Anatomical and biochemical studies of Spartium junceum infected by Xylella fastidiosa subsp. multiplex ST 87.

Spartium junceum L. is a typical species of Mediterranean shrubland areas, also grown in gardens and parks as an ornamental. In recent years in Europe, S. junceum has been recurrently found to be infected by different subspecies and genotypes of the quarantine regulated bacterium Xylella fastidiosa (Xf). This work presents for the first time the anatomy of S. junceum plants that we found, by means of genetic and immunochemistry analysis, to be naturally infected by Xf subsp. multiplex ST87 (XfmST87) in Monte Argentario (Grosseto, Tuscany, Italy), a new outbreak area within the EU. Our anatomical observations showed that bacteria colonized exclusively the xylem conductive elements and moved horizontally to adjacent vessels through pits. Interestingly, a pink/violet matrix was observed with Toluidine blue staining in infected conduits indicating a high content of acidic polysaccharides. In particular, when this pink-staining matrix was observed, bacterial cells were either absent or degenerated, suggesting that the matrix was produced by the host plant as a defense response against bacterial spread. In addition, a blue-staining phenolic material was found in the vessels and, at high concentration, in the pits and inter-vessels. SEM micrographs confirmed that polysaccharide and phenolic components showed different structures, which appear to be related to two different morphologies: fibrillary and granular, respectively. Moreover, our LM observations revealed bacterial infection in xylem conductive elements of green shoots and leaves only, and not in those of other plant organs such as roots and flowers.

Development of Three Molecular Diagnostic Tools for the Identification of the False Codling Moth (Lepidoptera: Tortricidae).

Three molecular protocols using qPCR TaqMan probe, SYBR Green, and loop-mediated isothermal amplification (LAMP) methods were set up for the identification of larvae and adults of an African invasive moth, Thaumatotibia leucotreta (Meyrick, 1913) (Lepidoptera: Tortricidae). The DNA extracts from larval and adult samples of T. leucotreta were perfectly amplified with an average Ct value of 19.47 ± 2.63. All assays were demonstrated to be inclusive for T. leucotreta and exclusive for the nontarget species tested; the absence of false positives for nontarget species showed a 100% of diagnostic specificity and diagnostic sensitivity for all assays. With the SYBR Green protocol, the Cq values were only considered for values less than 22 (cutoff value) to prevent false-positive results caused by the late amplification of nonspecific amplicons. The limit of detection (LoD) for the qPCR probe protocol was equal to 0.02 pg/µl while a value equal to 0.128 pg/µl for the qPCR SYBR Green assay and LAMP method were established, respectively. The intrarun variabilities of reproducibility and repeatability in all the assays evaluated as CV%, ranged between 0.21 and 6.14, and between 0.33 and 9.52, respectively; the LAMP values were slightly higher than other assays, indicating a very low interrun variability. In order for an operator to choose the most desirable method, several parameters were considered and discussed. For future development of these assays, it is possible to hypothesize the setup of a diagnostic kit including all the three methods combined, to empower the test reliability and robustness.

Worldwide Network for Blood and Marrow Transplantation (WBMT) recommendations for establishing a hematopoietic stem cell transplantation program in countries with limited resources (Part II): Clinical, technical and socio-economic considerations.

The development of hematopoietic stem cell transplantation (HSCT) programs can face significant challenges in most developing countries because such endeavors must compete with other government health care priorities, including the delivery of basic services. While this is may be a limiting factor, these countries should prioritize development of the needed expertise to offer state of the art treatments including transplantation, by providing financial, technological, legal, ethical and other needed support. This would prove beneficial in providing successful programs customized to the needs of their population, and potentially provide long-term cost-savings by circumventing the need for their citizens to seek care abroad. Costs of establishing HSCT program and the costs of the HSCT procedure itself can be substantial barriers in developing countries. Additionally, socioeconomic factors intrinsic to specific countries can influence access to HSCT, patient eligibility for HSCT and timely utilization of HSCT center capabilities. This report describes recommendations from the Worldwide Network for Blood and Marrow Transplantation (WBMT) for establishing HSCT programs with a specific focus on developing countries, and identifies challenges and opportunities for providing this specialized procedure in the resource constrained setting.

Serum biomarkers for sepsis in children with febrile neutropenia and cancer.

The National Institute for Health and Care Excellence (NICE) defines febrile neutropenia or "neutropenic sepsis" as a patient with an absolute neutrophil count (ANC) less than 0.5 x 109/L and temperature >38°C or signs and symptoms of sepsis.

Steam Sterilization of Equine Bone Block: Morphological and Collagen Analysis.

INTRODUCTION: The use of equine bone blocks is widely reported for bone augmentation techniques. The block must be shaped according to the form of the defect that should be regenerated. The shaping could be performed by hand before or during the surgery, in a sterile ambient, or using a CNC milling machine that could not be sterile. The aim of our study was to evaluate if a steam sterilization could provide a medical grade sterilization of the blocks and to evaluate if bone microstructure and collagen structures change after different steam sterilization protocols provided by mainstream autoclave. MATERIALS AND METHOD: Two blocks of equine bone were divided into 16 samples. 1 sample was used as control and not submitted to any treatment. 15 samples were infected with a Streptococcus faecalis bacterial culture. The samples were singularly packed, randomly divided into 3 groups, and submitted to autoclave sterilization on the same device. The groups were submitted to a sterilization cycle (Gr. A: 121°C, 1,16 bar for 20'; Gr. B:134°C, 2,16 bar for 4'; Gr. C: 134°C, 2,16 bar for 3.30 min.). 2 samples for each group were evaluated for the sterility. 3 samples for each group were observed at SEM to notice the macro- and microstructure modification and to confocal microscope to observe the collagen. RESULTS: All samples were sterile. The SEM evaluation showed, in all groups, a preserved morphological structure. Confocal microscope evaluation shows that the collagen structure appears to be more uniform and preserved in group C. CONCLUSION: Data show that autoclave steam sterilization could be reliable to obtain sterilization of equine bone blocks.

Molecular Typing of Bois Noir Phytoplasma Strains in the Chianti Classico Area (Tuscany, Central Italy) and Their Association with Symptom Severity in Vitis vinifera 'Sangiovese'.

Bois noir (BN) is the most widespread disease of the grapevine yellows complex in the Euro-Mediterranean area. BN is caused by 'Candidatus Phytoplasma solani' (BNp), transmitted from herbaceous plants to grapevine by polyphagous insect vectors. In this study, genetic diversity among BNp strains and their prevalence and possible association with grapevine symptom severity were investigated in a Sangiovese clone organic vineyard in the Chianti Classico area (Tuscany). Field surveys over 2 years revealed a range of symptom severity on grapevine and an increase of BN incidence. A TaqMan allelic discrimination assay detected only tufB type b among BNp strains, suggesting the prevalence of the bindweed-related ecology. Nucleotide sequence analyses of vmp1 and stamp genes identified 12 vmp1 and 16 stamp sequence variants, showing an overall positive selection for such genes. The prevalent genotype was Vm43/St10, reported for the first time in this study and closely related to strains identified only in the French Eastern Pyrenees. BNp strains identified in the examined vineyard and mostly grouped in separate bindweed-related phylogenetic clusters showed statistically significant differences in their distribution in grapevines exhibiting distinct symptom severity. These results suggest the possible occurrence of a range of virulence within BNp strain populations in the Chianti Classico area.

CPAP Treatment Adherence in Women with Obstructive Sleep Apnea.

Untreated obstructive sleep apnea (OSA) has numerous negative health-related consequences. Continuous positive airway pressure (CPAP) is generally considered the treatment of choice for OSA, but rates of nonadherence are high. It is believed that OSA is more prevalent among men; therefore understanding how OSA presents among women is limited and treatment adherence has received little research attention. For this study, 29 women were recruited from primary care offices. They completed a questionnaire battery and underwent a night of nocturnal polysomnography (PSG) followed by a visit with a sleep specialist. Women diagnosed with OSA were prescribed CPAP; 2 years later CPAP adherence was evaluated. Results show that approximately half the sample was adherent. There were no significant differences between adherent and nonadherent women on OSA severity; however CPAP adherent women had worse nocturnal and daytime functioning scores at the time of diagnosis. Moreover, when the seven nocturnal and daytime variables were used as predictors in a discriminant analysis, they could predict 87% of adherent and 93% of the nonadherent women. The single most important predictor was nonrefreshing sleep. We discuss the implications of the findings for identifying women in primary care with potential OSA and offer suggestions for enhancing treatment adherence.

Safety of outpatient autologous hematopoietic cell transplantation for multiple myeloma and lymphoma.

Autologous hematopoietic cell transplantation (Auto-HCT) is commonly an in-patient procedure. However, Auto-HCT is increasingly being offered on an outpatient basis. To better characterize the safety of outpatient Auto-HCT, we compared the outcome of 230 patients who underwent Auto-HCT on an in-patient vs outpatient basis for myeloma or lymphoma within a single transplant program. All outpatient transplants occurred in a cancer center day hospital. Hematopoietic recovery occurred earlier in the outpatient cohort, with median time to neutrophil recovery of 10 vs 11 days (P<0.001) and median time to platelet recovery of 19 vs 20 days (P=0.053). Fifty-one percent of the outpatient cohort never required admission, with this percentage increasing in later years. Grade 3-4 non-hematologic toxicities occurred in 29% of both cohorts. Non-relapse mortality at 1 year was 0% in the outpatient cohort and 1.5% in the in-patient cohort (P=0.327). Two-year PFS was 62% for outpatient vs 54% for in-patient (P=0.155). One- and two-year OS was 97% and 83% for outpatient vs 91% and 80% for in-patient, respectively (P=0.271). We conclude that, with daily outpatient evaluation and aggressive supportive care, outpatient Auto-HCT can result in excellent outcomes for myeloma and lymphoma patients.

Influence of TNF-α inhibition on oxidative stress of rheumatoid arthritis patients.

The aim of this study was to assess circulating levels of reactive oxygen metabolites (ROMs) as a marker of oxidative stress in rheumatoid arthritis (RA) patients during an anti-tumor necrosis factor alpha (TNF-α) treatment. We enrolled 40 patients with RA (36 females; age 53 ± 13 yrs) treated with different subcutaneously administered TNF-α inhibitors. The oxidative status was determined on the basis of plasma samples taken before, at 24 and 52 weeks of the anti-TNF-α treatment. Hydroperoxide levels were measured using the d-ROMs test, a useful clinically proven oxidative stress marker. During the anti-TNF-α therapy, we observed a significant reduction in serum ROMs levels in RA patients from 33.2 ± 10 mg H2O2/L at baseline to 29.5 ± 7 and 29.3 ± 9 mg H2O2/L, at 24 and 52 weeks, respectively (p<0.05). We also identified a significant correlation between the oxidative stress status and the disease activity score on 28 joints/C-reactive protein and health assessment questionnaire disability index. The results of our study demonstrate that a good control of the disease with anti-TNF-α agents can reduce oxidative stress in RA patients. However, further studies of larger patient cohorts are needed to confirm these preliminary data.

IgM peak independently predicts treatment-free survival in chronic lymphocytic leukemia and correlates with accumulation of adverse oncogenetic events.

We examined the significance of IgM peaks in chronic lymphocytic leukemia (CLL), including its association with newly reported MYD88, BIRC3, NOTCH1 and SF3B1 mutations. A total of 27, 25, 41 and 57 patients with monoclonal IgM or IgG peaks (IgM and IgG groups), hypogammaglobulinemia (Hypo-γ group) and normal immunoglobulin serum levels (normal-γ group) were, respectively, included. IgM peaks were mainly associated with Binet stage C and the del(17p). Biased usage of IGHV3-48 was shared by both IgM and IgG groups. IGHV3-74 and IGHV4-39 gene rearrangements were specific for IgM and IgG peaks, respectively. SF3B1, NOTCH1, MYD88 and BIRC3 mutation frequencies were 12%, 4%, 2% and 2%, respectively, being over-represented in IgM, IgG and Hypo-γ groups for SF3B1, and being equal between normal-γ and IgM groups for MYD88. Overall, 76%, 87%, 49% and 42% of cases from IgM, IgG, Hypo-γ and normal-γ groups had at least one intermediate or poor prognosis genetic marker, respectively. By multivariate analysis, IgM peaks were associated with shorter treatment-free survival independently from any other univariate poor prognosis biological parameters, including IgG peaks, Hypo-γ, IGHV status, SF3B1 mutations, cytogenetics and lymphocytosis. Therefore, as with IgG peaks, IgM peaks aggravated the natural course of CLL, with increased accumulation of adverse genetic events.

Effects of the administration of epidermal growth factor receptor specific inhibitor cetuximab, alone and in combination with cisplatin, on proliferation and apoptosis of Hep-2 laryngeal cancer cells.

BACKGROUND: Epidermal growth factor receptor (EGFR) overexpression and prognostic value in head and neck squamous cell cancer is the basis for targeting by anti-EGFR antibodies, which increase the efficacy of radiotherapy. In order to evaluate the best therapeutic schedule, the effects of cetuximab (C225) on Hep-2 cell proliferation, alone and in combination with cisplatin, were studied. METHODS: Hep-2 cells were treated with cetuximab alone or in combination with cisplatin. After determining cell viability with trypan blue, morphological features of apoptotic degeneration were analysed by fluorescence microscopy with Hoechst 33258 stain. RESULTS: Cetuximab alone mildly inhibited Hep-2 proliferation and showed no pro-apoptotic effects. When administered concomitantly with cisplatin, cetuximab synergistically increased inhibition of proliferation and apoptosis. CONCLUSION: The antiproliferative activity of cetuximab is consistent with its hypothesised role in inhibiting repopulation. However, the increase in the effects of pro-apoptotic agents induced by cetuximab may be even more relevant to its clinical effectiveness than the inhibition of repopulation.

First Report of the NA2 Lineage of Phytophthora ramorum from an Ornamental Rhododendron in the Interior of California.

In July 2012, we collected a rhododendron var. Trilby with twig dieback symptoms in the lower canopy, consistent with the disease "ramorum blight" caused by Phytophthora ramorum. The symptomatic plant had been planted a year earlier to replace a dead rhododendron in a landscape setting in Placer County, California (Lat: 39.036216°; Long: -120.999274°), Sierra Nevada foothills at ~600 m elevation. Isolations yielded a culture with a fast growth rate and overall morphology resembling the P. ramorum NA2 lineage described by Ivors et al. (4). DNA was extracted from the culture as described previously (4) and six SSR loci: MS18, MS39, MS43, MS45, MS64, MS145, were amplified (2,4). Allelic patterns were compared with those of three testers from each of the three lineages NA1, NA2, and EU1 known to be present in ornamental plants in North America, and they unambiguously confirmed the isolate belongs to the NA2 lineage of the pathogen. Although the symptomatic plant was confined to a landscape setting, it had been planted in that location for a year, providing a possible source of inoculum for the surrounding area. This is the first report of P. ramorum from the Sierra Nevada eco-region in the interior of California. It is also the first report of a NA2 isolate from a plant outside of commercial nurseries in California. The mating type of the isolate was not determined, but NA2 isolates are normally A2, the same mating type of NA1 isolates. The only other report of a NA2 isolate found outside of a nursery is from Washington State (1). Although there is no evidence the pathogen may have infected other plants, the infected rhododendron was found at a location situated over 100 km east of the closest known infestation (www.sodmap.org). Additionally, this is the first report of the pathogen outside the coast mountain range of California. Because the three lineages are genetically and phenotypically distinct (3), the escape of NA2 or EU1 isolates, both still absent from plants in natural settings, could have significant implications for California ecosystems. This finding highlights that introductions of P. ramorum via ornamental plants are still possible, in spite of current regulations. References: (1) G. Chastagner et al. Phytopathology 101:S32, 2011. (2) P. P. Croucher et al. Biol. Invasions 15:2281, 2013. (3) N. J. Grünwald et al. Trends Microbiol. 20:131, 2012. (4) K. Ivors et al. Mol. Ecol. 15:1493, 2006.

Household model of Chagas disease vectors (Hemiptera: Reduviidae) considering domestic, peridomestic, and sylvatic vector populations.

ABSTRACT Disease transmission is difficult to model because most vectors and hosts have different generation times. Chagas disease is such a situation, where insect vectors have 1-2 generations annually and mammalian hosts, including humans, can live for decades. The hemataphagous triatominae vectors (Hemiptera: Reduviidae) of the causative parasite Trypanosoma cruzi (Kinetoplastida: Trypanosomatidae) usually feed on sleeping hosts, making vector infestation of houses, peridomestic areas, and wild animal burrows a central factor in transmission. Because of difficulties with different generation times, we developed a model considering the dwelling as the unit of infection, changing the dynamics from an indirect to a direct transmission model. In some regions, vectors only infest houses; in others, they infest corrals; and in some regions, they also infest wild animal burrows. We examined the effect of sylvatic and peridomestic vector populations on household infestation rates. Both sylvatic and peridomestic vectors increase house infestation rates, sylvatic much more than peridomestic, as measured by the reproductive number R0. The efficacy of manipulating parameters in the model to control vector populations was examined. When R0 > 1, the number of infested houses increases. The presence of sylvatic vectors increases R0 by at least an order of magnitude. When there are no sylvatic vectors, spraying rate is the most influential parameter. Spraying rate is relatively unimportant when there are sylvatic vectors; in this case, community size, especially the ratio of houses to sylvatic burrows, is most important. The application of this modeling approach to other parasites and enhancements of the model are discussed.

IGHV gene features and MYD88 L265P mutation separate the three marginal zone lymphoma entities and Waldenström macroglobulinemia/lymphoplasmacytic lymphomas.

To clarify the relationships between marginal zone lymphomas (MZLs) and Waldenström macroglobulinemia/lymphoplasmacytic lymphomas (WM/LPLs), immunoglobulin heavy chain variable gene (IGHV) features were analyzed and the occurrence of MYD88 L265P mutations was identified in a series of 123 patients: 53 MZLs from the spleen (SMZLs), 11 from lymph nodes (NMZLs), 28 mucosa-associated lymphatic tissue (MALT) lymphomas and 31 WM/LPLs. SMZLs were characterized by overrepresentation of IGHV1-2 gene rearrangements with a canonical motif, without selection pressure and with long CDR3 segments. NMZLs had increased frequencies of IGHV3 genes. The IGHV gene was unmutated in most cases, often with long CDR3 segments. MALT lymphomas were usually associated with a mutated IGHV gene, but with the absence of selection pressure. WM/LPLs were associated with an IGHV3-23 overrepresentation and high IGHV mutation rate, with features of selection pressure and short CDR3 segments. MYD88 L265P mutations were almost restricted exclusively to WM/LPL patients. Taken all diagnoses together, all patients with MYD88 L265P mutations had an immunoglobulin M peak and almost all patients except one had bone marrow infiltration. These results demonstrate that the history of antigen exposure of the four entities studied was different and MYD88 L265P was specifically associated with WM/LPLs. WM/LPL may thus be functionally associated with constitutive nuclear factor-κB activation.

First Report of Knot Disease Caused by Pseudomonas savastanoi on Sweet Olive in Central Italy.

In April 2012 the presence of hyperplastic outgrowths on trunks, branches, and twigs of sweet olive plants, Osmanthus fragrans Lour (Fam. Oleaceae), was recorded in two ornamental hedges made up of five and four plants, respectively, in the city center of Montecatini (Pistoia-Italy). All sweet olive plants were seriously affected by the disease with outgrowths appearing either singly or close together, often forming a single mass that could extend up to 20 cm along the stems, occasionally surrounding the entire circumference. The symptoms observed on O. fragrans closely resembled those induced by the bacterium Pseudomonas savastanoi on Olea europea (common olive) and other plant species. Suspecting a bacterial origin of the disorder, young knots were collected from four diseased plants and used for bacterial isolation with standard techniques on nutrient sucrose agar medium (1). After 3 days of incubation at 26°C, non-levan forming colonies about 3 mm in diameter that were circular, convex, smooth, and cream colored with entire margins appeared on the surface of the agar medium. Purified isolates were gram negative, levan negative, oxidase negative, potato rot negative, arginine dihydrolase negative, showed a tobacco hypersensitive reaction, and tested positive to PCR screening for the presence of the iaaM (tryptophan-2-monooxygenase), iaaH (indoleacetamide hydrolase), ptz (isopentenyl transferase) (1) and iaaL (IAA-lysine synthethase) (3) genes. Three isolates were selected arbitrarily and further characterized by sequencing a fragment of the housekeeping genes rpoD (sigma factor 70) and pgi (phosphoglucose isomerase) (2). All sequenced gene fragments, of 620 bp and 552 bp for the rpoD and pgi genes, respectively, were identical to those of P. savastanoi pv. savastanoi strain NCPPB3335. The pathogenicity of the three isolates was verified on three O. fragrans plants and three Olea europea (cv. Frantoio) plants. Per each isolate, three 1-cm wounds were made on the branches of each plant using a sterile scalpel dipped in a bacterial suspension (1 × 108 CFU/ml). P. savastanoi pv. savastanoi PVFi-t2b isolated from olive was also inoculated as reference strain. After 30 days, all isolates including the reference strain induced typical knots on both plant species while no symptoms were observed on wounds inoculated with sterile water. Bacteria were reisolated from induced knots and Koch's postulates were confirmed. On the basis of biochemical tests, PCR screening, pathogenicity testing, and sequence analyses, the causal agent of knot disease on O. fragrans was identified as P. savastanoi. The potential susceptibility of O. aquifolium Sieb. to the causal agent of olive knot disease has been demonstrated in the past by means of artificial inoculations but interestingly, in the same trials, O. fragrans had tested negative (4). To the best of our knowledge, this is the world's first report of O. fragrans as natural host of P. savastanoi, which extends the growing list of cultivated and ornamental plant species affected by this phytopathogenic bacterium. References: (1) G. Marchi et al. Eur J. Plant Pathol. 112:101, 2005. (2) N. Parkinson et al. Plant Pathol. 60:338, 2011. (3) R. Penyalver et al. Appl. Environ. Microbiol. 66:2673, 2000. (4) C. O. Smith. Phytopathology 12:271, 1922.

Occurrence of Lily mottle virus on Lilium in Italy.

Lily mottle virus (LMoV), a member of the genus Potyvirus, is one of the main viruses infecting lily. Symptoms on lily differ according to the susceptibility and sensitivity of different cultivars and hybrids. They range from leaf mottle or mosaic, vein clearing, chlorotic and yellow streaking, leaf curling, and necrotic spots, to milder forms of leaf symptoms. Plants may even be symptomless at some stages of growth. A varietal collection of Lilium from the early 1990s is held in Pistoia Province (Tuscany, Italy) and is composed of Asian hybrids obtained from intraspecific breeding of commercial cultivars. During a survey conducted from May to June 2010, several plants showing vein clearing, leaf mottle, leaf mosaic, and reddish brownish necrotic spots were observed. Leaf samples from 60 symptomatic or symptomless lily plants, belonging to 20 cultivars, were collected and tested for the presence of LMoV. Samples were assayed by double-antibody sandwich (DAS)-ELISA and eight of them, belonging to four different cultivars, tested positive. Total RNA was extracted from 2 g of leaf tissue of every collected sample according to the protocol described earlier (3) and cDNA synthesis was performed with an iScript cDNA Synthesis Kit (Bio-Rad, Hercules, CA). Samples were tested by reverse transcription (RT)-PCR and real-time PCR assays using primers LMoV1 (5'-GCAAATGAGACACTCAATGCTG-3') and LMoV2 (5'-CGTGCGTGAAGTAACTTCATAG-3') designed to amplify 651 bp of the coat protein (CP) gene of LMoV (1). Results obtained with RT-PCR and real-time PCR exactly matched those achieved with ELISA assay, and the eight positive samples showed amplicons of the expected size. PCR products from five infected samples were directly sequenced from both directions and submitted in GenBank (Accessions Nos. JQ655106 to JQ655110). Our isolates share more than 99% nucleotide identity among each other. Comparison with other LMoV-CP gene sequences present in GenBank showed nucleotide identities ranging from 93 to 94% with LMoV isolates from South Korea (GenBank Accession Nos. GQ150683 to GQ150686), China (GenBank Accession Nos. EU348826, AJ748256, AJ564636, and AJ564637), Australia (GenBank Accession No. JN127341), and Japan (GenBank Accession No. AB570195). To our knowledge, this is the first report of LMoV on Lilium in Italy where this virus was already reported to infect escarole (2). Considering the economic importance of Lilium production as a flowering plant in Pistoia Province, and in several other areas of Italy, the report of LMoV present on lilies suggests the use of healthy propagation material and the adoption of preventive measures to avoid its diffusion. References: (1) J.-H. Lim et al. Korean J. Microbiol. 45:251, 2009. (2) V. Lisa et al. Plant Dis. 86:329, 2002. (3) D. J. MacKenzie et al. Plant Dis. 81:222, 1997.

Clinical approach and treatment of benign and malignant parotid masses, personal experience.

Parotid gland tumours account for 80% of all salivary gland neoplasms, 20% of these are malignant, but in daily clinical practice most parotid masses are operated on before obtaining the final histological diagnosis. This clinical setting further complicates the critical point of parotid surgery, which is the management of the facial nerve. In the present study, data were evaluated referring to 540 patients who underwent parotidectomy for a mass which was discovered to be a benign (470 cases) or a malignant (70 cases) neoplasm, between November 1994 and December 2007, at our Institution. The most significant single parameter in this series of malignancies regarding disease specific survival was the clinical involvement of the facial nerve at diagnosis (p = 0.006). Also for this reason, as there is no evidence that liberal VIIth nerve sacrifice improves prognosis, when it is not clinically involved, every attempt is made to dissect and preserve it. At present, the most complicated situation concerning nerve preservation may be, on the other hand, recurrence of a benign tumour, in particular pleomorphic adenoma, which, in our series, has a higher incidence (8.3%) of permanent facial dysfunction, than surgery with nerve preservation for malignancy (3.7%).