Subject(s)
COVID-19 , Antibodies, Viral , Humans , Immunocompromised Host , SARS-CoV-2 , SeroconversionABSTRACT
Transient osteoporosis of the hip (TOH) is usually reported in middle-aged men or during pregnancy as a benign self-limiting condition. Nevertheless, its impact on quality of life in terms of pain and disability is considerable. Also, it can lead to insufficiency fractures or, more rarely, evolve into osteonecrosis. This condition is anecdotally described in the pediatric age and very little is known about how it may affect the growing bone. We herein describe a case of TOH in a 10-year-old child treated at our pediatric rheumatology service and summarize the pediatric cases of TOH previously reported in literature. There are two points of interest in our case report, the first one being the unusual complication of TOH with a femoral physis fracture and the second the complete recovery after the off-label therapy with bisphosphonates. We suggest that interventional medical treatment could be considered in selected cases of juvenile TOH, to prevent any possible irreversible damage on the femoral physis. As far as we know, this is the first report of neridronate employment in children affected by TOH.
Subject(s)
Osteoporosis , Quality of Life , Child , Diphosphonates/therapeutic use , Female , Hip Joint , Humans , Magnetic Resonance Imaging , Male , Middle Aged , Osteoporosis/etiology , PregnancyABSTRACT
As the SARS-CoV-2 pandemic continues to rage worldwide, the emergence of numerous variants of concern (VOC) represents a challenge for the vaccinal protective efficacy and the reliability of commercially available high-throughput immunoassays. Our study demonstrates the administration of two doses of the BNT162b2 vaccine that elicited a robust SARS-CoV-2-specific immune response which was assessed up to 3 months after full vaccination in a cohort of 37 health care workers (HCWs). SARS-CoV-2-specific antibody response, evaluated by four commercially available chemiluminescence immunoassays (CLIA), was qualitatively consistent with the results provided by the gold-standard in vitro neutralization assay (NTA). However, we could not observe a correlation between the quantity of the antibody detected by CLIA assays and their neutralizing activity tested by NTA. Almost all subjects developed a SARS-CoV-2-specific T-cell response. Moreover, vaccinated HCWs developed a similar protective neutralizing antibodies response against the EU (B.1), Alpha (B.1.1.7), Gamma (P.1), and Eta (B.1.525) SARS-CoV-2 variants, while Beta (B.1.351) and Delta (B.1.617.2) strains displayed a consistent partial immune evasion. These results underline the importance of a solid vaccine-elicited immune response and a robust antibody titre. We believe that these relevant results should be taken into consideration in the definition of future vaccinal strategies.
Subject(s)
BNT162 Vaccine/immunology , COVID-19 Vaccines/immunology , COVID-19/immunology , COVID-19/prevention & control , SARS-CoV-2/immunology , Adult , Aged , Antibodies, Neutralizing/blood , Antibodies, Neutralizing/immunology , Antibodies, Viral/blood , Antibodies, Viral/immunology , BNT162 Vaccine/administration & dosage , BNT162 Vaccine/genetics , COVID-19/blood , COVID-19/virology , COVID-19 Vaccines/administration & dosage , COVID-19 Vaccines/genetics , Female , Humans , Immunity, Cellular , Immunity, Humoral , Immunoassay , Longitudinal Studies , Male , Middle Aged , Prospective Studies , SARS-CoV-2/genetics , T-Lymphocytes/immunology , Vaccination , Young AdultABSTRACT
BACKGROUND: Different concepts have been used to date (e.g. non-nursing tasks, organizational work) to define tasks performed by clinical nurses other than nursing care. However, the true essence of nursing work is still poorly understood mostly because nurses are lacking an appropriate lexicon to describe their practice. AIMS: To describe non-nursing tasks as experienced by nurses, exploring antecedents and consequences as perceived in daily practice. METHODS: A descriptive qualitative study from 2015 to 2016. A purposeful sample of nurses was approached. Semi-structured interviews were used, and content analysis was performed on audio-recorded and verbatim-transcribed interviews. FINDINGS: A total of 22 nurses participated, the majority of whom were female (16; 72.7%) and their average age was 42.6 years. The concept of 'Non-nursing tasks' is limited in describing what nurses experience in daily practice; the concept of 'Being out of the nursing role' emerged as being fully descriptive of the nurses' experience and this can occur in two dimensions: outside and inside the role of other healthcare professions. The first dimension includes administrative work separating nurses from patients. The second dimension was reported to happen in proximity to patients but in three different directions towards professions requiring: (a) less education (e.g. healthcare assistants), (b) the same amount of education at university level (e.g. physiotherapists), and (c) higher education at university level as compared to nurses, thus performing activities expected by physicians. Antecedents of 'Being out of the nursing role' have been identified at the organizational, individual and educational levels; their consequences have been reported at the patient, professional and organizational levels. CONCLUSION: Nurses play various non-nursing roles, below, above and in the horizontal levels, both inside and outside other healthcare professionals' role, mainly as a result of their felt moral obligation to offer the best to their patients, the organization's demand to nurses and the imprinting of nursing education. IMPLICATIONS FOR NURSING/HEALTH POLICY: Strategies at the nursing professional and policy levels are needed aimed at (a) supporting nurses in optimizing their professional identity, (b) sharing their sense of moral obligation towards patients with other healthcare professionals, and (c) implementing models of care based on interprofessional cooperation.
Subject(s)
Job Description , Nurse's Role , Task Performance and Analysis , Work Engagement , Workload/psychology , Adult , Female , Humans , Male , Middle Aged , Qualitative ResearchSubject(s)
Cytoskeletal Proteins/genetics , Growth Disorders/diagnosis , Growth Disorders/genetics , Intellectual Disability/diagnosis , Intellectual Disability/genetics , Microcephaly/diagnosis , Microcephaly/genetics , Syndactyly/diagnosis , Syndactyly/genetics , Facies , Growth Disorders/physiopathology , Humans , Infant , Intellectual Disability/physiopathology , Male , Microcephaly/physiopathology , Mutation , Syndactyly/physiopathologyABSTRACT
PURPOSE: To evaluate the efficacy and tolerability of Perampanel (PER) in children and adolescents with refractory epilepsies in daily clinical practice conditions. PATIENTS AND METHODS: This Italian multicenter retrospective observational study was performed in 16 paediatric epilepsy centres. Inclusion criteria were: (i) ≤18 years of age, (ii) history of refractory epilepsy, (iii) a follow-up ≥5 months of PER add-on therapy. Exclusion criteria were: (i) a diagnosis of primary idiopathic generalized epilepsy, (ii) variation of concomitant AEDs during the previous 4 weeks. Response was defined as a ≥50% reduction in monthly seizure frequency compared with the baseline. RESULTS: 62 patients suffering from various refractory epilepsies were included in this study: 53% were males, the mean age was 14.2 years (range 6-18 years), 8 patients aged <12 years. Mean age at epilepsy onset was 3.4 years and the mean duration of epilepsy was 10.8 years (range 1-16), which ranged from 2 seizures per-month up to several seizures per-day (mean number=96.5). Symptomatic focal epilepsy was reported in 62.9% of cases. Mean number of AEDs used in the past was 7.1; mean number of concomitant AEDs was 2.48, with carbamazepine used in 43.5% of patients. Mean PER daily dose was 7.1mg (2-12mg). After an average of 6.6 months of follow-up (5-13 months), the retention rate was 77.4% (48/62). The response rate was 50%; 16% of patients achieved ≥75% seizure frequency reduction and 5% became completely seizure free. Seizure aggravation was observed in 9.7% of patients. Adverse events were reported in 19 patients (30.6%) and led to PER discontinuation in 4 patients (6.5%). The most common adverse events were behaviour disturbance (irritability and aggressiveness), dizziness, sedation and fatigue. CONCLUSION: PER was found to be a safe and effective treatment when used as adjunctive therapy in paediatric patients with uncontrolled epilepsy.
Subject(s)
Anticonvulsants/therapeutic use , Drug Resistant Epilepsy/drug therapy , Pyridones/therapeutic use , Adolescent , Anticonvulsants/adverse effects , Child , Female , Follow-Up Studies , Humans , Italy , Male , Nitriles , Pyridones/adverse effects , Retrospective Studies , Seizures/drug therapy , Treatment OutcomeABSTRACT
Malignant mesothelioma (MM) is an aggressive, fatal tumor strongly associated with asbestos exposure. There is an urgent need to improve MM patient outcomes and this requires functionally validated pre-clinical models. Mesothelioma-derived cell lines provide an essential and relatively robust tool and remain among the most widely used systems for candidate drug evaluation. Although a number of cell lines are commercially available, a detailed comparison of these commercial lines with freshly derived primary tumor cells to validate their suitability as pre-clinical models is lacking. To address this, patient-derived primary mesothelioma cell lines were established and characterized using complementary multidisciplinary approaches and bioinformatic analysis. Clinical markers of mesothelioma, transcriptional and metabolic profiles, as well as the status of p53 and the tumor suppressor genes CDKN2A and NF2, were examined in primary cell lines and in two widely used commercial lines. Expression of MM-associated markers, as well as the status of CDKN2A, NF2, the 'gatekeeper' in MM development, and their products demonstrated that primary cell lines are more representative of the tumor close to its native state and show a degree of molecular diversity, thus capturing the disease heterogeneity in a patient cohort. Molecular profiling revealed a significantly different transcriptome and marked metabolic shift towards a greater glycolytic phenotype in commercial compared with primary cell lines. Our results highlight that multiple, appropriately characterised, patient-derived tumor cell lines are required to enable concurrent evaluation of molecular profiles versus drug response. Furthermore, application of this approach to other difficult-to-treat tumors would generate improved cellular models for pre-clinical evaluation of novel targeted therapies.
Subject(s)
Lung Neoplasms/metabolism , Mesothelioma/metabolism , Metabolome , Aged , Cyclin-Dependent Kinase Inhibitor p16/genetics , Cyclin-Dependent Kinase Inhibitor p16/metabolism , Cyclin-Dependent Kinase Inhibitor p18/genetics , Cyclin-Dependent Kinase Inhibitor p18/metabolism , Female , Genomic Instability , Humans , Lung Neoplasms/pathology , Male , Mesothelioma/pathology , Mesothelioma, Malignant , Middle Aged , Neurofibromin 2/genetics , Neurofibromin 2/metabolism , Oxygen Consumption , Principal Component Analysis , Tandem Repeat Sequences , Transcriptome , Tumor Cells, Cultured , Tumor Suppressor Protein p14ARF/genetics , Tumor Suppressor Protein p14ARF/metabolism , Tumor Suppressor Protein p53/genetics , Tumor Suppressor Protein p53/metabolism , Up-RegulationABSTRACT
BACKGROUND AND PURPOSE: Our aim was to describe the clinical and electrical features and the long-term evolution of childhood occipital epilepsy of Gastaut (COE-G) in a cohort of patients and to compare long-term prognosis between patients with and without other epileptic syndromes. METHODS: This was a retrospective analysis of the long-term outcome of epilepsy in 129 patients with COE-G who were referred to 23 Italian epilepsy centres and one in Austria between 1991 and 2004. Patients were evaluated clinically and with electroencephalograms for 10.1-23.0 years. The following clinical characteristics were evaluated: gender, patient age at seizure onset, history of febrile seizures and migraine, family history of epilepsy, duration and seizure manifestations, circadian distribution and frequency of seizures, history of medications including the number of drugs, therapeutic response and final outcome. RESULTS: Visual hallucinations were the first symptom in 62% and the only manifestation in 38.8% of patients. Patients were subdivided into two groups: group A with isolated COE-G; group B with other epileptic syndromes associated with COE-G. The most significant (P < 0.05) difference concerned antiepileptic therapy: in group A, 45 children responded to monotherapy; in group B only 15 children responded to monotherapy. At the end of follow-up, the percentage of seizure-free patients was significantly higher in group A than in group B. CONCLUSIONS: Childhood occipital epilepsy of Gastaut has an overall favourable prognosis and a good response to antiepileptic therapy with resolution of seizures and of electroencephalogram abnormalities. The association of typical COE-G symptoms with other types of seizure could be related to a poor epilepsy outcome.
Subject(s)
Anticonvulsants/pharmacology , Lennox Gastaut Syndrome , Occipital Lobe/physiopathology , Outcome Assessment, Health Care , Adolescent , Adult , Austria , Child , Child, Preschool , Electroencephalography , Female , Humans , Infant , Lennox Gastaut Syndrome/diagnosis , Lennox Gastaut Syndrome/drug therapy , Lennox Gastaut Syndrome/physiopathology , Male , Prognosis , Retrospective Studies , Young AdultABSTRACT
Many antiepileptic drugs (AEDs) are associated with hematological disorders that range from mild thrombocytopenia or neutropenia to anemia, red cell aplasia, until bone marrow failure. Fortunately, potentially fatal hematological disorders such as aplastic anemia are very rare. This review investigates hematological effects associated with classic and newer AEDs: a PubMed search indexed for MEDLINE was undertaken to identify studies in adults, children and animals using the name of all anticonvulsant drugs combined with the terms "hematological disease" and "hematological abnormalities" as key words. The most common hematological alterations occur with older AEDs than newer. Indeed, careful hematological monitoring is needed especially using carbamazepine, phenytoin and valproic acid. The pathogenetic mechanisms are still unknown: they seem to be related to an immunological mechanism, but drugs pharmacokinetics and pharmacodynamics interactions may also play an important role. Further research is needed to assess the real pathogenetic mechanism at the basis of hematological complications caused by AEDs.
Subject(s)
Anticonvulsants/therapeutic use , Epilepsy/drug therapy , Epilepsy/etiology , Hematologic Diseases/complications , Hematologic Diseases/drug therapy , HumansABSTRACT
OBJECTIVE: Available data on the efficacy of lacosamide in children with Lennox-Gastaut syndrome (LGS) are scarce and controversial. We present our experience with lacosamide therapy in children affected by LGS. MATERIAL AND METHODS: Medical charts of all children affected by LGS receiving oral lacosamide adjunctive therapy in six paediatric neurology centres were retrospectively evaluated. Efficacy was determined according to the frequency of countable seizures during the 4 weeks prior to treatment and the frequency in the last 4 weeks of observation. Patients whose seizure frequency was reduced by at least 50% were defined as responders. RESULTS: Eighteen children (mean age 12.3 years) were identified. After a mean follow-up period of 9 months, 33% of patients were responders. None of them was seizure-free during the study period. The overall seizure reduction rate was 29%. The percentage reductions from baseline in tonic seizures and drop-attacks rates were 31% and 20%, respectively. Adverse reactions occurred in 44% of patients. The drug was discontinued in four (22%) patients because of increased seizure frequency (three cases) and walking instability (another patient). CONCLUSIONS: A third of children with LGS were responders after lacosamide adjunctive therapy. Although caution is still necessary when the drug is used in children with LGS, our preliminary observations suggest that lacosamide might be effective and represent a possible therapeutic option in children affected by LGS.
Subject(s)
Acetamides/therapeutic use , Anticonvulsants/therapeutic use , Lennox Gastaut Syndrome/drug therapy , Acetamides/administration & dosage , Acetamides/adverse effects , Administration, Oral , Adolescent , Anticonvulsants/administration & dosage , Anticonvulsants/adverse effects , Child , Child, Preschool , Drug Therapy, Combination , Female , Follow-Up Studies , Humans , Lacosamide , Male , Retrospective Studies , Time Factors , Treatment OutcomeABSTRACT
Difficulties in accessing high-quality data on trace gas fluxes and performance of bioenergy/bioproduct feedstocks limit the ability of researchers and others to address environmental impacts of agriculture and the potential to produce feedstocks. To address those needs, the GRACEnet (Greenhouse gas Reduction through Agricultural Carbon Enhancement network) and REAP (Renewable Energy Assessment Project) research programs were initiated by the USDA Agricultural Research Service (ARS). A major product of these programs is the creation of a database with greenhouse gas fluxes, soil carbon stocks, biomass yield, nutrient, and energy characteristics, and input data for modeling cropped and grazed systems. The data include site descriptors (e.g., weather, soil class, spatial attributes), experimental design (e.g., factors manipulated, measurements performed, plot layouts), management information (e.g., planting and harvesting schedules, fertilizer types and amounts, biomass harvested, grazing intensity), and measurements (e.g., soil C and N stocks, plant biomass amount and chemical composition). To promote standardization of data and ensure that experiments were fully described, sampling protocols and a spreadsheet-based data-entry template were developed. Data were first uploaded to a temporary database for checking and then were uploaded to the central database. A Web-accessible application allows for registered users to query and download data including measurement protocols. Separate portals have been provided for each project (GRACEnet and REAP) at nrrc.ars.usda.gov/slgracenet/#/Home and nrrc.ars.usda.gov/slreap/#/Home. The database architecture and data entry template have proven flexible and robust for describing a wide range of field experiments and thus appear suitable for other natural resource research projects.
Subject(s)
Greenhouse Effect , Soil , Agriculture , Carbon , Crops, Agricultural , Fertilizers , Soil/chemistryABSTRACT
Urban pollution is a current problem, constituting a serious health risk. Many pollutants are present in the urban atmosphere, they are mainly anthropogenic, resulting from the combustion of coal. Several studies have shown the harmful effects of these pollutants, particularly on the respiratory system. Through a library review, we want to explore the effects on the health of general population and of outdoor workers exposed to the products resulting from the combustion of coal, in order to compare them with the estimated effects of the pollutants from the new generation coal plants, certainly less harmful to the environment. In recent years investments in "clean technologies" have resulted in a rapid and significant reduction of all polluting emissions: sulfur dioxide, dust, oxides of nitrogen, ash and gypsum resulting from the desulfurization process. To conclude in the light of the scientific literature we can say that the activation of coal plants based on technologies for the maximum containment and optimal treatment of the production cycle is essential to defend both the environment and the health of the population. Nevertheless it remains necessary to ensure a systematic monitoring of the environment of sanitation and food processing of the population living in the area, with a long-term follow-up.
Subject(s)
Coal/adverse effects , Environment , Environmental Exposure/adverse effects , Power Plants , Environmental Exposure/prevention & control , Humans , Risk FactorsABSTRACT
The resistance of hypoxic cells to radiotherapy and chemotherapy is a major problem in the treatment of cancer. Recently, an additional mode of hypoxia-inducible factor (HIF)-dependent transcriptional regulation, involving modulation of a specific set of micro RNAs (miRNAs), including miR-210, has emerged. We have recently shown that HIF-1 induction of miR-210 also stabilizes HIF-1 through a positive regulatory loop. Therefore, we hypothesized that by stabilizing HIF-1 in normoxia, miR-210 may protect cancer cells from radiation. We developed a non-small cell lung carcinoma (NSCLC)-derived cell line (A549) stably expressing miR-210 (pmiR-210) or a control miRNA (pmiR-Ctl). The miR-210-expressing cells showed a significant stabilization of HIF-1 associated with mitochondrial defects and a glycolytic phenotype. Cells were subjected to radiation levels ranging from 0 to 10 Gy in normoxia and hypoxia. Cells expressing miR-210 in normoxia had the same level of radioresistance as control cells in hypoxia. Under hypoxia, pmiR-210 cells showed a low mortality rate owing to a decrease in apoptosis, with an ability to grow even at 10 Gy. This miR-210 phenotype was reproduced in another NSCLC cell line (H1975) and in HeLa cells. We have established that radioresistance was independent of p53 and cell cycle status. In addition, we have shown that genomic double-strand breaks (DSBs) foci disappear faster in pmiR-210 than in pmiR-Ctl cells, suggesting that miR-210 expression promotes a more efficient DSB repair. Finally, HIF-1 invalidation in pmiR-210 cells removed the radioresistant phenotype, showing that this mechanism is dependent on HIF-1. In conclusion, miR-210 appears to be a component of the radioresistance of hypoxic cancer cells. Given the high stability of most miRNAs, this advantage could be used by tumor cells in conditions where reoxygenation has occurred and suggests that strategies targeting miR-210 could enhance tumor radiosensitization.
Subject(s)
Carcinoma, Non-Small-Cell Lung/genetics , Gene Expression Regulation, Neoplastic/radiation effects , Hypoxia-Inducible Factor 1/genetics , Hypoxia/genetics , Lung Neoplasms/genetics , MicroRNAs/genetics , Apoptosis/radiation effects , Carcinoma, Non-Small-Cell Lung/metabolism , Carcinoma, Non-Small-Cell Lung/pathology , Cell Hypoxia/genetics , Cell Hypoxia/radiation effects , Cell Line, Tumor , DNA Breaks, Double-Stranded , DNA Repair , Gamma Rays , Humans , Hypoxia/metabolism , Hypoxia/pathology , Hypoxia-Inducible Factor 1/metabolism , Lung Neoplasms/metabolism , Lung Neoplasms/pathology , MicroRNAs/metabolism , Mitochondria/genetics , Mitochondria/metabolism , Mitochondria/radiation effects , Radiation Tolerance , Signal Transduction/radiation effects , Transcription, Genetic/radiation effectsSubject(s)
von Willebrand Diseases/genetics , von Willebrand Factor/genetics , Adolescent , Alleles , Exons , Factor VIII/analysis , Factor VIII/therapeutic use , Female , Gene Frequency , Humans , Partial Thromboplastin Time , Polymorphism, Single Nucleotide , von Willebrand Diseases/diagnosis , von Willebrand Diseases/drug therapy , von Willebrand Factor/analysis , von Willebrand Factor/therapeutic useABSTRACT
BACKGROUND: To report on the first multicenter Italian experience with rufinamide as adjunctive drug in children, adolescents and young adults with refractory childhood-onset epileptic encephalopathies other than Lennox-Gastaut syndrome. METHODS: Thirty-eight patients (19 males, 19 females), aged between 4 and 34 (mean 13.7 ± 8.3, median 12.5), all affected by different types of childhood-onset refractory epileptic encephalopathies other than Lennox-Gastaut syndrome, were treated with rufinamide as adjunctive drug for a mean period of 11.4 months (range 3-26 months). RESULTS: Fifteen of 38 patients (39.5%) had a ≥ 50% seizure reduction in countable seizures. Complete seizure freedom was achieved in one of these patients (2.6%). Three patients (7.9%) had a 25-49% seizure reduction, whilst seizure frequency remained unchanged in 15 (39.5%) and increased in five patients (13.1%). Eleven patients (28.9%) reported adverse side effects. Vomiting was reported in five patients (13.1%); drowsiness, decreased appetite and irritability with migraine manifested in other four patients. They were transient and mild in all cases. CONCLUSION: Rufinamide may be an effective and well-tolerated adjunctive drug for the treatment of refractory childhood-onset epileptic encephalopathies other than Lennox-Gastaut syndrome. Rufinamide was most effective in patients with drop-attacks and (bi)frontal spike-wave discharges.
Subject(s)
Anticonvulsants/therapeutic use , Epilepsy/drug therapy , Triazoles/therapeutic use , Adolescent , Adult , Brain Diseases/complications , Brain Diseases/drug therapy , Child , Child, Preschool , Epilepsy/etiology , Female , Humans , Male , Young AdultABSTRACT
Following the identification of a set of hypoxia-regulated microRNAs (miRNAs), recent studies have highlighted the importance of miR-210 and of its transcriptional regulation by the transcription factor hypoxia-inducible factor-1 (HIF-1). We report here that miR-210 is overexpressed at late stages of non-small cell lung cancer. Expression of miR-210 in lung adenocarcinoma A549 cells caused an alteration of cell viability associated with induction of caspase-3/7 activity. miR-210 induced a loss of mitochondrial membrane potential and the apparition of an aberrant mitochondrial phenotype. The expression profiling of cells overexpressing miR-210 revealed a specific signature characterized by enrichment for transcripts related to 'cell death' and 'mitochondrial dysfunction', including several subunits of the electron transport chain (ETC) complexes I and II. The transcript coding for one of these ETC components, SDHD, subunit D of succinate dehydrogenase complex (SDH), was validated as a bona fide miR-210 target. Moreover, SDHD knockdown mimicked miR-210-mediated mitochondrial alterations. Finally, miR-210-dependent targeting of SDHD was able to activate HIF-1, in line with previous studies linking loss-of-function SDH mutations to HIF-1 activation. miR-210 can thus regulate mitochondrial function by targeting key ETC component genes with important consequences on cell metabolism, survival and modulation of HIF-1 activity. These observations help explain contradictory data regarding miR-210 expression and its putative function in solid tumors.
Subject(s)
Carcinoma, Non-Small-Cell Lung/genetics , Hypoxia-Inducible Factor 1, alpha Subunit/metabolism , Lung Neoplasms/genetics , MicroRNAs/metabolism , Mitochondria/pathology , Apoptosis , Carcinoma, Non-Small-Cell Lung/enzymology , Carcinoma, Non-Small-Cell Lung/pathology , Carcinoma, Non-Small-Cell Lung/ultrastructure , Caspase 3/metabolism , Caspase 7/metabolism , Cell Hypoxia/genetics , Cell Line, Tumor , Cell Survival/genetics , Down-Regulation/genetics , Gene Expression Profiling , Gene Expression Regulation, Neoplastic , Humans , Lung Neoplasms/enzymology , Lung Neoplasms/pathology , Lung Neoplasms/ultrastructure , MicroRNAs/antagonists & inhibitors , MicroRNAs/genetics , Mitochondria/enzymology , Mitochondria/ultrastructure , Mitochondrial Proteins/metabolism , Neoplasm Staging , Phenotype , RNA, Messenger/genetics , RNA, Messenger/metabolism , RNA, Small Interfering/metabolism , Succinate Dehydrogenase/metabolism , Up-Regulation/geneticsABSTRACT
From 1999 to 2010, withering of white willow was observed on trees growing along roads or irrigation canals in Torino, Alessandria, and Vercelli provinces of Italy, with incidence varying from 15, 25, and 30%, respectively. In spring and autumn 2008, six samples from withering branches with bark cankers were collected. On the bark surface near the cankers, iridescent traces of dried ooze were found. Tissues immediately below the cankers were dark with water-soaked, olive-colored edges. In some cases, the xylem appeared affected. Small fragments taken from the affected tissue on both edges of bark alterations and darkened vessels were crushed into mortars with sterile saline. Ten-fold serial dilutions (10-1, 10-2) were also performed. Aliquots of 0.1 ml were plated on nutrient agar and incubated at 25°C for 4 days. Bacterial colonies were ivory to white, circular, and bright, with a diameter of ~2 mm. Isolates were negative for Gram staining, presence of arginine dehydrolase, oxidase, phenylalanine deaminase, urease, hydrolysis of gelatin and starch, nitrate reduction, acidity from d-arabinose, cellobiose, lactose, maltose, trehalose, xylose, and pectinolytic activity on potato slices; positive for the presence of catalase and levan, fermentative metabolism of glucose, acid production from aesculin, l-arabinose, dextrose, d-galactose, inositol, d-mannitol, α-methylglucoside, raffinose, salicin and sucrose, H2S production from cysteine, and bright yellow pigment production on autoclaved potato tissue. They were not fluorescent on King's medium B and did not induce hypersensitivity reaction on tobacco leaf. Similar results were obtained with Brenneria salicis control strain, LMG 6089, except for acid production from α-methylglucoside (negative) and l-arabinose (negative). Acid production from α-methylglucoside has been reported for the Japanese strains of B. salicis, which do not produce acidity from inositol (4). Genomic DNA was extracted (1) from three isolates, and PCR reactions were performed with Es1A and Es4B primers (2) that amplify a 553-bp fragment from the 16S rDNA of B. salicis. The isolates showed a PCR product of expected size, like the positive control LMB 6089. On the basis of colony features, biochemical tests, and the PCR assay, we conclude that the isolates belong to B. salicis, a pathogen reported in Belgium, Germany, Great Britain, the Netherlands, Hungary, Japan, and New Zealand (2,3) but, as well as watermark disease symptoms, never previously reported in Italy. In summer 2009, pathogenicity tests were performed by inoculating young, white willow plants with B. salicis suspensions of ~1 to 2 × 109 CFU/ml placed with a syringe at the intersection of 1-year-old branches on the trunk. However, a year later, no symptoms of disease have been noted on the inoculated plants. According to the literature, pathogenicity tests rarely lead to the expected results because the bacterium can survive for many years in latent form, breaking out only when proper environmental conditions occur (3). Also the tests with B. salicis LMG 6089 gave negative results. Further investigation is necessary to clarify the relationships between this bacterium and the environment in causing withering of white willows in Piedmont. References: (1) W. P. Chen and T. T. Kuo. Nucleic Acids Res. 21:2260, 1993. (2) L. Hauben et al. Appl. Environ. Microbiol. 64:3966, 1998. (3) M. Maes et al. Environ. Microbiol. 11:1453, 2009. (4) Y. Sakamoto et al. Plant Pathol. 48:613, 1999.