Search

Cardiovascular diseases represent one of the most frequent diseases worldwide; among these, lower limb ischemia is a threatening condition, which can lead to permanent disability if not promptly and correctly diagnosed and treated. A patient’s clinical evaluation and diagnostic imaging (e.g., color-Doppler ultrasound, computed tomography angiography (CTA), and magnetic resonance imaging (MRI)) are mandatory to carefully assess arterial lesion extension and severity. Digital subtraction angiography (DSA) is a minimally invasive technique that represents the gold standard for percutaneous revascularization treatment of symptomatic patients who are refractory to medical management. However, when dealing with patients with lower limb terminal ischemia, the correct interpretation of diagnostic DSA findings is mandatory for treatment re-planning and to effectively evaluate post-treatment results and complications. The purpose of this review is to provide interventional radiologists and endovascular practitioners with an up-to-date practical guide to diagnostic angiography of the lower limbs, which is mandatory to address correct treatment decisions and post-treatment evaluation.

Gallbladder percutaneous tissue sampling is a not-so-common technique in cytohistological diagnosis of gallbladder tissue or masses, which can be useful in cases of surgically unresectable disease and unfeasible endoscopic assessment to address the most adequate chemotherapy course. Nonetheless, gallbladder percutaneous tissue sampling can be of great utility in the patient’s diagnostic and therapeutic work-up. This article summarizes the literature evidence on gallbladder biopsy techniques, complications, and technical precautions for a safe and effective sampling.

Background: More effective strategies are needed to promote poststroke functional recovery. Here, we evaluated the impact of bihemispheric transcranial direct current stimulation (tDCS) on forelimb motor function recovery and the underlying mechanisms in mice subjected to focal ischemia of the motor cortex. Methods: Photothrombotic stroke was induced in the forelimb brain motor area, and tDCS was applied once per day for 3 consecutive days, starting 72 hours after stroke. Grid-walking, single pellet reaching, and grip strength tests were conducted to assess motor function. Local field potentials were recorded to evaluate brain connectivity. Western immunoblotting, ELISA, quantitative real-time polymerase chain reaction, and Golgi-Cox staining were used to uncover tDCS-mediated stroke recovery mechanisms. Results: Among our results, tDCS increased the rate of motor recovery, anticipating it at the early subacute stage. In this window, tDCS enhanced BDNF (brain-derived neurotrophic factor) expression and dendritic spine density in the peri-infarct motor cortex, along with increasing functional connectivity between motor and somatosensory cortices. Treatment with the BDNF TrkB (tropomyosin-related tyrosine kinase B) receptor inhibitor, ANA-12, prevented tDCS effects on motor recovery and connectivity as well as the increase of spine density, pERK (phosphorylated extracellular signal-regulated kinase), pCaMKII (phosphorylated calcium/calmodulin-dependent protein kinase II), pMEF (phosphorylated myocyte-enhancer factor), and PSD (postsynaptic density)-95. The tDCS-promoted rescue was paralleled by enhanced plasma BDNF level, suggesting its potential role as circulating prognostic biomarker. Conclusions: The rate of motor recovery is accelerated by tDCS applied in the subacute phase of stroke. Anticipation of motor recovery via vicariate pathways or neural reserve recruitment would potentially enhance the efficacy of standard treatments, such as physical therapy, which is often delayed to a later stage when plastic responses are progressively lower.

In this retrospective study we share our single-center experience using a magnetic seed for the preoperative localization of non-palpable breast lesions. Patients who underwent a preoperative localization with Magseed® (Endomagnetics, Cambridge, UK) placement between 2020 and 2022 were enrolled. Indications to Magseed placement have been established during multidisciplinary meetings prior to surgery and all patients underwent breast-conserving surgery (BCS). 45 patients were included. Magnetic seeds have been introduced under ultrasound guidance in 40 patients (88.9%) and under stereotactic guidance in 5 patients (11.1%). We registered a highly successful placement rate (97.8%), with only one case of migration (2.2%). After BCS, all the magnetic seeds were recovered (100% retrieval rate). The re-excision rate for positive margins was 0%. Our experience, with a highly successful placement and retrieval rate and a re-excision rate equal to 0%, is consistent with the encouraging literature published on Magseed so far, suggesting this technique to be extremely effective. Moreover, our single case of seed migration supports the existing data stating that Magseed migration is rare. In conclusion, despite acknowledging Magseed limitations, we highly value the advantages linked to this technique, and we, therefore, uphold its use.

Targeted axillary dissection (TAD) is an axillary staging technique after NACT that involves the removal of biopsy-proven metastatic lymph nodes in addition to sentinel lymph node biopsy (SLNB). This technique avoids the morbidity of traditional axillary lymph node dissection and has shown a lower false-negative rate than SLNB alone. Therefore, marking positive axillary lymph nodes before NACT is critical in order to locate and remove them in the subsequent surgery. Current localization methods include clip placement with intraoperative ultrasound, carbon-suspension liquids, localization wires, radioactive tracer-based localizers, magnetic seeds, radar reflectors, and radiofrequency identification devices. The aim of this paper is to illustrate the management of axillary lymph nodes based on current guidelines and explain the features of axillary lymph node markers, with relative advantages and disadvantages.

Consistent body of evidence shows that transcranial direct-current stimulation (tDCS) over the primary motor cortex (M1) facilitates motor learning and promotes recovery after stroke. However, the knowledge of molecular mechanisms behind tDCS effects needs to be deepened for a more rational use of this technique in clinical settings. Here we characterized the effects of anodal tDCS of M1, focusing on its impact on glutamatergic synaptic transmission and plasticity. Mice subjected to tDCS displayed increased long-term potentiation (LTP) and enhanced basal synaptic transmission at layer II/III horizontal connections. They performed better than sham-stimulated mice in the single-pellet reaching task and exhibited increased forelimb strength. Dendritic spine density of layer II/III pyramidal neurons was also increased by tDCS. At molecular level, tDCS enhanced: 1) BDNF expression, 2) phosphorylation of CREB, CaMKII, and GluA1, and 3) S-nitrosylation of GluA1 and HDAC2. Blockade of nitric oxide synthesis by L-NAME prevented the tDCS-induced enhancement of GluA1 phosphorylation at Ser831 and BDNF levels, as well as of miniature excitatory postsynaptic current (mEPSC) frequency, LTP and reaching performance. Collectively, these findings demonstrate that anodal tDCS engages plasticity mechanisms in the M1 and highlight a role for nitric oxide (NO) as a novel mediator of tDCS effects.

Although association between hearing impairment and dementia has been widely documented by epidemiological studies, the role of auditory sensory deprivation in cognitive decline remains to be fully understood. To address this issue we investigated the impact of hearing loss on the onset and time-course of cognitive decline in an animal model of Alzheimer’s disease (AD), that is the 3×Tg-AD mice and the underlying mechanisms. We found that hearing loss induced by noise exposure in the 3×Tg-AD mice before the phenotype is manifested caused persistent synaptic and morphological alterations in the auditory cortex. This was associated with earlier hippocampal dysfunction, increased tau phosphorylation, neuroinflammation, and redox imbalance, along with anticipated memory deficits compared to the expected time-course of the neurodegenerative phenotype. Our data suggest that a mouse model of AD is more vulnerable to central damage induced by hearing loss and shows reduced ability to counteract noise-induced detrimental effects, which accelerates the neurodegenerative disease onset.

Rheumatoid arthritis (RA) flare is related to increased joint damage, disability, and healthcare use. The impact of short-term air pollution exposure on RA disease activity is still a matter of debate. In this cross-sectional study, we investigated whether short-term exposure to particulate matter (PM)10, PM2.5, nitrogen dioxide (NO2), and ozone (O3) affected RA disease activity (DAS28 and SDAI) in 422 consecutive RA residents in Lombardy, North of Italy. Air pollutant concentrations, estimated by Regional Environmental Protection Agency (Lombardy—Italy) at the municipality level, were used to assign short-term exposure from the day of enrolment, back to seven days. Some significant negative associations emerged between RA disease activity, PM10, and NO2, whereas some positive associations were observed for O3. Patients were also stratified according to their ongoing Disease-Modifying anti-Rheumatic Drugs (DMARDs) treatment: no DMARDs (n = 25), conventional synthetic DMARDs (n = 108), and biological or targeted synthetic DMARDs (n = 289). Therapy interaction seemed partially able to influence the relationship between short-term air pollution exposure and RA disease activity (PM2.5 levels and DAS28 at the day of the visit-O3 levels and disease activity scores for the seven days before the evaluation). According to our results, the impact of short-term air pollution exposure (seven days) minimally impacts disease activity. Moreover, our study suggests therapy could alter the response to environmental factors. Further evidence is needed to elucidate determinants of RA flare and its management.

The Valley of Sacco River (VSR) (Latium, Italy) is an area with large-scale industrial chemical production that has led over time to significant contamination of soil and groundwater with various industrial pollutants, such as organic pesticides, dioxins, organic solvents, heavy metals, and particularly, volatile organic compounds (VOCs). In the present study, we investigated the potential impact of VOCs on the spermatozoa of healthy young males living in the VSR, given the prevalent presence of several VOCs in the semen of these individuals. To accomplish this, spermiograms were conducted followed by molecular analyses to assess the content of sperm nuclear basic proteins (SNBPs) in addition to the protamine-histone ratio and DNA binding of these proteins. We found drastic alterations in the spermatozoa of these young males living in the VSR. Alterations were seen in sperm morphology, sperm motility, sperm count, and protamine/histone ratios, and included significant reductions in SNBP–DNA binding capacity. Our results provide preliminary indications of a possible correlation between the observed alterations and the presence of specific VOCs.

Environmental chemicals and contaminants coming from multiple external sources enter the human body, determining a potential risk for human health. Human biomonitoring (HBM), measuring the concentrations of biomarkers in human specimens, has become an emerging approach for assessing population-wide exposure to hazardous chemicals and health risk through large-scale studies in many countries. However, systematic mapping of HBM studies, including their characteristics, targeted hazardous pollutants, analytical techniques, and sample population (general population and occupationally exposed workers), has not been done so far. We conducted a systematic review of the literature related to airborne hazardous pollutants in biofluids to answer the following questions: Which main chemicals have been included in the literature, which bodily fluids have been used, and what are the main findings? Following PRISMA protocol, we summarized the publications published up to 4 February 2021 of studies based on two methods: gas-chromatography/mass spectrometry (GC/MS) and electronic noses (e-noses). We screened 2606 records and 117 publications were included in the analysis, the most based on GC/MS analysis. The selected HBM studies include measurements of biomarkers in different bodily fluids, such as blood, urine, breast milk, and human semen as well as exhaled air. The papers cover numerous airborne hazardous pollutants that we grouped in chemical classes; a lot of hazardous and noxious compounds, mainly persistent organic pollutants (POPs) and volatile organic compounds (VOCs), have been detected in biological fluids at alarming levels. The scenario that emerged from this survey demonstrates the importance of HBM in human exposure to hazardous pollutants and the need to use it as valid tool in health surveillance. This systematic review represents a starting point for researchers who focus on the world of pollutant biomonitoring in the human body and gives them important insights into how to improve the methods based on GC/MS. Moreover, it makes a first overview of the use of gas sensor array and e-noses in HBM studies.

PURPOSE: To compare peripapillary vessel density (VD) measured by spectral domain optical coherence tomography angiography (SD-OCT-A) with morphometric parameters assessing the neuroretinal rim based on Bruch’s membrane opening (BMO) by spectral domain optical coherence tomography (SD-OCT) in glaucoma. METHODS: In this non-interventional cohort study, 50 eyes of 25 consecutively enrolled patients with diagnosis of glaucoma underwent SD-OCT and SD-OCT-A imaging of the optic nerve head (ONH). BMO minimum rim width (BMO-MRW) and area (BMO-MRA) as well as peripapillary retinal nerve fiber layer (RNFL) thickness were compared to peripapillary VD in the RNFL layer around the ONH. RESULTS: Mean BMO-MRW was 221.46 ± 81.5 µm, mean BMO-MRA was 1.05 ± 0.04 mm(2), mean RNFL thickness was 72.46 ± 23.16 µm, and mean VD was 43.8 ± 11.4%. VD was significantly lower when morphometric parameters had lower thickness values (p 90 µm) showed less VD variance (σ(2) = 48.1) compared to eyes with highly reduced RNFL thickness (

Two areas in central-southern Italy Land of Fires in Campania and Valley of Sacco river in Lazio are known to be contaminated sites, the first due to illegal fly-tipping and toxic fires, and the second due to an intensive industrial exploitation done by no-scruple companies and crooked public administration offices with dramatic consequences for environment and resident people. The work is intended to contribute to Human BioMonitoring (HBM) studies conducted in these areas on healthy young male population by a semiconductor gas sensor array trained by SPME-GC/MS. Human semen, blood and urine were investigated. The fingerprinting of the Volatile Organic Compounds (VOCs) by a gas sensors system allowed to discriminate the different contamination of the two areas and was able to predict the chemical concentration of several VOCs identified by GC/MS.

Volatile Organic Compound (VOC) analysis is usually applied in pollution assessment by checking for toxic or harmful volatile compounds in air, water and soil samples. In this study, exogenous VOCs and their derivatives, metabolized by cells, were valued into specific body fluids. In particular, the VOC profiles of blood, urine and human semen samples collected from young men living in two high pollution areas in Italy, i.e. Land of Fires and Valley of Sacco River, were fingerprinted. The analysis is based on Headspace Solid Phase MicroExtraction (HS-SPME) followed by Gas Chromatography-Mass Spectrometric detection (GC-MS). The volatile composition of the three body fluids showed that some VOCs are in common between blood, urine and human semen samples, whereas others are present only in a body fluid. Some compounds, as well as also some chemical classes show a higher affinity for a specific body fluid. Statistical analysis allowed to discriminate the two contaminated areas and identify those compounds which significantly contribute to the two areas classification. Some of these compounds are toxic and found prevalently in Valley of Sacco River samples, correspondingly to sperm analysis results for young men living in this zona worse than those living in Land of Fires.

Objectives To evaluate the strength of association between maternal and pregnancy characteristics and the risk of adverse perinatal outcomes in pregnancies with laboratory confirmed COVID-19. Methods Secondary analysis of a multinational, cohort study on all consecutive pregnant women with laboratory-confirmed COVID-19 from February 1, 2020 to April 30, 2020 from 73 centers from 22 different countries. A confirmed case of COVID-19 was defined as a positive result on real-time reverse-transcriptase-polymerase-chain-reaction (RT-PCR) assay of nasal and pharyngeal swab specimens. The primary outcome was a composite adverse fetal outcome, defined as the presence of either abortion (pregnancy loss before 22 weeks of gestations), stillbirth (intrauterine fetal death after 22 weeks of gestation), neonatal death (death of a live-born infant within the first 28 days of life), and perinatal death (either stillbirth or neonatal death). Logistic regression analysis was performed to evaluate parameters independently associated with the primary outcome. Logistic regression was reported as odds ratio (OR) with 95% confidence interval (CI). Results Mean gestational age at diagnosis was 30.6±9.5 weeks, with 8.0% of women being diagnosed in the first, 22.2% in the second and 69.8% in the third trimester of pregnancy. There were six miscarriage (2.3%), six intrauterine device (IUD) (2.3) and 5 (2.0%) neonatal deaths, with an overall rate of perinatal death of 4.2% (11/265), thus resulting into 17 cases experiencing and 226 not experiencing composite adverse fetal outcome. Neither stillbirths nor neonatal deaths had congenital anomalies found at antenatal or postnatal evaluation. Furthermore, none of the cases experiencing IUD had signs of impending demise at arterial or venous Doppler. Neonatal deaths were all considered as prematurity-related adverse events. Of the 250 live-born neonates, one (0.4%) was found positive at RT-PCR pharyngeal swabs performed after delivery. The mother was tested positive during the third trimester of pregnancy. The newborn was asymptomatic and had negative RT-PCR test after 14 days of life. At logistic regression analysis, gestational age at diagnosis (OR: 0.85, 95% CI 0.8–0.9 per week increase; p Conclusions Early gestational age at infection, maternal ventilatory supports and low birthweight are the main determinants of adverse perinatal outcomes in fetuses with maternal COVID-19 infection. Conversely, the risk of vertical transmission seems negligible.

Volatile Organic Compound (VOC) analysis is usually applied in pollution assessment by checking for toxic or harmful volatile compounds volatiles in air, water and soil samples. In this study, exogenous VOCs and their derivatives, metabolized by cells, were valued into specific body fluids. In particular, the VOC profiles of blood, urine and human semen samples collected from young men living in two high pollution areas in Italy, i.e. Land of Fires and Valley of Sacco River, were fingerprinted. The analysis is based on Headspace Solid Phase Micro-extraction (HS-SPME) followed by Gas Chromatography-Mass Spectrometric detection (GC-MS). The volatile composition of the three body fluids showed that some VOCs are in common between blood, urine and human semen samples, whereas others are present only in a body fluid. Some compounds, as well as also some chemical classes show a higher affinity for a specific body fluid. Statistical analysis allowed to discriminate the two contaminated areas and identify those compounds which significantly contribute to the two areas classification. Some of these compounds are toxic and found prevalently in Valley of Sacco River samples, correspondingly to sperm analysis results for young men living in this zona worse than those living in Land of Fires.

Background Although the diffusion of novel synthetic opioids has become a worldwide phenomenon, their prevalence of use in Italy seems to be limited. Existing national data is mainly derived by anamnestic surveyslacking of toxicological validation and not always disclosing the use of these compounds, which might remain under-diagnosed. Methods an assessment of the metabolites of the main synthetic opioids on hair samples was carried out among patients admitted at the Addiction Treatment Unit of Trento. The analytical approach included: (a) screening by means of immunoenzymatic method for fentanyl, fentanyl analogs and oxicodone; (b) confirmation of the samples resulted positive for fentanyl and oxicodone by means of HPLC-MS/; (c) search and dosage detection of Tramadol by means of HPLC-MS/MS. Results 3 out of 309 analysed samples were found positive: one was positive to Fentanyl and two to 4-ANPP. In the same cohort, 6 samples were also found positive for Oxycodone . Tramadol was searched in 189 samples and 12 of them resulted positive. Discussion and conclusion Those found positive were mainly young adults engaging in dangerous patterns of use and lacking awareness of risks. The phenomenon requires further consideration by health professionals. Training and more evidence-based information on synthetic opioids as well as other Novel Psychoactive Substances (NPS) are urgently needed.

Air pollution is involved in many pathologies. The pollutants act through several mechanisms that can affect numerous physiological functions, including reproduction. In this work we used a double approach to analyse the Volatile Organic Compound (VOC) component in 4 different biosamples (whole blood, urine, human semen and hairs) of young men (about 18 years old) living in the sadly famous area called “Land of fires” in Campania region (South-Western Italy), known to contain the largest illegal waste dump in Europe, in order to explore hypothesized correlation between VOCs in biofluid and man infertility due to environmental pollution. The double approach is based on two different technologies: (a) SPME-GC/MS (Solid Phase Microextraction—Gas Chromatography and Mass Spectrometry) system and (b) array of gas sensors. The preliminary data seems to be compatible with the EcoFoodFertility Project hypothesis that human semen is a good biological fluid to study volatile component as bioaccumulator of both exogenous and endogenous VOCs.

Early diagnosis of Alzheimer's disease (AD) supposedly increases the effectiveness of therapeutic interventions. However, presently available diagnostic procedures are either invasive or require complex and expensive technologies, which cannot be applied at a larger scale to screen populations at risk of AD. We were looking for a biomarker allowing to unveil a dysfunction of molecular mechanisms, which underly synaptic plasticity and memory, before the AD phenotype is manifested and investigated the effects of transcranial direct current stimulation (tDCS) in 3×Tg-AD mice, an experimental model of AD which does not exhibit any long-term potentiation (LTP) and memory deficits at the age of 3 months (3×Tg-AD-3M). Our results demonstrated that tDCS differentially affected 3×Tg-AD-3M and age-matched wild-type (WT) mice. While tDCS increased LTP at CA3-CA1 synapses and memory in WT mice, it failed to elicit these effects in 3×Tg-AD-3M mice. Remarkably, 3×Tg-AD-3M mice did not show the tDCS-dependent increases in pCREB Ser133 and pCaMKII Thr286 , which were found in WT mice. Of relevance, tDCS induced a significant increase of plasma BDNF levels in WT mice, which was not found in 3×Tg-AD-3M mice. Collectively, our results showed that plasticity mechanisms are resistant to tDCS effects in the pre-AD stage. In particular, the lack of BDNF responsiveness to tDCS in 3×Tg-AD-3M mice suggests that combining tDCS with dosages of plasma BDNF levels may provide an easy-to-detect and low-cost biomarker of covert impairment of synaptic plasticity mechanisms underlying memory, which could be clinically applicable. Testing proposed here might be useful to identify AD in its preclinical stage, allowing timely and, hopefully, more effective disease-modifying interventions.

Summary Background Indocyanine green (ICG) lymphography gained widespread acceptance as a gold standard in lymphatic surgery, although this imaging modality has several limitations. Ultrasound is progressively emerging as a powerful tool in LVA preoperative planning, as it expands the available knowledge for both lymphatic channels and recipient venules. The purpose of this study is to evaluate the accuracy of ultrahigh frequency ultrasound (UHFUS) in detecting the lymphatic vessels and in evaluating their degeneration status, including wall and lumen characteristics, compared with direct histology observation. Patients and Methods From May 2018 to November 2018, 26 consecutive patients affected by extremities lymphedema who underwent LVA were prospectively assessed. For each patient, we evaluated the lymphatic vessels suitable for LVA by UHFUS preoperatively, intraoperatively, and by histology. Qualitative and quantitative data of lymphatic channels were analyzed for each evaluation including lymphatic outer diameter, inner diameter, and wall thickness. Quantitative outcome analysis after LVA was also performed. Results On a total of 26 patients, 14 were upper limb lymphedema (ULL) and 12 lower limb lymphedema (LLL). All cases were secondary lymphedema. The qualitative and the quantitative analysis showed significant correlation and agreement between ultrasound and histological measurements. Conclusions UHFUS preoperative and intraoperative analysis of lymphatic channel strongly correlates with their histology. By coupling this high detailed information on lymphatic channels with the ability to dynamically study preoperative recipient venules, UHFUS represents the quintessence in lymphatic surgery, and it should be considered as the new gold standard in the preoperative planning of LVA. Level of Evidence II (diagnostic).

Introduction The evaluation of volatile organic compounds(VOCs) emitted by human body offers a unique tool to set up new non-invasive devices for early diagnosis and long-lasting monitoring of most human diseases. However, their cellular origin and metabolic fate have not been completely elucidated yet, thus limiting their clinical application. Endothelium acts as an interface between blood and surrounding tissues. As such, it adapts its physiology in response to different environmental modifications thus playing a role in the pathogenesis of many metabolic and inflammatory diseases. Objectives Since endothelium specifically reshapes its physiologic functions upon environmental changes the objective of this study was to evaluate if and how pro-inflammatory stimuli affect VOC metabolism in endothelial cell in culture. Methods Gas chromatography with mass spectrometric detection was applied to profile VOCs in the headspace of cultured endothelial cells (EC) in the absence or presence of the pro-inflammatory stimulus lipopolysaccharide (LPS). Results We observed that, under resting conditions, EC affected the amount of 58 VOCs belonging to aldehyde, alkane and ketone families. Among these, LPS significantly altered the amount of 15 VOCs. ROC curves show a perfect performance (AUC = 1) for 10 metabolites including 1-butanol, 3-methyl-1-butanol and 2-ethyl-1-hexanol. Discussion The emission and uptake of the aforementioned VOCs disclose potential unexplored metabolic pathways for EC that deserve to be investigated. Overall, we identified new candidate VOC potentially exploitable, upon experimental confirm in in vivo model of disease, as potential biomarkers of sepsis and pro-inflammatory clinical settings.

It is estimated that about the 12% of world’s couples are infertile, and half of these infertility cases are due to male factors. The main conditions that can affect male fertility are asthenozoospermia and oligozoospermia. For the first time, we analysed semen headspace in order to characterize the pattern of Volatile Organic Compounds (VOCs) in human seminal fluid and to identify specific metabolites that may be potential biomarkers of pathological conditions. By headspace solid-phase microextrac- tion-gas chromatography-mass spectrometry (HS-SPME-GC-MS) method, we detected both endogenous and exogenous compounds that form a complex mix of metabolites with a high inter- individual biological variability. Nevertheless, it was possible to point out the interconnection between some VOCs and asthenozoospermia and/or oligozoospermia. Furthermore, this work emphasizes the importance of semen as matrix for metabolomic studies that usually focus on other biological fluids.

Infertility is one of principal health and social problems of this century. Male factors are involved in half of the cases and often the alteration concerns sperm motility. Seminogram is the gold standard technique for semen analysis, but it presents several limits. For this reason, we propose a new method for discriminating asthenozoospermic samples (low sperm motility) from normozoospermic ones (progressive motility > 32%) based on the never explored analysis of the volatile metabolites in the headspace of human semen sample by Gas Chromatograph (GC) equipped with two detectors: a Mass Spectrometer (MS) and a metal oxide based gas sensor sensitive (MOX) to Volatile Organic Compounds (VOCs). VOC sensor signal profiles (resistance vs. time) showed a higher sensitivity to specific organic classes such as aldehydes and ketones. The sensorgrams were preprocessed and analysed by PLS-DA. The results showed that sensorgrams analysis by suitable bioinformatics techniques has a good discrimination power and could support physiological parameters in human semen assessment. The analysis of the human semen Volatilome may be a proof-of-concept for the development of a novel micro-GC device with a sensor array detector, a potential candidate for infertility assessment in clinical practice.

The world is impacting with a drastic demographic change that is reflected in a progressive ageing population. If on the one side increasing health care for older people is important, stimulating the level of birth becomes decisive. The principal goal of this work is to set up of new method for early diagnosis of male infertility based on analysis of seminal Volatile Organic Compounds (VOCs), potentially biomarkers of infertility status. The identification of the volatile metabolite patterns in semen samples was done by an unconventional GC/[−MS + gas sensor] system. Once validate this approach could integrate and improve traditional semen analysis based on physiological parameters and addressed to the development of novel medical devices based on gas microsensors for male infertility screening.

In this work we proposed a novel tele-medicine system called Smart Breath Network (SBN) with the perspective to improve the life of patients affected by a well-known respiratory disease called Chronic Obstructive Pulmonary Disease (COPD). The main therapy for COPD is home-assisted ventilation that requires frequent home intervention by specialized hospital staff in order to adjust dispensing and to verify the effectiveness of the therapy. Moreover, healthcare services need to improve the care of patients with difficult access to services, particularly those in rural or remote areas without sustaining high costs of service maintenance. The SBN system tries to fill the gap between healthcare services, that need to take care of a growing number of home-assisted COPD patients due to the today’s alarming air pollution. The SBN network gathers all patient’s respiratory data from Smart Breath Analyzers (SBAs) devices that are low-cost, simple to use, compact-sized devices that fit to almost any ventilator with bi-tube capabilities.

In this work we developed a Smart Breath Analyzer device devoted to the tele-monitoring of exhaled air in patients suffering Chronic Obstructive Pulmonary disease (COPD) and home-assisted by mechanical ventilation. The device based on sensors allows remote monitoring of a patient during a ventilotherapy session, and transmit the monitored signals to health service unit by TCP/IP communication through a cloud remote platform. The aim is to check continuously the effectiveness of therapy and/or any state of exacerbation of the disease requiring healthcare. By preliminary experimental tests, the prototype was validated on a volunteer subject.

Impairment of adult hippocampal neurogenesis is an early event in Alzheimer's disease (AD), playing a crucial role in cognitive dysfunction associated with this pathology. However, the mechanisms underlying defective neurogenesis in AD are still unclear. Recently, the nucleoporin Nup153 has been described as a new epigenetic determinant of adult neural stem cell (NSC) maintenance and fate. Here we investigated whether Nup153 dysfunction could affect the plasticity of NSCs in AD. Nup153 expression was strongly reduced in AD-NSCs, as well as its interaction with the transcription factor Sox2, a master regulator of NSC stemness and their neuronal differentiation. Similar Nup153 reduction was also observed in WT-NSCs treated with amyloid-? (A?) or stimulated with a nitric oxide donor. Accordingly, AD-NSCs treated with either a ?-secretase inhibitor or antioxidant compounds showed higher Nup153 levels suggesting that both nitrosative stress and A? accumulation affect Nup153 expression. Of note, restoration of Nup153 levels in AD-NSCs promoted their proliferation, as assessed by BrdU incorporation, neurosphere assay, and stemness gene expression analysis. Nup153 overexpression also recovered AD-NSC response to differentiation, increasing the expression of pro-neuronal genes, the percentage of cells positive for neuronal markers, and the acquisition of a more mature neuronal phenotype. Electrophysiological recordings revealed that neurons differentiated from Nup153-transfected AD-NSCs displayed higher Na+ current density, comparable to those deriving from WT-NSCs. Our data uncover a novel role for Nup153 in NSCs from animal model of AD and point to Nup153 as potential target to restore physiological NSC behavior and fate in neurodegenerative diseases.

Purpose Activated platelet gel (PG) derived from adult peripheral blood (APB) has been extensively used for topical therapy of various clinical conditions. Conversely, few observations on PG from umbilical cord blood (CB) have been reported so far. Although PG preparations are known to contain a high concentration of a large number of biological factors involved in inflammation and tissue repair, their comprehensive characterization is still missing. The innovative goal of our research was to use proteomics technologies in order to profile biologically active components in these blood derivatives. Experimental design Supernatants recovered from three independent APB and CB-derived PGs, prepared using batroxobin, were enriched for low-abundance proteins with ProteoMiner and subsequently analyzed by GeLC-MS/MS. Results 751 and 760 proteins were identified in the APB and CB-derived PG releasates, respectively. A core dataset including only proteins found in 2/3 and 3/3 biological replicates was generated and functionally characterized by gene ontology. Searching against Vesiclepedia database showed that 33% of our dataset consists of novel releasate proteins. Comparison between the two types of PG secretomes revealed that 117 proteins are present only in the APB-derived samples, 104 proteins are distinctive of the CB-derived samples, and 229 are in common. Conclusions and clinical relevance Our study highlighted a differential content of proteins supporting tissue repair and regeneration between APB and CB-derived PGs. These findings may help better identifying future appropriate clinical applications. This article is protected by copyright. All rights reserved

Plants display a wide capacity range to survive cold and freezing conditions. Indeed, they are able to sense low, non-freezing temperatures and activate processes that lead to an increase in freezing tolerance, a phenomenon known as cold acclimation (CA). Metabolome analysis not only improves the recognition of the complex interactive nature of plant metabolic networks and their responses to environmental changes but also provides valuable information about plant abiotic stress resistance. Metabolomics is still an evolving field and has not reached adequate development, routine and coverage, such as other “omics” technologies. Volatilomics, a recently born “omics” science which allows to analyse the volatile component of the metabolome, has received considerable attention in abiotic stress studies. However, the integration of “omics” technologies is faced with some limitations, and, subsequently, of about 5000 primary and secondary metabolites that are present in a single plant metabolome, only a small portion has been identified. The main metabolome changes during cold stress include the production of osmoprotecting metabolites, which are involved in the regulation of cellular water relations and the reduction of cellular dehydration. Furthermore, they participate in the remodelling of membrane lipids and maintaining membrane integrity and energy sources. Cold-responsive metabolites have cryoprotective and scavenging activities and possibly also act as stabilisers of proteins and enzymes or as regulators of gene expression. Such cold-responsive metabolites particularly include soluble sugars, amino acids, betaines, organic acids, polyols, polyamines and lipids. Altogether, the accumulation of these functional metabolites is an important strategy for increasing plant survival under freezing temperatures. The analysis of metabolome dynamics in overwintering plants during the different stages of CA and plant development may provide valuable information for the improvement of cold tolerance in breeding processes. Interestingly, the majority of cold-responsive metabolites are also induced by other abiotic stresses, such as drought and salinity. In this chapter, we discuss the effects of cold stress on the metabolome and the roles of some functional metabolites during CA and overwintering.

Background Psoriasis is a chronic inflammatory skin disease. It is characterized by immune cell activation and altered epidermal differentiation. S100A7 (psoriasin) is overexpressed in psoriasis, suggesting a determinant role of this protein in inflammation and keratinocyte differentiation. Objective The purpose of this study was to investigate the expression of S100A7 in the skin from psoriatic patients undergoing biological therapy with adalimumab, etanercept or ustekinumab. Methods S100A7 expression and distribution were analyzed by immunohistochemistry. Results S100A7, overexpressed in epidermal keratinocytes of psoriatic lesions, was downregulated, under the biological therapy with adalimumab, etanercept or ustekinumab, only in patients achieving a PASI score Conclusions Dysregulation of S100A7 may represent a non-negligible player in the maintenance of psoriasis and the relative epidermal changes. Blockage of S100A7 may represent an additional therapeutic approach in the treatment of psoriasis.

One of the hallmarks of blood bank stored red blood cells (RBCs) is the irreversible transition from a discoid to a spherocyte-like morphology with membrane perturbation and cytoskeleton disorders. Therefore, identification of the storage-associated modifications in the protein–protein interactions between the cytoskeleton and the lipid bilayer may contribute to enlighten the molecular mechanisms involved in the alterations of mechanical properties of stored RBCs. Here we report the results obtained analyzing RBCs after 0, 21 and 35 days of storage under standard blood banking conditions by label free mass spectrometry (MS)-based experiments. We could quantitatively measure changes in the phosphorylation level of crucial phosphopeptides belonging to β-spectrin, ankyrin-1, α-adducin, dematin, glycophorin A and glycophorin C proteins. Data have been validated by both western blotting and pseudo-Multiple Reaction Monitoring (MRM). Although each phosphopeptide showed a distinctive trend, a sharp increase in the phosphorylation level during the storage duration was observed. Phosphopeptide mapping and structural modeling analysis indicated that the phosphorylated residues localize in protein functional domains fundamental for the maintenance of membrane structural integrity. Along with previous morphological evidence acquired by electron microscopy, our results seem to indicate that 21-day storage may represent a key point for the molecular processes leading to the erythrocyte deformability reduction observed during blood storage. These findings could therefore be helpful in understanding and preventing the morphology-linked mechanisms responsible for the post-transfusion survival of preserved RBCs. Copyright © 2015 John Wiley & Sons, Ltd.

Leukemia is a group of cancers that usually begin in the bone marrow and result in high numbers of abnormal and dysfunctional white blood cells. Many studies were carried out to investigate metabolism of these cells. Metabolome analysis has been successfully applied to leukemia disease and emerged as a powerful tool for obtaining information about the biological processes that occur in organisms, and as a useful platform for discovering new clinical biomarkers and make diagnosis of disease using different biofluids. Whatever has not been investigated in leukemic cells is volatile metabolic signature that in recent literature is called “volatilome”. Volatile organic compounds (VOCs) from the headspace of cultured THP1 cells and normal human peripheral blood mononuclear cells, were collected by headspace solid-phase microextraction (HS-SPME) and analyzed by gas chromatography combined with mass spectrometry (GC–MS), thus defining a volatile metabolomics signature. Styrene, cyclohexanol, cyclohexanone, 1-hexanol-2-ethyl, cyclohexane, 1,1’-(1,2-dimethyl-1,2-ethanediyl)bis-, benzene, 1,3-bis(1,1-dimethylethyl)- were present in higher amount in cultured THP1 cell than in PBMC, while 2-butanone has an opposite trend. Cell stimulation with lipopolysaccharide affected normal cells, but not leukemic cells. The establishment of the volatile fingerprint of THP1 cell lines presents a powerful approach to find endogenous VOCs that could be used to improve the diagnostic tools and explore the associated metabolomic pathways.

Dark rearing is known to delay the time course of the critical period for ocular dominance plasticity in the visual cortex. Recent evidence suggests that a period of dark exposure (DE) may enhance or reinstate plasticity even after closure of the critical period, mediated through modification of the excitatory–inhibitory balance and/or removal of structural brakes on plasticity. Here, we investigated the effects of a week of DE on the recovery from a month of monocular deprivation (MD) in the primary visual cortex (V1) of juvenile mice. Optical imaging of intrinsic signals revealed that ocular dominance in V1 of mice that had received DE recovered slightly more quickly than of mice that had not, but the level of recovery after three weeks was similar in both groups. Two-photon calcium imaging showed no significant difference in the recovery of orientation selectivity of excitatory neurons between the two groups. Parvalbumin-positive (PV+) interneurons exhibited a smaller ocular dominance shift during MD but again no differences in subsequent recovery. The percentage of PV+ cells surrounded by perineuronal nets, a structural brake on plasticity, was lower in mice with than those without DE. Overall, DE causes a modest enhancement of mouse visual cortex plasticity. This article is part of the themed issue ‘Integrating Hebbian and homeostatic plasticity’.

Cigarette smoking harms nearly every organ of the body and causes many diseases. The analysis of exhaled breath for exogenous and endogenous volatile organic compounds (VOCs) can provide fundamental information on active smoking and insight into the health damage that smoke is creating. Various exhaled VOCs have been reported as typical of smoking habit and recent tobacco consumption, but to date, no eligible biomarkers have been identified. Aiming to identify such potential biomarkers, in this pilot study we analyzed the chemical patterns of exhaled breath from 26 volunteers divided into groups of nonsmokers and subgroups of smokers sampled at different periods of withdrawal from smoking. Solid-phase microextraction technique and gas chromatography/mass spectrometry methods were applied. Many breath VOCs were identified and quantified in very low concentrations (ppbv range), but only a few (toluene, pyridine, pyrrole, benzene, 2-butanone, 2-pentanone and 1-methyldecyclamine) were found to be statistically significant variables by Mann-Whitney test. In our analysis, we did not consider the predictive power of individual VOCs, as well as the criterion of uniqueness for biomarkers suggests, but we used the patterns of the only statistically significant compounds. Probit prediction model based on statistical relevant VOCs-patterns showed that assessment of smoking status is heavily time dependent. In a two-class classifier model, it is possible to predict with high specificity and sensitivity if a subject is a smoker who respected 1 hour of abstinence from smoking (short-term exposure to tobacco) or a smoker (labelled "blank smoker") after a night out of smoking (long-term exposure to tobacco). On the other side, in our study "blank smokers" are more like non-smokers so that the two classes cannot be well distinguished and the corresponding prediction results showed a good sensitivity but low selectivity.

This paper reports the development of calibration models for quality control in the production of ethylene/propylene/1-butene terpolymers by the use of multivariate tools and FT-IR spectroscopy. 1-Butene concentration prediction is achieved in terpolymers by coupling FT-IR spectroscopy to multivariate regression tools. A dataset of 26 terpolymers (14 coming from a constrained experimental design for mixtures, plus 12 terpolymers used for external validation) was analysed by FT-IR spectroscopy. An internal method of "Polimeri Europa" plant, based on (13)C NMR spectroscopy is used to determine the percentage of 1-butene in the samples. Then, different multivariate tools are used for 1-butene concentration prediction based on the FT-IR spectra recorded. Different multivariate calibration methods were explored: principal component regression (PCR), partial least squares (PLS), stepwise OLS regression (SWR) and artificial neural networks (ANNs). The model obtained by back-propagation neural networks turned out to be the best one. The performances of the BP-ANN model were further improved by variable selection procedures based on the calculation of the first derivative of the network. The proposed approach allows the monitoring in real time of the polymer synthesis and the estimation of the characteristics of the product attainable from the concentration of 1-butene.

Caspase proteases are responsible for the regulated disassembly of the cell into apoptotic bodies during mammalian apoptosis. Structural homologues of the caspase family (called metacaspases) are involved in programmed cell death in single-cell eukaryotes, yet the molecular mechanisms that contribute to death are currently undefined. Recent evidence revealed that a programmed cell death process is induced by acetic acid (AA-PCD) in Saccharomyces cerevisiae both in the presence and absence of metacaspase encoding gene YCA1. Here, we report an unexpected role for the yeast metacaspase in protein quality and metabolite control. By using an “omics” approach, we focused our attention on proteins and metabolites differentially modulated en route to AA-PCD either in wild type or YCA1-lacking cells. Quantitative proteomic and metabolomic analyses of wild type and Δyca1 cells identified significant alterations in carbohydrate catabolism, lipid metabolism, proteolysis and stress-response, highlighting the main roles of metacaspase in AA-PCD. Finally, deletion of YCA1 led to AA-PCD pathway through the activation of ceramides, whereas in the presence of the gene yeast cells underwent an AA-PCD pathway characterized by the shift of the main glycolytic pathway to the pentose phosphate pathway and a proteolytic mechanism to cope with oxidative stress. Significance The yeast metacaspase regulates both proteolytic activities through the ubiquitin–proteasome system and ceramide metabolism as revealed by proteome and metabolome profiling of YCA1-knock-out cells during acetic-acid induced programmed cell death.

The yeast Saccharomyces cerevisiae expresses one member of the metacaspase Cys protease family, encoded by the YCA1 gene. Combination of proteomics and metabolomics data showed that YCA1 deletion down-regulated glycolysis, the TCA cycle and alcoholic fermentation as compared with WT cells. Δyca1 cells also showed a down-regulation of the pentose phosphate pathway and accumulation of pyruvate, correlated with higher levels of certain amino acids found in these cells. Accordingly, there is a decrease in protein biosynthesis, and up-regulation of specific stress response proteins like Ahp1p, which possibly provides these cells with a better protection against stress. Moreover, in agreement with the down-regulation of protein biosynthesis machinery in Δyca1 cells, we have found that regulation of transcription, co-translational protein folding and protein targeting to different subcellular locations were also down-regulated. Metabolomics analysis of the nucleotide content showed a significant reduction in Δyca1 cells in comparison with the WT, except for GTP content which remained unchanged. Thus, our combined proteome–metabolome approach added a new dimension to the non-apoptotic function of yeast metacaspase, which can specifically affect cell metabolism through as yet unknown mechanisms and possibly stress-response pathways, like HOG and cell wall integrity pathways. Certainly, YCA1 deletion may induce compensatory changes in stress response proteins offering a better protection against apoptosis to Δyca1 cells rather than a loss in pro-apoptotic YCA1-associated activity.

Psoriasis is a chronic inflammatory skin disease, characterized by hyperproliferation of keratinocytes and by skin infiltration of activated T cells. To date, the pathophysiology of psoriasis has not yet been fully elucidated. The Notch pathway plays a determinant role in cell fate determination, proliferation, differentiation, immune cell development and function. Many biological agents, used in the treatment of psoriasis, include TFN-α inhibitors, such as etanercept, adalimumab, and anti IL-12/IL-23 p40 antibody, such as ustekinumab. This study aimed to determine mRNA expression levels by real-time RT-PCR, and protein expression levels, analysed by Western blot and immunohistochemistry, of some components of the Notch pathway, such as NOTCH1, NOTCH2, JAGGED1, and HES1 after biological treatments in psoriatic patients. mRNA and protein levels of NOTCH1, NOTCH2, JAGGED1 and HES1 were upregulated in skin samples from untreated psoriatic patients compared with normal controls. Biological therapy showed to downregulate differently the protein expression levels of the molecules under study. Our study suggests that Notch pathway components might be a potential therapeutic target against psoriasis.

Piedmontese meat tenderness becomes higher by extending the ageing period after slaughter up to 44 days. Classical physical analysis only partially explain this evidence, so in order to discover the reason of the potential beneficial effects of prolonged ageing, we performed omic analysis in the Longissimus thoracis muscle by examining main biochemical changes through mass spectrometry-based metabolomics and proteomics. We observed a progressive decline in myofibrillar structural integrity (underpinning meat tenderness) and impaired energy metabolism. Markers of autophagic responses (e.g. serine and glutathione metabolism) and nitrogen metabolism (urea cycle intermediates) accumulated until the end of the assayed period. Key metabolites such as glutamate, a mediator of the appreciated umami taste of the meat, were found to constantly accumulate until day 44. Finally, statistical analyses revealed that glutamate, serine and arginine could serve as good predictors of ultimate meat quality parameters, even though further studies are mandatory.

Meat derives from a muscle that undergoes a great number of biochemical and physiological changes. The anoxic condition established from the moment of animal sacrifice forces muscle cells to a sort of reaction, resulting in methodical programmed cell death to avoid necrosis. The duality autophagy and/or apoptosis is at the center of the scientific debate about the biological processes driving the muscle to meat conversion. Here we report an omic time course overview carried on proteome, phosphoproteome and metabolome of Piedmontese longissimus thoracis muscle searching for clues helping us to extricate through the dilemma. The survey depicts a progressive physiological impairing and our evidences push towards the apoptotic behavior: the proteomic time course trend of annexin A2, RKIP, HSPB6, αB crystalline, adenylate kinase, DJ-1 and 31 kDa actin fragment; the 0–1 day increased phosphorylation of myosin 2 and synaptopodin and the metabolomic time course trend of key metabolic indicators, like GSH/GSSG ratio, taurine and nitrotyrosine. The employed techniques provide strong indications about the likely apoptotic behavior of aging meat in muscle-to-meat conversion process. Biological significance Our work underlines compelling evidences of the apoptotic behavior of Piedmontese beef muscle cells undergoing the muscle-to-meat process, whereas no autophagic clues are inferred from this omic investigation.

One of the hallmarks of blood bank stored red blood cells (RBCs) is the irreversible transition from a discoid to a spherocyte-like morphology with membrane perturbation and cytoskeleton disorders. Therefore, identification of the storage-associated modifications in the protein-protein interactions between the cytoskeleton and the lipid bilayer may contribute to enlighten the molecular mechanisms involved in the alterations of mechanical properties of stored RBCs. Here we report the results obtained analyzing RBCs after 0, 21 and 35 days of storage under standard blood banking conditions by label free mass spectrometry (MS)-based experiments. We could quantitatively measure changes in the phosphorylation level of crucial phosphopeptides belonging to β-spectrin, ankyrin-1, α-adducin, dematin, glycophorin A and glycophorin C proteins. Data have been validated by both western blotting and pseudo-Multiple Reaction Monitoring (MRM). Although each phosphopeptide showed a distinctive trend, a sharp increase in the phosphorylation level during the storage duration was observed. Phosphopeptide mapping and structural modeling analysis indicated that the phosphorylated residues localize in protein functional domains fundamental for the maintenance of membrane structural integrity. Along with previous morphological evidence acquired by electron microscopy, our results seem to indicate that 21-day storage may represent a key point for the molecular processes leading to the erythrocyte deformability reduction observed during blood storage. These findings could therefore be helpful in understanding and preventing the morphology-linked mechanisms responsible for the post-transfusion survival of preserved RBCs.