Apple trees of cv. Starking Delicious producing fruits with depressed green spots 3-4 mm in diameter, scattered on the whole fruit surface and merging around the calyx, were observed in the Bekaa ...valley (Lebanon). These symptoms resembled those of apple dimple fruit and dapple apple diseases induced by Apple dimple fruit viroid (ADFVd) and Apple scar skin viroid (ASSVd), respectively. Dot-blot hybridization using digoxigenin-labelled riboprobes, multiplex RT-PCR amplification assay specific for detecting these viroids in both single and mixed infections and sequencing of the generated amplicons showed that the symptoms were associated exclusively with the presence of ADFVd, whereas ASSVd was never detected. Full length ADFVd cDNAs from one symptomatic apple tree were also amplified and cloned. Sequencing of four independent clones identified ADFVd sequence variants composed by 307 nt with limited variability. Multiple sequence alignment of these variants with those reported previously in databank allowed the identification of one additional polymorphic position in the viroid genome. To our knowledge, this is the first report of ADFVd in Lebanon and the first in a country other than Italy. Tissue-printing hybridization was successfully used for a large-scale survey for the presence of ADFVd and ASSVd in Lebanon, the results of which are presented and discussed.
Fifty eight consecutive untreated patients with locally advanced cervical carcinoma (LACC) received neoadjuvant chemotherapy (NACT) with cisplatin (CDDP) 80 mg/sqm (day 1) + vinorelbine (VRL) 25 ...mg/sqm (day 1 and 8). Three cycles of chemotherapy were planned every 21 days. Within 28 days from the completion of chemotherapy patients in complete or partial response were submitted to radical hysterectomy with pelvic lymphadenectomy. On 155 cycles, hematologic toxicity was mild (G3-4 neutropenia and anemia occurred in 16% and 5%, respectively). Forty-seven patients (81%) were submitted to radical surgery; eight (14%) patients were deemed ineligible for surgery because of poor response to treatment, two (3%) for anesthesia contraindications and one (2%) refused surgery. At pathologic examination 12 patients (25%) had a complete response, one (2%) in-situ carcinoma, six (13%) residual microinvasive disease, and 28 (60%) a partial response. None had extracervical disease. Eight patients (14%) had microscopic lymph node metastases. The number of cycles administered was significantly associated with a good pathologic response. Association of CDDP and VRL as NACT in LACC appears safe and effective. Low cost and modest toxicity would support the initiation of a multicenter randomized phase III trial to compare this association with cisplatin alone.
A disease, named apple dimple fruit (ADF), was reported in Southern Italy first in trees of cultivar 'Starking Delicious' and later in 'Royal Gala' and 'Annurca'. The symptoms are roundish and ...depressed green spots, 3-4 mm in diameter, scattered on the fruit skin. In some cases, the spots coalesce and large discoloured skin areas appear predominantly around the calyx end. The disease is caused by Apple dimple fruit viroid (ADFVd) that has been experimentally transmitted to other commercial apple cultivars, in which it induces typical symptoms or no symptoms at all, and to the 'Fieud 37' pear indicator in which no symptoms develop. Here we report that dot- and northern-blot hybridizations of extracts from different apple and pear cultivars collected in several areas of Campania (Italy) revealed the presence of ADFVd in 'Starking Delicious' trees with typical fruit symptoms and in an 'Annurca' tree without symptoms. The distribution and spread of the viroid, monitored over a three-year period, showed a scattered pattern of infected field trees and lack of transmission between adjacent or remote trees. ADFVd was not detected in seedlings obtained from symptomatic fruits of an ADFVd-infected tree. Molecular hybridizations with specific probes failed to reveal the presence of Apple scar skin viroid (ASSVd), Pear blister canker viroid (PBCVd), Hop stunt viroid (HSVd) and Peach latent mosaic viroid (PLMVd) in the apple trees tested.
Preliminary surveys to investigate the presence and spread of pome fruit viroids in Bosnia and Herzegovina and in Malta were carried out by a tissue printing hybridization (TPH) method. More than 300 ...samples of apple and pear trees from Bosnia and Herzegovina were tested for the presence of Apple scar skin viroid (ASSVd), Apple dimple fruit viroid (ADFVd) and Pear blister canker viroid (PBCVd). Whereas ASSVd and ADFVd were not detected, almost 17% of assayed pear samples, belonging to 13 different cultivars, gave positive TPH signals when hybridized with a PBCVd-specific cRNA probe. In parallel experiments, a total of 113 pear samples (mainly cv Bambinella) from Malta were tested for PBCVd showing an infection rate of 12%. These results were largely confirmed by Northern-blot hybridization assays and by RT-PCR followed by cloning and sequencing of the amplified cDNAs. No symptoms were observed in infected field-grown trees. Seedlings of the pear indicator LA62 were graft-inoculated with bark tissues from several new PBCVd isolates from both countries. Six month post-inoculation, molecular hybridization assays and RT-PCR followed by direct sequencing of the amplicon detected the viroid in the inoculated plants which remained symptomless. Further molecular characterization of several of the new PBCVd isolates allowed the identification of previously unreported polymorphic positions in the viroid genome. Altogether, these data show for the first time that PBCVd infects pear trees in Bosnia and Herzegovina and in Malta, and validate the TPH method for large scale surveys of pome fruit viroid infections.
This paper describes numerical experiments conducted by using the meteorological model CALMET. The aim is to assess how the computational time of the model runs depends on user-defined parameters ...involved in the iterative procedure used by CALMET to solve the meteorological field. The influence of the weather conditions on the simulation time has been also evaluated.
The topography of and the area covered by tenascin, laminin and type IV collagen (all components of the subendothelial basement membrane), and the microvessel area (an index of angiogenesis), as ...evaluated with factor VIII, were investigated immunohistochemically in 61 B-cell non-Hodgkin's lymphomas (B-NHL) and 30 benign lymphadenopathies as controls. The three components were located in the microvessels and in a microvessel-bound stromal reticular network, the expression of tenascin being always more extended and finer than the other components. Of the lymphadenopathies, reactive and atypical lymphoid hyperplasias showed vessels and stromal network in the interfollicular zone only, whereas in Castleman's and angioimmunoblastic forms these structures were widely scattered in the tissue, and the area of the three components and that of the microvessels were significantly larger. Of the low-grade B-NHL, follicular subtypes had vessels and stromal network confined to the interfollicular inflammatory zone, but not in tumor follicles, whereas these structures were irregularly distributed throughout the small lymphocytic subtype. The levels of areas in low-grade B-NHL overlapped those of Castleman's and angioimmunoblastic lymphadenopathies. Among the intermediate-grade tumors, the follicular subtype resembled the follicular tumors, and the diffuse subtypes displayed vessels and stromal network throughout the tissue in close association with the neoplastic cells, and with significant increments of both the tenascin and the microvessel areas, but with a significant reduction of both the laminin and the type IV collagen areas. Distribution was similar in high-grade B-NHL, but tenascin and microvessel area variations, on the one hand, and those of laminin and type IV collagen areas were still more apparent than in the intermediate-grade. A high correlation was demonstrated in all groups of tissues between tenascin and microvessel area. In addition, in the diffuse intermediate-grade and high-grade B-NHL highly immature vessels were frequently detected by ultramicroscopy. The results show that tenascin expression and angiogenesis are closely related, and that both increase in function of tumor malignancy. Unlike laminin and type IV collagen, tenascin is associated with highly immature vessels in B-NHL. We suggest that tenascin expression and angiogenesis are governed by the B-NHL-associated inflammatory infiltrate, as well as by the B-NHL cells, particularly in more malignant tumors.
Peach latent mosaic viroid (PLMVd), a chloroplast replicating viroid, induces a wide variety of symptoms in peach, including a severe chlorosis (albinism) denoted peach calico (PC). PLMVd variants ...causing PC differ from those inciting mosaics or no symptoms at all because they contain an insertion of 12-13 nt wherein the PC molecular determinant maps. This domain has limited sequence variability, folds itself into a hairpin capped by a Urich loop, and appears to emerge sporadically de novo (Malfitano et al., 2003). We have now identified other natural PLMVd variants from two new PC isolates containing insertions similar to, or very different from those previously reported. These variants are unevenly distributed within field plants according to the symptoms they exhibit. Our studies show that: (i) the position, size and folding are structural constrains of the insertions; (ii) natural variants containing an inserted hairpin capped by a GA-rich loop are latent; (iii) the PC phenotype is determined by variants containing insertions with a Urich hairpin loop, although the adjacent stem appears to have some influence, and (iv) the insertion can be acquired and lost during infection, suggesting that a latent variant can evolve into a pathogenic one and viceversa.
Peach calico, a disease characterized by a severe chlorosis (albinism) of leaves, stems and fruits, is caused by Peach latent mosaic viroid (PLMVd) variants containing a molecular determinant located ...in a 12-13 nt insertion that adopts a hairpin conformation capped by a U-rich loop. Histological, cytological and molecular alterations of peach leaves expressing PC symptoms are described. Albino leaf areas lack a differentiated mesophyll cell layer and contain cells with altered proplastids instead of fully developed chloroplasts. Because PLMVd replicates actively in PC symptomatic areas, our findings suggest that PLMVd variants inducing PC block an early step of chloroplast biogenesis impairing plastid-to-nucleus signaling pathways involved in leaf differentiation, a condition resembling that of some albino mutants. The discovery that PLMVd invades the shoot apex, the site of chloroplast biogenesis, and that proplastids from the apex of PC-expressing shoots are structurally altered, strongly support this hypothesis. Because the discolored areas had reduced accumulation of chloroplast ribosomal RNAs and plastid-encoded proteins, the inhibition of chloroplast differentiation could result from impaired protein translation in proplastids caused by the viroid. An important implication of these findings is that all host proteins mediating PLMVd chloroplastic replication must be nucleus-encoded.
In this study, we propose a robuststatistical method to discern anomalous patternsin geoelectrical time series measured in a seismicarea of the Southern Apennine chain. First, a filteringprocedure to ...remove seasonal effects related tometeo-climatic fluctuations was carried out.Then, we selected an autoregressive model able todescribe the time fluctuations of geoelectricalsignals and propose a method to obtain an objectiveestimate of probability of occurrence for each extremeevent detected in the time series. Our applications inSouthern Italy allow us to hypothesize that theambiguity of short-term prediction is within thecomplicated dynamics of the physical processresponsible for electrical anomalies observed on theearth's surface.PUBLICATION ABSTRACT
