Different peripheral expression patterns of the nicotinic acetylcholine receptor in dementia with Lewy bodies and Alzheimer's disease.

BACKGROUND: The diffuse distribution of nicotinic cholinergic receptors (nAChRs) in both brain and peripheral immune cells points out their involvement in several pathological conditions. Indeed, the deregulated function of the nAChR was previously correlated with cognitive decline and neuropsychiatric symptoms in Alzheimer's disease (AD) and Dementia with Lewy bodies (DLB). The evaluation in peripheral immune cells of nAChR subtypes, which could reflect their expression in brain regions, is a prominent investigation area. OBJECTIVES: This study aims to evaluate the expression levels of both the nAChR subunits and the main known inflammatory cytokines in peripheral blood mononuclear cells (PBMCs) of patients with DLB and AD to better characterize their involvement in these two diseases. RESULTS: Higher gene expression levels of TNFα, IL6 and IL1ß were observed in DLB and AD patients in comparison with healthy controls (HC). In our cohort, a reduction of nAChRα4, nAChRß2 and nAChRß4 was detected in both DLB and AD with respect to HC. Considering nAChR gene expressions in DLB and AD, significant differences were observed for nAChRα3, nAChRα4, nAChRß2 and nAChRß4 between the two groups. Moreover, the acetylcholine esterase (AChE) gene expression was significantly higher in DLB than in AD. Correlation analysis points out the relation between different nAChR subtype expressions in DLB (nAChRß2 vs nAChRα3; nAChRα4 vs nAChRα3) and AD (nAChRα4 vs nAChRα3; nAChRα4 vs nAChRß4; nAChRα7 vs nAChRα3; nAChRα7 vs nAChRα4). CONCLUSIONS: Different gene expressions of both pro-inflammatory cytokines and nAChR subtypes may represent a peripheral link between inflammation and neurodegeneration. Inflammatory cytokines and different nAChRs should be valid and accurate peripheral markers for the clinical diagnosis of DLB and AD. However, although nAChRs show a great biological role in the regulation of inflammation, no significant correlation was detected between nAChR subtypes and the examined cytokines in our cohort of patients.

A chromosome-level genome assembly enables the identification of the follicule stimulating hormone receptor as the master sex-determining gene in the flatfish Solea senegalensis.

Sex determination (SD) shows huge variation among fish and a high evolutionary rate, as illustrated by the Pleuronectiformes (flatfishes). This order is characterized by its adaptation to demersal life, compact genomes and diversity of SD mechanisms. Here, we assembled the Solea senegalensis genome, a flatfish of great commercial value, into 82 contigs (614 Mb) combining long- and short-read sequencing, which were next scaffolded using a highly dense genetic map (28,838 markers, 21 linkage groups), representing 98.9% of the assembly. Further, we established the correspondence between the assembly and the 21 chromosomes by using BAC-FISH. Whole genome resequencing of six males and six females enabled the identification of 41 single nucleotide polymorphism variants in the follicle stimulating hormone receptor (fshr) consistent with an XX/XY SD system. The observed sex association was validated in a broader independent sample, providing a novel molecular sexing tool. The fshr gene displayed differential expression between male and female gonads from 86 days post-fertilization, when the gonad is still an undifferentiated primordium, concomitant with the activation of amh and cyp19a1a, testis and ovary marker genes, respectively, in males and females. The Y-linked fshr allele, which included 24 nonsynonymous variants and showed a highly divergent 3D protein structure, was overexpressed in males compared to the X-linked allele at all stages of gonadal differentiation. We hypothesize a mechanism hampering the action of the follicle stimulating hormone driving the undifferentiated gonad toward testis.

Effectiveness Of Reactive Oxygen Species In An Oil-Based Medication For Healing Burn Wounds: A Case Series.

Oxygen is considered a key element in the complex tissue repair process. O3-Oil antiseptics are obtained from the chemical reaction between ozone and unsaturated fatty acids of vegetable oils. The purpose of this study was to assess the effectiveness of a commercially available O3-Oil in the management of recalcitrant burn wounds. This study involved 20 patients with mid-deep and deep burn wounds (average TBSA approximately 23%, range 7-35%). Patient average age was 47 years (range 26-74 years), 8 were male. The presence of exudate, pain and delayed re-epithelialization, absence of tissue necrosis and/or sepsis were the inclusion criteria for the topical use of O3-Oil. In each patient two clinically similar areas were identified and treated every 48 hours with O3-Oil (Group 1) and conventional medication (Group 2). All the patients had positive bacterial culture results before treatment. A quicker reduction of exudates and pain and a progressive and faster bacterial load reduction was observed in Group 1. Negative cultures were obtained after 4 days on average in Group 1 (range 1-7 days) and 8 days in Group 2 (range 5-11 days). The present study demonstrated that Novox® provides a significant antibacterial effect, while stimulating reparative processes. According to our experience, the use of gel or pad is useful for re-epithelializing lesions, while the impregnated gauzes, which tend to adhere to the wound, are better used on lesions with granulation tissue.

L'oxygène est un élément- clé du processus complexe de réparation tissulaire. Les antiseptique O3-huile sont obtenus par réaction chimique entre l'ozone et des acides gras polyinsaturés d'origine végétale. Cette étude avait pour but d'évaluer l'efficacité d'un tel produit sur les brûlures d'évolution torpide. Elle a concerné 20 patients (dont 8 hommes) de 47 ans d'âge moyen (26- 74) ayant des brûlures intermédiaires à profondes sur 23% de SCT (7 à 35). Les indications de mise sous O3- huile pouvaient être la persistance d'exsudation, la douleur, l'absence d'épithélialisation, l'absence de séparation de l'escarre, l'infection (tous les patients ayant d'ailleurs une bactériologie cutanée positive avant le début du traitement). Chez chaque patient, deux zones similaires recevaient toutes les 48 h soit de l'huile ozonée (groupe 1) soit un traitement conventionnel (groupe 2). La réduction des exsudats, de la douleur et de la charge bactérienne était plus rapide dans le groupe 1, les cultures se négativant à J4 (1 à 7) dans ce groupe contre J8 (5 à 11) dans le groupe 2. Cette étude montre que le Novox® a un effet antibactérien significatif et stimule la cicatrisation. Dans notre expérience, l'utilisation de gel ou de plaques imprégnées sont plus efficaces pour promouvoir l'épithélialisation quand les compresses saturées, qui adhèrent à la plaie, sont préférables sur des tissus hyperbourgeonnants.

Mid-transverse colon cancer and extended versus transverse colectomy: Results of the Italian society of surgical oncology colorectal cancer network (SICO CCN) multicenter collaborative study.

INTRODUCTION: Transverse colon cancer (TCC) is poorly studied, and TCC cases are often excluded from large prospective randomized trials because of their complexity and their potentially high complication rate. The best surgical approach for TCC has yet to be established. The aim of this large retrospective multicenter Italian series is to investigate the advantages and disadvantages of both hemicolectomy and transverse colectomy in order to identify the best surgical approach. MATERIALS AND METHODS: This was a retrospective cohort study of patients with mid-transverse colon cancer treated with a segmental colon resection or an extended hemicolectomy (right or left) between 2006 and 2016 in 28 high-volume (more than 70 procedures/year) Italian referral centers for colorectal surgery. RESULTS: The study included 1529 patients, 388 of whom underwent a segmental resection while 1141 underwent an extended resection. A higher number of complications has been reported in the segmental group than in the extended group (30.1% versus 23.6%; p 0.010). In 42 cases the main complication was the anastomotic leak (4.4% versus 2.2%; p 0.020). Recovery outcomes also showed statistical differences: time to first flatus (p 0.014), time to first mobilization (p 0.040), and overall hospital stay (p < 0.001) were significantly shorter in the extended group. Even if overall survival were similar between the groups (95.1% versus 97%; p 0.384), 3-year disease-free survival worsened after segmental resection (78.1% versus 86.2%; p 0.001). CONCLUSIONS: According to our results, an extended right colon resection for TCC seems to be surgically safer and more oncologically valid.

Age of HIV Acquisition Affects the Risk of Multi-Morbidity after 25 Years of Infection Exposure.

INTRODUCTION: Understanding the intersection of HIV, aging and health is crucial due to the increasing number of people aging with HIV. OBJECTIVE: The objective of the study was to assess the prevalence of, and risk factors for individual comorbidities and multi-morbidity in people living with HIV with similar duration of HIV infection, notwithstanding a 25-year difference at the time of HIV acquisition. METHODS: In a cross-sectional multicentre retrospective study, we compared three match-control age groups. The "Young" were selected from Romania and included HIV-positive patients prenatally infected and assessed at the age of 25-30 years. The "Old" and the "Geriatric" were selected from Italy. These respectively included subjects infected with HIV at the age of 25 years and assessed at the age of 50-55 years, and those infected at the age of 50 years and assessed at the age of 75-80 years. Each group was sex and age matched in a 1:5 ratio with controls selected from the CINECA ARNO database from Italy. We described non-infectious comorbidities (NICM), including cardiovascular disease, hypertension, dyslipidaemia, diabetes, chronic kidney disease, and multi-morbidity (MM≥ 3 NICM). RESULTS: MM prevalence in the "Young" group compared to controls was 6.2% vs 0%, while in the "Geriatric" was "68.2% vs 3.6%. Using "Young" as a reference, in multivariate analyses, predictors for MM were as follows: HIV serostatus (OR=47.75, IQR 14.78-154.25, p<0.01) and "Geriatric" vs "Young" (OR=30.32, IQR 5.89-155.98, p<0.01). CONCLUSION: These data suggest that age at acquisition of HIV should be considered as a risk factor for NICM and MM.

Effectiveness of topical α-Tocopherol Acetate in burn infection treatment.

Infection following burn injury is critical, especially for patients with large total body surface area burns and in skin graft donor sites. Although various aspects of α-tocopherol acetate (α-TA) beneficial effects on wound healing have been validated, it appears that no study has specifically addressed its antimicrobial potential. The purpose of this study was to explore the therapeutic efficacy of topical application of α-TA in terms of bacterial load reduction. Between January 2018 and June 2018, 20 patients with mid-deep and deep burn wounds were included in the present study (average TBSA approximately 42%, range 25-67%). Patient average age was 47.75 years (range 25-72 years), 8 were male. In each patient two clinically similar areas were identified and treated every 24 hours with topical application of α-TA in the form of Filme Olio® (Group 1) and conventional medication (Group 2). All the patients had positive results for bacterial cultures before treatment. Despite the presence of initial bacterial infection, a quicker reduction of exudates and pain and a progressive and faster bacterial load reduction was observed in Group 1. Negative cultures were obtained after 3 days on average in Group 1 (range 1-6 days) and 8 days in Group 2 (range 5-9 days). This study clearly shows the importance of the therapeutic targeting of infection in the treatment of burns. α-TA may represent a safe, simple and inexpensive method for improving the healing of difficult wounds with local infection.

Les infections sont une complication majeure des brûlures, en particulier chez les patients largement atteints et au niveau des sites donneurs. Alors que ATA a été évalué, et considéré bénéfique, dans de nombreux aspects de la cicatrisation, aucune étude ne s'est penchée sur son potentiel antimicrobien. Cette étude a exploré l'effet de ATA topique sur la réduction de l'inoculum bactérien. Vingt patients dont 12 femmes, ayant des brûlures intermédiaires et profondes, ont été inclus dans cette étude, qui s'est déroulée entre janvier et juin 2018. Leur âge était en moyenne de 47,75 ans (25- 72), la surface brûlée de 42% (25- 67%). Chez tous les patients, deux zones similaires, à la culture bactérienne positive, étaient traitées par ATA (groupe 1) ou traitement conventionnel (groupe 2). La diminution des l'exsudation, de la douleur et de la charge bactérienne était plus rapide dans le groupe 1. Les cultures se négativaient en 3 jours (1- 6) dans le groupe 1 contre 8 (5- 9) dans le groupe 2. Cette étude montre l'intérêt du traitement local des infections de zones brûlées, ATA apparaissant dans ce cadre comme un traitement simple, sûr et peu onéreux.

Enzymatic escharolysis with nexobrid® on partial thickness burn wounds: pre- and post-debridement histological assessment.

Enzymatic escharolysis is an innovative, non-surgical treatment method for severe burn patients as it allows very early, nontraumatic removal of necrotic tissue even on patients whose overall clinical conditions would mandate delaying traditional surgical escharectomy. The aim of this work was to examine aspects related to the "quality" of enzymatic debridement, which is inherently different from surgical debridement. To this end, biopsies harvested from partial thickness burn wounds, before and after enzymatic treatment, were histologically assessed. As is well known, surgical escharectomy removes the necrosis as well as some of its neighbouring healthy tissue, sharply and radically, leaving a perfectly clean and viable wound bed. On the other hand, enzymatic escharolysis is more selective, as it completely wipes out the necrotic portion while sparing unharmed and partially damaged tissue. In this study, only mid-deep partial thickness wounds were examined, and it was observed that partially damaged dermis was always spared by the lytic action. This dermis, however, showed some "homogenization" characteristics, had few vital skin annexes in it, and therefore looked very similar to the scaffold of dermal matrices currently available on the market. This scaffold should be safeguarded with a view to possibly achieving a more complete and functional spontaneous tissue regeneration. Conversely, if this dermal portion is mismanaged, it could desiccate, thus leading to the formation of a neo-eschar with unpredictable clinical evolution. Understanding how escharolysis actually works allowed us to extrapolate fruitful usage suggestions to optimize the procedure and fully exploit its potential.

La détersion enzymatique est une technique innovante non chirurgicale permettant l'ablation très précoce et non traumatique des tissus nécrosés même chez des patients dont l'état général nécessiterait de repousser une excision chirurgicale. Le but de ce travail était d'évaluer la « qualité ¼ du débridement enzymatique, par essence différent du traitement chirurgical. À cette fin, nous avons examiné histologiquement des biopsies réalisées avant et après détersion. Il est bien connu que la chirurgie emporte totalement et radicalement la nécrose et une partie du tissu environnant, laissant en place un tissu parfaitement propre et viable. Le débridement enzymatique est plus sélectif, emportant tout le tissu nécrosé sans affecter les tissus sains ou viables. Cette étude ne s'est intéressée qu'aux brûlures intermédiaires et nous avons observé que les régions saines étaient toujours préservées. Ce derme restant apparaît toutefois homogénéisé, avec peu d'annexes viables ce qui fait penser aux matrices des dermes artificiels actuellement commercialisés. Il doit être préservé afin de promouvoir une régénération tissulaire complète et fonctionnelle. Ainsi, si ce derme restant n'est pas correctement pris en charge, il peut se dessécher et aboutir à la formation d'un nouvel escarre, d'évolution imprévisible. Le compréhension du mécanisme exact de la lyse de la brûlure permet de développer des protocoles d'optimisation de la technique de lyse enzymatique.

Modulation Instability Induced Frequency Comb Generation in a Continuously Pumped Optical Parametric Oscillator.

Continuously pumped passive nonlinear cavities can be harnessed for the creation of novel optical frequency combs. While most research has focused on third-order "Kerr" nonlinear interactions, recent studies have shown that frequency comb formation can also occur via second-order nonlinear effects. Here, we report on the formation of quadratic combs in optical parametric oscillator (OPO) configurations. Specifically, we demonstrate that optical frequency combs can be generated in the parametric region around half of the pump frequency in a continuously driven OPO. We also model the OPO dynamics through a single time-domain mean-field equation, identifying previously unknown dynamical regimes, induced by modulation instabilities, which lead to comb formation. Numerical simulation results are in good agreement with experimentally observed spectra. Moreover, the analysis of the coherence properties of the simulated spectra shows the existence of correlated and phase-locked combs. Our results reveal previously unnoticed dynamics of an apparently well assessed optical system, and can lead to a new class of frequency comb sources that may stimulate novel applications by enabling straightforward access to elusive spectral regions, such as the midinfrared.

Survival of Escherichia coli O157:H7 during the manufacture and ripening of Cacioricotta goat cheese.

The aim of this study was to assess the growth and survival of Escherichia coli O157:H7 during the manufacturing and ripening of Cacioricotta goat cheese. Goat milk was artificially contaminated with E. coli O157:H7 and the bacterial load was monitored from production up to 90 days of ripening. Goat milk was inoculated with 102 cfu ml-1 of E. coli O157:H7 and the bacterial count of the curd at time zero was 2.31 log10 cfu g-1. During the first day of ripening, the bacterial load has increased to 5.73 log10 cfu g-1 to more than 6.20 log10 cfu g-1 during the first week. The bacterial load remained constant up to 28 days and then slightly decreased until the end of ripening, with values of aw and pH of 0.88 and 5.41 respectively. The results of this study highlighted that E. coli O157:H7 is able to survive the manufacturing process and they suggest that the 90-day period of ripening alone is insufficient to remove E. coli O157:H7 in contaminated Cacioricotta goat cheese. Moreover, these results support the assumption that the presence of a low contamination of milk with E. coli O157:H7 could represent a potential source of infection and a threat to consumers.

Directionally induced quasi-phase matching in homogeneous AlGaAs waveguides.

We report on the experimental observation of quasi-phase matching in a homogeneous waveguide. By fabricating a monolithic snake-shaped suspended AlGaAs nanowire on a (001) GaAs wafer, we demonstrate the unraveled version of a χ(2) whispering-gallery-mode microdisk, obtaining second-harmonic generation in the optical telecom wavelength range. With a radius of curvature of 50 µm and four spatial oscillations along the (110) average direction, a splitting of the second-harmonic spectrum occurs around the phase-matching wavelength of the corresponding straight waveguide. This splitting, which increases as the radius of curvature decreases, provides a useful degree of freedom for the design of small-footprint nonlinear photonic devices on-chip.

Effects of d-cloprostenol on different layers and regions of the bovine uterus during the follicular and luteal phases.

INTRODUCTION: D-cloprostenol is a synthetic PGF2α, commonly used in bovine reproduction, that increases myometral contractility. However, little is known regarding its contractile behaviour and how it depends on the reproductive state and uterine topography (regions and muscular layers). HYPOTHESIS AND OBJECTIVES: These aspects would affect the action of d-cloprostenol on the uterus. Therefore, we hypothesize a possible use of this drug at the time of artificial insemination, to improve conception rates and, in the post-partum, in order to accelerate uterine involution in dairy cattle. The purpose of the present study was to investigates the modulatory effect of d-cloprostenol on contractility of the bovine uterine region (horn and corpus) and their muscle layers (circular and longitudinal), in follicular and luteal phases. To our knowledge, motility effects of d-cloprostenol on different regions from healthy bovine uterus have not been described up to now. MATERIALS AND METHODS: Uterine specimens were collected from uterine body and horn of cattle in the follicular (n = 28) and luteal phase (n = 32) of the estrous cycle at slaughter. Two strips were prepared from each regions corresponding to the circular and the longitudinal muscular layers, respectively. Samples were cultured in an organ bath, exposed to synthetic prostaglandin (1 µM d-cloprostenol) and their contractile activity was monitored for 10 min. The functionality of the strip throughout the experiment was tested by a dose of carbachol (10-5 M). RESULTS: The mean basal amplitude of contractions was higher in the follicular compared to the luteal phase in uterine horn samples, but not in muscles collected from the uterine body. The amplitude of contractions increased after d-cloprostenol administration in all tissues with a greater increase in samples from cattle in the follicular phase. The frequency of contractions increased after d-cloprostenol administration in longitudinal but not in circular fibrees. CONCLUSION: The contractile responses to d-cloprostenol in both horn and corpus were strongest in the circular muscles but weak in the longitudinal muscles.

"Leaching or not leaching": an alternative approach to antimicrobial materials via copolymers containing crown ethers as active groups.

In this work, new copolymers containing either MMA and 18C6 crown-ether pendants, or PEG, MMA and 18C6 crown-ether pendants were synthesized to test the idea that sequestering structural alkali-earth ions from the bacterial outer membrane (OM) may lead to bacterial death. The copolymers were obtained either via uncontrolled radical polymerization or ATRP; the latter approached allowed us to produce not only linear copolymers but also branched Y-like structures. After checking for the capability of complexing magnesium and calcium ions, the antimicrobial activity of all copolymers was tested placing their casted plaques in contact with pure water E. coli suspensions. All plaques adsorbed alkali-earth ions and killed bacteria, albeit with different antimicrobial efficiencies. Differences in the latter characteristic were attributed to different plaque roughness. The role of the 18C6 crown-ether pendants was elucidated by pre-saturating plaques with Mg/Ca ions, the marked reduction in antimicrobial efficiency indicating that losing the latter from OM due to surface complexation does play an important role in killing bacteria at short (<5 h) contact times. At longer times, the mode of action is instead related to the poly-cationic nature acquired by the plaques due to ion sequestering.

Production of human pro-relaxin H2 in the yeast Pichia pastoris.

BACKGROUND: Initially known as the reproductive hormone, relaxin was shown to possess other therapeutically useful properties that include extracellular matrix remodeling, anti-inflammatory, anti-ischemic and angiogenic effects. All these findings make relaxin a potential drug for diverse medical applications. Its precursor, pro-relaxin, is an 18 kDa protein, that shows activity in in vitro assays. Since extraction of relaxin from animal tissues raises several issues, prokaryotes and eukaryotes were both used as expression systems for recombinant relaxin production. Most productive results were obtained when using Escherichia coli as a host for human relaxin expression. However, in such host, relaxin precipitated in the form of inclusion bodies and, therefore, required several expensive recovery steps as cell lysis, refolding and reduction. RESULTS: To overcome the issues related to prokaryotic expression here we report the production and purification of secreted human pro-relaxin H2 by using the methylotrophic yeast Pichia pastoris as expression host. The methanol inducible promoter AOX1 was used to drive expression of the native and histidine tagged forms of pro-relaxin H2 in dual phase fed-batch experiments on the 22 L scale. Both protein forms presented the correct structure, as determined by mass spectrometry and western blotting analyses, and demonstrated to be biologically active in immune enzymatic assays. The presence of the tag allowed to simplify pro-relaxin purification obtaining higher purity. CONCLUSIONS: This work presents a strategy for microbial production of recombinant human pro-relaxin H2 in Pichia pastoris that allowed the obtainment of biologically active pro-hormone, with a final concentration in the fermentation broth ranging between 10 and 14 mg/L of product, as determined by densitometric analyses.

Hybrid complexes of high and low molecular weight: evaluation using an in vitro model of osteoarthritis.

Hyaluronan (HA) is central in joint and cartilage functions and to restore synovial fluid viscosity. In patients with osteoarthritis (OA), molecular weight (MW) and concentration of hyaluronic acid (HA) are reduced, diminishing joint lubrication. IL-1ß treatment was used to mimic osteoarthritis in a chondrocytes based in vitro model. The aim of our research, using this model and human chondrocytes was to assess the anti-inflammatory effect of H/L-HA hybrid complexes (SINOVIAL-HL®) in comparison with HA at high (H-HA) and low molecular weight (L-HA) separately used, through the evaluation of specific biomarkers involved in cartilage degradation and correlated to osteoarthritis. Specifically, TNF-α and IL-6 mRNA were evaluated by qRT-PCR. Cytokines levels were measured using Bio-plex assays and COMP-2 through immunofluorescence staining and western blot. H/L-HA significantly reduced inflammation biomarkers respect to both L-HA or H-HA separately considered at transcriptional and protein level.

Incidence, prevalence, costs and quality of care of type 1 diabetes in Italy, age 0-29 years: The population-based CINECA-SID ARNO Observatory, 2002-2012.

BACKGROUND AND AIMS: To assess temporal trend in incidence (2003-12) and prevalence (2002-12) of type 1 diabetes in children and young adults, direct costs and selected indicators of quality of care under the coverage of the universalistic Italian National Health System (NHS). METHODS AND RESULTS: The ARNO Observatory, a healthcare monitoring system based on administrative data, identified a population-based multiregional cohort of subjects aged 0-29 years. Type 1 diabetes was defined by at least two prescriptions of insulin over 12 months and continuous insulin-treatment in the following year. Indicators of quality of care and directs costs were assessed in persons with diabetes and in people without diabetes, individually matched for age, gender and health unit (1:4 ratio). We identified 2357 incident cases of type 1 diabetes aged 0-29 years (completeness of ascertainment, 99%). Incidence rates were similar in ages 0-14 (15.8, 95% CI 14.9-16.8) and 15-29 years (16.3, 15.4-17.2), with no significant trend. Prevalence increased from 137 to 166.9/100,000, particularly in the age 15-29 years. Direct costs accounted for € 2117 in persons with diabetes and € 292 in control individuals. A statistically significant decreasing trend in hospitalization for acute complications was evident (p < 0.001), which was almost completely due to ketoacidosis. People with at least one HbA1c measurement over the year were 48.5%. CONCLUSION: We showed high incidence and increasing prevalence of type 1 diabetes in young adults in Italy, which impact on direct costs under the universalistic coverage of the NHS.

The 1,4 benzoquinone-featured 5-lipoxygenase inhibitor RF-Id induces apoptotic death through downregulation of IAPs in human glioblastoma cells.

BACKGROUND: Embelin is a potent dual inhibitor of 5-lipoxigenase (5-LOX) and microsomal prostaglandin E2 synthase (mPGES)-1 that suppresses proliferation of human glioma cells and induces apoptosis by inhibiting XIAP and NF-κB signaling pathway. Synthetic structural modification yielded the derivative 3-((decahydronaphthalen-6-yl)methyl)-2,5-dihydroxycyclohexa-2,5-diene-1,4-dione (RF-Id), an embelin constrained analogue, with improved efficiency against 5-LOX in human neutrophils and anti-inflammatory activity in vivo. Taking into account that lipoxygenase (LOX) metabolites, from arachidonic acid and linoleic acid, have been implicated in tumor progression, here, we determined whether RF-Id was able to hinder glioblastoma (GBM) cancer cell growth and the related mechanisms. METHODS: U87MG and LN229 cells were plated in 96-wells and treated with increasing concentrations of RF-Id. Cell viability was evaluated by MTT assay. The effects of the compounds on cell cycle, apoptosis, oxidative stress and autophagy were assessed by flow cytometry (FACS). The mode of action was confirmed by Taqman apoptosis array and evaluating caspase cascade and NFκB pathway by western blotting technique. RESULTS: Here, we found that RF-Id induced a stronger inhibition of GBM cell growth than treatment with embelin. Flow cytometry analysis showed that RF-Id induced about 30 % apoptosis and a slight increase of autophagy after 72 h on U87-MG cells. Moreover, the compound induced an increase in the percentage of cells in G2 and S phase that was paralleled by an increase of p21 and p27 expression but no significant changes of the mitochondrial membrane potential; array analysis showed a significant upregulation of CASP8 and a downregulation of IAP family and NFκB genes in cells treated with RF-Id. RF-Id induced a significant cleavage of caspases 8, 9, 3 and 7, blocked c-IAP2/XIAP interaction by inducing XIAP degradation and inhibited NFκB pathway. CONCLUSIONS: RF-Id induced a caspase-dependent apoptosis in GBM cells by inhibiting IAP family proteins and NFκB pathway and represents a promising lead compound for designing a new class of anti-cancer drugs with multiple targets.

The relative burden of diabetes complications on healthcare costs: The population-based CINECA-SID ARNO Diabetes Observatory.

BACKGROUND AND AIMS: In the present population-based study, we aimed to describe the per patient annual healthcare cost of people with diabetes in 2007-2012, to assess the relative burden of diabetes complications and other potential determinants on healthcare costs in the 2012 cohort, and to describe and analyse the determinants of the cost of incident cases diagnosed in 2012. METHODS AND RESULTS: We analysed data from a retrospective cohort of residents in four Italian areas that were served by Local Health Units participating in the ARNO Observatory. Per patient annual healthcare costs (Euros) were estimated as the sum of all the resources supplied during that year (drugs, outpatient care, and hospitalisations). The mean per patient annual healthcare cost increased from €2752 in 2007 to €3191 in 2010, before decreasing to €2791 in 2012. The largest component of these costs was represented by hospitalisations (around €1550, on average; 51.7% of total cost), followed by outpatient care (€422; 14.6%) and drugs (€973; 33.7%). In 2012, the most relevant cost determinants were chronic diabetes complications, with an additional cost due to nephropathy/end stage renal disease (€4683), amputations (€5042), lower extremity revascularization (€4808), and cerebrovascular diseases (€3861). Costs associated with incidence cases were higher than those associated with prevalent. CONCLUSION: The present study provides evidence on the excess of healthcare costs due to diabetes complications in both prevalent and incident cases.

Adherence to alendronic or risedronic acid treatment, combined or not to calcium and vitamin D, and related determinants in Italian patients with osteoporosis.

PURPOSE: Osteoporosis is a chronic disease and an important health and social burden due to its worldwide prevalence. Literature and clinical experience report incomplete adherence to the therapy. This retrospective observational study aimed at assessing the adherence to first-line antiosteoporosis drugs (AODs; reimbursed by the National Health System, according to the Italian Medicine Agency recommendation number 79), alendronate or risedronate, with or without calcium and/or vitamin D supplements, in a real, Italian clinical setting. PATIENTS AND METHODS: Analyses were carried out on data present in the ARNO Observatory, a population-based patient-centric Italian database. From a population of 5,808,832 inhabitants with available data, a cohort of 3.3 million of patients aged ≥40 years was selected. New users of first-line AODs as monotherapy (accrual period, 2007-2009) were followed up over 3 years to assess adherence at 6, 12, and 36 months to AODs and to supplements and related determinants. RESULTS: Approximately 40,000 new users were identified: mostly women, aged on average (standard deviation) 71±10 years. Alendronate was the most prescribed (38.2% of patients), followed by risedronate (34.9%) and alendronate with colecalciferol as a fixed-dose combination (25.8%). Adherence at the 6-month follow-up was 54%, and this constantly and significantly decreased after 1 year to 46%, and after 3 years to 33% (P<0.01). Adherence to the fixed-dose combination was higher than to plain alendronate throughout the follow-up period. Similarly, adherence to supplements constantly decreased with the duration of treatment. Women and patients aged >50 years were more likely to adhere to treatment regimen (P<0.001). The use of drugs for peptic ulcer and gastroesophageal reflux disease and of corticosteroids for systemic use were significantly associated with high adherence at different times. Polytherapy (>5 drugs), cardiovascular, and neurological therapies were significantly associated with low adherence throughout the follow-up period. CONCLUSION: In a huge clinical practice sample, this study highlights suboptimal adherence to first-line AODs and to supplements and important determinants, such as concomitant therapies.

Walk-Off-Induced Modulation Instability, Temporal Pattern Formation, and Frequency Comb Generation in Cavity-Enhanced Second-Harmonic Generation.

We derive a time-domain mean-field equation to model the full temporal and spectral dynamics of light in singly resonant cavity-enhanced second-harmonic generation systems. We show that the temporal walk-off between the fundamental and the second-harmonic fields plays a decisive role under realistic conditions, giving rise to rich, previously unidentified nonlinear behavior. Through linear stability analysis and numerical simulations, we discover a new kind of quadratic modulation instability which leads to the formation of optical frequency combs and associated time-domain dissipative structures. Our numerical simulations show excellent agreement with recent experimental observations of frequency combs in quadratic nonlinear media [Phys. Rev. A 91, 063839 (2015)]. Thus, in addition to unveiling a new, experimentally accessible regime of nonlinear dynamics, our work enables predictive modeling of frequency comb generation in cavity-enhanced second-harmonic generation systems. We expect our findings to have wide impact on the study of temporal and spectral dynamics in a diverse range of dispersive, quadratically nonlinear resonators.