Features of basidiomycetous yeasts from grapes and apples associated with crop environment and fermenting juice.

AIMS: The characterization of grape and apple yeasts was carried out to investigate the ecology of basidiomycetes associated with crop environment and fermenting juice. METHODS AND RESULTS: A total of 15 basidiomycetous strains were analysed for plant-growth promoting properties, sensitivity to fungicides and features related to their survival in fermenting juice (low pH, SO2 and ethanol sensitivity). Only one strain displayed 1-aminocyclopropane-1-carboxylate deaminase activity, whereas other strains were able to produce ammonia and indole-3-acetic acid, solubilize calcium phosphate, and display catalase activity and antagonism against Botrytis cinerea. Strains presented great variability in their sensitivity to fungicides. Rhodotorula mucilaginosa Yl26 and Sporobolomyces agrorum PYCC 8108T displayed low sensitivity to all fungicides, with high tolerance to SO2 and ethanol, and were able to survive in fermenting grape and apple juice. CONCLUSIONS: This study revealed the diversity of basidiomycetous yeasts in the important physiological traits that affect their growth, either in the crop environment or in fermenting juice. SIGNIFICANCE AND IMPACT OF THE STUDY: Identify the possibility of selective effects of fungicide treatments on basidiomycetous yeasts that could offer benefits for grapevines and apple trees, as well as the survival of strains that are better adapted to fermenting juice and that potentially have a role in the aroma of beverages.

Filamentous fungi associated with natural infection of noble rot on withered grapes.

The effects of noble rot infection of grapes on the characteristics of different types of wine, including Italian passito wine, are well known. Nevertheless, there is still little information on filamentous fungi associated with noble-rotten grapes. In this study, withered Garganega grapes for passito wine production, naturally infected by noble rot, were analyzed and compared to sound grapes. Skin morphology and fungal population on berry surfaces were analyzed. Scanning electron microscopy analysis revealed microcracks, germination conidia and branched hyphae on noble-rotten berries. Penicillium, Aureobasidium and Cladosporium were the most frequent genera present. Analysis of single berries displayed higher heterogeneity of epiphytic fungi in those infected by noble-rot than in sound berries. Penicillium adametzoides, Cladosporium cladospoirioides and Coniochaeta polymorpha were recovered. These, to the best of our knowledge, had never been previously isolated from withered grapes and, for C. polymorpha, from grapevine. This study provided novel data on noble rot mycobiota and suggests that fungi that co-habit with B. cinerea could have an important role on grape and wine quality.

Polymorphism and phylogenetic species delimitation in filamentous fungi from predominant mycobiota in withered grapes.

Filamentous fungi are the main pathogens of withered grapes destined for passito wine production. Knowledge of which species inhabit these post-harvest fruits and their pathogenicity is essential in order to develop strategies to control infection, but is still scarce. This study investigated the predominant mycobiota of withered grapes through a cultivation-dependent approach. Strain and species heterogeneity was evidenced on examining isolates collected over three consecutive years. Colony morphology and PCR-restriction fragment length polymorphism (PCR-RFLP) analysis revealed the occurrence of several phenotypes and haplotypes, respectively. Strains were phylogenetically analyzed based on sequence typing of different genes or regions (e.g. calmodulin, ß-tubulin and internal transcribed spacer region). Beside the most common necrotrophic-saprophytic species of Penicillium, Aspergillus, Alternaria and Botrytis species responsible for fruit rot, other saprobic species were identified (e.g. Trichoderma atroviride, Sarocladium terricola, Arthrinium arundinis and Diaporthe eres) generally not associated with post-harvest fruit diseases. Species such as Penicillium ubiquetum, Cladosporium pseudocladosporioides, Lichtheimia ramosa, Sarocladium terricola, Diaporthe nobilis, Bipolaris secalis, Paraconiothyrium fuckelii and Galactomyces reessii that had never previously been isolated from grapevine or grape were also identified. Moreover, it was not possible to assign a species to some isolates, while some members of Didymosphaeriaceae and Didymellaceae remained unclassified even at genus level. This study provides insights into the diversity of the epiphytic fungi inhabiting withered grapes and evidences the importance of their identification to understand the causes of fruit diseases. Finally, phylogenetic species delimitation furnished data of interest to fungal taxonomy.

Isolation of Neofusicoccum parvum from withered grapes: strain characterization, pathogenicity and its detrimental effects on passito wine aroma.

AIMS: There is scarce information on the occurrence of several fungi that infect withered grapes to produce passito wine. Isolation and characterization of Neofusicoccum parvum strains and evaluation of their effects on withered grape and wine were carried out. METHODS AND RESULTS: Nine isolates were phenotypically characterized by colony morphology and genetically discriminated by molecular methods. Two representative strains were identified as N. parvum according to the phylogenetic analysis of internal transcribed spacer (ITS), and a part of translation elongation factor 1-alfa (TEF) and ß-tubulin DNA sequences. The pathogenicity of both strains on grape berries varied according to the inoculation and incubation conditions. Under withering conditions, infected berries showed browning and shrivelling and some berries showed pycnidial development on the surface. The infection affected laccase, esterase, ß-glucosidase and tannase on grape juice as well as the content of several aroma molecules on resulting wines. Strain-specific effects on wine composition were also observed. CONCLUSIONS: Neofusicoccum parvum occurred in withered grapes and was able to infect grapes under withering condition changing the aroma wine. SIGNIFICANCE AND IMPACT OF THE STUDY: This study reports for the first time the N. parvum isolation in fruit-drying rooms and indicates its important role on postharvest grape infection.

A tool for measuring the bending length in thin wires.

Great effort is currently being put into the development and construction of the second generation, advanced gravitational wave detectors, Advanced Virgo and Advanced LIGO. The development of new low thermal noise suspensions of mirrors, based on the experience gained in the previous experiments, is part of this task. Quasi-monolithic suspensions with fused silica wires avoid the problem of rubbing friction introduced by steel cradle arrangements by directly welding the wires to silica blocks bonded to the mirror. Moreover, the mechanical loss level introduced by silica (φfs ∼ 10(-7) in thin fused silica wires) is by far less than the one associated with steel. The low frequency dynamical behaviour of the suspension can be computed and optimized, provided that the wire bending shape under pendulum motion is known. Due to the production process, fused silica wires are thicker near the two ends (necks), so that analytical bending computations are very complicated. We developed a tool to directly measure the low frequency bending parameters of fused silica wires, and we tested it on the wires produced for the Virgo+ monolithic suspensions. The working principle and a set of test measurements are presented and explained.

Postharvest grape infection of Botrytis cinerea and its interactions with other moulds under withering conditions to produce noble-rotten grapes.

AIMS: To investigate the interactions between Botrytis cinerea and other moulds during grape withering and postharvest infection to obtain noble-rotten grapes. METHODS AND RESULTS: Strains of Botrytis cinerea, Penicillium expansum, Penicillium crustosum, Aspergillus niger, Fusarium verticilloides and Alternaria alternata, isolated from naturally withered grapes and identified by molecular tools, were used to infect Garganega and Corvina grapes. Individually sterilized berries were infected by a single inoculation of each strain or a simultaneous inoculation of B. cinerea together with one of each of the other moulds. Withering kinetics, glycerol, gluconic acid, total polyphenols, total anthocyanins and laccase activity greatly varied among each strain and also in respect to untreated berries. Successful noble rot settlement was ascertained by an additional infection assay carried out on nonsterilized berries. CONCLUSIONS: The suitability of inducing noble rot infection during grape withering and the improvement of the health of noble-rotten grapes have been demonstrated. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides insights on the effects of mould interactions on withered grape quality. Implementing noble rot induction by postharvest infection in winery drying fruit rooms to standardize the level of grape botrytization is encouraged.

A "gentle" nodal suspension for measurements of the acoustic attenuation in materials.

Loss angle measurements in ultralow mechanical loss materials is normally affected by a large systematic error due to the excess losses introduced by the suspension system used to hold the samples. Crystals such as sapphire and silicon or amorphous materials such as fused silica can have loss angles in the range of 10(-10)-10(-7); such materials are of extreme interest in the detection of small displacements as it is required in quantum measurements, frequency stabilization, Micro Electro-Mechanical Systems (MEMS), and gravitational wave research. In the system proposed here the sample is suspended in equilibrium on top of a sphere, touching on one of the nodal points of vibration. The advantage of this system, as compared to others used so far, is twofold: (i) one surface only of the sample is touched and the contact surface is minimized because of the absence of applied forces; (ii) some relevant parameters of the suspension can be measured and eventually varied, giving the experimentalist the possibility to identify whether the measured loss is limited by the suspension system in use or it is an intrinsic property of the sample under investigation. The measurements of a 75 mm diameter and 3 mm thickness disk of Suprasil 311 gave a loss angle phi of 5x10(-8).

Improvement of posture stability by vibratory stimulation following anterior cruciate ligament reconstruction.

Surgical reconstruction of the anterior cruciate ligament (ACL) may reduce, but it does not always eliminate, knee and body instability because of a persisting proprioceptive deficit. In order to enhance body stability, a new protocol of treatment has been proposed consisting of mechanical vibration (100 Hz frequency and < 20 microm amplitude) of the quadriceps muscle in the leg that has undergone ACL reconstruction. In our trials, stimulation was performed when the quadriceps muscle was kept isometrically contracted. Treatment was started one month after surgery. Vibration was applied for short periods over three consecutive days. Nine months after treatment, postural stability was re-evaluated with the subjects standing on one leg with open and with closed eyes. The postural stability of the subjects having undergone vibration treatment, standing on the operated leg was significantly improved one day after treatment when evaluated as mean of speed and elliptic area of the center of pressure. The improvement persisted and increased during the following weeks. Peak torques of the operated leg extensor muscles also increased and reached values close to that of the leg, which had not been operated. Conversely, the balance of the untreated subjects standing on the operated leg did not improve and the restoration of the extensor muscle peak torque was poor. It is concluded that short lasting proprioceptive activation by vibration may lead to a faster and more complete equilibrium recovery probably by permanently changing the network controlling knee posture.

Giant basal cell carcinoma of the thoracic wall: a case report and review of the literature.

Giant basal cell carcinoma is a rare skin tumour with aggressive biological behaviour, and deep invasion and metastasis have been reported. The authors describe a giant basal cell carcinoma involving the anterior chest wall. The lesion infiltrated the mediastinum, occluding the left brachiocephalic vein. Vascular invasion caused venous occlusion affecting the left upper limb. Neither surgical treatment nor radiotherapy were practicable.

Can a post-operative brace in slight hyperextension prevent extension deficit after anterior cruciate ligament reconstruction? A prospective randomised study.

It has been our observation that post-operative anterior cruciate ligament (ACL) braces together with the post-operative bandages do not always allow the knee to reach full extension. In ten uninjured knees with known hyperextension, the knees were bandaged in the same way as after an ACL-reconstruction. The knees were then studied radiologically in a Hypex brace set at 0 degrees, -5 degrees and -10 degrees of knee extension. Not a single knee was found to be straight in the brace set at 0 degrees. At -5 degrees most of the knees were straight or in slight hyperextension. It took -10 degrees to get all knees straight or in hyperextension. In a prospective randomised study 44 patients who underwent an arthroscopic ACL-reconstruction with a bone patellar tendon bone graft were randomised to use either a brace set at -5 degrees or a straight brace (0 degrees ) for at least the first three postoperative weeks. Before and three months after surgery range of motion was determined, using a goniometer with long arms, and sagittal knee laxity was measured with a KT-2000 arthrometer at manual max. Pre- and post-operative pain was evaluated with the Visual Analogue Scale (VAS). The same examiner (blindfolded to what type of brace was used) performed all the measurements. At three months, two of the 22 patients with the brace set at -5 degrees and twelve of the 22 patients with the straight brace had a loss of full extension of 2 degrees or more ( p<0.001). No significant differences were found between the groups in terms of knee flexion, sagittal knee laxity or post-operative pain. Although extension deficit after ACL-reconstruction can be prevented also in other ways, a Hypex brace set at -5 degrees seems to be an easy way of ensuring full knee extension.

Predictive carcinogenicity: a model for aromatic compounds, with nitrogen-containing substituents, based on molecular descriptors using an artificial neural network.

A back-propagation neural network to predict the carcinogenicity of aromatic nitrogen compounds was developed. The inputs were molecular descriptors of different types: electrostatic, topological, quantum-chemical, physicochemical, etc. For the output the index TD50 as introduced by Gold and colleagues was used, giving a continuous numerical parameter expressing carcinogenicity. From the tens of descriptors calculated, principal component analysis enabled us to restrict the number of parameters to be used for the artificial neural network (ANN). We used 104 molecules for the study. An Rcv2 = 0.69 was obtained. After removal of 12 outliers, a new ANN gave an Rcv2 of 0.82.

Successful reconstruction of refractory posttraumatic skull base defects with free radial forearm flap.

After our experience with two patients who had persistent posttraumatic defects of the anterior skull base, we recommend using a deepithelialized free radial forearm flap for one-stage reconstruction. The flap provides a thin, water-tight barrier between the cranial cavity and the subcranial spaces for effective prevention of cerebrospinal fluid leakage and ascending infection. It is particularly useful when local flap options are not available.

Studies on 6-aminoquinolones: synthesis and antibacterial evaluation of 6-amino-8-methylquinolones.

The 6-aminoquinolone had previously been identified as a new class of quinolone antibacterial agents. To continue our structure-activity relationship (SAR) study in this series, novel 6-amino-8-methylquinolone derivatives have now been synthesized and evaluated for in vitro antibacterial activity. We have shown that the coupled presence of a methyl group at the C-8 position with an amino group at C-6 is effective for enhancing antibacterial activity, particularly against Gram-positive bacteria. The SARs associated with the N-1, C-6, and C-7 are discussed. The 1,2,3,4-tetrahydroisoquinolinyl derivative 19v showed the highest antibacterial activity with MIC values on Gram-positive bacteria superior to that of ciprofloxacin, especially against Staphylococcus aureus strains, including those strains which are methicillin-and ciprofloxacin-resistant.

[Intra-oral cancer: immediate reconstruction using a free radial forearm flap]. / Le neoplasie del cavo orale: ricostruzione immediata con lembo libero antibrachiale.

The radial forearm flaps is ideal for intra-oral reconstruction. In that it offers thin, pliable, hairless skin to replace oral mucosa removed during wide resection in surgical treatment of cancer. The vascularity of the area allows considerable variation in design and size of the flap and offers the possibility of covering structures on different levels and, when bone is included, of carrying out immediate reconstruction of the mandible. The Authors used this flap in 24 patients for intra-oral reconstruction. The ages of the subjects ranged from 30 to 70 years. The arteries and the veins of the flap used for microsurgical anastomosis were radial arteries and superficial veins of the elbow. Neck vessels were superior arteries of the thyroid and jugularis externa vein. Operating time was slightly prolonged when compared to repair employing a myocutaneous island flap. This reconstruction method was completely successful in all 24 patients; in one patient a DIC partially compromised the vascularity of the flap on day 8. Morbidity was observed, resouled, however, with minor surgical procedures. Our results support the choice of this method for intraoral reconstruction.

Autoantibodies in 16 patients with extensive burns and a review of the literature.

Serum samples were taken during the first 3 weeks and 1 year after discharge from hospital of 16 patients with burns covering more than 35 per cent of the body surface area. The sera were tested for the presence of antinuclear, antimitochondrial, anti smooth muscle, anti brush border and anti gastric parietal cell autoantibodies using the technique of indirect immunofluorescence. Compared with measurements for the same autoantibodies, in sera from 25 healthy blood donors, the burned patients' sera showed the presence of anti smooth muscle autoantibodies in 81.2 per cent, anti brush border autoantibodies in 56.2 per cent, antimitochondrial autoantibodies in 31.2 per cent and antinuclear autoantibodies in 12.5 per cent of the sera. Autoantibodies to gastric parietal cells were not detected. A correlation was noted between the presence of antimitochondrial autoantibodies and alterations in liver function. In contrast, no correlations were found between the raised levels of the various autoantibodies and resuscitative fluid or antibiotic therapy, intercurrent infections, serum immunoglobulin levels, surgical treatment or the duration of stay in hospital. The high titre of autoantibodies to anti smooth muscle could have arisen from tissue protein degradation associated with the raised requirements for energy by the hypermetabolic patients.

The use of Omiderm, a new skin substitute, in a burn unit.

The use of a new synthetic material for temporary dressing of burn surfaces and skin donor sites is reported. Forty-six patients whose burns were divided into superficial dermal, deep dermal and full thickness post-escharectomy areas and donor sites are presented. For each group of patients the evolution of the wounds and the final results are described.