Cancer prevalence in the subcategories of the indeterminate class III (AUS/FLUS) of the Bethesda system for thyroid cytology: a meta-analysis.

PURPOSE: The indeterminate cytologic report represents a major challenge in the field of thyroid nodule. The indeterminate class III of the Bethesda classification system (i.e., AUS/FLUS) includes a heterogeneous group of subcategories characterized by doubtful nuclear and/or architectural atypia. The study aim was to  conduct a systematic review and meta-analysis to evaluate the rate of malignancy in each subcategory of Bethesda III. METHODS: PubMed, CENTRAL, and Scopus databases were searched until April 2020. Original articles reporting data on the subcategories of Bethesda III were included. The histological diagnosis was the reference standard to classify true/false negative and true/false positive cases. RESULTS: The pooled cancer prevalence in each subcategory of Bethesda III was estimated using a random-effects model. Twenty-three papers with 4241 nodules were included. Overall, 1163 (27.4%) were malignant. The cancer rate observed in the subcategories ranged from 15%, in "Hürthle cell aspirates with low risk pattern", to 44%, in "Focal cytologic atypia". CONCLUSIONS: The overall cancer rate found in the Bethesda III ranged more largely than that originally estimated (10-30%) and varied among any scenarios. These evidence-based data represent a reference for the clinical management of these patients.

Preoperative Assessment of TERT Promoter Mutation on Thyroid Core Needle Biopsies Supports Diagnosis of Malignancy and Addresses Surgical Strategy.

In the last decade, several molecular markers have been proposed to improve the diagnosis of thyroid nodules. Among these, mutations in the telomerase reverse transcriptase (TERT) promoter have been correlated to malignant tumors, characterized by highest recurrence and decreased patients' survival. This suggests an important role of TERT mutational analysis in the clinical diagnosis and management of thyroid cancer patients. The aim of the study was to demonstrate the adequacy of core needle biopsy (CNB) for the preoperative assessment of TERT mutational status, to reach a more accurate definition of malignancy and a more appropriate surgical planning. Indeed, CNB is gaining momentum for improving diagnosis of thyroid nodules deemed inconclusive by fine needle aspirate (FNA). The study included 50 patients submitted to CNB due to inconclusive FNA report. TERT mutational status was correlated with BRAF mutation, definitive histology, and post-operative TNM staging of the neoplasia. C228T mutation of the TERT promoter was reported in 10% of the papillary carcinomas (PTC) series. When compared with final histology, all cases harboring TERT mutation resulted as locally invasive PTCs. The prevalence of TERT mutated cases was 17.6% among locally advanced PTCs. TERT analysis on CNB allows the assessment of the pathological population on paraffin sections before DNA isolation, minimizing the risk of false negatives due to poor sampling that affects FNA, and gathering aggregate information about morphology and TERT mutational status. Data indicating a worse outcome of the tumor might be used to individualize treatment decision, surgical option, and follow-up design.

Immunohistochemistry for BRAF(V600E) antibody VE1 performed in core needle biopsy samples identifies mutated papillary thyroid cancers.

BRAF(V600E) is the most frequent genetic mutation in papillary thyroid cancer (PTC) and has been reported as an independent predictor of poor prognosis of these patients. Current guidelines do not recommend the use of BRAF(V600E) mutational analysis on cytologic specimens from fine needle aspiration due to several reasons. Recently, immunohistochemistry using VE1, a mouse anti-human BRAF(V600E) antibody, has been reported as a highly reliable technique in detecting BRAF-mutated thyroid and nonthyroid cancers. The aim of this study was to test the reliability of VE1 immunohistochemistry on microhistologic samples from core needle biopsy (CNB) in identifying BRAF-mutated PTC. A series of 30 nodules (size ranging from 7 to 22 mm) from 30 patients who underwent surgery following CNB were included in the study. All these lesions had had inconclusive cytology. In all cases, both VE1 and BRAF(V600E) genotypes were evaluated. After surgery, final histology demonstrated 21 cancers and 9 benign lesions. CNB correctly diagnosed 20/20 PTC and 5/5 adenomatous nodules. One follicular thyroid cancer and 4 benign lesions were assessed at CNB as uncertain follicular neoplasm. VE1 immunohistochemistry revealed 8 mutated PTC and 22 negative cases. A 100% agreement was found when positive and negative VE1 results were compared with BRAF mutational status. These data are the first demonstration that VE1 immunohistochemistry performed on thyroid CNB samples perfectly matches with genetic analysis of BRAF status. Thus, VE1 antibody can be used on thyroid microhistologic specimens to detect BRAF(V600E)-mutated PTC before surgery.

First Report of the Southern Root-Knot Nematode Meloidogyne incognita on the Invasive Weed Araujia sericifera in Italy.

Moth plant, Araujia sericifera, is native to South America and was exported to many other countries as an ornamental plant. However, it is now considered an invasive, perennial, noxious weed in Italy. Because of the ability of this plant to spread rapidly and invade natural ecosystems, A. sericifera has been included on the Alert list by the European and Mediterranean Plant Protection Organization (EPPO). In September 2013, numerous plants of A. sericifera with chlorotic leaves and large root-galls were observed in agricultural fields, gardens, and uncultivated locations in Nocera Inferiore, Salerno Province, Italy. Ten samples were collected from a vegetable farm (40°45'40.8″ N, 14°38'18.4″ E) and nematodes were extracted from soil and root samples using standard procedures (1). Meloidogyne sp. was found in all soil and root samples, with nematode population densities ranging from 420 to 1,270 eggs and J2s/10 cm3 of soil and 84 to 2,200 eggs and J2s/5 g of fresh roots. The morphological identification of the nematode was based on characterization of second-stage juveniles (J2s), males, eggs, and females (2). Measurements of J2s (n = 30) include: mean body length (L) = 403 ± 3.7 µm; L/maximum body width = 27.5 ± 0.3; L/esophageal length = 6.2 ± 0.1; stylet length = 12.8 ± 0.1 µm; L/tail length = 9.5 ± 0.1; tail length = 42.5 ± 0.3 µm. Males (n = 10): L = 1,491 ± 0.04 µm; L/maximum body width = 44.4 ± 0.8; L/esophageal length = 16.1 ± 0.3; stylet length = 22.1 ± 0.7 µm; spicules length = 30.1 ± 0.8 µm. Eggs (n = 30): length = 96.6 ± 1 µm; width = 45.1 ± 0.5 µm; length/width ratio = 2.1. Females (n = 20): L = 909.5 ± 38.4 µm; body width = 588.3 ± 19.3 µm; stylet length = 17.0 ± 0.2 µm. Perineal patterns of females had a high dorsal arch with wavy striae bending toward the lateral lines and the absence of distinct lateral line incisures. All measurements conformed to the description of Meloidogyne incognita (Kofoid & White, 1919) Chitwood 1949. DNA was extracted from five individual adult females from each sample and morphological identification was confirmed by a sequence-characterized amplified region (SCAR)-PCR technique using species-specific primers. The amplified product obtained was 1.2 kb in length, demonstrating proper amplification of the species-specific, length variant SCAR marker (3). Weeds are known to serve as hosts for nematodes in the absence of crop plants and to affect the success of nematode management programs. Meloidogyne spp. have been reported to survive and even thrive on weeds; among them, M. incognita is considered the most economically important agricultural nematode pest worldwide as it causes severe yield losses on many hosts. Thus, the invasive plant A. sericifera can be a potential reservoir for M. incognita in Italy and elsewhere. To our knowledge, this is the first report of M. incognita parasitizing A. sericifera. References: (1) K. R. Barker. Page 19 in: An Advanced Treatise on Meloidogyne. Vol. II, Methodology. K. R. Barker et al., eds. North Carolina State University Graphics, Raleigh, 1985. (2) J. D. Eisenback et al. A Guide to the Four Most Common Species of Root-Knot Nematodes (Meloidogyne spp.), with a Pictorial Key. North Carolina State University, Raleigh, 1981. (3) C. Zijlstra et al. Nematology 2:847, 2000.

CONTROL OF VIRAL DISEASES TRANSMITTED IN A PERSISTENT MANNER BY THRIPS IN PEPPER (TOMATO SPOTTED WILT VIRUS).

Tomato spotted wilt disease is caused by Tomato Spotted Wilt Virus (TSWV) (Tospovirus, Bunyaviridae), a virus that severely damages and reduces the yield of many economically important plants worldwide and actually it is a major disease affecting the production of tomato and pepper in Italy. Due to the non-predictive nature of its outbreaks combined with the lack of forecasting, adoption of preventive measures have not always been practical, in fact the disease cycle has proven to be extremely difficult to break because of the wide and often overlapping host range of both the virus and the thrips vectors, which transmit the virus in a persistent, circulative, and propagative manner. Moreover recently, resistance breaking (RB) isolates of TSWV that overcome the resistance conferred by the Tsw gene in different pepper hybrids have been recovered in different locations in Italy and also in Brazil, USA, Spain and Australia, and this occurrence raises the question on the importance of a new approach of integrated pest management for TSWV management, including both control of its insect vector and the induction of the plant's resistance against viral infection. In this perspective, a study was performed in 2012 and 2013 with the purpose of evaluating the efficacy of the insecticide Cyantraniliprole alone or combined with Acibenzolar-S-Methyl (ASM), inducer of systemic acquired resistance, in the control of tomato spotted wilt disease in pepper. The experiment was performed in laboratory, in a thermo-conditioned greenhouse, into separate insect-proof cages and consisted of 5 treatments and 2 applications (plus a pre-transplant application for treatments were ASM was used. Variables were the mode of application of ASM in pre-transplant (by foliar or by drench) and the duration of the exposure time of the treated plants to viruliferous insects. Pepper cv. Corno di Toro, devoid of any resistance to TSWV, was used. Plants were observed daily to record any symptom induced by TSWV and/or of phytotoxicity. The possible TSWV infection was verified by DAS-ELISA using antisera supplied by Agdia Biofords. It was shown that the combination of ASM and Cyantraniliprole allowed the best control of TSWV transmission by thrips and of the development of the infection in treated pepper plants, regardless of the exposure time to thrips infestation. The application via drench of the combination seems to be the most effective. The efficacy of Cyantraniliprole used alone was comparable to that obtained with its combination with ASM only when plants were exposed to viruliferous insects for 24 h after product application, and the effectiveness of protection decreased with increasing time of exposure to viruliferous thrips. Treatment with ASM alone determined a good protection to plants regardless of the length of the exposure timing to viruliferous insects, with a smaller number of infected plants and milder symptoms compared to the check. Results of our current experiments raise the interesting possibility to combine in one product an insecticide and a non-specific (to various biotic and abiotic stresses) resistance inducer, particularly against diseases caused by viruses, and offer great opportunity for the integrated pest management (IPM) strategy for the control of pepper diseases.

Resistance breaking tomato spotted wilt virus isolates on resistant pepper varieties in Italy.

In spring 2012, resistance breaking (RB) isolates of tomato spotted wilt virus (TSWV) that overcome the resistance conferred by the Tsw gene in different pepper hybrids have been recovered in different locations in southern Italy (Campania and Apulia regions) in protected cultivation, about one month after transplant. The percentage of symptomatic plants was 5-10% and, only in particular cases of advanced stage of cultivation, it reached 30-50% at the end of cycle. All TSWV isolates induced similar systemic symptoms in all resistant infected pepper hybrids: yellowing or browning of apical leaves, which later become necrotic, long necrotic streakson stems, extending to the terminal shoots, complete necrosis of younger fruits and large necrotic streaks and spots on fruits formed after infection. On ripe fruits, yellow spots with concentric rings or necrotic streaks could be observed. Leaf extracts of these samples were tested in ELISA for the detection of TSWV, Cucumber mosaic virus (CMV), Tobacco mosaic virus (TMV), Tomato mosaic virus (ToMV), Impatiens necrotic spot virus (INSV), Potato virus Y (PVY), Alfalfa mosaic virus (AMV), Pepper mild mottle virus (PMMoV) and Pepper Mottle Virus (PepMoV). Only TSWV was detected in all the field samples tested. The correspondent virus isolates were inoculated mechanically and by Frankliniella occidentalis on to a set of different pepper and tomato hybrids, as well as on some herbaceous test plants, in order to investigate for their ability to overcome the resistance genes Tsw and Sw5, respectively. Tomato hybrids carrying the Sw5 gene were uninfected by all RB isolates, whereas all resistant pepper hybrids became systemically infected. RB isolates did not differ noticeably in transmission efficiency when they were tested with the thrips F. occidentalis. Obtained results demonstrate that evolved strains of TSWV have emerged, that they are able to overcome the Tsw resistance gene in pepper plants experimentally inoculated both by mechanical infection and by the vector F. accidentalis. Moreover, evolved strains, here indicated as TSWV-RB-PI (Pepper, Italy), are not able to infect tomato plants carrying the Sw5 gene, confirming that distinct viral gene products control the outcome of infection in plants carrying Sw5 and Tsw. The simultaneous presence of evolved TSWV strains, showing the same biological characteristics (such as host range and symptomatology on different hosts) in two geographic areas of southern Italy, very far each other, let us hypothesize a unique focus of infection. Only subsequently, infected plants, through trade of plant material, have been diffused in locations so far apart. Further molecular studies are underway to assess the possible correlation between the different isolates in different geographical areas.

New report of Phoma glomerata on Coriandrum sativum L.

In 2011, coriander plants (Coriandrum sativum L.), showing symptoms of greyish to hell brown lesions with dark brown border between discoloured and asymptomatic tissues, were observed in Bulgaria. Pycnidia with small unicellular conidia appeared in the pale centre of the lesions. In this work, the identification of the causal agent of this disease was made applying Koch's postulates. Moreover, for a rapid and unambiguous identification of the fungal species, the internal transcribed spacers and the 5.8 rDNA gene (ITS1-5.8-ITS2) were amplified by PCR from DNA extracted from the isolated colonies. The amplicon was sequenced and analyzed using BLASTn, and showed a homology of 100% with a corresponding sequence of Phoma glomerata (accession number DQ093699). The fungus isolated, after the morphological and molecular characterization, was ascribed to the species Phoma glomerata (Corda) Wollenweber and Hochapfel. This is the first report of P. glomerata as agent of stem rot of coriander in Bulgaria and elsewhere.

Evaluation of efficacy and effect of application timing of a new herbicide, a.i. propoxy-carbazone + iodosulfuron + mefenpyr on Triticum durum.

A study was performed from February to May 2010 by Bioagritest according to EPPO guidelines and Principles of Good Experimental Practice (GEP), in the land of Altamura (BA), in the core of Murgia, Apulia Region (Italy). The purpose of the study was to evaluate the efficacy and effect of two different application times of the herbicide SIT90 (propoxycarbazone + iodosulfuron + mefenpyr) on Triticum durum for weeds' control. Sit90 was applied alone or in combination with the herbicide Dicuran (a.i. chlortolorun) in early post-emergence, and in late post emergence once more alone or in combination with the adjuvant Biopower or with the commercial formulate Atlantis WG (mesosulfuron+ iodosulfuron+ mefenpyr) + Biopower. T. durum cultivar was "Iride". The study has given sufficient results on the use of SIT90, in the conditions foreseen by the protocol and in consideration of the present weeds, which represented the species mostly diffused in the Murgia, mainly Avena ludoviciana and Papaver rhoaes. Treatments with the SIT90 alone (even in combination with Chortoluron), applied in early or in late post-emergence, were very ineffective on grass weeds and dicotyledonous, highlighting the importance of the adjuvant Biopower to enhance the expression of the herbicide. In fact, the presence of the adjuvant allowed the expression of a clear and good overall herbicide activity of SIT90 for the control of P. rhoaes and other dicotyledonous, but also a discrete activity against grasses. More reliable was the treatment with SIT90 mixed with Atlantis, against the entire community of grass weeds and on F. officinalis. The performance of all the treatments was insufficient on P. rhoaes, because of the clear resistance shown by the weed.

Chlorantraniliprole/lambda-cyhalothrin, a new insecticide mixture to control Tuta absoluta and Spodoptera littoralis in tomato.

A study was performed in the period May-July 2011 by Bioagritest test facility according to EPPO guidelines and Principles of Good Experimental Practice (GEP), in the land of Eboli (SA), southern Italy, with the purpose to test a new insecticide mixture in the defense strategies of processing tomato against Tuta absoluta and Spodoptera littoralis. The insecticide mixture Chlorantraniliprole 100 g/lambda-Cyhalothrin 50 g/l 0,4 It/ha was applied in a tomato field in four (A-B-C-D) application timings (a.t.) alone (treatment 2) and in two a.t. (A-B) combined with emamectin benzoate 0.5%, 1,5 Kg/ha (a.t. C-D) (treatment 3). Its effect in containing insect populations was compared with that exerted by emamectin benzoate 0.5%, 1,5 Kg/ha (a.t. A-B-C-D) (treatment 4) and the combination of chlorantraniliprole 35%, 0.115 Kg/ha (a.t. A-B) and indoxacarb 30% 0.125 Kg/ha (a.t. C-D) (treatment 5). Application of treatments (trt.) and mode of assessment (2 assessments), recording and measurements followed the guidelines foreseen by EPPO Standards PP1/150 and PP1/275. The lepidoptera infestation in the tomato crop was very high on both the leaves and fruits, and this enabled us to evaluate the efficacy of the tested products in conditions of severe pests infestation. If we consider the leaf damage, either as number of mines by T. absoluta or as % of leaf erosion by S. litteralis, all the treatments values were statistically lower than control, but with no difference among them. But when using as an infestation threshold the percentage of damaged fruits (much more relevant in economic terms), we observed statistically significant differences among treatments. Actually, treatment with chlorantraniliprole-lambda-cyhalothrin mixture was significantly more effective than the one with emamectin benzoate in reducing the attack of T. absoluta on tomato fruits, in both dates of assessments. As concerns damage by S. littoralis on the fruits, at the first assessment, trt. 2 and trt. 5 significantly reduced the percentage of infested berries in comparison with the control, whereas the infestation was not significantly reduced by trt. 3 and trt. 4. At the second assessment, no statistically significant difference between the four different treatments was observed. Since the infestation of tomato fruits by S. littoralis was much lower than the one caused by T. obsoluta, the overall performance of the four insecticide treatments was mainly due to the control effects towards T. obsoluta.

Morphological and molecular characterisation of fungal populations possibly involved in the biological alteration of stones in historical buildings.

The deterioration process of historical building is progressive and irreversible, and the timing and mode of impact are different depending on the characteristics of building materials used, local microclimate, air pollution, presence of specific flora and fauna. The chemical and microbiological characterisation of building materials is mandatory in preventing and eventually recovering degradation effects. Ideally, the analysis of structural stones should be complete, efficient, rapid, and non destructive when dealing with a precious or unique construction. The investigation has been performed on a private historical building made using calcarenite stones and sited between the archaeological site of Lavello, a little town located in the Basilicata Region (South Italy), and the industrial area surrounding this town. To study in progress the degradation of stone materials, a new building sample (ca. 1 m3) was constructed by using the same stones (33 x 15cm), collected from a local quarry. The intact calcarenite stone was characterised by using different methods of surface analysis (XRD, XPS, SEM), and exposed to outdoor conditions. The analyses of the stone material were repeated after three and six months to early evaluate the progression of alterations and the forward modifications of calcarenite structure. After only three months of the new building sample exposure, the adopted analytical methods were able to provide a series of data, which allowed the assessment of the incipient modification of the stone surfaces. The degradation appeared worsened performing the same observations on sixth month replicates, suggesting that environmental conditions modified the structure and the compactness of stones and favoured the biological colonization of surfaces especially in the South-East direction of prevailing winds. For this reason the presence of fungi on the stones' surface was investigated and a morphological and molecular characterization of sampled fungi was performed. Several genera and species of fungi, possibly, involved in degradation were found. The most frequent colonies belonged to Alternaria (A. infectoria, A. citri and Alternaria sp.), Coprinopsis sp., Penicillium piceum, Fusatrium equiseti and Scytalidium termophilus.

Phenotypic changes of the thyrocyte membrane in papillary thyroid carcinoma. A three-dimensional study.

Aim of the study was to assess the presence of structural changes in the complex carbohydrate chains of thyroid epithelia undergoing neoplastic transformation. We investigated thyroid cells from neoplastic lesions using a panel of lectins with specific affinity for distinct carbohydrate residues. Sixty samples of thyroid tissue, including normal, hyperplastic and neoplastic lesions were obtained from surgical specimens and blindly evaluated with lectin stains. Confocal microscopy was used to obtain three-dimensional (3-D) images of the samples with a positive reaction. Wheat germ agglutinin (WGA) was consistently positive on the apical membrane of papillary thyroid carcinomas (PTC), was weakly expressed in follicular carcinomas (FC) and resulted negative in normal thyrocytes and in benign conditions. The 3-D microscopy model showed that the WGA staining pattern in light microscopy corresponds to a continuous layer on the luminal surface of both papillary and tubular structures of PTC cells. The other lectins under evaluation did not provide any significant result. In conclusion, in PTC the apical border of thyrocytes showed a strong, specific and consistent staining with WGA. These findings may be related to a modified interaction of thyroglobulin molecule with thyroid cell membrane and with the expression of molecules that are involved in the process of tumorigenesis and tumor progression.

Evaluation of the efficacy and selectivity of Oberon (Spiromesifen) for the control of Tetranychus urticae on strawberry.

A study was performed in the period June to July 2009 by Bioagritest test facility according to EPPO guidelines and Principles of Good Experimental Practice (GEP), in the campaign of Tursi (MT), southern Italy, in order to evaluate the efficacy and selectivity of Spiromesifen on strawberry for the control of Tetranyichus urticae. Two different dosages of OBERON (a.i. Spiromesifen)--45 and 60 ml/hl--were compared with a unique dosage of two commercial formulates: VERTIMEC (a.i. Abamectine, Syngenta Crop Protection), 60 ml/hl, and MAGISTER (a.i. Fenazaquin, Dow AgroSciences), 60 ml/hl. The study has achieved the purpose of evaluating/measuring with a single application the activity of Spiromesifen (Oberon) to control T. urticae on strawberry and its selectivity on phytoseiids. All the treatments differed significantly from the checks and showed high ability to control mites, on all stages of the population. About selectivity on phytoseiids, only Spiromesifen (at both doses) has demonstrated a good selectivity, while Abamectine (in part) and Fenazaquin (totally), have limited the population of the predator. The study confirmed the usefulness, indeed the need for the new compounds, to get confirmation of their selectivity against the useful entomofauna. The experiment has allowed to demonstrate the suitability of Spiromesifen to be included in strategies for strawberry integrated pest management.

Evaluation of the efficacy of Oberon (Spiromesifen), to contain infestations of mites and whiteflies on Capsicum annuum L.

Following the EPPO guidelines and Principles of Good Experimental Practice (GEP), an experiment was realised in autumn 2008 for evaluating the efficacy of Oberon applied by foliar treatments to contain infestations of mites and whiteflies on Capsicum annuum L.. Two different dosages of OBERON (a.i. Spiromesifen)--45 and 60 g/hl--were compared with a unique dosage of two commercial formulates: VERTIMEC (a.i. Abamectine, Syngenta Crop Protection), 60 g/hl, and MAGISTER (a.i. Fenazaquin, Dow AgroSciences), 110 g/hl. Oberon resulted very effective in the control of phytopathogenic mites at both doses of 45 and 60 g/hl. Its effectiveness demonstrated to be remarkable for approximately one month after application. By contrast, Vertimec and Magister have proven their effectiveness for a much lower period of time (about the first 15 days post application). About the efficacy against whiteflies, even 36 days after the foliar application Oberon showed a strong containment of the populations of aleurodides. There were no phenomena of phytotoxicity nor on leaves nor on flowers and fruits, in none of the treatments. About the phytotoxicity on the useful entomofauna, the assessments made on the different treatments have highlighted the lack of harmful effects on predators and on parasitoids of insects and mites.

Integrated approach to TYLCD management in Sardinia (Italy).

In Sardinia (Italy) Tomato yellow leaf curl disease (TYLCD) re-emerged in 2003, after a period of decline, producing severe yield losses in protected tomato crops. This epidemic outbreak highlighted the inadequacy of the approach to TYLCD management based chiefly on the chemical control of its vector, the whitefly Bemisia tabaci, and the use of 40-mesh nets for greenhouse screening. To evaluate the reliability of alternative practices for implementation within IP&DM programmes, in 2007 and 2008 we carried out two field experiments on greenhouse tomato crops planted in summer. In both trials we tested the use of UV-reflective mulches (UVRM) or floating row covers (NWRC), against an uncovered control on clear mulch. Furthermore, we assessed the effectiveness of acibenzolar-S-methyl, a SAR elicitor, as subplot factor in a split plot design. In 2007 because of the low incidence of the disease it was not possible to assess differences among the treatments. However a significant increase in plant growth, and production was observed in UVRM plots. By contrast, in 2008 the study crop was seriously affected by TYLCD. In the plots not treated with acibenzolar-S-methyl, the disease progression was initially slowed down in NWRC and, to a lesser extent, in UVRM plots compared to control plots but, during the second month of the cropping period, the differences among treatments became statistically not significant. In the plots treated with acibenzolar-S-methyl the dynamics of TYLCD infection during the first five weeks exhibited the same trend as in the untreated plots. Thereafter, the combination of UVRM and, to a lesser extent, of NWRC with the SAR elicitor resulted in a reduction of disease incidence with respect to the control. In conclusion, all the control measures tested in the present work showed the potential for TYLCD management in greenhouse tomatoes. However, the most promising results were achieved using UV-reflective mulch and non-woven row cover in combination with acibenzolar-S-methyl. In view of its positive impact on plant growth and yield in summer planted tomato crops, as well as on TYLCD infection, the use of UV-reflective mulch warrants further investigation.

First record of tomato yellow leaf curl Sardinia virus (TYLCSV) on pepper in Italy.

During a survey in summer 2007, a disease of pepper (Capsicum annuum) under plastic tunnels was observed in Policoro (Matera), on the Ionic coast of Basilicata Region, with a disease incidence in some cases of more than 50%. Affected cultivars were Eppo and Almund (S Et G). The diseased plants exhibited light mosaic or mottling, leaf distortion, interveinal and marginal leaf chlorosis, upward curling of leaf margins of older leaves. The causal pathogen was suspected to be a begomovirus due to the large population of the whitefly Bemisia tabaci observed on the crop. Detection assays for Tomato yellow leaf curl Sardinia virus (TYLCSV) and Tomato yellow leaf curl virus (TYLCV) were used. In DAS-ELISA, positive results (178 plants resulted positive over 200 symptomatic plants assayed) were obtained using a "broad-spectrum" reagent combination (distributed by Bioreba AG) detecting TYLCV, TYLCSV, and other begamoviruses. A couple of synthetic oligonucleotides allowing the amplification of the whole coat protein (CP) gene of TYLCSV and TYLCV was used for PCR of ELISA positive samples in order to perform the molecular characterisation of the viral isolate responsible of the disease. RFLP analysis performed on the PCR product, 1008 bp long, showed the presence of only TYLCSV in the infected pepper plants. The same couple of primers allowed the detection of the virus also in symptomless pepper plants. To test whitefly transmission, adults of B. tabaci allowed to feed on naturally infected pepper plants were transferred on 10 healthy Eppo pepper seedlings (15 whiteflies/plant). Insects were killed 2 days later using an insecticide. Twenty days post exposition 10 plants/10 resulted positive in ELISA, and showed the same symptoms observed in natural infection. TYLCSV was not reported before on pepper in the surveyed area, but it was recorded with severe outbreaks on tomato, both in protected and in open field crops. This species was probably the primary source of infection from which subsequent diffusion by way of the vector B. tabaci followed on pepper. To our knowledge this is the first time that a natural infection of TYLCSV on pepper is recorded in Italy, with serious implications for the epidemiology of TYLCSV in our country.

PD-1 /PD-L1 checkpoint in hematological malignancies.

Programmed cell death protein 1 (PD-1), is a cell surface receptor with an important role in down-regulating the immune system and promoting self-tolerance by suppressing T cell inflammatory activity. PD-1/PDL1 axis represents a checkpoint to control immune responses and it is often used as a mechanism of immune escaping by cancers and infectious diseases. Many data demonstrate its important role in solid tumors and report emerging evidences in lymphoproliferative disorders. In this review, we summarized the available data on the role of PD-1/PD-L1 checkpoint in lymphoproliferative diseases and the therapeutics use of monoclonal blocking antibodies.

Differences between ATA, AACE/ACE/AME and ACR TI-RADS ultrasound classifications performance in identifying cytological high-risk thyroid nodules.

OBJECTIVE: Thyroid ultrasound is crucial for clinical decision in the management of thyroid nodules. In this study, we aimed to estimate and compare the performance of ATA, AACE/ACE/AME and ACR TI-RADS ultrasound classifications in discriminating nodules with high-risk cytology. DESIGN: Cross-sectional study. METHODS: 1077 thyroid nodules undergoing fine-needle aspiration were classified according to ATA, AACE/ACE/AME and ACR TI-RADS ultrasound classifications by an automated algorithm. Odds ratios (ORs) and receiver operating characteristic (ROC) curves for high-risk cytology categories (TIR3b, TIR4 and TIR5) were calculated for the different US categories and compared. RESULTS: Cytological categories of risk increased together with all US classifications' sonographic patterns (P < 0.001). The diagnostic performance (C-index) of ACR TI-RADS and AACE/ACE/AME significantly improved when adding clinical data as gender and age in the regression model (P < 0.001). A significant difference in the final model C-index between the three US classification systems was found (P < 0.029), with the ACR TI-RADS showing the highest nominal C-index value, significantly superior to ATA (P = 0.008), but similar to AACE/ACE/AME (P = 0.287). ATA classification was not able to classify 54 nodules, which showed a significant 7 times higher risk of high-risk cytology than the 'very low suspicion' nodules (OR: 7.20 (95% confidence interval: 2.44-21.24), P < 0.001). CONCLUSIONS: The ACR TI-RADS classification system has the highest area under the ROC curve for the identification of cytological high-risk nodules. ATA classification leaves 'unclassified' nodules at relatively high risk of malignancy.

Detecting N-RAS Q61R Mutated Thyroid Neoplasias by Immunohistochemistry.

Recently, the immunohistochemistry (IHC) for N-RAS Q61R has been developed and commercialized for clinical practice. Here, we investigated the reliability of IHC to identify N-RAS Q61R mutated thyroid neoplasia. A series of 24 consecutive thyroid lesions undergone surgery following indeterminate cytology were enrolled. Paraffin sections were stained for IHC using the rabbit monoclonal anti-human N-RAS Q61R, clone SP174. N-RAS mutations in codon 61 were also investigated by automated sequencing. At histology, 12 cases of follicular carcinoma, cytologically defined as follicular lesions, 1 papillary cancer, 7 follicular adenomas, and 4 hyperplastic nodules were found. Of these, 4 showed a positive IHC for anti N-RAS antibody where N-RAS expression was detected mainly at cytoplasmic level with similar intensity of reaction. The remaining cases had negative IHC. A 100% concordance between IHC and molecular analysis for N-RAS Q61R was observed. In conclusion, this study shows high reliability of IHC to identify N-RAS Q61R mutated thyroid lesions with high cost-effectiveness. These data indicate the reliability of IHC to identify N-RAS Q61R mutated thyroid neoplasia and suggest to adopt this approach for a more accurate management of patients, when indicated.

Typing of tomato yellow leaf curl viruses and their vector in Italy.

A molecular survey of TYLCV/TYLCSV and their associated vector Bemisia tabaci, was performed during 2004-2005 in five regions of Southern Italy: i.e. Sardinia (one locations), Sicily (one location), Calabria (three locations), Campania (two locations) and Basilicata (one location). A total of 71 tomato samples were checked for virus infection and for the presence of the vector. Degenerate primers allowing the amplification of the coat protein gene of both TYLCSV and TYLCV isolates were designed. PCR fragments were then digested with restriction endonuclease Ava II, which was expected to cut TYLCSV differently from TYLCV. Results clearly suggested that in all the inspected Italian regions the two viruses are widespread and present in single plant both alone and in mixed infections. The identity of the two viruses was confirmed by total or partial sequencing of field isolates. Concerning the populations of the B. tabaci associated with TYLCD epidemics, the molecular characterization of COI gene (citocrome oxidase I) indicated that Q biotype was the most prevalent biotype. This fact might be the result of the large use of some insecticides against which Q biotype populations easily develop resistances, as already confirmed in some countries of Mediterranean basin.