Antimicrobial resistance in wildlife: detection of antimicrobial resistance genes in Apennine wolves (Canis lupus italicus Altobello, 1921) from Central Italy.

The aim of this study was to molecularly investigate the presence of antimicrobial resistance genes (ARGs) in organ samples from 11 Apennine wolves (Canis lupus italicus) collected in Central Italy. Samples from lung, liver, spleen, kidney, tongue and intestine were investigated by PCRs targeting the following genes: tet(A), tet(B), tet(C), tet(D), tet(E), tet(G), tet(K), tet(L), tet(M), tet(O), tetA(P), tet(Q), tet(S), tet(X), sul1, sul2, sul3, blaCTX-M, blaSHV, blaTEM and mcr-1. A PCR positivity was highlighted for 13 out of the 21 tested genes; no positive results were obtained for tet(C), tet(D), tet(E), tet(G), sul3, blaCTX, blaSHV and mcr-1 genes. All 11 animals sampled showed positivity for one or more resistance genes. The results confirm the potential role of the wolf as an indicator and/or vector of antimicrobial-resistant bacteria or ARGs.

Antimicrobial resistance genes in a golden jackal (Canis aureus L. 1758) from Central Italy.

In recent years an increasing interest has been focused on the contribution of wildlife in ecology and evolution of the antimicrobial resistance (AMR). The aim of this study was to molecularly investigate the presence of antimicrobial resistance genes (ARGs) in organ samples from a golden jackal (Canis aureus) found dead in the Marche region (Central Italy). Samples from lung, liver, spleen, kidney, and intestine were investigated by PCRs targeting the following genes: tet(A), tet(B), tet(C), tet(D), tet(E), tet(G), tet(K), tet(L), tet(M), tet(O), tet(S), tet(P), tet(Q), tet(X), sul1, sul2, sul3, blaCTX-M, blaSHV, blaTEM, and mcr-1 to mcr-10. One or more ARGs were detected in all organs tested, except the spleen. Specifically, the lung and liver were positive for tet(M) and tet(P), the kidney for mcr-1 and the intestine for tet(A), tet(L), tet(M), tet(O), tet(P), sul3 and blaTEM-1. These results, according to the opportunistic foraging strategy of the jackal, confirm its potential role as a good bioindicator of AMR environmental contamination.

Restoration of energy homeostasis by SIRT6 extends healthy lifespan.

Aging leads to a gradual decline in physical activity and disrupted energy homeostasis. The NAD+-dependent SIRT6 deacylase regulates aging and metabolism through mechanisms that largely remain unknown. Here, we show that SIRT6 overexpression leads to a reduction in frailty and lifespan extension in both male and female B6 mice. A combination of physiological assays, in vivo multi-omics analyses and 13C lactate tracing identified an age-dependent decline in glucose homeostasis and hepatic glucose output in wild type mice. In contrast, aged SIRT6-transgenic mice preserve hepatic glucose output and glucose homeostasis through an improvement in the utilization of two major gluconeogenic precursors, lactate and glycerol. To mediate these changes, mechanistically, SIRT6 increases hepatic gluconeogenic gene expression, de novo NAD+ synthesis, and systemically enhances glycerol release from adipose tissue. These findings show that SIRT6 optimizes energy homeostasis in old age to delay frailty and preserve healthy aging.

Bioactivity of volatile organic compounds by Aureobasidium species against gray mold of tomato and table grape.

Aureobasidium strains isolated from diverse unconventional environments belonging to the species A. pullulans, A. melanogenum, and A. subglaciale were evaluated for Volatile Organic Compounds (VOCs) production as a part of their modes of action against Botrytis cinerea of tomato and table grape. By in vitro assay, VOCs generated by the antagonists belonging to the species A. subglaciale showed the highest inhibition percentage of the pathogen mycelial growth (65.4%). In vivo tests were conducted with tomatoes and grapes artificially inoculated with B. cinerea conidial suspension, and exposed to VOCs emitted by the most efficient antagonists of each species (AP1, AM10, AS14) showing that VOCs of AP1 (A. pullulans) reduced the incidence by 67%, partially confirmed by the in vitro results. Conversely, on table grape, VOCs produced by all the strains did not control the fungal incidence but were only reducing the infection severity (< 44.4% by A. pullulans; < 30.5% by A. melanogenum, and A. subglaciale). Solid-phase microextraction (SPME) and subsequent gas chromatography coupled to mass spectrometry identified ethanol, 3-methyl-1-butanol, 2-methyl-1-propanol as the most produced VOCs. However, there were differences in the amounts of produced VOCs as well as in their repertoire. The EC50 values of VOCs for reduction of mycelial growth of B. cinerea uncovered 3-methyl-1-butanol as the most effective compound. The study demonstrated that the production and the efficacy of VOCs by Aureobasidium could be directly related to the specific species and pathosystem and uncovers new possibilities for searching more efficient VOCs producing strains in unconventional habitats other than plants.

Primordial odontogenic tumour: A systematic review of the common but also unusual features of this novel entity.

BACKGROUND: Primordial odontogenic tumour (POT) is a novel entity that was described in 2014 and that is included in the group of benign mixed epithelial and mesenchymal odontogenic tumours. In recent years, several papers have added new cases with some clinical and histopathological aspects that slightly differ from those described in the original report. The aim of this systematic review is to update all available data on POT published in the literature and to identify those features of the neoplasm that require further investigation. MATERIALS AND METHODS: A systematic review of literature was conducted using PubMed, Embase, Web of Science and Scopus. Additional sources were also checked. Publications reporting cases with enough clinicopathological information were included, without any time or language restrictions. Histopathological or radiological studies were considered for qualitative analysis. RESULTS: A total of 30 publications were included. Seventeen papers were used for quantitative analysis while 13 papers were used only for qualitative analysis. A total of 18 cases of POT were identified. Some clinical, radiographic, histopathological and therapeutic features were common in all reported cases, while other aspects of the neoplasm were inconsistent through published cases. This inconsistency was particularly remarkable when dealing with the histopathological features of the neoplasm. DISCUSSION: Some issues about POT remain unclear and deserve to be clarified by future reports. The description of the odontogenic epithelium covering the ectomesenchyme is often contradictory, while it remains debatable whether peripheral ameloblastic epithelial islands or hard dental tissue deposition can occasionally occur within the tumour.

Effect of Aureobasidium pullulans strains against Botrytis cinerea on kiwifruit during storage and on fruit nutritional composition.

Kiwifruit, wounded at the equator or by pedicel removal, to simulate the stem end wound, were treated with Aureobasidium pullulans (L1 and L8 strains) and subsequently inoculated with conidia of Botrytis cinerea. Fruits were stored at -1 °C in normal refrigeration (NR) or in controlled atmosphere (CA) (2% O2; 4.5% CO2). After 4 months, both antagonists significantly reduced the disease in all experiments, L1 better than L8. In NR, their efficacy was higher than 80%. In CA, the disease reduction was lower: between 30% (L1) and 60% (L8). The ability of both strains to compete with the pathogen for nutrients was tested in kiwifruit juice (0.5%) by in vitro experiments. Antagonists significantly reduced pathogen conidia germination in water and in juice. An HPLC analysis was performed to define the amino acid composition of kiwifruit juice upon L1 and L8 treatment. L1 and L8 greatly increased the concentration of both glutamic and aspartic acids and stimulated the production of new amino acids, although at low concentrations. Each amino acid displayed an antifungal effect against mycelium growth of B. cinerea. Finally, L1 and L8, cold tolerant and active strains in CA, can be effectively applied to control the stem end rot of kiwifruit in long storage.

Genome sequence analysis of a distinctive Italian infectious bursal disease virus.

In a recent study, an emerging infectious bursal disease virus (IBDV) genotype (ITA) was detected in IBDV-live vaccinated broilers without clinical signs of infectious bursal disease (IBD). VP2 sequence analysis showed that strains of the ITA genotype clustered separately from vaccine strains and from other IBDV reference strains, either classic or very virulent. In order to obtain a more exhaustive molecular characterization of the IBDV ITA genotype and speculate on its origin, genome sequencing of the field isolate IBDV/Italy/1829/2011, previously assigned to the ITA genotype, was performed, and the sequences obtained were compared to the currently available corresponding sequences. In addition, phylogenetic and recombination analyses were performed. Interestingly, multiple amino acid (AA) sequence alignments revealed that the IBDV/Italy/1829/2011 strain shared several AA residues with very virulent IBDV strains as well as some virulence markers, especially in the VP1 protein. Nevertheless, sequence analysis demonstrated the presence of several residues typical of IBDV strains at a low degree of virulence in the IBDV/Italy/1829/2011 strain. Although homologous recombination and reassortant phenomena may occur naturally among different IBDV strains, no evidence of those events was found in the genome of the IBDV/Italy/1829/2011 strain, which was confirmed to be a genetically distinctive IBDV genotype.

Odontomas: review of the literature and case reports.

Odontomas are one of the most common Odontogenic Tumors of the jaw. The exact etiology of odontomas is unknown. Histologically they are composed of various formations of dental tissue (enamel, dentin, cementum and sometimes pulp). In the WHO classification, they are divided into complex odontoma and compound odontoma. Clinically, odontomas are generally asymptomatic and only in rare cases cause swelling, pain, suppuration or bony expansion. Radiologically, the tumor is initially lucent, but with time, it develops small calcifications, which eventually coalesce to form a radiodense lesion with a lucent rim. Surgical resection is the treatment of choice and there is no recurrence. The aim of this paper is to define the principal characteristics and the treatment of these lesions, based on literature and personal experience.

Antifungal effect of volatile organic compounds produced by Bacillus amyloliquefaciens CPA-8 against fruit pathogen decays of cherry.

The present work focuses on the antifungal effect of volatile organic compounds (VOCs) produced by Bacillus amyloliquefaciens CPA-8 against Monilinia laxa, M. fructicola and Botrytis cinera, three postharvest fruit pathogens of sweet cherry fruit. VOCs were evaluated with a double petri dish assay against mycelial and colony growth of target pathogens. For this purpose, CPA-8 was grown on different media and cultured for 24 and 48 h at 30 °C before assays. Data showed that mycelial growth inhibition was higher when CPA-8 was grown on Tryptone Soya Agar (TSA) while no differences were generally observed when CPA-8 was cultured for either, 24 and 48 h. Moreover, no effects were observed on colony growth. The main volatile compounds emitted by CPA-8 were identified by solid-phase microextraction (SPME)-gas chromatography as 1,3 pentadiene, acetoin (3-hydroxy-2-butanone) and thiophene. Pure compounds were also tested in vitro on mycelial growth inhibition and their EC50 values against the three pathogens were estimated. Thiophene was the most effective VOC, showing more than 82% suppression of mycelial growth at the highest concentration (1.35 µL/mL headspace) and EC50 values ranging from 0.06 to 6.67 µL/mL headspace. Finally, the effectiveness of thiophene and CPA-8 VOCs was evaluated against artificially inoculated cherry fruits. Among the target pathogens, M. fructicola was clearly controlled by CPA-8 with less than 25% of rotten fruits compared to the control (65% disease incidence) and for all pathogens, less than 37.5% of CPA-8 treated decayed fruits produced spores (disease sporulation). Otherwise, pure thiophene showed no effect against any pathogen on disease incidence and disease sporulation. The results indicated that VOCs produced by B. amyloliquefaciens CPA-8 could develop an additive antifungal effect against postharvest fruit pathogens on stone fruit.

Continuous intraarticular and periarticular levobupivacaine for management of pain relief after total knee arthroplasty: A prospective randomized, double-blind pilot study.

BACKGROUND: Total knee arthroplasty (TKA) can result in major postoperative pain which can impact the recovery and rehabilitation of patients and for this reason the use of a pain-control infusion pumps (PCIP) enhances analgesia for TKA. PURPOSE: To investigate whether a PCIP of levobupivacaine would reduce pain in patients following TKA. METHODS: This was a prospective, randomized, controlled study conducted in 55 patients. Criteria for participation were unilateral TKA for osteoarthritis and no allergies to levobupivacaine. The primary outcomes measured were postoperative pain intensity on Visual Analogue Scale (VAS) score measured at 24 h and 48 h. Other measures included amount of narcotics, presence of adverse events, and length of hospital stay. RESULTS: PCIP-treated patients (n = 28) showed significant reductions in VAS score at any time versus control (p < 0.01). Amount of narcotics, presence of adverse events, and length of hospital stay were significantly less with the PCIP versus control (each p < 0.01). CONCLUSION: The use of a mix of levobupivacaina, ketoral-trometamina, and adrenalin provides a safe and effective means in post-operative pain relief in patients undergoing TKA. LEVEL OF EVIDENCE: Level II therapeutic study.

Resistance to the atypical retinoid ST1926 in SK-N-AS cells selected the subline rAS-ST with enhanced sensitivity to ATRA mediated by not conventional mechanisms: DNA damage, G2 accumulation and late telomerase inhibition.

BACKGROUND AND PURPOSE: 13-cis-Retinoic acid represents a well-established clinical strategy for the management of minimal residual disease of high risk neuroblastoma (NB) patients. However, the clinical efficacy on the overall survival of these patients remains limited, addressing the issue of better understanding the molecular mechanisms and intracellular pathways mediating Retinoic Acid (RA) clinical effects. EXPERIMENTAL APPROACH: This work investigates the mechanism underlying the sensitivity/resistance to RA in NB by taking advantage of the paired SK-N-AS/rAS-ST cells showing different responsivity to ATRA. The subline rAS-ST was selected by inducing resistance to the novel retinoid ST1926 in the NB SK-N-AS cell line. KEY RESULTS: Resistance to ST1926 was neither dependent on cellular uptake nor on multi-drug resistance phenotype. Rather, both delayed/lower DNA damage and apoptosis appeared involved in reduced sensitivity of rAS-ST cells to ST1926. This subline showed enhanced responsivity to ATRA compared to the wt counterpart, that was associated with enhanced RARα/ß expression, DNA damage, G2 accumulation, PI3K/AKT pathway inhibition, cellular differentiation and delayed telomerase inhibition, without involvement of either p27/p53 or caspase-mediated apoptosis. CONCLUSIONS AND IMPLICATIONS: The present data add important information to the understanding of RA sensitivity in NB, providing further insights towards a more efficacious clinical use of this drug.

Western blot expression of 5-lipoxygenase in the brain from striped dolphins (stenella coeruleoalba) and bottlenose dolphins (tursiops truncatus) with or without encephalitis/meningo-encephalitis of infectious nature.

Dolphin Morbillivirus (DMV), Toxoplasma gondii and Brucella ceti are pathogens of major concern for wild cetaceans. Although a more or less severe encephalitis/meningo-encephalitis may occur in striped dolphins (Stenella coeruleoalba) and bottlenose dolphins (Tursiops truncatus) infected by the aforementioned agents, almost no information is available on the neuropathogenesis of brain lesions, including the neuronal and non-neuronal cells targeted during infection, along with the mechanisms underlying neurodegeneration. We analyzed 5-lipoxygenase (5-LOX) expression in the brain of 11 striped dolphins and 5 bottlenose dolphins, affected or not by encephalitic lesions of various degrees associated with DMV, T. gondii and B. ceti. All the 8 striped dolphins with encephalitis showed a more consistent 5-LOX expression than that observed in the 3 striped dolphins showing no morphologic evidence of brain lesions, with the most prominent band intensity being detected in a B. ceti-infected animal. Similar results were not obtained in T. gondii-infected vs T. gondii-uninfected bottlenose dolphins. Overall, the higher 5-LOX expression found in the brain of the 8 striped dolphins with infectious neuroinflammation is of interest, given that 5-LOX is a putative marker for neurodegeneration in human patients and in experimental animal models. Therefore, further investigation on this challenging issue is also needed in stranded cetaceans affected by central neuropathies.

Prevalence of Simkania negevensis in chlorinated water from spa swimming pools and domestic supplies.

AIMS: This study aimed to investigate the prevalence of Simkania negevensis in the chlorinated water of spa swimming pools and domestic network systems. METHODS AND RESULTS: A total of 10 and 36 samples were collected from two domestic water supplies and four spa facilities, respectively. Simkania negevensis was isolated in Acanthamoeba castellanii monolayers and detected by Gimenez staining and immunofluorescence test. Simkania negevensis DNA, extracted from the positive samples, was confirmed by a nested PCR assay followed by sequencing. Simkania negevensis was detected in 41·3% of samples (domestic water: 50%; untreated spa supply water: 25%; chlorinated spa pool water: 42·9%) from all the examined water systems in successive samplings performed in 1 year. The presence of S. negevensis was not correlated with the counts of Heterotrophic Bacteria and Pseudomonas aeruginosa. Unlike Legionella spp., which were never isolated from the swimming-pool water samples, S. negevensis was also detected in chlorinated spa water. CONCLUSIONS: This investigation shows that Simkania is widespread in natural and man-made aquatic environments, which may represent possible sources of infection: in the swimming pools, in particular, the aerosol generated by the water movement could increase the risk of inhalation of infected particles. SIGNIFICANCE AND IMPACT OF THE STUDY: This study represents the first evidence of the presence of S. negevensis in spa pool water. Sero-epidemiological surveys on spa users could help to clarify its transmission in this environment.

Profesionales de la Salud y Formación Docente: Una propuesta de Curriculum integrado / Profissionais de saúde e formação de professores: uma proposta de currícula integrada / Health Professionals And Teacher Training: Integrated Syllabus Proposal

La presente investigación tiene el propósito de describir el proceso de formación en investigación-acción efectuada en la integración realizada por dos cátedras de la Carrera del Profesorado Universitario: Investigación educativa y Didáctica Especial producto de siete años de trabajo.Dicho proceso se sostiene a partir del reconocimiento de problemáticas planteadas por los alumnos-profesionales,en su trabajo como docentes. La construcción del problema de investigación requiere de diferentes estrategias pedagógicas. La metodología se basó en el enfoque de investigación-acción. Los resultados confirman que la integración inter cátedras un elemento clave para transformar el quehacer universitario y es una alternativa para fomentar y desarrollar la investigación científica que contribuya al mejoramiento y transformación de la sociedad, como uno de los objetivos de la Universidad Católica de Córdoba a la par que se constituye en una estrategia para el acuerdo, los consensos, la innovación e investigación que están en consonancia con la misión institucional...

The purpose of this research is to describe the training process in research-action that tookplace in the integration of Educational Research and Special Didactics; two chairs of thecourse of studies University Teaching, result of seven-year-work. Said process arises from theacknowledgment of issues proposed by students-professionals in their work as teachers. Theconstruction of the research problem requires different pedagogic strategies. Methodologywas based on the research-action approach. Results confi rm that inter-chair integrationis a key factor to transform university tasks and an alternative to encourage and developscientifi c research helping to improve and transform society as one of the objectives on theCatholic University of Cordoba. At the same time, it is becoming a strategy for agreement,consensus, innovation and investigation, consistent with the institutional mission...

First Report of Asiatic Brown Rot Caused by Monilinia polystroma on Peach in Italy.

Monilinia spp. are well-known pathogens causing brown rot of fruit trees in many fruit production areas worldwide. In Italy, three Monilinia species are particularly significant with regard to fruit trees, causing blossom and twig blight and brown rot in fruits: Monilinia laxa (Aderhold and Ruhland) Honey, M. fructicola (Winter) Honey, and M. fructigena (Aderhold and Ruhland). In 2009, a new species, M. polystroma, was distinguished from M. fructigena based on morphological and molecular characteristics in Europe (3). M. polystroma is not known to occur in Italy and to date has been reported from the Czech Republic (1), Hungary (3), Poland (4), Serbia (5), and Switzerland (2). In July 2013, during a survey for fungal postharvest pathogens, stored peaches (Prunus persica (L.) Batsch) belonging to different cultivars showing brown rot symptoms were observed in the Emilia Romagna and Sardinia regions of Italy. Typical decay spots were circular and brown, tending toward black, and 5% of peaches presented a large number of yellowish or buff-colored stromata and firm decayed tissues, the symptoms originated by M. polystroma. The pathogen was isolated on V8 agar (V8A) and culture plates were incubated at 25°C in darkness for 5 days. A conidial suspension was spread on malt extract agar (MEA) and single spores were selected. M. polystroma colonies grown on potato dexstrose agar (PDA) were yellowish in color. Irregular black stromatal crusts occurred on the edges of the colonies after 10 to 12 days of incubation and on the margin was present sporogenous tissue slightly elevated above the colony surface, color buff/pale luteous (1). The conidia were one-celled, ovoid or limoniform, smooth and hyaline, and 12 to 20 × 8 to 12 µm in distilled water when grown on V8A at 22°C. The ribosomal ITS1-5.8S-ITS2 region was PCR-amplified from genomic DNA obtained from mycelium using primers ITS1 and ITS4. A BLAST search in GenBank revealed the highest similarity (99%) to M. polystroma sequences (GenBank Accession No. GU067539). Pathogenicity was confirmed using surface-sterilized mature 'Red Heaven' peaches. The fruits were wounded (2 × 2 × 2 mm) twice with a sterile needle and inoculated with 2-mm plugs of 7-day-old mycelia from fungal colony margins. The sample unit was represented by 10 fruits. Control fruits were inoculated with PDA. After 7 days of incubation at 20°C in plastic containers with high humidity, typical symptoms of brown rot developed on both the wounds of all inoculated fruits, while control fruits remained symptomless. By the 14th day, all fruits had rotted and the yellowish exogenous stromata appeared on the surface of infected peaches. The fungus isolated from inoculated fruit exhibited the same morphological and molecular features of the original isolates; the molecular analysis performed using the primers by Petroczy (3) confirmed the result of the PCR with ITS1 and ITS4 primers. To our knowledge, this is the first report of M. polystroma on peach in Italy. This is relevant because the new pathogen could spread into other European countries that are main peach producers (such as Spain), causing economic losses. Bringing it to the attention of the scientific community allows the arrangement of research studies for assessing potential resistances with a significant impact on disease control management. Further studies are necessary to determine geographic distribution, prevalence, and economic importance of this organism in Italy. References: (1) EPPO Reporting Service. 2011/134: First reports of Monilinia polystroma in Hungary and the Czech Republic. No. 6, 2011. (2) M. Hilber-Bodmer et al. Plant Dis. 96:146, 2012. (3) M. Petroczy and L. Palkovics. Eur. J. Plant Pathol. 125:343, 2009. (4) A. Poniatowska et al. Eur. J. Plant Pathol. 135:855, 2013. (5) M. Vasic et al. Plant Dis. 97:145, 2013.

Pelvic sacral and hemi lumbar spine resection of low grade pelvic chondrosarcoma: a multistage procedure involving vascular bypass, spine fixation and vascular exclusion.

Peripheral chondrosarcoma is a rare tumor particularly insidious when arising from the pelvis, becoming symptomatic later in time when surgery may be too difficult and dangerous due to this complex area. In the present case, the tumor arose from an exostosis located on the medial surface of the left iliac wing. Its diameter was 25 cm × 20 cm × 15 cm, adhering to the last three vertebrae, involving the left iliac vein and artery, displacing the left ureter. In a similar case, a hindquarter amputation is indicated but, if the patient refuses, a resection remains possible. In this paper, we describe a multistage technique consisting of an extra-anatomic vascular bypass, a lumbar stabilization, a neurovascular bundles anterior isolation and a postero-lateral resection of this mass. After a five-year follow-up, the patient is alive and able to stand and walk with support, after undergoing twice lung metastasis removal.

A cyanoacrylate and silastic patch to reduce the risk of opening of the tumor: technical note.

BACKGROUND: Wide surgery is the main factor influencing survival in muscular skeletal tumor. Sometimes the margin can be very thin and the contamination risk can be very high because of manipulation of the mass. MATERIALS AND METHODS: A patch of cyanoacrylate and a silastic mesh are applied on tumor surface. In order to demonstrate the tumor sealing an histologic exam was performed. DISCUSSION: The application of protective patch can decrease the risk of accidental tumor rupture and neoplastic cells spreading.

Endocannabinoid signaling in Alzheimer’s disease: current knowledge and future directions.

The importance of the endocannabinoid system (ECS) in the modulation functions of the central nervous system has been extensively investigated during the last few years. In particular, accumulated evidence has implicated ECS in the pathophysiology of Alzheimer’s disease (AD), that is a progressive, degenerative, and irreversible disorder characterized by the accumulation in the brain of beta-amyloid fragments forming insoluble plaques, and of intracellular neurofibrillary tangles (NTFs) associated with synaptic and neuronal loss. In all the processes involved in the formation of both plaques and NFTs, the key-role played by the ECS has been documented. Here, we review current knowledge and future directions of ECS modulation both in animal models of AD and in human tissues, underlying the role of endocannabinoid signaling in the development of AD hallmarks. Overall, the available data suggest that next generation therapeutics might target distinct ECS elements, for instance CB2 receptor or fatty acid amide hydrolase, as a promising approach to halt or at least to slow down disease progression.