Bidimensional Tandem Mass Spectrometry for Selective Identification of Nitration Sites in Proteins.

Nitration of protein tyrosine residues is very often regarded as a molecular signal of peroxynitrite formation during development, oxidative stress, and aging. However, protein nitration might also have biological functions comparable to protein phosphorylation, mainly in redox signaling and in signal transduction. The major challenge in the proteomic analysis of nitroproteins is the need to discriminate modified proteins, usually occurring at substoichiometric levels from the large amount of nonmodified proteins. Moreover, precise localization of the nitration site is often required to fully describe the biological process. Existing methodologies essentially rely on immunochemical techniques either using 2D-PAGE fractionation in combination with western blot analyses or exploiting immunoaffinity procedures to selectively capture nitrated proteins. Here we report a totally new approach involving dansyl chloride labeling of the nitration sites that rely on the enormous potential of MSn analysis. The tryptic digest from the entire protein mixture is directly analyzed by MS on a linear ion trap mass spectrometer. Discrimination between nitro- and unmodified peptide is based on two selectivity criteria obtained by combining a precursor ion scan and an MS3 analysis. This new procedure was successfully applied to the identification of 3-nitrotyrosine residues in complex protein mixtures

Involvement of pro-inflammatory cytokines and growth factors in the pathogenesis of Dupuytren's contracture: a novel target for a possible future therapeutic strategy?

Dupuytren's contracture (DC) is a benign fibro-proliferative disease of the hand causing fibrotic nodules and fascial cords which determine debilitating contracture and deformities of fingers and hands. The present study was designed to characterize pro-inflammatory cytokines and growth factors involved in the pathogenesis, progression and recurrence of this disease, in order to find novel targets for alternative therapies and strategies in controlling DC. The expression of pro-inflammatory cytokines and of growth factors was detected by immunohistochemistry in fibrotic nodules and normal palmar fascia resected respectively from patients affected by DC and carpal tunnel syndrome (CTS; as negative controls). Reverse transcription (RT)-PCR analysis and immunofluorescence were performed to quantify the expression of transforming growth factor (TGF)-β1, interleukin (IL)-1β and vascular endothelial growth factor (VEGF) by primary cultures of myofibroblasts and fibroblasts isolated from Dupuytren's nodules. Histological analysis showed high cellularity and high proliferation rate in Dupuytren's tissue, together with the presence of myofibroblastic isotypes; immunohistochemical staining for macrophages was completely negative. In addition, a strong expression of TGF-β1, IL-1β and VEGF was evident in the extracellular matrix and in the cytoplasm of fibroblasts and myofibroblasts in Dupuytren's nodular tissues, as compared with control tissues. These results were confirmed by RT-PCR and by immunofluorescence in pathological and normal primary cell cultures. These preliminary observations suggest that TGF-β1, IL-1β and VEGF may be considered potential therapeutic targets in the treatment of Dupuytren's disease (DD)

Real time detection of cache-based side-channel attacks using hardware performance counters

Cache-based side-channel attacks are increasingly exposing the weaknesses of many cryptographic libraries and tools by showing that, even though the algorithms might be considered strong, their implementations often lead to unexpected behaviors that can be exploited to obtain sensitive data, usually encryption keys. In this study we analyze three methods to detect cache-based side-channel attacks in real time, preventing or limiting the amount of leaked information. We focus our efforts on detecting three attacks on the well-known OpenSSL library: one that targets AES, one that targets RSA and one that targets ECDSA. The first method is based on monitoring the involved processes and assumes the victim process is known. By collecting and correlating the monitored data we find out whether there exists an attacker and pinpoint it. The second method uses anomaly detection techniques and assumes the benign processes and their behavior are known. By treating the attacker as a potential anomaly we understand whether an attack is in progress and which process is performing it. The last method is based on employing a neural network, a machine learning technique, to profile the attacker and to be able to recognize when a process that behaves suspiciously like the attacker is running. All the three of them can successfully detect an attack in about one fifth of the time required to complete it. We could not experience the presence of false positives in our test environment and the overhead caused by the detection systems is negligible. We also analyze how the detection systems behave with a modified version of one ofthe spy processes. With some optimization we are confident these systems can be used in real world scenarios

What determines the complication rate in high-risk chronic obstructive pulmonary disease patients: surgery or pulmonary function?

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HIV-1 Tropism Test Evaluation: Assessment and Clinical Implications

CCR5 and CXCR4 chemokines receptors are critical coreceptors for the binding of HIV to specific host cells. Guidelines recommend its assessment in case of virological failure or before prescription of CCR5 inhibitors. Strategies to assess viral tropism may be divided into phenotypic and genotypic assays; registrative trials of CCR5 inhibitors used phenotypic assay, but recently genotypic ones have been used in clinical practice. The presence of CXCR4 is increasing in naïve patients, with both acute and chronic HIV-1 infections; this coreceptor usage is associated with CD4 depletion. The assessment of viral tropism should be considered in every stage of HIV-1 infection

High incidence of vascular complication after computed tomography guided lung biopsy : what's matter , the patient or the tecnique?

We read with interest the article of Anzidei on a predicitive score of complication after lung computed tomography -guided biopsy . We think there is a percentage of complications especially pneumotohorax which are impossible to avoid , but it would be useful to predict in which cases they occur with hiogh frequency

Is Post-Operative Lung Ultrasonography Effective in Lung Abnormalities Analysis?

We read with interest the article of Dr. Patella (1) about the use of chest ultrasonography after thoracic surgery for pneumothorax evaluation and management. We completely agree about the use of ultrasound in the post-operative evaluation with the possibility to reduce the number of chest X-ray (CXR). In fact, it is well known that in many cases the postoperative CXR, especially when daily performed, represents a possible waste of time and resources(2)

SICOB-endorsed national Delphi consensus on obesity treatment optimization: focus on diagnosis, pre-operative management, and weight regain/insufficient weight loss approach

Purpose: Overweight and obesity affects 60% of adults causing more than 1.2 million deaths across world every year. Fight against involved different specialist figures and multiple are the approved weapons. Aim of the present survey endorsed by the Italian Society of Bariatric Surgery (SICOB) is to reach a national consensus on obesity treatment optimization through a Delphi process. Methods: Eleven key opinion leaders (KOLs) identified 22 statements with a major need of clarification and debate. The explored pathways were: (1) Management of patient candidate to bariatric/metabolic surgery (BMS); (2) Management of patient not eligible for BMS; (3) Management of patient with short-term (2 years) weight regain (WR) or insufficient weight loss (IWL); (4) Management of the patient with medium-term (5 years) WR; and (5) Association between drugs and BMS as WR prevention. The questionnaire was distributed to 65 national experts via an online platform with anonymized results. Results: 54 out of 65 invited panelists (83%) respond. Positive consensus was reached for 18/22 statements (82%); while, negative consensus (s20.4; s21.5) and no consensus (s11.5, s17) were reached for 2 statements, respectively (9%). Conclusion: The Delphi results underline the importance of first-line interdisciplinary management, with large pre-treatment examination, and establish a common opinion on how to properly manage post-operative IWL/WR. Level of evidence v: Report of expert committees