation reduces plant primary metabolism but enhances secondary metabolism, one can infer that the slow larval development arises from lower leaf nutritive value and/or higher plant defence. As an attempt to study the first alternative, we examined the tomato-T. absoluta interaction in terms of resource supply by leaves and intake by larvae. Tomato plants were raised under controlled conditions on N-sufficient vs. N-limited complete nutrient solutions. Plants were kept healthy or artificially inoculated with larvae for seven days. Serial harvests were taken and the N, C, dry mass and water contents were determined in roots, stems and leaves. Leaf and mine areas were also measured and the N, C, dry mass and water surface densities were calculated in order to characterize the diet of the larvae. The infestation of a specific leaf lessened its local biomass by 8-26%, but this effect was undetectable at the whole plant scale. Infestation markedly increased resource density per unit leaf area (water, dry mass, C and N) suggesting that the insect induced changes in leaf composition. Nitrogen limitation lessened whole plant growth (by 50%) and infested leaflet growth (by 32-44%). It produced opposite effects on specific resource density per unit area, increasing that of dry mass and C while decreasing water and N. These changes were ineffective on insect mining activity, but slowed down larval development. Under N limitation, T. absoluta consumed less water and N but more dry mass and C. The resulting consequences were a 50-70% increase of C:N stoichiometry in their diet and the doubling of faeces excretion. The observed limitation of larval development is therefore consistent with a trophic explanation caused by low N and/or water intakes.

Modeling the diversion of primary carbon flux into secondary metabolism under variable nitrate and light/dark conditions.

In plants, the partitioning of carbon resources between growth and defense is detrimental for their development. From a metabolic viewpoint, growth is mainly related to primary metabolism including protein, amino acid and lipid synthesis, whereas defense is based notably on the biosynthesis of a myriad of secondary metabolites. Environmental factors, such as nitrate fertilization, impact the partitioning of carbon resources between growth and defense. Indeed, experimental data showed that a shortage in the nitrate fertilization resulted in a reduction of the plant growth, whereas some secondary metabolites involved in plant defense, such as phenolic compounds, accumulated. Interestingly, sucrose, a key molecule involved in the transport and partitioning of carbon resources, appeared to be under homeostatic control. Based on the inflow/outflow properties of sucrose homeostatic regulation we propose a global model on how the diversion of the primary carbon flux into the secondary phenolic pathways occurs at low nitrate concentrations. The model can account for the accumulation of starch during the light phase and the sucrose remobilization by starch degradation during the night. Day-length sensing mechanisms for variable light-dark regimes are discussed, showing that growth is proportional to the length of the light phase. The model can describe the complete starch consumption during the night for plants adapted to a certain light/dark regime when grown on sufficient nitrate and can account for an increased accumulation of starch observed under nitrate limitation.

Comparison of Immediate With Delayed Stenting Using the Minimalist Immediate Mechanical Intervention Approach in Acute ST-Segment-Elevation Myocardial Infarction: The MIMI Study.

BACKGROUND: Delayed stent implantation after restoration of normal epicardial flow by a minimalist immediate mechanical intervention aims to decrease the rate of distal embolization and impaired myocardial reperfusion after percutaneous coronary intervention. We sought to confirm whether a delayed stenting (DS) approach (24-48 hours) improves myocardial reperfusion, versus immediate stenting, in patients with acute ST-segment-elevation myocardial infarction undergoing primary percutaneous coronary intervention. METHODS AND RESULTS: In the prospective, randomized, open-label minimalist immediate mechanical intervention (MIMI) trial, patients (n=140) with ST-segment-elevation myocardial infarction ≤12 hours were randomized to immediate stenting (n=73) or DS (n=67) after Thrombolysis In Myocardial Infarction 3 flow restoration by thrombus aspiration. Patients in the DS group underwent a second coronary arteriography for stent implantation a median of 36 hours (interquartile range 29-46) after randomization. The primary end point was microvascular obstruction (% left ventricular mass) on cardiac magnetic resonance imaging performed 5 days (interquartile range 4-6) after the first procedure. There was a nonsignificant trend toward lower microvascular obstruction in the immediate stenting group compared with DS group (1.88% versus 3.96%; P=0.051), which became significant after adjustment for the area at risk (P=0.049). Median infarct weight, left ventricular ejection fraction, and infarct size did not differ between groups. No difference in 6-month outcomes was apparent for the rate of major cardiovascular and cerebral events. CONCLUSIONS: The present findings do not support a strategy of DS versus immediate stenting in patients with ST-segment-elevation infarction undergoing primary percutaneous coronary intervention and even suggested a deleterious effect of DS on microvascular obstruction size. CLINICAL TRIAL REGISTRATION: URL: http://www.clinicaltrials.gov. Unique identifier: NCT01360242.

Tomato response traits to pathogenic Pseudomonas species: Does nitrogen limitation matter?

Induced chemical defence is a cost-efficient protective strategy, whereby plants induce the biosynthesis of defence-related compounds only in the case of pest attack. Plant responses that are pathogen specific lower the cost of defence, compared to constitutive defence. As nitrogen availability (N) in the root zone is one of the levers mediating the concentration of defence-related compounds in plants, we investigated its influence on response traits of tomato to two pathogenic bacteria, growing plants hydroponically at low or high N supply. Using two sets of plants for each level of N supply, we inoculated one leaf of one set of plants with Pseudomonas syringae, and inoculated the stem of other set of plants with Pseudomonas corrugata. Tomato response traits (growth, metabolites) were investigated one and twelve days after inoculation. In infected areas, P. syringae decreased carbohydrate concentrations whereas they were increased by P. corrugata. P. syringae mediated a redistribution of carbon within the phenylpropanoid pathway, regardless of N supply: phenolamides, especially caffeoylputrescine, were stimulated, impairing defence-related compounds such as chlorogenic acid. Inoculation of P. syringae produced strong and sustainable systemic responses. By contrast, inoculation of P. corrugata induced local and transient responses. The effects of pathogens on plant growth and leaf gas exchanges appeared to be independant of N supply. This work shows that the same genus of plant pathogens with different infection strategies can mediate contrasted plant responses.

Temporal changes of bacterial communities in the Tuber melanosporum ectomycorrhizosphere during ascocarp development.

Ectomycorrhizae create a multitrophic ecosystem formed by the association between tree roots, mycelium of the ectomycorrhizal fungus, and a complex microbiome. Despite their importance in the host tree's physiology and in the functioning of the ectomycorrhizal symbiosis, detailed studies on ectomycorrhiza-associated bacterial community composition and their temporal dynamics are rare. Our objective was to investigate the composition and dynamics of Tuber melanosporum ectomycorrhiza-associated bacterial communities from summer to winter seasons in a Corylus avellana tree plantation. We used 16S ribosomal RNA (rRNA)-based pyrosequencing to compare the bacterial community structure and the richness in T. melanosporum's ectomycorrhizae with those of the bulk soil. The T. melanosporum ectomycorrhizae harbored distinct bacterial communities from those of the bulk soil, with an enrichment in Alpha- and Gamma-proteobacteria. In contrast to the bacterial communities of truffle ascocarps that vastly varies in composition and richness during the maturation of the fruiting body and to those from the bulk soil, T. melanosporum ectomycorrhiza-associated bacterial community composition stayed rather stable from September to January. Our results fit with a recent finding from the same experimental site at the same period that a continuous supply of carbohydrates and nitrogen occurs from ectomycorrhizae to the fruiting bodies during the maturation of the ascocarps. We propose that this creates a stable niche in the ectomycorrhizosphere although the phenology of the tree changes.

Thin strut bare metal stents in patients with atrial fibrillation: Is there still a need for BMS?

OBJECTIVES: This observational study assessed the 9-month clinical outcomes in an « all comers ¼ population with a focus on patients with atrial fibrillation (AF) after thin strut bare metal stenting. BACKGROUND: Drug eluting stent (DES) implantation is the treatment of choice for coronary artery disease (CAD) leaving only marginal indications for the use of bare metal stents (BMS). However, selected treatment populations with DES contraindications such as patients who cannot sustain 6-12 months of dual antiplatelet therapy (DAPT) remain candidates for BMS implantations. METHODS: Thin strut bare metal stenting in a priori defined subgroups were investigated in a non-randomized, international, multicenter «all comers¼ observational study. Primary endpoint was the 9-month TLR rate whereas secondary endpoints included the 9-month MACE and procedural success rates. RESULTS: A total of 783 patients of whom 98 patients had AF underwent BMS implantation. Patient age was 70.4 ± 12.8 years. Cardiovascular risk factors in the overall population were male gender (78.2%, 612/783), diabetes (25.2%, 197/783), hypertension (64.1%, 502/783), cardiogenic shock (4.9%, 38/783) and end stage renal disease (4.9%, 38/783). In-hospital MACE was 4.1% (30/783) in the overall population. The 9-month TLR rate was 4.5% (29/645) in the non-AF group and 3.3% (3/90) in the AF group (P = 0.613). At 9 months, the MACE rate in the AF-group and non-AF group was not significantly different either (10.7%, 69/645 vs. 6.7%, 6/90; P = 0.237). Accumulated stroke rates were 0.3% (2/645) in the non-AF subgroup at baseline and 1.1% (1/90) in the AF subgroup (P = 0.264). CONCLUSION: Bare metal stenting in AF patients delivered acceptably low TLR and MACE rates while having the benefit of a significantly shorter DAPT duration in a DES dominated clinical practice. © 2015 Wiley Periodicals, Inc.

Black truffle-associated bacterial communities during the development and maturation of Tuber melanosporum ascocarps and putative functional roles.

Although truffles are cultivated since decades, their life cycle and the conditions stimulating ascocarp formation still remain mysterious. A role for bacteria in the development of several truffle species has been suggested but few is known regarding the natural bacterial communities of Périgord Black truffle. Thus, the aim of this study was to decipher the structure and the functional potential of the bacterial communities associated to the Black truffle in the course of its life cycle and along truffle maturation. A polyphasic approach combining 454-pyrosequencing of 16S rRNA gene, TTGE, in situ hybridization and functional GeoChip 3.0 revealed that Black truffle ascocarps provide a habitat to complex bacterial communities that are clearly differentiated from those of the surrounding soil and the ectomycorrhizosphere. The composition of these communities is dynamic and evolves during the maturation of the ascocarps with an enrichment of specific taxa and a differentiation of the gleba and peridium-associated bacterial communities. Genes related to nitrogen and sulphur cycling were enriched in the ascocarps. Together, these data paint a new picture of the interactions existing between truffle and bacteria and of the potential role of these bacteria in truffle maturation.

Desorption kinetics of PAHs from aged industrial soils for availability assessment.

Persistent organic pollutants (POPs), such as polycyclic aromatic hydrocarbons (PAHs), may be found in high concentrations in soils of former industrial sites including manufactured gas plants or coking plants. Techniques using moderate solvent extraction, biological tests or solid phase extraction have proved useful for pollution availability estimation. However, more accurate and reliable measurement tools specifically adapted to low concentrations are still needed. Based on a solid-liquid extraction using a Tenax® resin, we suggest a protocol to assess the bioavailability of PAHs, dedicated to aged industrial wasteland soils. Desorption kinetics were measured on three representative contaminated industrial soils. Results were modeled using a first order two-compartment model that provided an estimate of the rapidly desorbing fraction, which was considered to be available, over a 30 h extraction period. In conclusion, this method, allowing the measurement of the available fraction, might prove more relevant than the total concentration value when assessing soil contamination related risks. It may also predict achievable bioremediation performances.

Is it mandatory to carry out completion arteriography after carotid endarterectomy with patch angioplasty?

BACKGROUND: This study compares the neurologic evolution and number of restenoses between 2 groups of patients who underwent internal carotid endarterectomy with patch angioplasty (CEP): one group with systematic intraoperative completion arteriography (CA) and another group without. METHODS: This monocentric retrospective study was performed from January 2000 to December 2008 on 559 consecutive CEPs; 179 were controlled with CA and 380 were not. Surgery was chosen for patients with greater than 50% symptomatic or greater than 60% asymptomatic stenosis (North American Symptomatic Carotid Endarterectomy Trial criteria). Each patient's neurologic and ultrasound status was followed-up postoperatively, and at 3 months in the presence of neurologic symptoms, and at the end of the follow-up. Residual stenosis and restenosis were defined by a reduction in diameter of greater than 50% or a maximal systolic velocity greater than 150 cm/s. The surgical team included 4 vascular surgeons: 2 experienced surgeons who did not perform CA, 1 experienced surgeon monitoring surgical trainees, and 1 who had little experience performing CA. RESULTS: In both groups, the patients were comparable in terms of cardiovascular risks, except for age, gender, statin treatment, and neoplastic medical history. During the first 30 postoperative days, the mean rate of cerebrovascular accident (CVA) and death was 0.4%. In the CA group, 1 incidence (0.6%) of partial monocular blindness occurred, and in the no-CA group, 1 incidence (0.2%) of hemorrhagic CVA and 3 (0.5%) transient ischemic attacks (TIAs) occurred. One of these TIAs resulted from a residual stenosis. Mean follow-up was 41 months. During this time, 6 ipsilateral neurologic accidents occurred in the CA group (1 ischemic CVA, 3 TIAs, 1 partial monocular blindness, and 1 hemorrhagic CVA), whereas 10 occurred in the no CA group (1 ischemic CVA, 4 TIAs, 3 hemorrhagic CVAs, 1 partial monocular blindness, and 1 recurrence of vertebrobasilar insufficiency). The combined morbidity and mortality rates in the 2 groups were 3.3% and 2.8%, respectively (P > 0.05). Two residual stenoses (0.5%) and 6 significant restenoses (1.6%) were diagnosed in the no-CA group; 4 significant restenoses (2.2%) occurred in the CA group (P > 0.05). CONCLUSIONS: When performed by experienced surgeons, systematic intraoperative CA does not reduce the incidence of ipsilateral neurologic complications after CEP. Systematic intraoperative arteriography enables surgical trainees to obtain results as satisfactory as those of experienced surgeons.

Carbon transfer from the host to Tuber melanosporum mycorrhizas and ascocarps followed using a 13C pulse-labeling technique.

Truffles ascocarps need carbon to grow, but it is not known whether this carbon comes directly from the tree (heterotrophy) or from soil organic matter (saprotrophy). The objective of this work was to investigate the heterotrophic side of the ascocarp nutrition by assessing the allocation of carbon by the host to Tuber melanosporum mycorrhizas and ascocarps. In 2010, a single hazel tree selected for its high truffle (Tuber melanosporum) production and situated in the west part of the Vosges, France, was labeled with (13)CO2. The transfer of (13)C from the leaves to the fine roots and T. melanosporum mycorrhizas was very slow compared with the results found in the literature for herbaceous plants or other tree species. The fine roots primarily acted as a carbon conduit; they accumulated little (13)C and transferred it slowly to the mycorrhizas. The mycorrhizas first formed a carbon sink and accumulated (13)C prior to ascocarp development. Then, the mycorrhizas transferred (13)C to the ascocarps to provide constitutive carbon (1.7 mg of (13)C per day). The ascocarps accumulated host carbon until reaching complete maturity, 200 days after the first labeling and 150 days after the second labeling event. This role of the Tuber ascocarps as a carbon sink occurred several months after the end of carbon assimilation by the host and at low temperature. This finding suggests that carbon allocated to the ascocarps during winter was provided by reserve compounds stored in the wood and hydrolyzed during a period of frost. Almost all of the constitutive carbon allocated to the truffles (1% of the total carbon assimilated by the tree during the growing season) came from the host.

Fine-scale spatial genetic structure of the black truffle (Tuber melanosporum) investigated with neutral microsatellites and functional mating type genes.

The genetic structure of ectomycorrhizal (ECM) fungal populations results from both vegetative and sexual propagation. In this study, we have analysed the spatial genetic structure of Tuber melanosporum populations, a heterothallic ascomycete that produces edible fruit bodies. Ectomycorrhizas from oaks and hazels from two orchards were mapped and genotyped using simple sequence repeat markers and the mating type locus. The distribution of the two T. melanosporum mating types was also monitored in the soil. In one orchard, the genetic profiles of the ascocarps were compared with those of the underlying mycorrhizas. A pronounced spatial genetic structure was found. The maximum genet sizes were 2.35 and 4.70 m in the two orchards, with most manifesting a size < 1 m. Few genets persisted throughout two seasons. A nonrandom distribution pattern of the T. melanosporum was observed, resulting in field patches colonized by genets that shared the same mating types. Our findings suggest that competition occurs between genets and provide basic information on T. melanosporum propagation patterns that are relevant for the management of productive truffle orchards.

Identification and characterization of novel PDYN mutations in dominant cerebellar ataxia cases.

We have recently identified missense mutations in prodynorphin (PDYN), the precursor to dynorphin opioid peptides, as the cause for spinocerebellar ataxia (SCA23) in Dutch ataxia cases. We report a screen of PDYN for mutations in 371 cerebellar ataxia cases, which had a positive family history; most are of French origin. Sequencing revealed three novel putative missense mutations and one heterozygous two-base pair deletion in four independent SCA patients. These variants were absent in 400 matched controls and are located in the highly conserved dynorphin domain. To resolve the pathogenicity of the heterozygous variants, we assessed the peptide production of the mutant PDYN proteins. Two missense mutations raised dynorphin peptide levels, the two-base pair deletion terminated dynorphin synthesis, and one missense mutation did not affect PDYN processing. Given the outcome of our functional analysis, we may have identified at least two novel PDYN mutations in a French and a Moroccan SCA patient. Our data corroborates recent work that also showed that PDYN mutations only account for a small percentage (~0.1 %) of European SCA cases.