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The use of megaprostheses or custom-made devices is an established treatment for or- thopedic malignancies, but their indication has been expanded to some non-neoplastic conditions such as complex cases of re-revision surgery or failed osteosynthesis. A con- secutive series of 5 patients were treated with megaprostheses as a solution for complex cases of re-revision surgery around the knee joint. Each patient was assessed clinically and radiographically at 1, 3, 6, and 12 months after surgery. One patient died for intestinal infarc- tion after surgery. The 4 remaining patients resumed gait and knee function. We did not observe infections or mobilizations of the implants, and the only complication we described was patellar dislocation of the extensor mechanism when the implant with proximal tibia resection was used. Complex re-revision surgery after TKA represents a major challenge for the orthopedic surgeon due to poor bone stock and the presence of prosthetic revision components. The implant of megaprostheses or custom-made devices can play a crucial role in these rare but complex cases of non-oncological orthopedic surgery ensuring early functional recovery.

Background Some periprosthetic femoral fractures (PFFs) present history and radiographic aspect consistent with an atypical femoral fracture (AFF), fulfilling the criteria for AFF except that PFFs by themselves are excluded from the diagnosis of AFFs. The aim of this study is to evaluate in a single institution series of PFFs if any of them could be considered a periprosthetic atypical femoral fracture (PAFF), and their prevalence. Methods Surgical records were searched for PFFs around a primary hip stem from January 2013 to December 2019. Cases were classified according to Vancouver classification. Demographic and medical history was extracted. Fisher’s exact test was used for statistical analysis. Results One hundred fifteen PFFs were identified, 59 of them were type B1 and 16 were type C. Radiographs and medical records were available for all patients. Twenty-four patients (32%) have been treated with bisphosphonates (BPs) for longer than 4 years. Four patients presented a fracture with characteristics of PAFF. When enlarged to all PFFs of the series, no other PAFF was found: prevalence of PAFFs was 5.3% for type B1 and C cases and 3.5% for all surgically treated PFFs. Statistical significative difference between PAFFs and PFFs was found for prolonged BP assumption and for the level of fracture clear of the stem. Conclusion Fracture with characteristics of AFFs can also happen over a prosthetic stem, configuring themselves as PAFFs, and they are related to prolonged BP use. As a correct diagnosis is mandatory for proper treatment, a revision of criteria for AFFs should be considered, accepting that PAFFs exist.

Chayote (Sechium edule (Jacq.) Sw.) is a vigorous perennial and climbing cucurbits, native to Mesoamerica, and cultivated for alimentary purposes in the American continent, Australia, New Zealand, South Europe, Asia and Africa. During spring 2019, some chayote plants showing bright yellow vein banding rings and lines were observed in a private garden in South Italy (Campania region). Symptoms coalesced in some leaves, covering almost the whole foliar area. Double-stranded RNAs were extracted from symptomatic leaves of a single chayote plant and reverse-transcribed, randomly amplified, and submitted to Illumina sequencing (Marais et al., 2018). Reads were assembled using CLC Genomics Workbench 11.1 (http://www.clcbio.com). Contigs were then annotated by Blastn and Blastx comparison with the Genbank database, which allowed the identification of eight contigs of between 380 and 980 nucleotides sharing significant identity with alfalfa mosaic virus (AMV) genomic RNAs. No other viral contigs were identified. Mapping of reads on AMV genomic RNAs identified 4,209 AMV reads (1.26% of total reads) and allowed the scaffolding of the contigs into three scaffolds corresponding to the three AMV genomic RNAs. To complete the sequence of the AMV chayote isolate genome (named See-1), primers were designed from the contig sequences and used to amplify RACE PCR products spanning the 5' and 3' terminal regions of the three genomic RNAs using the SMARTer™ RACE cDNA Amplification Kit (Clontech, China). All amplicons were cloned into the pGEM-T vector (Promega, USA) and sequenced (three clones for each amplicon) by Microsynth Seqlab (Microsynth AG, Switzerland). Finally, the complete genomic sequences of the three RNAs were assembled by MacVector 17.5 (MacVector Inc., USA). The RNA1, RNA2 and RNA3 of See-1 are 3,643, 2,593 and 2,037 nt respectively (GenBank accession Nos. MT093209 to MT093211), and share the highest nt sequence identity with the RNA1 and RNA3 of AMV isolate (HZ) from tobacco (99.5% for RNA1, HQ316635; 98.7% for RNA3, HQ316637) and with the RNA2 of isolate AMV-Gym from Gynostemma pentaphyllum (98.1%, MH332898), both from China. AMV isolate See-1 was classified as belonging to subgroup I based on the presence of a BamH I and two AvaII sites in the CP ORF (Parrella et al., 2000). Reverse transcription polymerase chain reaction, using primers targeting the CP gene (Parrella et al., 2000), confirmed AMV infection in three symptomatic cayote plants including that used for Illumina sequencing, with 100% of nt sequence identity of amplicons. Three plants each of Chenopodium amaranticolor, Nicotiana benthamiana and Solanum lycopersicon were mechanically inoculated with sap from isolate See-1 infected plant, leading to the appearance of typical AMV symptoms in all three hosts ten days post-inoculation (Jaspars & Bos, 1980). This note describes the first detection of AMV in cayote in Italy and, to the best of our knowledge, in the world. In some areas of Southern Italy, climatic conditions are favorable enough to allow chayote development in the wild. Further studies would be desirable to determine the distribution and incidence of AMV in chayote and to understand the possibility that this species may play a role in AMV epidemiology, representing a threat to other susceptible crops.

Tomato leaf curl New Delhi virus (ToLCNDV) is a bipartite begomovirus affecting tomato cultivation on the Indian subcontinent. Recently, however, a new strain of the virus, named ToLCNDV-ES, has spread to Mediterranean countries such as Spain, Italy, and Tunisia, and occurred in Cucurbita crops, causing economic damage. Although ToLCNDV is spread by the sweet potato whitefly (Bemisia tabaci), like other begomoviruses, it has not been clear how ToLCNDV suddenly spread from the Indian subcontinent to the Mediterranean region. In 2017, ToLCNDV was diagnosed in young seedlings germinated naturally from fruits fallen in the prior year on a farm located in Giugliano in Campania, Naples, Italy, suggesting a possible role of the seeds in vertical transmission of the virus. Because sweet potato whiteflies were widespread naturally in that region, it was necessary to verify that in an artificial insect vector-free condition. Seeds were harvested from two ToLCNDV-infected zucchini squash cultivars in Naples in 2017 and 2018 to examine whether ToLCNDV can be transmitted from zucchini squash seeds to young plants. Viral DNA was amplified from these seeds and 1- to 3-week-old seedlings germinated from them with a ToLCNDV-specific primer set. According to PCR results, viral contamination was confirmed from all harvested seeds and dissemination was proven from 61.36% of tested seedling samples. Mechanical transmission from seed-borne virus-infected seedlings to healthy zucchini squash plants was also succesful, demonstrating that seedlings from ToLCNDV-infected seeds did act as inoculum. This is the first report demonstrating that ToLCNDV is a seed-transmissible virus in zucchini squash plants in Italy.

Tomato leaf curl New Delhi virus (ToLCNDV), a bipartite begomovirus (genus Begomovirus, family Geminiviridae), was originally described in tomato in India in 1995 (Padidam et al. 1995). The virus apparently remained confined to the Asian continent for about 20 years and has only recently been discovered in Europe. In Italy, ToLCNDV was detected for the first time in 2015 in zucchini squash in Sicily (Panno et al. 2016) and later on in other regions of continental Italy (Panno et al. 2019). Recently, epiphytotics of ToLCNDV were reported in central and south Italy, associated with yellowing and leaf curling symptoms in pepper crops (Luigi et al. 2019). In 2016, a large-scale survey was conducted to assess the distribution and the genetic diversity of the viral isolates spreading in Italy. Samples collected at that time included cucurbits and solanaceous plants, including five leaf samples from five distinct eggplant (Solanum melongena L.) plants of cultivar Violetta di Napoli that showed yellowing and light curling of the apical leaves and noticed in a cultivation located in Campania region (Castel Volturno municipality). A few Bemisia tabaci individuals were noticed associated to the cultivation. All five leaf samples tested positive for ToLCNDV with ImmunoStrip (Agdia, Elkhart, IN). These results were confirmed by PCR using ToLCNDV-specific primers TLCNDVCP1/TLCNDVCP2 (Parrella et al. 2018) on the total DNA extracted from each sample using an E.Z.N.A. Plant DNA kit (Omega Bio-tek, Norcross, GA). Amplicons of the expected size (∼1.0 kb) were obtained only from the five symptomatic plants, and the nucleotide sequences of these isolates were identical. One representative sample (Som-166/16) was selected for full-length amplification of the genome (DNA-A and DNA-B-like sequences) using the rolling circle amplification (RCA) method with an Illustra TempliPhi amplification kit (GE Healthcare, Piscataway, NJ), in accordance with the manufacturer’s instructions. RCA products were digested with different restriction endonucleases to obtain a 2.8-kb linear DNA fragment. Among the different enzymes tested, BamHI resulted in a maximum DNA fragment length of 2.8 kb, which was cloned into a BamHI-linearized pUC19 plasmid. The ligated products were transformed into a competent DH5α strain of Escherichia coli, and the positive clones were sequenced in both orientations at Microsynth Seqlab (Gottingen, Germany). The obtained full-length DNA-A (2,738 nt; GenBank MN782303) and DNA-B (2,686 nt; GenBank MN782304) sequences showed the highest percentage of nucleotide identity with the ToLCNDV Italian isolates Caa-164/16 (GenBank MK732932) for the DNA-A (99.82%) and Cum-45/16 (GenBank MF688671) for the DNA-B (99.48%). Eggplant leaf yellowing caused by a ToLCNDV variant has been previously described in India (HQ264185 for DNA-A and HQ264186 for DNA-B) (Pratap et al. 2011), but the Italian and the Indian isolates exhibited only 90.20 and 79.00% nucleotide identity with respect to the nucleotide sequences of DNA-A and DNA-B. The Som-166/16 isolate was very similar at the molecular level to previously reported isolates from Mediterranean countries and belonged to the European strain ToLCNDV-ES, because the percentage of nucleotide identity was 98.83% for DNA-A with the ToLCNDV-ES strain A-MU.13.ME/4.3 (GenBank MH577751) and 97.88% for DNA-B with the ToLCNDV-ES strain B-AL.15.ZU/2.1 (GenBank MH577658) (Moriones et al. 2017; Panno et al. 2019). The ToLCNDV-ES strain evolved from ToLCNDV isolates of Asian origin and adapted to infect cucurbits (Moriones et al. 2017). Nevertheless, this report presents further evidence, in addition to a previous report (Luigi et al. 2019), that ToLCNDV-ES isolates can potentially pose a threat not only to cucurbits but also to solanaceous crops as well, because this is the first finding of ToLCNDV in eggplant in Italy. Control measures against ToLCNDV are limited and mainly based on vector control, cultivation in protected environments, and the prompt elimination of infected materials.

During 2016, a survey was carried out to investigate the presence and distribution of Tomato leaf curl New Delhi virus (ToLCNDV) in Campania region (Southern Italy). In three fields in the Castel Volturno and Giugliano in Campania municipalities (Caserta and Napoli provinces respectively), an unusual disease was noted affecting pumpkin (Cucurbita moschata) plants of the variety “Lunga di Napoli”, a traditional local ecotype. Symptoms consisted of stunting and severe foliar yellow mosaic, which, in some cases, resulted in completely yellow leaves. The affected plants produced less flowers and fruits. The incidence of the disease ranged from 80 to 100% of the plants. The insect vector of ToLCNDV, Bemisia tabaci, was found in each sampled field and the genotype of twenty specimens for each population was identified as previously described (Parrella et al., 2012 and 2014). The Q1 and Q2 variants of the Mediterranean species were identified with a percentage ranging from 11-14% and 86-89% respectively. Four sy...

During spring-summer 2012, 2013, and 2015, 10 samples of bell pepper (Capsicum annuum) plants showing basal bright rings and mosaic of the leaves with some necrotic patches, necrotic stems, and fruits with brown patches and corky rings, were identified in the Campania region (southern Italy), in a family garden (four plants in 2012 and three plants in 2015 of the Friariello ecotype) and in a commercial greenhouse (two plants of the Papaccella ecotype in 2012 and one plant of the Eppo hybrid in 2013). Symptomatic samples were tested at the Institute for Sustainable Plant Protection in Portici (Napoli, Italy). Screening of leaf and fruit symptomatic tissues by double antibody sandwich (DAS)-ELISA with commercial kits of antisera (Bioreba AG, Switzerland) to Cucumber mosaic virus(CMV), Alfalfa mosaic virus (AMV), Tomato spotted wilt virus (TSWV), Tomato mosaic virus(ToMV), Pepper mild mottle virus (PMMoV), and by indirect plate trapped antigen (PTA)-ELISA against potyviruses (potygroup test), gave negative results, whereas all samples tested positive in antigen-coated-plate (ACP)-ELISA with antisera, kindly provided by Dr. P. Roggero (Plant Virology Institute, Torino, Italy), produced against a tomato isolate (namely TI1 isolate) of Parietaria mottle virus (PMoV). The results from indirect ELISA were subsequently confirmed with reverse transcription (RT)-PCR using PMoV specific primers designed to amplify the entire gene of the movement protein (MP): PMoVMP1a (5?-AGGGTTTGAAATGGCATTTGTTC-3?) and PMoVMP2b (5?-CTTAGTGGACATTCACGAATATC-3?). A healthy pepper plant was used as a negative control both in ELISA and RT-PCR assays. RT-PCR products of 916 bp, which covered approximately 40.4% of PMoV RNA3 (isolate T32 as reference, accession number KT005245), were obtained in all symptomatic samples and one amplicon for each pepper ecotype and year of sampling was cloned and sequenced. The nucleotide sequences of the MP gene of three Italian PMoV isolates were deposited in GenBank under accession numbers LT160068 (isolate Fri-1, from the Friariello ecotype, in 2012), LT160069 (isolate Pap-1, from the Papaccella ecotype, in 2012), and LT160070 (isolate Fri-2, from the Friariello ecotype, in 2015). Isolates Fri-1 and Pap-1 showed 98.4 and 97.6% identity with the Spanish PMoV isolate JBT1 from tomato (accession number AM182747), respectively, while Fri-2 showed 96.7% identity with the Spanish PMoV isolate AC1 from tomato (accession number AM182743). To our knowledge, this is the first report of PMoV infecting bell pepper in Italy. In 2004, PMoV was identified in pepper crops in Spain (Janssen et al. 2005). The virus is also known to infect economically important plants as tomato in Italy, France, Spain, and Greece (Aramburu 2001;Marchoux et al. 1999; Roggero et al. 2000), as well as the ornamental Mirabilis jalapa (Parrella 2002). This new finding of PMoV in pepper in Italy confirms the tendency of the virus to broaden its range of natural hosts, particularly concerning species of economic interest. The symptoms induced by PMoV on pepper can be visually confused with those caused by other viruses infecting pepper (e.g., TSWV, CMV). Therefore, it is necessary to monitor regularly for the presence of PMoV in pepper crops in order to know the real incidence of the disease in the field with the aim of developing efficient and durable disease control strategies.