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Background Complex proximal humeral fractures (cPHFs) represent an important public health concern, and reverse shoulder arthroplasty (RSA) has emerged as a feasible treatment option in the elderly with high functional demands. Recent studies have shown that tuberosity healing leads to better clinical outcomes and an improved range of motion. However, the best surgical technique for the management of the tuberosities is still a topic of debate. The purpose of this retrospective observational study is to report the radiographic and clinical outcomes of a consecutive series of patients who underwent RSA for cPHFs using a novel “7 sutures and 8 knots” technique. Materials and methods A consecutive series of 32 patients (33 shoulders) were treated with this technique by a single surgeon from January 2017 to September 2021. Results at a minimum follow-up of 12 months and a mean ± SD follow-up of 35.9 ± 16.2 (range 12–64) months are reported. Results The tuberosity union rate was 87.9% (29 out of 33 shoulders), the mean Constant score was 66.7 ± 20.5 (range 29–100) points, and the mean DASH score was 33.4 ± 22.6 (range 2–85) points. Conclusions The “7 sutures and 8 knots” technique, which relies on three sutures around the implant and five bridging sutures between the tuberosities, is a relatively simple procedure which provides a reliable means for anatomic restoration of the tuberosities and allows functional recovery of the shoulder in elderly patients with cPHFs treated with RSA. Level of evidence: IV; retrospective atudy. Trial registration: At our institution, no institutional review board nor ethical committee approval is necessary for retrospective studies.

Background Delayed bone healing and nonunions represent a great challenge for the orthopedic surgeon. In addition to traditional surgical approaches, increasing attention is being given to the use of systemic anabolic therapy with Teriparatide, whose efficacy in preventing osteoporotic fractures is widely validated and whose application as a promoter of bone healing has been described but it is still debated. The aim of the study was to evaluate bone healing in a series of patients with delayed unions or nonunions treated with Teriparatide in conjunction with eventual appropriate surgical procedure. Methods Twenty patients with an unconsolidated fracture that were treated at our Institutions from 2011 to 2020 with Teriparatide were retrospectively included into the study. The pharmacological anabolic support was used off-label with a planned duration of 6 months; radiographic healing was evaluated at 1-, 3- and 6-months follow-up outpatient visits over plain radiographs. Also, eventual side-effects were registered. Results Radiographic signs indicative of favorable evolution of the bone callus were observed as early as at 1 month of therapy in 15% of cases; at 3 months, healing progression was appreciated in 80% of cases and complete healing in 10%; at 6 months, 85% of delayed and nonunions had healed. In all patients, the anabolic therapy was well tolerated. Conclusions In accordance to Literature, this study suggests that Teriparatide plays a potentially important role in the treatment of some forms of delayed unions or nou-nions, even in the presence of failure of hardware. The results suggest a greater effect of the drug when associated with a condition in which the bone is in an active phase of callogenesis, or with a “revitalizing” treatment which represents a local (mechanical and/or biological) stimulus to the healing process. Despite the small sample size and the variety of cases, the efficacy of Teriparatide in treating delayed unions or nonunions emerged, highlighting how this anabolic therapy can represent a useful pharmacological support in the treatment of such a pathology. Although the results obtained are encouraging, further studies, particularly prospective and randomized, are needed to confirm the efficacy of the drug, and define a specific treatment algorithm.

Tomato leaf curl New Delhi virus-ES (ToLCNDV-ES), a high threat to cucurbits in the Mediterranean Basin, is listed as a different strain from the Asian ToLCNDV isolates. In this study, the infectivity of two clones previously isolated from Italy and Pakistan were compared in cucumbers, which resulted in the opposite symptom appearance. The swapping subgenome was processed; however, the mechanisms related to the disease phenotype remain unclear. To identify the disease-associated genes that could contribute to symptom development under the two ToLCNDV infections, the transcriptomes of ToLCNDV-infected and mock-inoculated cucumber plants were compared 21 days postinoculation. The number of differentially expressed genes in ToLCNDV-India-infected plants was 10 times higher than in ToLCNDV-ES-infected samples. The gene ontology (GO) and pathway enrichment were analyzed using the Cucurbits Genomics Database. The flavonoid pathway-related genes were upregulated in ToLCNDV-ES, but some were downregulated in ToLCNDV-India infection, suggesting their role in resistance to the two ToLCNDV infections. The relative expression levels of the selected candidate genes were validated by qRT-PCR under two ToLCNDV-infected conditions. Our results reveal the different infectivity of the two ToLCNDVs in cucumber and also provide primary information based on RNA-seq for further analysis related to different ToLCNDV infections.

The tomato leaf curl New Delhi virus (ToLCNDV), a bipartite begomovirus (family Geminiviridae), poses a significant threat to various horticultural crops in many Asian and Mediterranean countries. Since its identification, the Asian strain of this virus has exhibited severe infectivity and caused high yield loss in tomato and cucurbit production in the Indian subcontinent and other parts of Asia. ToLCNDV-ES, a different strain of ToLCNDV, emerged recently in the Mediterranean Basin and caused significant outbreaks in Cucurbitaceae species but has shown low adaptation to tomatoes. In a field survey, tomato plants infected with this Mediterranean strain were not discovered. Nevertheless, the same field survey revealed that ToLCNDV-ES occurred in natural double infection with tomato yellow leaf curl virus (TYLCV) in tomato plants with an infection ratio of up to 50%. Moreover, results obtained from experiments where tomato plants agro-inoculated simultaneously with infectious clones of ToLCNDV-ES and TYLCV showed that ToLCNDV-ES was detected in tomatoes while synergized with TYLCV with infection ratios similar to those found under field conditions. Quantitative PCR data indicated the highest amount of ToLCNDV in co-infected plants and no significant change in TYLCV titers among the different mixed infections. Moreover, it was ascertained that not all begomoviruses can enhance the infectivity of Mediterranean ToLCNDV isolates in tomato plants. Our study reports a new finding regarding the ToLCNDV-ES response in tomato while synergized with TYLCV with evidence from both field and laboratory conditions.

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.

Introduction The COVID-19 pandemic has affected and is still deeply affecting all aspects of public life. World governments have been forced to enact restrictive measures to stem the contagion which have led to a decrease in the movement of people within national territory and to a redirection of health care resources with a suspension of non-urgent procedures. In Italy, a lockdown was imposed from March 9th to May 3rd, 2020. As a result, a significant reduction in the overall operative volume of orthopedic trauma was expected, but it was not possible to predict a similar trend regarding fragility fractures of the proximal femur in the elderly. Methods The aim of this paper was to examine the impact of COVID-19 on the operating volume for trauma surgeries and to determine how the pandemic affected the management of fragility hip fractures (FHFs) in non-COVID patients at a single Institution. Results The first result was a statistically significant reduction in the overall operative volume of orthopedic trauma during the period of the first lockdown and an increase in the mean age of patients undergoing surgery, as expected. As regard to the second aim, the incidence of FHFs remained almost unchanged during the periods analysed. The population examined were superimposable in terms of demographics, comorbidities, type of fracture, peri-operative complications, percentage of operations performed within 48 hours from hospitalization and 1-year outcome. Discussion Our results are in line with those already present in the Literature. Conclusions Our study revealed a significant impact of the restrictive anti-contagion measures on the overall orthopedic surgical volume, but, at the same time, we could affirm that the pandemic did not affect the management of FHFs in non-COVID patients, and their results.

Charcot Foot (CF), part of a broader condition known as Charcot Neuro-Osteoarthropathy (CNO), is characterized by neuropathic arthropathy with a progressive alteration of the foot. CNO is one of the most devastating complications in patients with diabetes mellitus and peripheral neuropathy but can also be caused by neurological or infectious diseases. The pathogenesis is multifactorial; many studies have demonstrated the central role of inflammation and the Receptor Activator of NF-κB ligand (RANKL)-Receptor Activator of NF-κB (RANK)-Osteoprotegerin (OPG) pathway in the acute phase of the disease, resulting in the serum overexpression of RANKL. This overexpression and activation of this signal lead to increased osteoclast activity and osteolysis, which is a prelude to bone destruction. The aim of this narrative review is to analyze this signaling pathway in bone remodeling, and in CF in particular, to highlight its clinical aspects and possible therapeutic implications of targeting drugs at different levels of the pathway. Drugs that act at different levels in this pathway are anti-RANKL monoclonal antibodies (Denosumab), bisphosphonates (BP), and calcitonin. The literature review showed encouraging data on treatment with Denosumab, although in a few studies and in small sample sizes. In contrast, BPs have been re-evaluated in recent years in relation to the high possibility of side effects, while calcitonin has shown little efficacy on CNO.

In this study, a new virus was identified in French hydrangea plants, exhibiting chlorotic vein banding and necrotic ring spots on older leaves. The virus was mechanically transmitted to herbaceous hosts, in which it induced local and systemic or only local symptoms. The genome of the new virus was characterized and consisted of three RNA sequences that were 3422 (RNA 1), 2905 (RNA 2) and 2299 (RNA 3) nucleotides long, with five predicted open reading frames; RNA2 was bicistronic and contained conserved domains and motifs typical of ilarviruses. The phylogenetic analysis of the predicted proteins—p1, p2a, p3a and p3b—revealed its close relationship to recognized members of subgroup 2 within the genus Ilarvirus. Homologous antiserum was effective in the detection of the virus in plant extracts and no cross reactions with two other distinct members of subgroup 2 were observed. Overall, the biological features, phylogenetic relationships and serological data suggest that this virus is a new member of the genus, for which we propose the name hydrangea vein banding virus (HdVBV).

Tomato leaf curl New Delhi virus (ToLCNDV) became an alerting virus in Europe from 2017 to 2020 because of its significant damage to Cucurbitaceae cultivation. Until now, just some cucurbit crops including sponge gourd, melon, pumpkin, and cucumber were reported to be resistant to ToLCNDV, but no commercial cultivars are available. In this study, a new isolate of ToLCNDV was identified in Pakistan and analyzed together with ToLCNDV-ES which was previously isolated in Italy. Furthermore, infectious clones of two ToLCNDV isolates were constructed and agroinoculated into different cucurbit crops to verify their infectivity. Results showed that both isolates exhibited severe infection on all tested cucurbit (>70%) except watermelon. Thus, those cultivars may be good candidates in the first step of screening genetic resources for resistance on both Southeast Asian and Mediterranean ToLCNDV isolates. Additional, comparison pathogenicity of different geographical ToLCNDV isolates will be aided to understand viral characterization as such knowledge could facilitate breeding resistance to this virus.

Introduction An increasing number of patients is annually undergoing total hip arthroplasty (THA), and a significant proportion of these patients are elderly and consequently at a higher risk of complications because of age, osteoporosis, and medical comorbidities. Periprosthetic femoral fractures (PFFs) are one of the worst complications of THA associated with high rates of unfavorable prognosis. Besides, in the last decade, a new independent disease entity called “atypical femoral fracture” (AFF) has been identified and defined by the American Society for Bone and Mineral Research (ASBMR) task force. Some PFFs present clinical history and radiographic aspect consistent with an AFF, meeting the ASBMR criteria for the diagnosis of AFF except that PFFs by themselves are an exclusion criterion for AFF. However, there is an increasing number of published studies suggesting that periprosthetic atypical femoral fractures (PAFFs) exist and should not be excluded by definition. Significance Nowadays, although there is an increasing interest in PAFFs, there are still very few studies published on the topic and a lack of consensus regarding their treatment. This narrative literature review aims to introduce this new emerging topic to a wider readership describing the characteristics of PAFFs and the state-of-the-art in their management. Conclusions Many authors agree that PAFFs should be considered as a subgroup of PFFs that have atypical characteristics; they also show a significant correlation with prolonged bisphosphonate use. A correct diagnosis is paramount for proper treatment of the disease that requires both surgical and medical actions to be taken.

Parietaria mottle virus (PMoV) is an emerging virus in Mediterranean countries, responsible for severe disease in tomato and pepper crops in the field and protected cultivation. The principal wild reservoir of PMoV is Parietaria officinalis, and only few additional wild plants have been described as natural reservoirs of the virus. During field survey in southern Italy, several plants of Phytolacca americana showing virus-like symptoms were collected. Serological and molecular assays showed that these plants were infected by PMoV. Sequence comparison of the movement protein gene of the PMoV isolate from P. americana showed the greatest similarity to the corresponding sequence from tomato plants growing nearby. These results indicate that P. ameriacana is a new natural host of PMoV, and further investigation is warranted to establish the potential of this host as reservoir of the virus in the field.

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.

Aim To evaluate the efficacy of systemic plus local tranexamic acid (TXA) in reducing post-operative bleeding, haemoglobin loss and the need for allogeneic blood transfusion (ABT) in total knee arthroplasty (TKA). Methods All patients undergoing TKA between January 2017 and September 2019 were retrospectively evaluated. Exclusion criteria were cardiovascular comorbidities, diabetes and the assumption of any anticoagulant/antiaggregant therapy in the pre-operative period. All patients received the same prosthesis with the same surgical technique and were operated on by the same surgeon. Twenty patients were found (group A) that received intra-operative TXA (20 mg/kg intravenous 10 minutes before deflating tourniquet and 1g intra-articular after capsular suture). A control group of 26 patients not receiving TXA was matched for demographics (group B). Results Two (10%) patients in group A and 16 (61.5%) in group B needed ABT in the post-operative period (p=0.0004). Each patient in group A received 2 red blood cells (RBCs) units, while in group B 2 patients received one RBCs unit and one patient 4 RBCs units, for a total of 4 and 32 RBCs units in group A and B, respectively (p=0.0006). The minimum haemoglobin level was observed at 48 hours post-operatively in both groups: mean decrease was 3.54 and 4.64 g/dL in group A and B, respectively (p=0.0126). Conclusion The association of systemic and local TXA administration seems to significantly reduce post-operative bleeding and the need for RBCs transfusions after TKA in patients not assuming any anticoagulant / antiaggregant therapy and without cardiovascular and diabetic morbidities.

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.

Virus-like symptoms consisting of light mosaic, chlorotic spots and oak chlorotic line patterns were observed on lilac plants (Syringa vulgaris L.) growing in a public garden in Imola (Emilia Romagna region, Italy). The causal agent was identified as cucumber mosaic virus (CMV) on the basis of biological, serological and nucleotide sequence properties of partial coat protein and movement protein genes, and the isolate was designated SYV. The CMV-SYV isolate caused mosaic symptoms on indicator plants of Nicotiana tabacum cv. Xanthi-nc, N. rustica and Cucumis sativus, while symptoms of local necrotic spots or pin points were observed on inoculated leaves of Vigna unguiculata and Vicia faba. To assess genetic differences between CMV-SYV and other known CMV isolates, phylogenetic analyses were carried out using 16 nucleotide sequences of coat protein and movement protein genes, including for SYV. The CMV-SYV isolate was most related to CMV subgroup IA isolates, and had 85.1-100% nucleotide sequence similarity to subgroup I isolates. This is the first report of CMV infecting S. vulgaris.

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.

Introduction. The ever-expanding indications for total hip arthroplasty are leading to more implants being placed in younger as well as in older patients with high functional demand. Also, prolonged life expectancy is contributing to an overall increment of periprosthetic femoral fractures. The Vancouver classification has been the most used for guiding the surgeon choice since its proposal in 1995. Fractures occurring over a hip femoral implant can be divided into intra-operative and post-operative PFFs, and their treatment depends on factors that may severely affect the outcome: level of fracture, implant stability, quality of bone stock, patients’ functional demand, age and comorbidities, and surgeon expertise. There are many different treatment techniques available which include osteosynthesis and revision surgery or a combination of both. The goals of surgical treatment are patients’ early mobilization, restoration of anatomical alignment and length with a stable prosthesis and maintenance of bone stock. Significance. The aim of this review is to describe the state-of-the-art treatment and outcomes in the management of PFFs. We performed a systematic literature review of studies reporting on the management of PFFs around hip stems and inter-prosthetic fractures identifying 45 manuscripts eligible for the analysis. Conclusions. PFFs present peculiar characteristic that must be considered and special features that must be addressed. Their management is complex due to the extreme variability of stem designs, the possibility of having cemented or uncemented stems, the difficulty in identifying the “real” level of the fracture and the actual stability of the stem. As a result, the definition of a standardized treatment is unlikely, thereby high expertise is fundamental for the surgical management of PPFs, so this kind of fractures should be treated only in specialized centres with both high volume of revision joint arthroplasty and trauma surgery.

Tomato leaf curl New Delhi virus (ToLCNDV) is a bipartite begomovirus (family Geminiviridae) identified for the first time in 1995 in Asia, from where it spread into several countries of the Mediterranean basin. ToLCNDV was found in Spain in 2012, and subsequently in Tunisia and Italy. The first outbreak in Italy occurred at the end of 2015 in Trapani province (Sicily) on zucchini squashes. Then in 2016, ToLCNDV was found in infected zucchini plants in Campania, Lazio and Sardinia regions, and in 2017 in Calabria. This study addressed the dispersion and genetic diversity of ToLCNDV isolates in Italy. A total of 1400 plants were analysed. Phylogenetic analysis showed low variability among the Italian isolates, probably as a consequence of the recent introduction and rapid spread of this virus in Italy. Two statistically significant clusters were reported: one grouping only Italian isolates and the other grouping Italian, Spanish, Tunisian and Moroccan isolates. Furthermore, the highest incidence of ToLCNDV was observed in Sicily, although the disease also appears to be critical in other Italian regions. In this work, a high efficiency of ToLCNDV mechanical transmission into Cucurbita pepo, Cucumis melo inodorus and Cucumis melo cantalupensis has been demonstrated. The rapid spread of ToLCNDV in the Mediterranean basin represents a threat for horticultural production, thus it is very important to develop suitable crop management practices, applying genetic resistance strategies and more restrictive phytosanitary measures.

Peroneus longus tendon (PLT) dislocation associated with inferior peroneal retinaculum (IPR) tear is an extremely rare injury. Only 6 cases are described in English literature to date, and its diagnosis is always delayed from the initial trauma. Swelling at the lateral face of the heel and a snapping during the movement of the foot and ankle are typical signs that can induce the suspect of this injury. Operative treatment is required as it typically affects young adults with high functional demands and tendon instability causes pain and functional limitation; also, a neglected dislocated or instable PTL may eventually end up with a rupture. In the few cases reported in literature, different operative techniques have been described, all with excellent results. A case of an initially overlooked dynamic dislocation of the PLT due to IPR tear in a young professional soccer player is described. The lesion was treated with a not yet described operative technique that consists in groove deepening and subsequent IPR repair through transosseous sutures, instead of peroneal tubercle resection as mostly advocated. Final follow up at 15 months showed complete recovery, with a Foot and Ankle Outcome Score of 100%.

IntroductionThere is lack of consensus regarding best operative fixation strategy for periprosthetic femoral fractures (PFFs) around a stable stem. Evidence exists that some patterns of fracture around a stable stem are better treated with revision surgery than with standard fixation. Anyway, a more aggressive surgical procedure together with medical treatment could allow for stem retention, and reduced risk of nonunion/hardware failure, even in these cases.SignificanceThis paper is placed in a broader context of lack of studies on the matter, and its aim is to shed some light on the management of PFFs around a stable stem, when peculiar mechanical and biological aspects are present.ResultsBased on our casuistry in the treatment of nonunions after PFF successfully treated with original stem retention, and on review of Literature about risk factors for fixation failure, an algorithm is proposed that can guide in choosing the ideal surgical technique even for first-time PFFs with a stable stem, without resorting to revision. Mechanical (major and minor) and biological (local and systemic) factors that may influence fracture healing, leading to nonunion and hardware failure, and subsequent need for re-operation, are considered. The proposed surgical technique consists of rigid fixation with absolute stability (using a plate and structural allograft) plus local biological support (structural allograft and autologous bone marrow concentrate over a platelet-rich plasma-based scaffold) at fracture site. Systemic anabolic treatment (Teriparatide) is also administered in the post-operative period.ConclusionMechanical factors are not the only issues to be considered when choosing the surgical approach to PFFs over a stable stem. Systemic and local biological conditions should be taken into account, as well. A therapeutic algorithm is proposed, given the prosthetic stem to be stable, considering mechanical and biological criteria.

In July 2019, disease outbreaks were observed in two tomato fields in South Sardinia. The first field located in Samassi was established with tomato spotted wilt virus (TSWV)-resistant cultivars Taylor and Docet. The second field located in Serramanna was established with TSWV-resistant cultivars Dask and SV051, and the TSWV-susceptible cultivar Creso. The two fields were about 4.0 km away from each other. Symptoms resembling those elicited by TSWV, consisting in leaf and stem necrosis, and necrotic rings and patches on the fruits, were noticed on most of the plants in the two fields. Preliminary screening of two symptomatic plants of each cultivar with a specific TSWV Lateral Flow (LF) assay (Forsite Diagnostics, UK) showed that none of the samples, with the exception of Creso, reacted positively to TSWV. Then RT-PCR was conducted with total RNA extracted from non-necrotic tissue of the same plants tested in LF using the E.Z.N.A.® Plant RNA kit (Omega Bio-tek, USA) and orthotospovirus universal primers BR60/BR65 and primers PMoVMP1a/PMoVMP2b specific to parietaria mottle virus (PMoV) (Parrella et al. 2016). Amplicons of the expected size (916 bp) for PMoV were obtained from all symptomatic leaf samples tested, whereas TSWV amplicons (453 bp) were obtained only from the Creso samples. No amplicons were obtained from asymptomatic samples or healthy samples used as controls. The PMoV PCR products from three tomato plants were cloned and sequenced. Sequences of the PMoV isolates were identical (GenBank accession number MN782302) and shared 96% nucleotide identity with the Spanish isolate RAMS1 from tomato (AM182748). Further, PMoV was detected in symptomatic leaf samples by dot-blot hybridization assay using a PMoV-specific riboprobe (Parrella 2002). Using this assay, the PMoV incidence was around 80% in both tomato fields (52 of 67 plants were positive in the first field and 41 of 51 plants were positive in the second field). PMoV has been previously reported in protected and open field tomato cultivation in Spain only with low incidence (Soler et al. 2010). This is the first report of a significant outbreak of PMoV in tomato in the Mediterranean basin

Tomato leaf curl New Delhi virus (ToLCNDV), a bipartite begomovirus (family Geminiviridae) with two circular ssDNA genome components (DNA-A and DNA-B), is transmitted in a circulative nonpropagative manner by the whitefly Bemisia tabaci (Gennadius). Although it was first reported in Asia on tomato and other solanaceous crops such as eggplant, potato, and chilli pepper in the Mediterranean basin, this virus was mainly detected on cucurbits and only sporadically on tomato and on two wild solanaceous species, Datura stramonium L. and Solanum nigrum L. (Juárez et al. 2019). In 2018, separate surveys were carried out in protected cultivations of sweet pepper (Capsicum annuum L.) in two Italian regions: Lazio and Campania. The greenhouses were in areas with high density of B. tabaci and where ToLCNDV outbreaks occurred on cucurbits since 2016 (Panno et al. 2019). Some plants showing symptoms of yellowing and leaf curling were found in both regions, whereas fruit symptoms were neither observed nor reported by farmers. This disease syndrome, known as yellow leaf curl disease (YLCD), can be caused in pepper by several begomoviruses, as reported recently in a review listing the viruses causing YLCD in peppers in Thailand (Chiemsombat et al. 2018). Symptomatic leaves were collected during late summer 2018 from different pepper plants as well as from the neighboring zucchini cultivations, showing the typical symptomatology induced by ToLCNDV. Total DNA was extracted (DNeasy Plant Mini kit, Qiagen, Germany), and the presence of ToLCNDV was ascertained by PCR with the specific primers ToLCNDV-CP1 and ToLCNDV-CP2 (Panno et al. 2019; Parrella et al. 2018). ToLCNDV infection was further ascertained in three symptomatic leaf samples from Campania by using specific ToLCNDV ImmunoStrips (Agdia, Elkhart, IN). Successively, one symptomatic pepper sample from each greenhouse was selected and amplified by rolling circle amplification technique (RCA; Inoue-Nagata et al. 2004). The amplicons were cloned, and the DNA-A and DNA-B were full-length sequenced. The sequences were deposited in GenBank NCBI database (MK732932 DNA-A and MK732933 DNA-B, pepper sample from Campania; MK756106 DNA-A and MK756107 DNA-B, pepper sample from Lazio). The RCA analysis was performed also on a ToLCNDV-infected zucchini sample collected in the same area in Lazio region (MK756108 DNA-A and MK756109 DNA-B). The analysis of the ToLCNDV sequences showed a low level of genetic variability between the two pepper isolates from Lazio and Campania regions (rate of substitutions: 0.016 for DNA-A and 0.023 for DNA-B). A high genetic similarity was recorded between the zucchini isolate and both the pepper isolates from Campania (0.019 for DNA-A and 0.023 for DNA-B) and Lazio (0.003 for both DNA-A and B). The three characterized isolates showed a high sequence homology also with both the DNA-A (MH577751 from a melon isolate) and DNA-B (MH577673 from a zucchini isolate) of the ToLCNDV-ES genotype (Fortes et al. 2016), which differed in 15 and 13 nucleotide substitutions from pepper sample from Lazio, 29 and 51 substitutions from Campania sample, and 10 and 5 substitutions from zucchini sample. High homology was also identified compared with the other Spanish isolates collected since the first appearance of the virus (2014) and to the Tunisian (2015) and Moroccan (2018) isolates, confirming the hypothesis that the Mediterranean population of ToLCNDV is highly conserved (Juárez et al. 2019). To our knowledge, this is the first report of ToLCNDV infection on pepper in Europe and indicates that sweet pepper could also act as a reservoir of the virus for further spread to other solanaceous plants and cucurbits.

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...

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C3 and C4 plant carbonic anhydrases (CAs) are zinc-enzymes that catalyze the reversible hydration of CO2. They are sub-divided in three classes: α, β and γ, being distributed between both photosynthetic subtypes. The C4 dicotyledon species Flaveria bidentis (L.) “Kuntze” contains a small gene family encoding three distinct β-CAs, named FbiCA1, FbiCA2 and FbiCA3. We have expressed and purified recombinant FbiCA1, which is localized in the chloroplast where it is thought to play a role in lipid biosynthesis and antioxidant activity, and biochemically characterized it by spectroscopic and inhibition experiments. FbiCA1 is a compact octameric protein that is moderately inhibited by carboxylate molecules. Surprisingly, pyruvate, but not lactate, did not inhibit FbiCA1 at concentrations up to 10 mM, suggesting that its capacity to tolerate high pyruvate concentration reflects the high concentration of pyruvate in the chloroplasts of bundle-sheath and mesophyll cells involved in C4 photosynthesis.

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.