Surfaceome profiling enables isolation of cancer-specific exosomal cargo in liquid biopsies from pancreatic cancer patients.

Background: Detection of circulating tumor DNA can be limited due to their relative scarcity in circulation, particularly while patients are actively undergoing therapy. Exosomes provide a vehicle through which cancer-specific material can be enriched from the compendium of circulating non-neoplastic tissue-derived nucleic acids. We carried out a comprehensive profiling of the pancreatic ductal adenocarcinoma (PDAC) exosomal 'surfaceome' in order to identify surface proteins that will render liquid biopsies amenable to cancer-derived exosome enrichment for downstream molecular profiling. Patients and methods: Surface exosomal proteins were profiled in 13 human PDAC and 2 non-neoplastic cell lines by liquid chromatography-mass spectrometry. A total of 173 prospectively collected blood samples from 103 PDAC patients underwent exosome isolation. Droplet digital PCR was used on 74 patients (136 total exosome samples) to determine baseline KRAS mutation call rates while patients were on therapy. PDAC-specific exosome capture was then carried out on additional 29 patients (37 samples) using an antibody cocktail directed against selected proteins, followed by droplet digital PCR analysis. Exosomal DNA in a PDAC patient resistant to therapy were profiled using a molecular barcoded, targeted sequencing panel to determine the utility of enriched nucleic acid material for comprehensive molecular analysis. Results: Proteomic analysis of the exosome 'surfaceome' revealed multiple PDAC-specific biomarker candidates: CLDN4, EPCAM, CD151, LGALS3BP, HIST2H2BE, and HIST2H2BF. KRAS mutations in total exosomes were detected in 44.1% of patients undergoing active therapy compared with 73.0% following exosome capture using the selected biomarkers. Enrichment of exosomal cargo was amenable to molecular profiling, elucidating a putative mechanism of resistance to PARP inhibitor therapy in a patient harboring a BRCA2 mutation. Conclusion: Exosomes provide unique opportunities in the context of liquid biopsies for enrichment of tumor-specific material in circulation. We present a comprehensive surfaceome characterization of PDAC exosomes which allows for capture and molecular profiling of tumor-derived DNA.

Another possible risk for the Mediterranean Sea? Aspergillus sydowii discovered in the Port of Genoa (Ligurian Sea, Italy).

Aspergillus sydowii is a cosmopolitan fungus that has been responsible for the mass destruction of coral in the Caribbean Sea over the last 15years. To our knowledge, this study has found the first case of A. sydowii in the Mediterranean Sea, in marine-bottom sediments, water and calcareous shells of bivalve molluscs sampled during a campaign to characterise the mycobiota in the Port of Genoa (Italy). The area is characterised by adverse environmental conditions (high turbidity, organic pollution and high concentrations of phosphorus and nitrogen compounds). These parameters, in combination with a rising temperature, could contribute to A. sydowii bloom and dispersal. This fungal strain may have been imported into the Port of Genoa in the bilge water of vessels or by torrent input. This work represents the first step in the implementation of a monitoring programme to safeguard calcareous sponges and sea fan corals endemic in the Mediterranean Sea.

Environmental complexity of a port: Evidence from circulation of the water masses, and composition and contamination of bottom sediments.

Ports are complex environments due to their complicated geometry (quays, channels, and piers), the presence of human activities (vessel traffic, shipyards, industries, and discharges), and natural factors (stream and torrent inputs, sea action, and currents). Taking these factors into consideration, we have examined the marine environment of a port from the point of view of the circulation of the water masses, hydrological characteristics, distribution of the sediment grain-size, mineralogical characteristics, and metal concentrations of the bottom sediments. Our results show that, in the case of the Port of Genoa (north-western Italy), the impact of human activities (such as a coal power-plant, oil depots, shipyards, dredging of the bottom sediments, etc.), natural processes (such as currents, fresh water and sediment inputs from the torrents), and the morphology of the basin, are important factors in the sediment, water, and metal distributions that have given rise to a complex environment.

A mycological baseline study based on a multidisciplinary approach in a coastal area affected by contaminated torrent input.

Fungi include a vast group of eukaryotic organisms able to colonise different natural, anthropised and extreme environments, including marine areas contaminated by metals. The present study aims to give a first multidisciplinary characterisation of marine bottom sediments contaminated by metals (Cd, Co, Cr, Cu, Ni, and Zn), originating in the water leakage from an abandoned Fe-Cu sulphide mine (Libiola, north-western Italy), and evaluate how the chemical and physical parameters of water and sediments may affect the benthic fungal communities. Our preliminary results showed the high mycodiversity of the marine sediments studied (13 genera and 23 species of marine fungi isolated), and the great physiological adaptability that this mycobiota evolved in reaction to the effects of the ecotoxic bottom sediment contamination, and associated changes in the seawater parameters.

Hydrogen bonds vs. π-stacking interactions in the p-aminophenolp-cresol dimer: an experimental and theoretical study.

The gas phase structure and excited state lifetime of the p-aminophenolp-cresol heterodimer have been investigated by REMPI and LIF spectroscopy with nanosecond laser pulses and pump-probe experiments with picosecond laser pulses as a model system to study the competition between π-π and H-bonding interactions in aromatic dimers. The excitation is a broad and unstructured band. The excited state of the heterodimer is long lived (2.5 ± 0.5) ns with a very broad fluorescence spectrum red-shifted by 4000 cm-1 with respect to the excitation spectrum. Calculations at the MP2/RI-CC2 and DFT-ωB97X-D levels indicate that hydrogen-bonded (HB) and π-stacked isomers are almost isoenergetic in the ground state while in the excited state only the π-stacked isomer exists. This suggests that the HB isomer cannot be excited due to negligible Franck-Condon factors and therefore the excitation spectrum is associated with the π-stacked isomer that reaches vibrationally excited states in the S1 state upon vertical excitation. The excited state structure is an exciplex responsible for the fluorescence of the complex. Finally, a comparison was performed between the π-stacked structure observed for the p-aminophenolp-cresol heterodimer and the HB structure reported for the (p-cresol)2 homodimer indicating that the differences are due to different optical properties (oscillator strengths and Franck-Condon factors) of the isomers of both dimers and not to the interactions involved in the ground state.

Population assessment of tropical tuna based on their associative behavior around floating objects.

Estimating the abundance of pelagic fish species is a challenging task, due to their vast and remote habitat. Despite the development of satellite, archival and acoustic tagging techniques that allow the tracking of marine animals in their natural environments, these technologies have so far been underutilized in developing abundance estimations. We developed a new method for estimating the abundance of tropical tuna that employs these technologies and exploits the aggregative behavior of tuna around floating objects (FADs). We provided estimates of abundance indices based on a simulated set of tagged fish and studied the sensitivity of our method to different association dynamics, FAD numbers, population sizes and heterogeneities of the FAD-array. Taking the case study of yellowfin tuna (Thunnus albacares) acoustically-tagged in Hawaii, we implemented our approach on field data and derived for the first time the ratio between the associated and the total population. With more extensive and long-term monitoring of FAD-associated tunas and good estimates of the numbers of fish at FADs, our method could provide fisheries-independent estimates of populations of tropical tuna. The same approach can be applied to obtain population assessments for any marine and terrestrial species that display associative behavior and from which behavioral data have been acquired using acoustic, archival or satellite tags.

Marine sediment contamination and dynamics at the mouth of a contaminated torrent: The case of the Gromolo Torrent (Sestri Levante, north-western Italy).

In this study we have examined the currents and hydrological characteristics of the water column off the mouth of the Gromolo Torrent (north-western Italy) in relation to the grain-size, mineralogical characteristics and metal distribution in the marine sediment sampled. Our purpose was to quantify and map the contamination that was carried out to sea from the abandoned Libiola Fe-Cu sulphide mine that has heavily impacted the torrent. Our results show high concentrations of Cu and Zn, and relatively high concentrations of Cd and Ni inside the bay into which the Gromolo Torrent flows. However, high concentrations of As, Cr, Hg, Mn, Pb, and V found in the northern and/or eastern parts of the study area originated from other sources. The subdivision of study stations in terms of metal and mineral contents in the bottom sediments highlighted the clear influence of the currents on their dispersion and distribution in the area.

Evaluation of the boundary condition influence on PAH concentrations in the water column during the sediment dredging of a port.

The mobilisation of sediments and related contaminants connected to dredging activities is one of the most critical issues to the environmental risk and exposure assessment of a dredging project. The aim of this paper was an investigation of the mobilisation of polycyclic aromatic hydrocarbons (PAHs) due to the dredging of the Port of Genoa (Italy) to identify the temporal and spatial extent of the contaminant transport, and the influence of the dredging and the boundary conditions on it. The results showed relatively low background PAH concentrations in the water column and confirmed the dredging as the primary rising factor of concentrations in the water column, but also showed a complex scenario in which the different environmental and dredging factors forced the concentrations at different levels and moments. The post dredging phase showed PAH values close to the background conditions and the concentrations remained relatively high only for a few PAHs.

Simulations of dredged sediment spreading on a Posidonia oceanica meadow off the Ligurian coast, Northwestern Mediterranean.

The sandy deposits from dredging can have negative effects on the environment such as increase in suspended solids in the water column and their consequent transport. An experimental study was conducted to characterize water masses, dynamics, and sedimentation rates on the Ligurian continental shelf (Italy), where both a sand deposit, that could be used for beach nourishment, and a nearby Posidonia oceanica meadow coexist. The environmental plan provides a mathematical simulation of the sediment-dispersion to evaluate the possible impact on the meadow. It has been calculated that the dredging could double the concentration of suspended particles, but its scheduling will preclude a sediment accumulation. All the information obtained from this work will be used to study the environmental feasibility of the sand deposit exploitation and as starting point for drawing up the monitoring plan in case of dredging.

Seasonal and spatial variability of water quality parameters in the Port of Genoa, Italy, from 2000 to 2007.

This paper presents the results of the statistical analysis of a set of physico-chemical and biological water quality parameters, monthly collected from 2000 to 2007 in the Genoa Harbour area (Ligurian Sea). We applied multivariate methods, such as principal component analysis (PCA) and dynamic factor analysis (DFA) for investigating the spatial and temporal variability and for providing important background information on pollution problems in the region. PCA evidenced the role of the sewage and river discharges and of the exchanges with the open sea in determining the harbour water quality. DFA was used to estimate underlying common trends in the time series. The DFA results partly show a general improvement of water quality over the 8-years period. However, in other areas, we found inter-annual variations but no significant multi-annual trend. Furthermore, we included meteorological variables in our statistical analyses because of their potential influence on the water quality parameters. These natural forcings explain part of the variability in water quality parameters that are superimposed on the dominating anthropogenic pollution factors.

Consensus: the follow-up of the treated patient.

This is the first consensus document on the follow-up of the treated patient with non-small cell lung cancer that has been written by this group. The document has been drawn up by doctors coming from many different cultures and philosophical backgrounds. It acknowledges that there are published guidelines on the follow-up particularly those in trials, and does not wish to contradict these. There is lack of evidence-based medicine to recommend a strong general policy in this area. For those patients who were treated with curative intent the initial follow-up will depend upon the toxicity that is evident from the treatment given. Thereafter the interval between follow-up visits should be every 3 months for the first two years, then every 6 months for up to five years. Rapid and easy access to the multidisciplinary team should be available. Full examination and chest X-ray should be carried out on each visit but other investigations should be determined by clinical need. For those patients treated with palliative intent the interval between follow-up visits once the acute reactions have settled will depend upon the adequacy of the control of the symptom and the availability of separate palliative care teams. At all times the patient should have rapid access to the multidisciplinary team and in general frequent follow-up, that is at intervals of one to two months, may be appropriate during the first six months. Follow-up constitutes an important part of lung cancer management. Efforts should be made to gain clinical material to give us evidence-based guidelines.

Undiagnosed rupture of right hemidiaphragm--hepatothorax: a case report.

A case of late diagnosis of right diaphragmatic rupture due to blunt trauma is presented. Traumatic diaphragmatic rupture is an uncommon but severe problem in a patient with multiple injuries. One third of left-sided ruptures and half of right-sided ruptures have been undetected for several days. Traumatic diaphragmatic rupture should be suspected on the basis of an abnormal chest X-ray in patients with multiple injuries. Imaging studies like computed tomography, scan and magnetic resonance imaging can be helpful for differential diagnosis. Right-sided injuries occur more commonly than previously thought and often require thoracotomy.

[Ludwig's angina]. / Angine de Ludwig.

Ludwig's angina is a rare infectious condition originating in the oropharyngeal region, most commonly from the teeth. The principal symptoms consist of cervical pain, dyspnea, dysphagia, symmetrical neck swelling and fever. Serious complications include respiratory collapses, mediastinitis, pleural empyema, pericarditis and pericardial tamponade, and may result in the death of the patient. The most useful investigations in patients with suspected Ludwig's angina are clinical assessment, a cervico-thoracic CT-scan to determine the extent of the lesions, microbiological examination and panoramic radiography to detect possible dental focuses of infection. Treatment consists of ensuring adequate ventilation by securing the upper airways, broad spectrum antibiotic therapy, eradication of the source of infection, and often early surgical decompression may be necessary. Rapid diagnosis and appropriate treatment can reduce the mortality rate to 10%.

Estudio fenotípico familiar en el déficit de Alfa 1 Antitripsina / Familiar phenotypic study in Alpha 1 antitrypsin deficiency

El déficit de alfa 1 antitripsina (DA1AT), es una enfermedad hereditaria autosómica de expresión condominante, con una provalencia aproximada de 1 caso cada 3000 individuos. Fenotipos anormales de DA1AT particularmente el PiZ se asocian con el riesgo de padecer patología hepática en la infancia y enfisema pulmonar en la edad adulta. Las manifestaciones y el pronóstico en esta enfermedad hepática hereditaria es extremadamente variable. Se presenta un caso de DA1AT con signos de colestasis neonatal

Frequency of upper gastrointestinal lesions in patients with liver cirrhosis.

The frequency of gastroduodenal lesions has been investigated in 142 patients with liver cirrhosis of various degrees of severity and in 63 patients with mild liver disease (controls) in whom liver biopsy excluded nodular regeneration. Cirrhotic patients were subdivided in three groups according to the Pugh modification of the Child-Turcotte criteria. Although the frequency of peptic ulcer was not different, gastroduodenal erosions were observed more frequently in cirrhotics than in controls (29.6% vs 11.1%, P less than 0.01). The occurrence of erosions was related to the severity of the disease: in Child A and B patients their frequency was 21 and 26% respectively, but rose to 48.4 (15 of 31 vs 7 of 63 in controls, P less than 0.001) in the Child C group. Both mild and severe gastroduodenitis occurred more frequently, although not significantly, in patients with liver cirrhosis. All together one or more endoscopic lesions were observed in almost 60% of cirrhotics but only in 25.4% of controls (P less than 0.001). In conclusion, our data do not show an increased prevalence of peptic ulcer in cirrhotic patients; in contrast, liver cirrhosis is significantly associated with the endoscopic finding of gastroduodenal erosions, especially in the more advanced stages of the disease. These findings would suggest a cautious use, in cirrhotic patients, of drugs which may damage the gastroduodenal mucosa; moreover, long-term administration of antacids or of other drugs with a protective effect on gastroduodenal mucosa might be taken into consideration for Child C patients.