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PURPOSE No liquid biomarkers are approved in metastatic renal cell carcinoma (mRCC) despite the need to predict and monitor response noninvasively to tailor treatment choices. Urine and plasma free glycosaminoglycan profiles (GAGomes) are promising metabolic biomarkers in mRCC. The objective of this study was to explore if GAGomes could predict and monitor response in mRCC. PATIENTS AND METHODS We enrolled a single-center prospective cohort of patients with mRCC elected for first-line therapy (ClinicalTrials.gov identifier: NCT02732665 ) plus three retrospective cohorts (ClinicalTrials.gov identifiers: NCT00715442 and NCT00126594 ) for external validation. Response was dichotomized as progressive disease (PD) versus non-PD every 8‐12 weeks. GAGomes were measured at treatment start, after 6-8 weeks, and every third month in a blinded laboratory. We correlated GAGomes with response and developed scores to classify PD versus non-PD, which were used to predict response at treatment start or after 6-8 weeks. RESULTS Fifty patients with mRCC were prospectively included, and all received tyrosine kinase inhibitors (TKIs). PD correlated with alterations in 40% of GAGome features. We developed plasma, urine, and combined glycosaminoglycan progression scores that monitored PD at each response evaluation visit with the area under the receiving operating characteristic curve (AUC) of 0.93, 0.97, and 0.98, respectively. For internal validation, the scores predicted PD at treatment start with the AUC of 0.66, 0.68, and 0.74 and after 6-8 weeks with the AUC of 0.76, 0.66, and 0.75. For external validation, 70 patients with mRCC were retrospectively included and all received TKI-containing regimens. The plasma score predicted PD at treatment start with the AUC of 0.90 and at 6-8 weeks with the AUC of 0.89. The pooled sensitivity and specificity were 58% and 79% at treatment start. Limitations include the exploratory study design. CONCLUSION GAGomes changed in association with mRCC response to TKIs and may provide biologic insights into mRCC mechanisms of response.

International audience; We study the impact of competition on morals using a dynamic variant of the die-under-the-cup task. Players can repeatedly misreport the outputs of consecutive die rolls to earn more money, either under an individual piece-rate pay scheme or in a two-player tournament. In this dynamic setting, we disentangle the effect of the incentive scheme and the effect of information provision about one's relative performance, by comparing settings with continuous vs. final ex post feedback on the counterpart's reported outcome. We find that individuals lie more under competitive rather than non-competitive incentive schemes, but only if both players can cheat in the tournament. Continuous feedback on the counterpart's reports does not increase cheating in the tournament, while it does under the piece-rate scheme. These findings shed light on the effects that different competition and information policies have on morals in occupational settings.

Abstract Mucopolysaccharidosis type II (MPS II) is a neurometabolic disorder, due to the deficit of the lysosomal hydrolase iduronate 2-sulfatase (IDS). This leads to a severe clinical condition caused by a multi-organ accumulation of the glycosaminoglycans (GAGs/GAG) heparan- and dermatan-sulfate, whose elevated levels can be detected in body fluids. Since 2006, enzyme replacement therapy (ERT) has been clinically applied, showing efficacy in some peripheral districts. In addition to clinical monitoring, GAG dosage has been commonly used to evaluate ERT efficacy. However, a strict long-term monitoring of GAG content and composition in body fluids has been rarely performed. Here, we report the characterization of plasma and urine GAGs in Ids knock-out (Ids-ko) compared to wild-type (WT) mice, and their changes along a 24-week follow-up, with and without ERT. The concentration of heparan-sulfate (HS), chondroitin-sulfate (CS), and dermatan-sulfate (DS), and of the non-sulfated hyaluronic acid (HA), together with their differentially sulfated species, was quantified by capillary electrophoresis with laser-induced fluorescence. In untreated Ids-ko mice, HS and CS + DS were noticeably increased at all time points, while during ERT follow-up, a substantial decrease was evidenced for HS and, to a minor extent, for CS + DS. Moreover, several structural parameters were altered in untreated ko mice and reduced after ERT, however without reaching physiological values. Among these, disaccharide B and HS 2s disaccharide showed to be the most interesting candidates as biomarkers for MPS II. GAG chemical signature here defined provides potential biomarkers useful for an early diagnosis of MPS II, a more accurate follow-up of ERT, and efficacy evaluations of newly proposed therapies. Key messages Plasmatic and urinary GAGs are useful markers for MPS II early diagnosis and prognosis. CE-LIF allows GAG structural analysis and the quantification of 17 different disaccharides. Most GAG species increase and many structural features are altered in MPS II mouse model. GAG alterations tend to restore to wild-type levels following ERT administration. CS+DS/HS ratio, % 2,4dis CS+DS, and % HS 2s are potential markers for MPS II pathology and ERT efficacy.

Cancer mortality is exacerbated by late-stage diagnosis. Liquid biopsies based on genomic biomarkers can noninvasively diagnose cancers. However, validation studies have reported ~10% sensitivity to detect stage I cancer in a screening population and specific types, such as brain or genitourinary tumors, remain undetectable. We investigated urine and plasma free glycosaminoglycan profiles (GAGomes) as tumor metabolism biomarkers for multi-cancer early detection (MCED) of 14 cancer types using 2,064 samples from 1,260 cancer or healthy subjects. We observed widespread cancer-specific changes in biofluidic GAGomes recapitulated in an in vivo cancer progression model. We developed three machine learning models based on urine ( N urine = 220 cancer vs. 360 healthy) and plasma ( N plasma = 517 vs. 425) GAGomes that can detect any cancer with an area under the receiver operating characteristic curve of 0.83–0.93 with up to 62% sensitivity to stage I disease at 95% specificity. Undetected patients had a 39 to 50% lower risk of death. GAGomes predicted the putative cancer location with 89% accuracy. In a validation study on a screening-like population requiring ≥ 99% specificity, combined GAGomes predicted any cancer type with poor prognosis within 18 months with 43% sensitivity (21% in stage I; N = 121 and 49 cases). Overall, GAGomes appeared to be powerful MCED metabolic biomarkers, potentially doubling the number of stage I cancers detectable using genomic biomarkers.

Analytical approaches utilized for the characterization of polyphenols from propolis useful for the determination of its quality is investigated in this study. A qualitative and quantitative evaluation of propolis bioactive molecules is of interest in medicine and nutraceuticals. Recent powerful analytical techniques are of great utility to separate and quantify polyphenols in extracts and finished products due to their capacity to produce typical fingerprints and a reliable identification of many components. According to this, an HPLC-UV-MS procedure was validated and applied for the characterization and quantification of bioactive substances in propolis and for an accurate assessment of their content in extract samples. By using this analytical approach, we obtained specific compositions related to brown propolis acquired from different geographic areas (and preparations and treatment). This is more important by considering the scientific opinion of European Food Safety Authority (EFSA) which provided a negative response related to health claims of propolis and its polyphenols. These results prove that HPLC-MS is an attractive tool for the standardization and quality control of propolis and may be realistically applied to screen raw material and to evaluate finished commercial preparations and nutraceutical benefits. Key words: Propolis, polyphenols, flavonoids, phenolic acids, high-performance liquid chromatography-mass spectrometry (HPLC-MS), quality control, standardization

Plasma and urine glycosaminoglycans (GAGs) are long, linear sulfated polysaccharides that have been proposed as potential noninvasive biomarkers for several diseases. However, owing to the analytical complexity associated with the measurement of GAG concentration and disaccharide composition (the so-called GAGome), a reference study of the normal healthy GAGome is currently missing. Here, we prospectively enrolled 308 healthy adults and analyzed their free GAGomes in urine and plasma using a standardized ultra-high-performance liquid chromatography coupled with triple-quadrupole tandem mass spectrometry method together with comprehensive demographic and blood chemistry biomarker data. Of 25 blood chemistry biomarkers, we mainly observed weak correlations between the free GAGome and creatinine in urine and hemoglobin or erythrocyte counts in plasma. We found a higher free GAGome concentration - but not a more diverse composition - in males. Partitioned by gender, we also established reference intervals for all detectable free GAGome features in urine and plasma. Finally, we carried out a transference analysis in healthy individuals from two distinct geographical sites, including data from the Lifelines Cohort Study, which validated the reference intervals in urine. Our study is the first large-scale determination of normal free GAGomes reference intervals in plasma and urine and represents a critical resource for future physiology and biomarker research.

In response to the pandemic of COVID-19 and in lack of pharmaceutical solutions, many countries have introduced social and physical distancing regulations to contain the transmission of the virus. These measures are effective insofar as they are able to quickly change people's habits. This is achieved by changing the monetary incentives of rule violators but also by shifting people's perception regarding the appropriateness of socialization. We studied the effect of introducing, and then lifting, distancing regulations on the perceived norm regarding social encounters. We conducted an online incentivized experiment in France where we elicited the same participants' perceived norm and social distancing behavior every week for three months. We found that people shifted behavior and norm perception as soon as the government introduced or removed distancing measures. This effect was fast acting and long lasting. This is informative for future interventions, especially in light of a possible COVID-19 recurrence.

Plasma and urine glycosaminoglycans (GAGs) are long linear sulfated polysaccharides recognized as potential non-invasive biomarkers for several diseases. However, owing to the analytical complexity associated with the measurement of GAG concentration and disaccharide composition, the so-called GAGome, a reference study of the normal healthy GAGome is currently missing. Here, we prospectively enrolled 308 healthy adults and analyzed their urine and plasma free GAGomes using a standardized ultra-high-performance liquid chromatography coupled with triple-quadrupole tandem mass spectrometry (UHPLC-MS/MS) method together with comprehensive demographic and blood chemistry biomarker data. Of 25 blood chemistry biomarkers, we mainly observed weak correlations between the free GAGome and creatinine in urine, and hemoglobin or erythrocyte counts in plasma. We found higher free GAGome concentration – but not composition - in males. Partitioned by gender, we established reference intervals for all detectable free GAGome features in urine and plasma. We carried out a transference analysis in healthy individuals from two distinct geographical sites, including the Lifelines Cohort Study, which validated the reference intervals in urine. Our study is the first large-scale determination of normal plasma and urine free GAGomes reference intervals and represents a critical resource for physiology and biomarker research.

Background Viral infections are among the most common problems in health-care practice. Natural products offer great promise as potentially effective antiviral drugs. Propolis is a honeybee product with biological properties and therapeutic applications. We aimed to investigate the antiviral activity of different extracts of Standardized Propolis Preparations (M.E.D.®) with glycol, ethanol, glycerol and soya oil, against herpes simplex type-1 (HSV-1) and type 2 (HSV-2) viruses. Methods Chemical composition and antiviral activity of each extract were determined. The selective index (SI=CC50/EC50) was determined as a parameter to indicate the in vitro antiviral activity of the extracts compared with acyclovir as the control. Results SI values of glycol, ethanol, glycerol, soya oil extracts and acyclovir were determined as 6.8, 4.1, 2.2, 3.3 and 6.3 against HSV-1, and as 6.4, 7.7, 1.9, 4.2 and 2.9 against HSV-2, respectively. Glycolic propolis extract was found to possess a greater antiviral activity than acyclovir for both HSV-1 and 2, while glycolic, ethanolic and soya oil preparations were found to have more significant activity than acyclovir for HSV-2. Conclusions It was determined that standardized propolis preparations have antiviral bioactivity against HSV.

Pistacia lentiscus L. (PL) is an evergreen shrub from which it is derived a precious aromatic resin called mastic gum or mastic. This extract possesses numerous proprieties, such as antimicrobial, anti-inflammatory, anticancer, etc. The aim of this study was to investigate the biological activities of a patented PL Chios mastic extract in a human small intestine mucosa model, the Caco-2 cells. PL Chios mastic extract showed no toxic effect in Caco-2 cells treated with lower concentrations (

Objectives. We determined the polyphenol content in a defined volume of chemically characterized and standardized propolis sample to evaluate its effectiveness in patients with chronic periodontitis.Materials and methods. After having determined the polyphenol content of a given volume of propolis extract and characterized the molecular profile, 150 patients were enrolled, randomly divided into three groups and subjected to three different treatments, Scaling and Root planing (SRP) associated with propolis, SRP with 1% chlorhexidine gel and SRP only. Before the treatments, Full Mouth Plaque Score (FMPS), Pocket Depth at Probing (PPD), Full Mouth Bleeding Score (FMBS), Impaired furcations according to the Hamp Classification, Mobility, Gingival recession and Clinical Attack Level (CAL) were evaluated. Results. A significant reduction of all the studied variables was observed in the three examined groups even if in the group treated with propolis, after 6 weeks, the reduction, and therefore the improvement, was higher than SRP treatment alone and in the presence of 1% chlorhexidine gel. Finally, there were no significant differences in the reduction of PPD and CAL between the groups treated with chlorhexidine and the group treated with SRP alone.Conclusion. Compared to the other two groups, Subjects treated with 10.4 mg propolis showed a significant improvement in all four variables. Clinical relevance. This study shows that propolis could be used as a natural adjuvant in the treatment of periodontal disease.

Propolis is important in complementary and alternative medicine having well-known therapeutic applications. Artepillin C, a main component of Brazilian (green) propolis, has attracted great attention for its anticancer action. Consequently, the synthesis of artepillin C has been reported but, due to the limited yield and elevated costs, this biomolecule is largely produced from Brazilian propolis. We report the capillary electrophoresis (CE) separation of artepillin C in Brazilian propolis also comparing the results with those of HPLC-UV-MS. Optimal separation was obtained with a simple buffer constituted of sodium tetraborate 30 mM pH 9.2 and detection at 210 nm. Artepillin C and the polyphenols of propolis were fully separated with a voltage gradient of 30 to 8 kV and a current of 300 μA for a total run of 50 min. The sensitivity of CE-UV was 22 times greater than HPLC-UV and 100 times more than HPLC-MS with also a stronger reduction in the run time and a greater robustness and reproducibility. The development of CE as an effective and reliable method for the analysis of artepillin C is desired as the standardized quality controls are essential before propolis or its biomolecules can be adopted routinely in nutraceuticals, food ingredients and therapeutic applications.

Complex (hetero)polysaccharides, glycosaminoglycans (GAGs), are fundamental biomacromolecules for all living organisms having important biological and pathophysiological roles. Moreover, in the form of native or depolymerized biomolecules, they are active pharmaceutical and nutraceutical agents after extraction and purification from animal sources. Capillary electrophoresis (CE) is applied in many different biological, pharmacological, and nutraceutical fields, clearly showing evidence of the importance of this analytical method in glycosciences. Thanks to its versatility in separation and detection modes, CE may be applied for the analysis and quantification of intact high molecular mass heteropolysaccharides as well as their low molecular weight derivatives up to derived oligosaccharides, disaccharides, and monosaccharides, as single species but also in mixtures. As discussed in the present review and largely illustrated in the current scientific literature, CE may be one of the analytical techniques most useful in quality control laboratories of pharmaceutical and nutraceutical companies for the determination of GAGs’ purity and quality in raw material and finished products.

Glycosaminoglycans (GAGs) are long linear sulfated polysaccharides implicated in processes linked to disease development such as mucopolysaccharidosis, respiratory failure, cancer, and viral infections, thereby serving as potential biomarkers. A successful clinical translation of GAGs as biomarkers depends on the availability of standardized GAG measurements. However, owing to the analytical complexity associated with the quantification of GAG concentration and structural composition, a standardized method to simultaneously measure multiple GAGs is missing. In this study, we sought to characterize the analytical performance of a ultra-high-performance liquid chromatography coupled with triple-quadrupole tandem mass spectrometry (UHPLC-MS/MS)-based kit for the quantification of 17 GAG disaccharides. The kit showed acceptable linearity, selectivity and specificity, accuracy and precision, and analyte stability in the absolute quantification of 15 GAG disaccharides. In native human samples, here using urine as a reference matrix, the analytical performance of the kit was acceptable for the quantification of CS disaccharides. Intra- and inter-laboratory tests performed in an external laboratory demonstrated robust reproducibility of GAG measurements showing that the kit was acceptably standardized. In conclusion, these results indicated that the UHPLC-MS/MS kit was standardized for the simultaneous measurement of GAG disaccharides allowing for comparability of measurements and enabling translational research.SummaryAnalytical performance of a kit for standardized GAG measurements, based on an established UHPLC-MS/MS method

Anise hyssop, Agastache foeniculum, is a widely used medicinal herb with known antioxidant properties. We studied how dietary supplementation with dried A. foeniculum leaf powder affected physiological and metabolic traits as well as activities of antioxidant enzymes and markers of oxidative stress in Drosophila melanogaster. Dietary hyssop extended the lifespan in a sex and genotype independent manner over a broad range of concentrations up to 30 mg/ml. Dietary supplementation with the herb significantly increased fecundity, resistance to oxidative stress and starvation. Higher transcript levels of Drosophila insulin-like peptide (dilp2) and decreased dilp3 and dilp6 transcripts together with increased levels of glycogen and triacylglycerols support an alteration of insulin signaling by the plant extract. Increased enzymatic activities of superoxide dismutase and aconitase as well as elevated protein and low molecular mass thiols also supported an alteration of free radical process in flies treated with dietary A. foeniculum leaf powder. Thus, physiological and metabolic traits as well as free radical processed may be affected by active compounds detected in extracts of anise hyssop leaves and contribute to the increased lifespan and reproductive (egg-laying) activity observed.

Significance Using large-scale incentivized online experiments, we tested two possible origins of individuals’ forgetting about their past cheating behavior in a mind game. We found that purely hedonic considerations, such as the maintenance of a positive self-image, are not sufficient to motivate unethical amnesia, but the addition of an instrumental value to forgetting triggers such amnesia. Individuals forget their past lies more when amnesia can serve as an excuse not to engage in future morally responsible behavior. These findings shed light on the interplay between dishonesty and memory and suggest further investigations of the cost function of unethical amnesia. A policy implication is that improving ethics requires making unethical amnesia more difficult for individuals., Humans care about morality. Yet, they often engage in actions that contradict their moral self. Unethical amnesia is observed when people do not remember or remember less vividly these actions. This paper explores two reasons why individuals may experience unethical amnesia. Forgetting past unethical behavior may be motivated by purely hedonic or affective reasons, such as the willingness to maintain one’s moral self-image, but also by instrumental or strategic motives, in anticipation of future misbehavior. In a large-scale incentivized online experiment (n = 1,322) using a variant of a mind game, we find that hedonic considerations are not sufficient to motivate the forgetting of past cheating behavior. This is confirmed in a follow-up experiment (n = 1,005) in which recalls are elicited the same day instead of 3 wk apart. However, when unethical amnesia can serve as a justification for a future action, such as deciding on whether to keep undeserved money, motivated forgetting is more likely. Thereby, we show that motivated forgetting occurs as a self-excuse to justify future immoral decisions.

Background The most common symptoms of mild upper respiratory tract infections (URTIs) are sore throat, muffled dysphonia, and swelling and redness of the throat, which result from the inflammation process following acute bacterial or viral infection. Hypothesis/purpose As propolis is a natural resinous substance traditionally used to maintain oral cavity and upper respiratory tract health due to its antimicrobial and anti-inflammatory properties, the aim of this study is to evaluate the efficacy of an oral spray based on poplar-type propolis extract with a known and standardized polyphenol content, on the remission of the symptoms associated with mild uncomplicated URTIs. Study design A monocentric, randomized, double-blind, placebo-controlled clinical trial was performed. Methods This study was conducted in 122 healthy adults who had perceived mild upper respiratory tract infections. Participants, randomly assigned to receive either propolis oral spray (N = 58) or placebo (N = 64), underwent four visits (baseline = t0, after 3 days = t1 and after 5 days = t2 and after a follow-up of 15 days = t3) in an outpatient setting. Propolis oral spray total polyphenol content was 15 mg/ml. The dosage was 2-4 sprays three times/day (corresponding to 12-24 mg of polyphenols/day), for five days. The duration of the study was 8 weeks. Results After 3 days of treatment, 83% of subjects treated with propolis oral spray had remission of symptoms, while 72% of subjects in the placebo group had at least one remaining symptom. After five days, all subjects had recovered from all symptoms. This means that resolution from mild uncomplicated URTIs took place two days earlier, instead of taking place in five days as recorded in the control group. There was no relationship between the ingestion of propolis oral spray or placebo and adverse reactions. Conclusion Propolis oral spray can be used to improve both bacterial and viral uncomplicated URTI symptoms in a smaller number of days without the use of pharmacological treatment, leading to a prompt symptom resolution.

We study a dynamic variant of the die-under-the-cup task where players can repeatedly misreport the outcomes of consecutive die rolls to earn more money, either under a non- competitive piece rate scheme or in a two-player competitive tournament. In this dynamic setting we test (i) whether giving continuous feedback (vs. final ex post feedback) on the opponent's reported outcome to both players encourages cheating behavior, and (ii) to what extent this influence depends on the incentive scheme in use (piece rate vs. tournament). We also vary whether the opponent is able to cheat or not. We find that people lie more when placed in a competitive rather than a non-competitive setting, but only if both players can cheat in the tournament. Continuous feedback on the counterpart's reports increases cheating under the piece-rate scheme but not in a competitive setting. Our results provide new insights on the role that feedback plays on cheating behavior in dynamic settings under different payment schemes, and shed light on the origins of the effect of competition on dishonesty.

Deterrence institutions are widely used in modern societies to discourage rule violations but whether they have an impact beyond their immediate scope of application is usually ignored. Using a quasi-experiment, we found evidence of spillover effects across contexts. We identified fraudsters and non-fraudsters on public transport who were or not exposed to ticket inspections by the transport company. We then measured the intrinsic honesty of the same persons in a new, unrelated context where they could misappropriate money. Instead of having an expected educative effect across contexts, the exposure to deterrence practices increased unethical behavior of fraudsters but also, strikingly, of non-fraudsters, especially when inspection teams were larger. Learning about the prevailing norm is the most likely channel of this spillover effect.

In response to the pandemic of COVID-19 and in lack of pharmaceutical solutions, many countries have introduced social and physical distancing regulations to contain the transmission of the virus. These measures are effective insofar as they are able to quickly change people’s habits. This is achieved by changing the monetary incentives of rule violators but also by shifting people’s perception regarding the appropriateness of socialization. We studied the effect of introducing, and then lifting, distancing regulations on the perceived norm regarding social encounters. We conducted an online incentivized experiment in France where we elicited the same participants’ perceived norm and social distancing behavior every week for three months. We found that people shifted behavior and norm perception as soon as the government introduced or removed distancing measures. This effect was fast acting and long lasting. This is informative for future interventions, especially in light of a possible COVID-19 recurrence.

Chondroitin sulfate was extracted from the cartilage of smooth hound (CSSH) and then purified by anion exchange chromatography. The structual characteristic of CSSH was evaluated by acetate cellulose electrophoresis, FTIR, 13C NMR and SAX-HPLC. Molecular weight of CSSH was average 68.78 KDa. Disaccharide analysis indicated that CSSH was predominately composed of monosulfated disaccharides in position 6 and 4 of the N-acetylgalactosamine (45.34% and 32.49%, respectively). CSSH was tested for in vitro anticoagulant activity using the three classical coagulation assays (activated partial thromboplastin time (aPTT), prothrombine time (TT) and thrombin time (PT) tests). The finding showed that CSSH prolonged significatively (p CSSH demonstrated hopeful antiproliferative and anticoagulant potential, which may be used as a novel and effective drug.

Objective: In this pilot study, we report the composition, structure and properties of glycosaminoglycans (GAG) present in milk samples of various countries and ethnicities.Methods: Fifty samples o...

3034 Background: Non-invasive liquid biopsies promise to enable early cancer detection and improve patient outcomes. However, virtually all liquid biopsies rely on genomic biomarkers, with limited sensitivity to early-stage tumors and poor detection of cancers shedding little cell-free DNA, like genitourinary or brain tumors. Here, we explored the use of plasma and urine glycosaminoglycan (GAGs) profiles, or GAGomes, as biomarkers reflective of tumor metabolism to serve as an alternative pan-cancer liquid biopsy. Methods: In this case-control study, we enrolled retrospective and prospective cohorts from Sweden and Italy. Included cases were treatment-naïve early-stage/low-grade cancers or metastatic/high-grade cancers across 14 histological types. Included controls were healthy 22-78 y/o adults with no history of cancer. We measured GAGomes – encompassing 17 chondroitin sulfate (CS), heparan sulfate (HS), and hyaluronate (HA) disaccharides - using a standardized UHPLC-MS/MS-based kit in a central blind laboratory. We tested the top GAGome features different in cancer using Bayesian estimation. These were used to design one plasma and one urine GAG score for the binary classification of cancer vs. control in a discovery set. We computed the area-under-the-curve (AUC), and sensitivity at 98% specificity of each GAG score in the validation set. A subset analysis was performed in early-stage/low-grade cancers only. In the subset of cases with survival records, we used multivariable Cox regression to estimate the hazard ratio (HR) for overall survival (OS) on each GAG score adjusted for cancer type, age, and gender. Results: GAGomes were measured in 753 plasma samples (460 cancers across 14 types, median age = 66 y/o, 51% female vs. 293 healthy adults, median age = 58 y/o, 57% female) and 559 urine samples (219 cancers across 5 types, median age = 69 y/o, 23% female vs. 340 healthy adults, median age = 56 y/o, 60% female). In the discovery set, the urine GAG score had an AUC = 0.80 (95% CI: 0.74-0.85, 124 cancers across 5 types vs. 184 controls) while the plasma GAG score had an AUC = 0.82 (95% CI: 0.78-0.86, 153 cancers across 14 types vs. 282 controls). In the validation set, the urine GAG score had an AUC = 0.78 (95% CI: 0.71-0.84, 95 cancers across 5 types vs. 156 controls) with 35% sensitivity at 98% specificity. The plasma GAG score had an AUC = 0.84 (95% CI: 0.79-0.88, 178 cancers across 14 types vs. 140 controls) with 41% sensitivity at 98% specificity. In the subset of early-stage/low-grade cancers, the AUC was 0.78 and 0.72 in plasma and urine, respectively. The plasma and urine GAG scores were independent predictors of OS regardless of cancer type (HR = 1.39, p = 0.005 in plasma [ N = 283, 11 types, 67 deaths, median follow-up 17 months] and HR = 1.53, p = 0.016 in urine [ N = 161, 4 types, 32 deaths, median follow-up 15 months]). Conclusions: GAGomes were sensitive non-invasive metabolic biomarkers for any-stage cancer, including genitourinary and brain tumors.

Hyaluronic Acid (HA), or hyaluronan, is a polysaccharide composed of a repeated disaccharide unit of β-1,4-D-glucuronic acid - β-1,3-N-acetyl-D-glucosamine linked by glycosidic bonds.

Deterrence institutions are widely used in modern societies to discourage rule violations but whether they have an impact beyond their immediate scope of application is usually ignored. Using a quasi-experiment, we found evidence of spillover effects across contexts. We identified fraudsters and non-fraudsters on public transport who were or not exposed to ticket inspections by the transport company. We then measured the intrinsic honesty of the same persons in a new, unrelated context where they could misappropriate money. Instead of having an expected educative effect across contexts, the exposure to deterrence practices increased unethical behavior of fraudsters but also, strikingly, of non-fraudsters, especially when inspection teams were larger. Learning about the prevailing norm is the most likely channel of this spillover effect.

Dishonesty in daily life is widespread. Fare-dodging in public transport illustrates ordinary dishonesty. This chapter investigates how the size of fraud affects the behavior of actual fare-dodgers and non-fare-dodgers in a framed lab experiment. In our public transportation game, participants make successive decisions between buying and not buying a ticket for various distances. Between subjects, we vary whether gains are increasing or not in the distance traveled. Within subjects, we vary the probability of being inspected and the amount of the fine. We show that, in the game, fare-dodgers in real life fraud more than non-fare-dodgers. The fraud rate is lower for longer distances even in the absence of control, suggesting an increasing moral cost of fraud which does not depend on the relative payoff gained from cheating. Finally, both fare-dodgers and non- fare-dodgers respond to changes in the probability of being inspected and the severity of the punishment.

The pharmacokinetic profile of a new 800-mg tablet of nonanimal chondroitin sulfate (CS) (Mythocondro®, 800-mg tablets, Gnosis S.p.A., Italy) was investigated vs an animal CS in healthy volunteers for a total period of 48 hours. After a single 2400-mg dose of the test and the reference formulation, total CS, the compositional disaccharides (ΔDi6S, ΔDi4S and ΔDi0S), and the overall charge density were quantified in plasma. The safety and tolerability profile after a single dose of this new nonanimal CS tablets was excellent. After baseline-corrected concentrations, an overall greater plasma concentration was observed after 24 hours of ∼44% and after 48 hours of ∼45% from administration of nonanimal when compared to animal-derived CS. Moreover, nonanimal CS increases the specific sulfation in the 6-position of N-acetyl-galactosamine in human plasma CS and, as a consequence, the overall charge density, reaching double values (0.91), after 48 hours compared to bovine CS and to endogenous CS. In conclusion, nonanimal CS, possessing a lower molecular weight than an animal-derived sample, produces a greater CS concentration for a more prolonged period of time in plasma and an increase in charge density and specific 6-sulfation of endogenous plasma CS.

Antimicrobial activity is a well-known property of propolis, making it a candidate for antimicrobial surfaces in biomedical devices. Nevertheless, large-scale use of propolis as an anti-infective agent is limited by the heterogeneity of its chemical composition and consequent variation in antimicrobial activity. The aim of this study was to demonstrate that the multi dynamic extraction (M.E.D.) method produces standardized polyphenolic mixtures from poplar-type propolis, with reproducible chemical composition and anti-microbial activity, independently from the chemical composition of the starting raw propolis. Three raw propolis samples, from Europe, America, and Asia, were analyzed for their polyphenol chemical composition by means of HPLC&ndash, UV and then combined to obtain three mixtures of propolis, which werme submitted to the M.E.D. extraction method. The chemical composition and the antimicrobial activity of M.E.D. propolis against bacteria and fungi were determined. The three M.E.D. propolis showed similar chemical compositions and antimicrobial activities, exhibiting no relevant differences against antibiotic-susceptible and antibiotic-resistant strains. The batch-to-batch reproducibility of propolis extracts obtained with the M.E.D. method encourages the design of drugs alternative to traditional antibiotics and the development of anti-infective surface-modified biomaterials.

We report results on the structure, physicochemical characteristics and purity of chondroitin sulfate (CS) samples derived from three largely available and common biological sources such as bovine and porcine trachea and chicken keel bones with the aim to define their structural signatures. Many lots of CS produced by a manufacturer at industrial scale were characterized with a view to assess the reproducibility of the process as not controlled extractive procedures may produce final products with variable structure and biological contaminants as well as not constant clinical efficacy and safety. By using standardized source animal tissues and manufacturing procedure, highly pure CS (∼92 %) products with constant structure and characteristics were obtained. Bovine CS showed a lower molecular weight (MWw of ∼21,500 Da) than porcine (MWw of ∼26,000 Da) and chicken (MWw of ∼35,900 Da) products with a CV% of ∼2.0-7.5 and a polydispersity variability of 0.7-2.7 %. The ratio between the sulfate groups main located in position 4 and 6 of N-acetyl-galactosamine (4/6 ratio) was ∼1.70 for bovine CS versus a value of 3.60 for porcine and ∼2.70 for chicken samples with a overall charge density of 0.92-0.93 and a CV% of 2.1-2.5. The final products also showed the presence of a very low and constant content of other co-purified bio(macro)molecules (hyaluronic acid, keratan sulfate, dermatan sulfate, heparan sulfate, nucleic acids and proteins), calcium and sodium, and the absence of versican. Finally, a high reproducibility of molecular weight values, disaccharide composition, specific optical rotation and particle dimension was observed. The observed parameters are structural signatures useful to specifically identify the origin of CS and obtained by a standardized and highly reproducible manufacturing process. The compositional profile determined from this study provides a measure of the norm and range of variation in CS samples of terrestrial origin produced under standardized production protocol to which future pharmaceutical/nutraceutical final products can be compared. Moreover, the physicochemical properties including molecular weight, disaccharide composition, presence of natural contaminants and particle dimension were characterized to provide the basis of CS of high quality for application as pharmaceutical/nutraceutical active agents.

Mucopolysaccharidosis type IIIA (MPS IIIA, Sanfilippo A) is a neurodegenerative lysosomal storage disorder caused by the deficiency of sulphamidase enzyme (SGSH) leading to accumulation of heparan sulfate (HS). We quantitatively and structurally characterize primary stored HS and other glycosaminoglycans (GAGs) possibly accumulated through a secondary storage in brain, liver, kidney and lung of MPS IIIA mouse model. This analysis was also performed in MPS IIIA mice upon the intravenous treatment with an engineered human sulphamidase (chimeric hSGSH) capable to increase its secretion from the liver and to cross the blood-brain barrier. MPS IIIA animals showed a huge accumulation of HS, from ~15 up to ~24-times higher than wild type and also of hyaluronic acid (HA) (from 2.5 up to ~5.0-times more) and chondroitin sulfate (CS)/dermatan sulfate (DS) (from ~2 up to ~5-times more) in all studied organs. We also observed a significant increase in the overall HS charge density and in particular of 2-O-sulfation in MPS IIIA mice organs. 8 months after a systemic treatment with an engineered SGSH, the enzyme was highly efficient in the reduction of all accumulated GAGs in liver, brain and lung up to values of wild type mice. On the contrary, even if reduced, GAGs levels still remained significantly elevated in kidney. Overall data obtained by this detailed analysis of GAGs in the different organs of affected and treated animals with chimeric hSGSH may have implications for the evaluation of an effective therapeutic option of MPS IIIA and for the reduction of related neuropathology.

The concentration of high frequency controls in a limited period of time (" crackdowns ") constitutes an important feature of many law-enforcement policies around the world. In this paper, we offer a comprehensive investigation on the relative efficiency and effectiveness of various crackdown policies using a lab-in-the-field experiment with real passengers of a public transport service. We introduce a novel game, the daily public transportation game, where subjects have to decide, over many periods, whether to buy or not a ticket knowing that there might be a control. Our results show that (a) concentrated crackdowns are less effective and efficient than random controls; (b) prolonged crackdowns reduce fare-dodging during the period of intense monitoring but induces a burst of fraud as soon as they are withdrawn; (c) pre-announced controls induces more fraud in the periods without control. Overall, we also observe that real fare-dodgers fraud more in the experiment than non-fare-dodgers.

Chondroitin sulfate (CS) is a linear heteropolysaccharide of repeating disaccharide units bearing sulfate groups in various positions, commonly at C4 and/or C6 of galactosamine. CS plays important roles in various (patho)physiological processes also performing intriguing biological and therapeutical activities. Plasmatic CS is mainly composed of nonsulfated and 4-sulfated disaccharides. To obtain samples for the determination of CS amount and composition in blood/plasma, dried blood spot (DBS) could be used. DBSs have many advantages over other laboratory methods, allowing for large-scale population screening. Many analytical techniques may be used for the determination of CS. In particular, CE has proved to be a very attractive alternative separation technique for complex polysaccharide characterization. In this work, we compared CS levels between plasma and DBS samples, using CE equipped with the highly sensitive laser-induced fluorescence detector. CS from DBS differs from plasma CS owing to the high content of disaccharides sulfated in C4 and C6. This is due to the presence of the more sulfated CS derived from blood cellular fraction, in particular leukocytes. The identification and quantification of CS in blood plasma could be a useful prognostic and diagnostic tool in pathological conditions and for pharmacological applications.

Despite the great number of investigations, the common scientific approaches to study biological activities of propolis present some limitations due to the high natural variability of propolis and different extraction methods used. Therefore, the results obtained so far are often not comparable each other and are poorly reproducible. The aim of this work is the development of a new extraction method to obtain standardized propolis extracts to be studied in vitro for the determination of anti-inflammatory, antioxidant and antibacterial activities. The standardized extraction method was set and posited as patent (Multi Dynamic Extraction method M.E.D.®) and the extracts obtained were characterized; they shown similar chemical profiles (HPLC-ESI-MSn) even if formulated differently (dry, glyceric, glycolic, hydroalcoholic and oily extracts)1. The antioxidant activity of formulated extracts was firstly determined in vitro (TROLOX) and then, to better clarify the intracellular mechanisms behind the antioxidant and anti-inflammatory activities, miRNA, mRNA (RT-qPCR) and their protein validated target (ELISA) changes were evaluated2. Propolis M.E.D.® extracts showed similar antioxidant TROLOX values related to the amount of polyphenols; moreover, they are able to reduce the oxidative stress and inflammation level in HaCat cells acting on expression levels of mRNAs coding for NFE2L2, GPX2 and TNF-α and NFE2L2 protein2. These results highlight a possible molecular mechanism of action of Propolis M.E.D.® behind the antioxidant and anti-inflammatory activity. To test antibacterial activity, different propolis M.E.D.® extracts were tested (MIC) against several strains (ATCC, antibiotics resistant and susceptible) and showed comparable MIC values related to the amount of standardized polyphenols complex.

Chondroitin sulfate/dermatan sulfate (CS/DS) were isolated and purified for the first time from the bone of corb (Sciaena umbra) (CBG) and their chemical composition and anticoagulant activity were assessed. Infrared spectrum and agarose-gel electrophoresis for extracted CS/DS were also investigated. The results showed that the purified CS/DS obtained at a yield of 10% contains about 31.28% sulfate and an average molecular mass of 23.35 kDa. Disaccharide analysis indicated that CBG was composed of monosulfated disaccharides in positions 6 and 4 of the N-acetylgalactosamine (8.6% and 40.0%, respectively) and disulfated disaccharides in different percentages. The charge density was 1.4 and the ratio of 4:6 sulfated residues was equal to 4.64. Chondroitinase AC showed that the purified CS/DS contained mainly 74% CS and 26% DS. Moreover, the new CS/DS extracted from bone of corb showed a strong anticoagulant effect through activated partial thrombosis time (aPTT), thrombin time (TT) and prothrombin time (PT). In fact, CBG prolonged significantly (p 0.05), aPTT and PT about 2.62 and 1.26 fold, respectively, greater than that of the negative control at a concentration of 1000 μg/mL. However, TT assay of CBG was prolonged 3.53 fold compared with the control at 100 μg/mL. The purified CS/DS displayed a promising anticoagulant potential, which may be used as a novel and soothing drug.

Chondroitin sulfate/dermatan sulfate (CS/DS) was extracted from Atlantic bluefin tuna (

The goal of the present work was the extraction and structural characterization of novel sulfated glycosaminoglycans from European eel skin. The recovered glycosaminoglycans were physicochemically characterized and the uronic acid and sulfate contents were 35.12 ± 2.13% and 16.32 ± 0.4%, respectively. Cellulose acetate electrophoresis for extracted glycosaminoglycans was also investigated. Molecular weight of these sulfated glycosaminoglycans was determined (~37 kDa) by the gradient PAGE. Glycosaminoglycans obtained from the European eel were composed of non-sulfated, mono- and disulfated disaccharides. These sulfated glycosaminoglycans were evaluated for their in vitro anticoagulant activity using activated partial thromboplastin time, thrombin time and prothrombin time tests. The result showed that the recovered glycosaminoglycans exhibited interestingly anticoagulant activity. These glycosaminoglycans did not show haemolytic activity towards human erythrocytes. Furthermore, these bioactive substances can be explored as a functional food with antithrombotic function or used as source of anticoagulant drugs.