Search

Aim. It is known that children and adolescents with obesity are more prone to obstructive sleep apnea syndrome (OSAS) and that their lung function may show some disturbance. Literature is scarce about potential associations; therefore, we aimed to study the relationship between OSAS, lung function, and adiposity in a population of children suspected of OSAS. Material and Methods. We performed home respiratory polygraphy and spirometry in all subjects. The relationships between body mass index z -score (zBMI), polygraphy, and spirometry data were analyzed. Results. We recruited 81 subjects aged between 5 and 16 years, 63% being obese. 43.2% of subjects were diagnosed with OSAS (32.1% mild, 4.9% moderate, and 6.2% severe). We found no correlation between respiratory polygraphy and the zBMI. The mean spirometric value FEV1, FVC, and FEV1/FVC ratio z ’s were normal in all subjects, whereas FVC z ’s and FEV1/FVC ratio z ’s were significantly positively related for obesity and negatively for normal weight ( p < 0.05 ). FEV1 z ’s was inversely correlated to the percentage of analyzed time passed below 90% of SpO2 ( r = − 0.224 , p = 0.044 ). All subjects with FEV1 ( n = 8 ) and/or FVC ( n = 9 ) z ’s below the lower limit for normal (LLN) had an AHI ≥ 1 (FEV1: p = 0.001 ; FVC: p < 0.001 ), especially subjects with normal weight (FEV1: p = 0.003 ; FVC: p = 0.010 ). Conclusion. When comparing normal-weight children and adolescents with obesity, the prevalence of OSAS but not spirometric values was strongly related to BMI z -score, probably because obesity engenders advanced puberty and an accelerated growth spurt. FEV1 was more frequently z ’s and FEF25-75% z ’s. Moreover, all subjects with abnormal spirometric values were suffering from at least mild OSAS, again more frequently in normal-weight subjects.

Funding Acknowledgements Type of funding sources: None. Background Highly localized impedance (LI) measurements in combination with a catheter-tissue contact force (CF) during atrial fibrillation (AF) ablation may improve tissue characterization and lesion prediction during pulmonary vein isolation (PVI). Purpose This analysis explores the relationship between LI parameters, ablation spot locations and procedural success during ablation of PVs in a large population of consecutive AF patients. Methods Two-hundred twelve consecutive patients from 16 European centers undergoing de novo AF radiofrequency (RF) catheter ablation with a novel open-irrigated tip catheter enabled by CF and LI measurement capabilities (Stablepoint) were included. Ablation was guided by the magnitude and time-course of LI drop during RF delivery. The interlesion distance between each ablation spot was set ≤6 mm. First pass isolation (FPI) was defined as successful PVI at or before completion of the first encircling lesion set regardless of visual gaps. Procedural endpoint was the achievement of the PVI as assessed by entrance and exit block. Data are reported as mean±SD. Results A total of 13891 ablation spots performed around PVs were analyzed (baseline LI=161.2±19 Ω; LI drop=21.9±9 Ω; LI drop rate=3.1±2 Ω/s; RF Delivery time=9.2±4 s; CF=12.5±7 g). LI drop was predicted by baseline LI (r=0.56, 95%CI:0.55 to 0.57, p20Ω (Sensitivity=56.8%, Specificity=93.2%, PPV=99.7%, Area under the ROC curve=0.7841, p21Ω at anterior sites and >18Ω at posterior sites. No steam pops or complications were reported during the procedures. All PVs were successfully isolated in all study patients Conclusions In a large, international population of consecutive de novo AF cases, an ablation strategy guided by LI and CF information results in a very high first pass isolation rate. LI drop is predictive of PV segment isolation. A regional approach to RF ablation guided by LI information may be useful in patients with AF.

Objective Sub-motor threshold 5 Hz repetitive paired associative stimulation (5 Hz-rPAS25ms) produces a long-lasting increase in corticospinal excitability. Assuming a spike-timing dependent plasticity-like (STDP-like) mechanism, we hypothesized that 5 Hz-rPAS at a shorter inter-stimulus interval (ISI) of 15 ms (5 Hz-rPAS15ms) would exert a lasting inhibitory effect on corticospinal excitability. Methods 20 healthy volunteers received two minutes of 5 Hz-rPAS15ms. Transcranial magnetic stimulation (TMS) was applied over the motor hotspot of the right abductor pollicis brevis muscle at 90% active motor threshold. Sub-motor threshold peripheral electrical stimulation was given to the left median nerve 15 ms before each TMS pulse. We assessed changes in mean amplitude of the unconditioned motor evoked potential (MEP), short-latency intracortical inhibition (SICI), intracortical facilitation (ICF), short-latency afferent inhibition (SAI), long-latency afferent inhibition (LAI), and cortical silent period (CSP) before and for 60 minutes after 5-Hz rPAS15ms. Results Subthreshold 5-Hz rPAS15ms produced a 20–40% decrease in mean MEP amplitude along with an attenuation in SAI, lasting at least 60 minutes. A follow-up experiment revealed that MEP facilitation was spatially restricted to the target muscle. Conclusions Subthreshold 5-Hz rPAS15ms effectively suppresses corticospinal excitability. Together with the facilitatory effects of subthreshold 5-Hz rPAS25ms (Quartarone et al., J Physiol 2006;575:657–670), the results show that sub-motor threshold 5-Hz rPAS induces STDP-like bidirectional plasticity in the motor cortex. Significance The results of the present study provide a new short-time paradigm of long term depression (LTD) induction in human sensory-motor cortex.

Funding Acknowledgements Type of funding sources: None. Background Recently, a novel technology able to measure local tissue impedance (LI) and contact force (CF) aimed at validating ablation endpoints has become available for clinical use. Purpose This analysis explores the relationship between LI parameters, substrate characteristics and AF type during ablation in AF patients. Methods A novel ablation catheter with dedicated algorithm was used to measure LI at the distal electrode. Each ablation point was characterized in terms of RF delivery time, baseline LI and LI drop during ablation according to different voltages (above or below 0.5 mV) of the atrial substrate. Ablation endpoint was PVI as assessed by entrance and exit block with local capture. Data are reported as mean±SD. Results A total of 2032 RFC deliveries from 34 consecutive patients were analyzed (71% paroxysmal AF, 29% persistent AF). All PVs were successfully isolated with an overall procedure time of 128±33min (fluoroscopy time=15.6±9min, RF delivery time=9.6±4s, CF=11.1±7g). The baseline LI was 155±17Ω prior to ablation and 136±14Ω after ablation (p0.5mv, n=1240, 61%). Baseline LI and LI drops were both higher when underlying atrial voltages were >0.5mV than when 0.5mV and Conclusions LI measurements well distinguished contact with high-voltage areas from low-voltage areas and were able to detect significant differences in LI drop during ablation according to the voltage level.

Funding Acknowledgements Type of funding sources: None. Background Permanent cardiac stimulation is increasingly necessary in very old patients, due to the augmented life expectancy. This raise concerns in physicians, patients and caregivers, because pacemaker (PM) implantation can be perceived as too invasive for frail patients with significant comorbidities. Purpose The aim of the present study is to provide data about peri-operative complications, short and long-term survival and their predictors in this specific setting. Materials and methods All consecutive patients aged ≥90 years undergoing pacemaker implantation in our Centre from 2008 to 2020 were enrolled in the registry. Their baseline and follow up data were recorded in a specific database. Results Among the 2437 patients undergoing PM implantation in a 13-year period, 153 were nonagenarians (6.3%). Mean age was 92.7±2.3 years. Most of the patients (66.7%) received a ventricular single chamber PM. High-degree atrio-ventricular block was the first indication for implant (59.5%), followed by sick sinus syndrome or atrial fibrillation with slow ventricular response (30.1%). 15.7% patients needed temporary pacing at presentation. 4 patients (2.6%) experienced acute complications: 3 pneumothorax, 1 ventricular lead displacement with need of repositioning. Mean follow up duration was 2.5±2.0 years. 117 patients (78.5%) died during the follow up. Death occurred after a mean of 2.47 years after PM implantation (range 1-3272 days). Survival rates were 70.0%, 35.3% and 10.0% at 1, 3 and 5 years respectively. No pacemaker-related long-term complication was recorded during the follow up. 2 patients underwent generator replacement due to battery end of life. Chronic kidney insufficiency (p = 0.030) and congestive heart failure (p = 0.021) but not age were predictors of precocious mortality (within 1 month). Age at implant and longer length of hospital stay before PM implantation were predictors of long-term death (p = 0.001 and p = 0.043 respectively). Conclusions Extremely old age should not discourage PM implantation, when indicated; age is obviously predictor of long-term death but is not predictor of precocious mortality. Acute periprocedural complication rate is low and generally comparable to that of younger populations.

Background Permanent cardiac stimulation is increasingly necessary in very old patients, due to the augmented life expectancy. This raise concerns in physicians, patients and caregivers, because pacemaker (PM) implantation can be perceived as too invasive for frail patients with significant comorbidities. The aim of the present study is to provide data about peri–operative complications, short and long–term survival and their predictors in this specific setting. Materials and Methods All consecutive patients aged ≥ 90 years undergoing pacemaker implantation in our Centre from 2008 to 2020 were enrolled in the registry. Their baseline and follow up data were recorded in a specific database. Results Among the 2437 patients undergoing PM implantation in a 13–year period, 153 were nonagenarians (6.3%). Mean age was 92.7 ± 2.3 years. Most of the patients (66.7%) received a ventricular single chamber PM. High–degree atrio–ventricular block was the first indication for implant (59.5%), followed by sick sinus syndrome or atrial fibrillation with slow ventricular response (30.1%). 15.7% patients needed temporary pacing at presentation. 4 patients (2.6%) experienced acute complications: 3 pneumothorax, 1 ventricular lead displacement with need of repositioning. Mean follow up duration was 2.5 ± 2.0 years. 117 patients (78.5%) died during the follow up. Death occurred after a mean of 2.47 years after pacemaker implantation (range 1 – 3272 days). Survival rates were 70.0%, 35.3% and 10.0% at 1, 3 and 5 years respectively. No pacemaker–related long–term complication was recorded during the follow up. 2 patients underwent generator replacement due to battery end of life. Chronic kidney insufficiency (p = 0.030) and congestive heart failure (p = 0.021) but not age were predictors of precocious mortality (within 1 month). Age at implant and longer length of hospital stay before PM implantation were predictors of long–term death (p = 0.001 and p = 0.043 respectively).

Background In Switzerland, from March 15th to May 11th 2020, schools and most shops were closed nationwide due to the COVID-19-related lockdown. This cessation of activities may have impacted weight gain in children and adolescents. The aims of our study were to evaluate the effects of the COVID-19 lockdown on the BMI of children and adolescents in treatment for obesity, and to compare its evolution to that of the previous year at the same time, as well as to that of normal-weight children. Methods This retrospective study gathered demographic and anthropometric data from subjects aged 6–18 years both with normal weight and with obesity, who attended our hospital clinics at four time points: before and after the lockdown period in 2020, and at the same times of the year in 2019. We used paired t-tests to assess weight, BMI and BMI z-score changes, linear and standard multiple regressions, independent Student’s t-tests or Chi-square tests to compare groups, and Pearson correlation coefficient when appropriate. Results Forty-seven children with obesity and 18 normal-weight subjects had complete data for the 4 visits. The mean BMI increased in both groups during the lockdown (obese: + 0.96 ± 1.5 vs. control: + 0.51 ± 0.1), however the increase was significantly more important in the subjects with obesity compared to the same period in 2019 (2019: + 0.33 ± 1.0; mean difference between 2019 and 2020: + 0.63 ± 2.0 p = 0.034). Conclusion The COVID-19 lockdown had a negative impact on the BMI of youth with obesity. Interestingly we observed extreme changes in this population, which was not the case in normal-weight children. Therefore, families with a child with obesity must be actively supported during these stressful and obesogenic periods of confinement.

Symptoms of red stripe disease caused by Acidovorax avenae subsp. avenae in Louisiana between 1985 and 2010 were limited to the leaf stripe form, which caused no apparent yield loss. During 2010, the more severe top rot form was observed, and a study was initiated to investigate the distribution of red stripe in the field and determine its effects on cane and sugar yields. Soil properties data, red stripe incidence, and sugarcane yields were all highly variable and were not randomly distributed in the field. Combined harvest data showed a negative correlation between yield components and red stripe incidence, with the strongest relationship between sucrose per metric ton and disease incidence. Red stripe incidence was positively correlated with several soil properties, including phosphorus, potassium, zinc, and calcium. Red stripe incidence also was found to increase with increasing nitrogen rate, with the greatest effects in heavy soils. Results also indicated that using red-stripe-infected cane as a seed source can significantly decrease shoot emergence, stalk population, and subsequent cane and sugar yields. These combined data suggest that red stripe disease can exhibit a highly variable rate of infection in commercial sugarcane fields and may also significantly decrease sugar yields.

In April 2017, uredinial lesions typical of orange rust caused by Puccinia kuehnii E.J. Butler were observed on the experimental sugarcane clone HoCP 14-801 (a complex hybrid of Saccharum L. spp.) in a cultivar trial in Hidalgo County, Texas. Additional observations were made in cultivar trials at three other sites in Hidalgo, Cameron, and Willacy Counties during the following 5 months of 2017. Orange rust symptoms were observed on 12 experimental clones and one commercial check cultivar, CP 89-2143, at one or more of the three trial locations. Cultivar trials were embedded in commercial fields. Uredinial lesions were orange and linear, measuring 650 to 850 × 26 to 32 µm. The orange to light cinnamon brown urediniospores were mostly obovoid or pyriform and variable in size (30 to 60 × 25 to 30 µm). Walls were 1 to 2 µm thick on the sides with a pronounced apical thickening of 5 µm or more and were echinulate with evenly distributed spines 2 to 4.5 µm apart. These morphology features were consistent with those described by Ryan and Egan (1989) and others for Puccinia kuehnii. Telia and teliospores were not observed. Leaf samples containing rust lesions were collected from cultivar CP 89-2143 and experimental clones HoCP 14-801, HoCP 14-874, and CP 11-2013. Identification of the species causing the orange rust lesions was verified by PCR assays conducted independently in two laboratories (Sugarcane Research Unit, ARS, USDA, Houma, LA, and Soybean/Maize Germplasm, Pathology, and Genetics Research Unit, USDA-ARS, Urbana, IL). Primers PkPmF and PkPmR were used that amplify fragments of 606 bp from P. kuehnii and 585 bp from P. melanocephala, the causal agent of brown rust of sugarcane (Glynn et al. 2010). Only the 606-bp fragment corresponding to P. kuehnii was amplified from each sample. No fragments were amplified when a PCR analysis with primers AR-F2 and AR-R1, which amplify Macruropyxis fulva, the pathogen causing tawny rust on sugarcane (Martin et al. 2017), was conducted on the samples. Sequences of the fragments were identical to each other, and a BLAST search of GenBank revealed 100% identity to previously reported isolates of P. kuehnii including an isolate from Florida, U.S.A. (GenBank accession no. EU176008). Sequences from P. kuehnii infecting CP 11-2013 (GenBank accession no. MN547621) and HoCP 14-874 (GenBank accession no. MN547622) were submitted to GenBank with voucher collection maintained at the USDA, ARS, Sugarcane Research Unit in Houma, LA. This is the first report of orange rust in the sugarcane production area of Texas. In 2018, approximately 50% of the sugarcane production area in Texas was planted with cultivar CP 89-2143. Although this cultivar was known to be susceptible to orange rust, it was introduced and expanded prior to the observation of orange rust in the industry. Orange rust was observed in commercial fields of cultivar CP 89-2143; however, disease severity was less than that observed in some experimental clones. In Florida, overall losses from orange rust have been estimated to be 10% with reductions as high as 43% in a highly susceptible cultivar (Rott et al. 2014). As researchers attempt to identify new sugarcane cultivars that are adapted to the Texas production area and provide diversity among the cultivars, susceptibility to orange rust must be considered.

Figure S1. Autophagy and apoptosis levels in PBMCs isolated from responding (A) and non-responding (B) patients to anti-TNF drugs alone or anti-TNF drugs plus methotrexate. Figure S2. Relation between autophagy and citrullination in RA patients. A. Levels of anti-cyclic citrullinated peptide antibodies (anti-CCP Abs) in patients with RA before and after treatment with anti-TNF drugs. Values are expressed as means ± sd. B. Correlation between anti-CCP antibodies levels (U/ml) and spontaneous autophagy (expressed as LC3-II) and apoptosis (expressed as percentage of Annexin V-positive cells). C. Changes in autophagy and apoptosis before and after treatment with anti-TNF drugs in relation to anti-CCP in RA responding patients. Figure S3. Flow cytometry gate strategy for purification of CD4+, CD8+ T lymphocytes and B lymphocytes from patients with RA. Allophycocyanin = APC; Peridinin chlorophyll protein = PerCP; Phycoerythrin = PE; Fluorescein isothiocyanate = FITC. Figure S4. Effect of TNFα on apoptosis. Flow cytometry analysis of apoptosis in PBMCs after treatment with TNFα. Apoptosis is expressed as percentage of AV-positive cells. Representative dot plots (PI on y axis vs. AV on x axis), chosen as representative of five experiments, are also shown. Table S1. Clinical, demographic and serological characteristics of patients with RA enrolled for sorting experiments (n = 8). (PDF 673 kb)

Endothelial dysfunction (ED) predisposes to venous thrombosis (VT) and post-thrombotic syndrome (PTS), a long-term VT-related complication. Sulodexide (SDX) is a highly purified glycosaminoglycan with antithrombotic, pro-fibrinolytic and anti-inflammatory activity used in the treatment of chronic venous disease (CVD), including patients with PTS. SDX has recently obtained clinical evidence in the "extension therapy" after initial-standard anticoagulant treatment for the secondary prevention of recurrent deep vein thrombosis (DVT). Herein, we investigated how SDX counteracts ED.Human umbilical vein endothelial cells (HUVEC) were used. Metabolic and non metabolic-induced ED was induced by treating with methylglyoxal (MGO) or irradiation (IR), respectively. Bafilomycin A1 was used to inhibit autophagy. The production of reactive oxygen species (ROS), tetrazolium bromide (MTT) assay for cell viability, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay for cell apoptosis, Real-time PCR and Western blot analysis for gene and protein expression were used.SDX protected HUVEC from MGO- or IR-induced apoptosis by counteracting the activation of the intrinsic and extrinsic caspase cascades. The cytoprotective effects of SDX resulted from a reduction in a) ROS production, b) neo-synthesis and release of pro-inflammatory cytokines (TNFα, IL1, IL6, IL8), c) DNA damage induced by MGO or IR. These effects were reduced when autophagy was inhibited.Data herein collected indicate the ability of SDX to counteract ED induced by metabolic or non-metabolic stresses by involving the intracellular autophagy pathway. Our experience significantly increases the knowledge of the mechanisms of action of SDX against ED and supports the use of SDX in the treatment of CVD, PTS and in the secondary prevention of recurrent DVT.

Background. Surround inhibition is a system that sharpens sensation by creating an inhibitory zone around the central core of activation. In the motor system, this mechanism probably contributes to the selection of voluntary movements, and it seems to be lost in dystonia. Objectives. To explore if sensory information is abnormally processed and integrated in focal hand dystonia (FHD) and if surround inhibition phenomena are operating during sensory-motor plasticity and somatosensory integration in normal humans and in patients with FHD. Methods. We looked at the MEP facilitation obtained after 5 Hz repetitive paired associative stimulation of median (PAS M), ulnar (PAS U), and median + ulnar nerve (PAS MU) stimulation in 8 normal subjects and 8 FHD. We evaluated the ratio MU/(M + U) ∗ 100 and the spatial and temporal somatosensory integration recording the somatosensory evoked potentials (SEPs) evoked by a dual nerve input. Results. FHD had two main abnormalities: first, the amount of facilitation was larger than normal subjects; second, the spatial specificity was lost. The MU/(M + U) ∗ 100 ratio was similar in healthy subjects and in FHD patients, and the somatosensory integration was normal in this subset of patients. Conclusions. The inhibitory integration of somatosensory inputs and the somatosensory inhibition are normal in patients with focal dystonia as well as lateral surrounding inhibition phenomena during sensory-motor plasticity in FHD.

Nonalcoholic fatty liver disease (NAFLD) ranges from simple steatosis to steatohepatitis, which may progress to fibrosis, and cirrhosis, leading eventually to hepatocarcinoma development. Recently, cases of hepatocarcinoma have been diagnosed in steatotic patients without nonalcoholic steatohepatitis (NASH) and cirrhosis. The p53 protein, besides its function as tumor suppressor, is emerging as an important regulator of cellular metabolism, but its role in steatosis remains unclear. We induced steatosis in HepG2 (wt-p53) and Huh7.5.1 (Y220C-mutant p53) cells using free fatty acids. We observed a different modulation of p53, different intracellular lipid content, and similar down-regulation of the de novo lipid synthesis genes but opposite modulation of the fatty acid β-oxidation pathway between HepG2 and Huh7.5.1. Accordingly, we found a diverse amount of apoptosis and reactive oxygen species between the two cell lines. Transfection of the wt-p53 in Huh7.5.1 cells reverted the different lipid metabolism behavior observed in these cells. In conclusion, unlike the wt-p53, the Y220C mutant provides a specific protection against steatosis and potentially against its progression. Our findings highlight for the first time an unknown role of a p53 mutant in the setting of steatosis. Being this mutation very frequent in human cancers, this study could be a breakthrough in explaining the occurrence of hepatocarcinoma in steatotic patients without NASH and cirrhosis.

Surround inhibition is a system that sharpens sensation by creating an inhibitory zone around the central core of activation. In the motor system, this mechanism probably contributes to the selection of voluntary movements and seems to be lost in dystonia. To explore if sensory information is abnormally processed and integrated in focal dystonia and if surround inhibition phenomena are operating during sensory-motor plasticity and sensory-motor integration in normal humans and in patients with focal hand dystonia. We looked at motor evoked potential (MEP) facilitation, in the APB and ADM muscles, obtained after 5 Hz repetitive paired associative stimulation, stimulating median (PAS M), ulnar (PAS U) and median + ulnar nerve (PAS MU) in 8 normal subjects and 8 dystonic patients. We evaluated the ratio MU/(M + U) × 100. Moreover, we tested the spatial and temporal somatosensory integration recording SEPs evoked by a dual nerve input. Our data confirmed that in dystonic patients the amount of facilitation was larger than normal subjects and the spatial specificity was lost. The MU/(M + U) × 100 ratio was similar in healthy subjects and in dystonic patients and the somatosensory integration was normal in this subset of patients. These data suggest that lateral surround inhibition phenomena during sensory-motor plasticity are normal in focal hand dystonia.

Intracloud (IC) and cloud-to-ground (CG) lightning flashes produce transient changes in the electric field (E) above a thundercloud which drive transient currents in the global electric circuit (GEC). Using in-cloud and above-cloud E data from balloons, ground-based E data, and Lightning Mapping Array data, the above-cloud charge transfers due to lightning transients are estimated for five IC and five CG flashes from four thunderstorms that occurred above the mountains in New Mexico, USA, in 1999. For the five CG flashes (which transferred -4 to -13 C to the ground), the transient currents moved +1 to + 5 C of charge upward from cloudtop toward the ionosphere, with an average transient charge transfer ofabout 35% of the charge transferred to ground. For the five IC flashes (which neutralized 6 to 21 C inside the cloud), the transient currents moved -0.7 to -3 C upward, with an average transient charge transfer of about 12% of the lightning charge. Estimates for three thunderstorms indicate that the transient currents made only a small GEC contribution compared to the quasi-stationary Wilson currents because of the offsetting effects of IC and CG flashes in these storms. However, storms with extreme characteristics, such as high flash rates or predominance of one flash type, may make a significant GEC contribution via lightning transients.

The lightning data that are recorded with a three-dimensional lightning mapping array (LMA) are compared with data from an electric field change sensor (in this case a flat-plate antenna operated both as a “slow” and a “fast” antenna). The goal of these comparisons is to quantify any time difference that may exist between the initial responses of the two instruments to a lightning flash. The data consist of 136 flashes from two New Mexico thunderstorms. It is found that the initial radiation source detected by the LMA usually precedes the initial response of both the slow and fast antennas. In a small number of cases, the flat-plate antenna response precedes the initial LMA source, but by no more than 2 ms. The observations of such a close time coincidence suggest that the first LMA radiation source of each flash was located at or very near the flash-initiation point. Thus, the first LMA radiation source and the initial sequence of sources from a lightning flash can be used as remote sensing tools to give information about the magnitude of the electric field (relative to lightning-initiation thresholds) and the direction of the electric field at the initiation location.

Nonalcoholic fatty liver disease (NAFLD) ranges from simple steatosis to steatohepatitis, which may progress to fibrosis, and cirrhosis, leading eventually to hepatocarcinoma development. Recently, cases of hepatocarcinoma have been diagnosed in steatotic patients without nonalcoholic steatohepatitis (NASH) and cirrhosis. The p53 protein, besides its function as tumor suppressor, is emerging as an important regulator of cellular metabolism, but its role in steatosis remains unclear. We induced steatosis in HepG2 (wt-p53) and Huh7.5.1 (Y220C-mutant p53) cells using free fatty acids. We observed a different modulation of p53, different intracellular lipid content, and similar down-regulation of the de novo lipid synthesis genes but opposite modulation of the fatty acid ?-oxidation pathway between HepG2 and Huh7.5.1. Accordingly, we found a diverse amount of apoptosis and reactive oxygen species between the two cell lines. Transfection of the wt-p53 in Huh7.5.1 cells reverted the different lipid metabolism behavior observed in these cells. In conclusion, unlike the wt-p53, the Y220C mutant provides a specific protection against steatosis and potentially against its progression. Our findings highlight for the first time an unknown role of a p53 mutant in the setting of steatosis. Being this mutation very frequent in human cancers, this study could be a breakthrough in explaining the occurrence of hepatocarcinoma in steatotic patients without NASH and cirrhosis. © 2014 Wiley Periodicals, Inc.

Hepatitis C virus (HCV) infection is associated with hepatic and extrahepatic manifestations, including immunological disorders. Chronic Hepatitis C (CHC) is often characterized by cholesterol and lipid metabolism alterations, leading to hepatic steatosis. Cholesterol metabolism, in fact, is crucial for the viral life cycle. Recent works described that a higher dietary cholesterol intake is associated with the progression of HCV-related liver disease. CHC patients have increased levels of T helper 17 (Th17)-cells, a lymphocytic population involved in the pathogenesis of liver inflammation and autoimmune hepatitis. The balance between Th17 and regulatory T (Treg) cells is crucial for chronic inflammation and autoimmunity. Th17-cell differentiation is deeply influenced by the activation LXRs, nuclear receptors modulating cholesterol homeostasis. Moreover, HCV may affect these nuclear receptors, and cholesterol metabolism, through both direct and indirect mechanisms. On these bases, we hypothesized that modulation of cholesterol levels through Normocaloric Low Cholesterol Diet (NLCD) may represent an innovative strategy to reduce the progression of HCV infection, through the modulation of peripheral Th17/Treg balance. To this end, we performed a pilot study to investigate whether a Normocaloric Low Cholesterol Diet may be able to modulate Th17/Treg balance in patients affected by chronic HCV infection. After 30 days of NLCD CHC patients showed a significant reduction in Th17 cells frequency, which correlated with strong reduction of IL-17 and IL-22 serum levels. At the same time, we appreciated an increase in the percentage of Treg cells, thus improving Treg/Th17balance. Moreover, we observed an increased expression of LXRs and their target genes: SREBP-1c and ABCA-1. In conclusion, NLCD finely regulates Th17/Treg balance, improving immune system response in CHC patients. This study could pave the way for new treatments of CHC patients, suggesting that change in lifestyle could support the management of these patients, promoting well-being and possibly hindering disease progression. Trial Registration ClinicalTrials.gov NCT02038387

The use of the products derived from the olive tree on human health dates back centuries. In several civilizations, the olive tree had and still has a very strong cultural and religious symbolism. Notably, the official seal and emblem of the World Health Organization features the rod of Asclepius over a world map surrounded by olive tree branches, chosen as a symbol of peace and health. Recently, accumulating experimental, clinical and epidemiological data have provided support to the traditional beliefs of the beneficial effect provided by olive derivates. In particular, the polyphenols present in olive leaves, olives, virgin (unrefined) olive oil and olive mill waste are potent antioxidant and radical scavengers with anti-tumor and anti-inflammatory properties. Here, we review the positive impact on human health of oleuropein, the most prevalent polyphenol present in olives. In addition, we provide data collected in our laboratory on the role of oleuropein in counteracting lipid accumulation in a mouse model of non-alcoholic fatty liver disease.

The liver is crucial for human life, and the health of this organ often mirrors the health of the individual. The liver can be the target of several diseases, the most prevalent of which, as a consequence of development and changes in human lifestyles, is the nonalcoholic fatty liver disease (NAFLD). NAFLD is a multifactorial disease that embraces many histo-pathologic conditions and is highly linked to metabolic derangements. Technological progress and industrialization have also had the consequence of releasing pollutants in the environment, for instance pesticides or solvents, as well as by-products of discharge, such as the particulate matter. In the last decade, a growing body of evidence has emerged, shedding light on the potential impact of environmental pollutants on liver health and, in particular, on NAFLD occurrence. These contaminants have a great steatogenic potential and need to be considered as tangible NAFLD risk factors. There is an urgent need for a deeper comprehension of their molecular mechanisms of action, as well as for new lines of intervention to reduce their worldwide diffusion. This review wishes to sensitize the community to the effects of several environmental pollutants on liver health.

Most G protein-coupled receptors contain a series of highly conserved proline and tryptophan residues within their hydrophobic transmembrane domains (TMD I-VII). To study their potential role in ligand binding and receptor function, the rat m3 muscarinic acetylcholine receptor was used as a model system. A series of mutant receptors in which the conserved proline and tryptophan residues were individually replaced with alanine and phenylalanine, respectively, was created and transiently expressed in COS-7 cells. [3H]N-methylscopolamine ([3H]NMS) saturation binding studies showed that three of the seven mutant receptors studied (Pro242-->Ala, TMD V; Pro505-->Ala, TMD VI; Pro540-->Ala, TMD VII) were expressed at 35-100 times lower levels than the wild-type receptor while displaying 'm3-like' antagonist binding affinities. Pro201-->Ala (TMD IV) showed drastically reduced binding affinities (up to 450-fold) for both muscarinic agonists and antagonists. Whereas most mutant receptors retained strong functional activity, Pro540-->Ala (TMD VII) was found to be severely impaired in its ability to stimulate carbachol-induced phosphatidyl inositol hydrolysis (Emax approximately 25% of wild type m3). Interestingly, this mutant receptor bound muscarinic agonists with 7- to 19-fold higher affinities than the wild type receptor. The Trp-->Phe substitutions (Trp192-->Phe, TMD IV; Trp503-->Phe, TMD VI; Trp530-->Phe, TMD VII) resulted in less pronounced changes (compared with the Pro-->Ala mutant receptors) in both ligand binding and receptor function. Our data indicate that the proline residues that are highly conserved across the entire superfamily of G protein-coupled receptors play key roles in receptor expression, ligand binding and receptor activation.

[1] Data from ground-based and balloon-borne electric field meters are used in conjunction with lightning mapping array data to estimate the amount of charge transferred and energy liberated by 40 lightning flashes in four New Mexico mountain thunderstorms. Two models are developed to estimate the lightning charge transfer: point charges at the geometric center of two or more lightning charge distributions, and uniform charge distributions placed along the lightning channels. Results indicate that flashes with substantial horizontal extent, primarily intracloud (IC) flashes during the storm's mature stage, are better modeled with uniform charge distributions. Less extensive flashes, both cloud-to-ground (CG) and early-stage IC flashes, are better modeled with point charges. The amount of charge transferred by 29 IC flashes ranged from ±3.9 to ±85.8 C, with an average of ±17.6 C. Mature-stage IC flashes transfer significantly more charge, on average, than early-stage IC flashes. The 11 CG flashes transferred negative charge to the ground; the average amount of charge lowered was −8.7 C. All but one CG flash also had substantial in-cloud charge transfers (average ±9.3 C, maximum ±44 C) in addition to the charge moved to the ground. Individual lightning charge transfers are combined with concurrent in-cloud potentials to estimate the electrical energy released by each flash. The IC flashes released an average of 1.6 GJ of energy per flash, while the average for CG flashes was about half as much energy per flash. Using average charge and potential values for the group of storms yields lightning energy estimates of 2 × 108 to 7 × 109 J, depending on flash type and storm stage.

[1] The electric field (E) at the ground beneath a thunderstorm often exhibits an end-of-storm oscillation (EOSO) during the storm's decay phase in which E typically undergoes three polarity changes over a period of 30–75 min. To determine why the surface E oscillates, the timing of the EOSO polarity changes of a multicellular storm in New Mexico has been compared to a series of balloon, radar, and lightning data. Three in situ balloon soundings provide vertical profiles of E and inferred charge regions just before and during the EOSO. These data have been compared to the evolving precipitation structure determined from orthogonal range-height scans of two radars and to several other parameters including lightning mapping array data. Before the EOSO began, the storm had the typical electrical structure of the late mature stage. Estimates of the total charge magnitudes in each of the principal regions before and during the EOSO ranged from 7 to 33 C. The comparisons indicate that surface E polarity changes during the EOSO were (1) primarily owing to the successive fallout of three of the four principal charge regions in the storm's mature stage charge structure, modified by (2) the growth or decay of the charge regions as they descended, and (3) changes in the screening charges at the upper and lower cloud boundaries. More limited data from two other decaying storms indicate that similar in-cloud evolution was probably occurring during their EOSOs.

[1] This study examines the way in which different types of lightning, both cloud-to-ground (CG) and intracloud (IC) flashes, drive current in the global circuit. A numerical model of the transient electric field due to CG and IC flashes and their Maxwell relaxation (slow transients) is developed. The electric field (E) and current distributions, the decay time of E, and the total charge transferred to the ionosphere and the ground are calculated. Because of the slow transients, only a portion of the charge neutralized by a flash contributes to the global circuit, with the efficiency depending on the altitudes of the lightning charges. Typical CG flashes have efficiencies of 55–75%, and typical IC flashes have 5–15%. An example from balloon E data has been used to verify the theory. Total current estimations of the combined CG and IC slow transient processes in the global electric circuit range from 50–400 A.

[1] In situ electric field (E) measurements and inferred lightning initiation locations of three cloud-to-ground flashes are used to identify a thunderstorm region in which the preflash E exceeded the threshold for runaway breakdown. The maximum measured E in the region was 186 kV m−1 at 5.77 km altitude, which for runaway electrons is equivalent to 370 kV m−1 at sea level; this E value is ∼130% of the estimated threshold for an avalanche of runaway electrons. In addition, the volume where E exceeded the runaway threshold was estimated to be 1–4 km3, with a vertical depth of about 1000 m. At least within part of this volume the characteristic scale height for exponential growth of runaway electrons was 100 m or less. Thus for these three flashes the electric field conditions necessary for runaway breakdown existed, and runaway breakdown could have initiated the flashes.

Aim of the present study was to investigate motor cortex synaptic plasticity in both hemispheres of patients affected by brain tumors. We employed an electrophysiological protocol, named repetitive paired associative stimulation (rPAS), involving repetitive transcranial magnetic stimulation (rTMS) paired with electrical stimulation of the contralateral median nerve, that modifies cortico-spinal excitability. Ten right-handed patients (3 women), aged 49–70 years (mean age: 54.8 years), participated in the study. Motor evoked potentials (MEP) were measured at baseline before and for up to 30 min after 5 Hz-rPAS in abductor pollicis brevis (APB). rPAS consisted of 600 pairs of transcranial magnetic stimuli, at a rate of 5 Hz for 2 min, coupled with electrical median nerve stimulation preceding TMS over the contralateral M1 at an interstimulus interval of 25 ms. In both hemispheres of patient with brain tumors rPAS did not induce any significant changes in MEP amplitudes, compared with aged matched healthy controls. Synaptic plasticity is impaired in both hemispheres of patients affected by brain tumors.

In this study we investigated cortical excitability, inhibitory and excitatory intracortical circuitries, by using transcranial magnetic stimulation, in both hemispheres of patients affected by brain tumors. Cortical excitability was probed with transcranial magnetic stimulation in 10 right-handed patients (3 women), aged 49–70 years (mean age: 54.8 years) and 10 age-matched healthy controls. We tested motor thresholds (MT), motor evoked potential (MEP) recruitment curve, cortical silent period (CSP), short and long intracortical inhibition (SICI and LICI), intracortical facilitation (ICF), short and long afferent inhibition (SAI and LAI) and inter-hemispheric inhibition (IHI). RMT and AMT were higher in both hemispheres of patients with brain tumors than healthy controls. The slope of MEP recruitment curve was deeper in both hemispheres of patients with brain tumors. SICI, LICI, SAI and LAI are reduced in patients with brain tumors compared with aged matched healthy controls. Cortical excitability, specially inhibitory intracortical circuitries, are impaired in both hemispheres of patients affected by brain tumors.

We have previously shown that systemic injection of (-)nicotine produces a selective up-regulation of fibroblast growth factor (FGF)-2 mRNA levels in rat striatum. Because (-)nicotine can increase striatal release of dopamine and glutamate, in the present study we have investigated the contribution of these neurotransmitters in the modulation of FGF-2 expression. We found that coinjection of dopaminergic D1 (SCH23390) or D2 (haloperidol) receptor antagonists prevents nicotine-induced elevation of FGF-2 expression. However, injection of the NMDA receptor antagonist MK-801 produced a significant increment of FGF-2 mRNA and protein levels in rat striatum similar to the effect produced by (-)nicotine alone. Interestingly this effect of MK-801 could also be prevented by D1 or D2 receptor antagonists, suggesting that an elevation of dopamine levels may be required for the regulation of the trophic molecule. Accordingly we found that the non-selective dopaminergic agonist apomorphine can similarly increase striatal FGF-2 mRNA levels. Despite the observation that both D1 and D2 receptors appear to contribute to the modulation of FGF-2 expression, only a direct activation of D2 receptors, through quinpirole administration, was able to mimic the effect of apomorphine. On the basis of FGF-2 neurotrophic activity, these results suggest that direct or indirect activation of dopaminergic system can be neuroprotective and might reduce cell vulnerability in degenerative disorders.

By means of the expression of two chimeric receptors, alpha(2)/M(3) and M(3)/alpha(2), in which the carboxy-terminal receptor portions, containing transmembrane domains VI and VII, were exchanged between the alpha(2C)-adrenergic and the M(3) muscarinic receptor, it has been shown that G protein-coupled receptors are able to interact functionally with each other at the molecular level to form (hetero)dimers. In the present study, we tested the hypothesis that interaction between two different muscarinic receptor subtypes can lead to the formation of a heterodimeric muscarinic receptor with a new pharmacological profile. Initially, muscarinic M(2) or M(3) wild-type receptors were expressed together with gene fragments originating from M(3) or M(2) receptors, respectively. Antagonist binding, performed with pirenzepine and tripitramine, revealed the presence of two populations of binding sites: one represents the wild-type M(2) or M(3) receptors, the other the heterodimeric M(2)/M(3) receptor. In another set of experiments, we constructed a point mutant M(2) receptor M(2) (Asn404--Ser), in which asparagine 404 was replaced by serine. Although this receptor alone did not show any binding for N-[(3)H]methylscopolamine (up to 2 nM), when cotransfected with M(3), it resulted in the rescue of a high-affinity binding for tripitramine. These findings demonstrate that M(2) and M(3) muscarinic receptor subtypes can cross-interact with each other and form a new pharmacological heterodimeric receptor.

Apomorphine is a potent non selective agonist at the D1 and D2 dopamine receptors acting both pre- and post-synaptically. In this report we describe a novel function of apomorphine, independent from its dopaminergic activity. Apomorphine inhibits Chinese hamster ovary (CHO)-K1 cell proliferation in a dose-dependent manner. The EC50 of apomorphine-induced inhibition of CHO-K1 cell proliferation determined by cell counting was 3.24 +/- 0.07 microM. Remarkably, the dose-response curve obtained by measuring the incorporation of [3H]thymidine was practically identical to the previous one giving an EC50 of 3.52 +/- 0.04 microM. The dopaminergic antagonists SCH23390 and spiperone at a concentration of 10 microM (well beyond their Kd values for the dopamine D1- and D2-like receptors respectively) were not able to antagonize the effect of apomorphine on CHO-K1 cell proliferation. Apomorphine exerts its effect early during incubation; CHO-K1 cells exposed to apomorphine for a period as short as 1 h and then allowed to grow for three days were significantly reduced in number with respect to untreated control cells. After four hours of exposition to apomorphine (10 microM) the antiproliferative effect was similar to that seen when this compound was present in the bath for all three days. Concentrations of apomorphine higher than 10 microM induced cell death, and the colony was completely destroyed at 50 microM. Cytometric analyses showed a significant accumulation of CHO-K1 cells in the G2/M phase.