Treatment of chronic viral hepatitis in woodchucks by prolonged intrahepatic expression of interleukin-12

Chronic hepatitis B is a major cause of liver-related death worldwide. Interleukin-12 (IL-12) induction accompanies viral clearance in chronic hepatitis B virus infection. Here, we tested the therapeutic potential of IL-12 gene therapy in woodchucks chronically infected with woodchuck hepatitis virus (WHV), an infection that closely resembles chronic hepatitis B. The woodchucks were treated by intrahepatic injection of a helper-dependent adenoviral vector encoding IL-12 under the control of a liver-specific RU486-responsive promoter. All woodchucks with viral loads below 10(10) viral genomes (vg)/ml showed a marked and sustained reduction of viremia that was accompanied by a reduction in hepatic WHV DNA, a loss of e antigen and surface antigen, and improved liver histology. In contrast, none of the woodchucks with higher viremia levels responded to therapy. The antiviral effect was associated with the induction of T-cell immunity against viral antigens and a reduction of hepatic expression of Foxp3 in the responsive animals. Studies were performed in vitro to elucidate the resistance to therapy in highly viremic woodchucks. These studies showed that lymphocytes from healthy woodchucks or from animals with low viremia levels produced gamma interferon (IFN-gamma) upon IL-12 stimulation, while lymphocytes from woodchucks with high viremia failed to upregulate IFN-gamma in response to IL-12. In conclusion, IL-12-based gene therapy is an efficient approach to treat chronic hepadnavirus infection in woodchucks with viral loads below 10(10) vg/ml. Interestingly, this therapy is able to break immunological tolerance to viral antigens in chronic WHV carriers

Evaluación físico-química de aceite pigmentado obtenido de la cabeza de camarón

In this work the proximal analysis, physicochemical characterization, fatty acid profile and astaxanthin content of pigmented oil obtained by fermentation shrimp heads are presented. Lipids are the major components in the oil (95%). The saponification number is 178.62 mg KOH/g, iodine value 139.8 cg iodine/g, and the peroxide value was not detected. Density and viscosity were 0.92 mg/ml and 64 centipoises, respectively. The highest contents of fatty acids were linoleic (C18:2n6), oleic (C18:1n9) and palmitic (C16:0). Eicosapentaenoic acid (C20:5n3, EPA) and docosahexaenoic acid (C22:6n3, DHA) account for 9% of the total. The content of astaxanthin was 2.72 mg/g dry weight. The pigmented oil is a dietary source of nutrients with high value such as astaxanthin.En este trabajo se presenta el análisis proximal, caracterización físico-química, perfil de ácidos grasos y contenido de astaxantina en aceite pigmentado aislado por fermentación láctica de los residuos de camarón. Los lípidos son los componentes mayoritarios (95%). El índice de saponificación es 178.62 mg KOH/g, el de yodo 139.8 cg yodo/g, y los peróxidos no fueron detectados. La densidad y la viscosidad fueron de 0.92 mg/ml y 64 centipoises, respectivamente. Los ácidos grasos en mayor cantidad fueron el linoleico (C18:2n6), oleico (C18:1n9) y palmítico (C16:0). El ácido eicosapentaenoico (C20:5n3, EPA) y el docosahexaenoico (C22:6n3, DHA) suman el 9% del total. El contenido promedio de astaxantina fue de 2.72 mg/g base seca. El aceite pigmentado es una fuente dietética de nutrientes con alto valor como la astaxantina

Changes in parental smoking during pregnancy and risks of adverse birth outcomes and childhood overweight in Europe and North America: An individual participant data meta-analysis of 229,000 singleton births

Background. Fetal smoke exposure is a common and key avoidable risk factor for birth complications and seems to influence later risk of overweight. It is unclear whether this increased risk is also present if mothers smoke during the first trimester only or reduce the number of cigarettes during pregnancy, or when only fathers smoke. We aimed to assess the associations of parental smoking during pregnancy, specifically of quitting or reducing smoking and maternal and paternal smoking combined, with preterm birth, small size for gestational age, and childhood overweight. Methods and findings. We performed an individual participant data meta-analysis among 229,158 families from 28 pregnancy/birth cohorts from Europe and North America. All 28 cohorts had information on maternal smoking, and 16 also had information on paternal smoking. In total, 22 cohorts were population-based, with birth years ranging from 1991 to 2015. The mothers’ median age was 30.0 years, and most mothers were medium or highly educated. We used multilevel binary logistic regression models adjusted for maternal and paternal sociodemographic and lifestyle-related characteristics. Compared with nonsmoking mothers, maternal first trimester smoking only was not associated with adverse birth outcomes but was associated with a higher risk of childhood overweight (odds ratio [OR] 1.17 [95% CI 1.02–1.35], P value = 0.030). Children from mothers who continued smoking during pregnancy had higher risks of preterm birth (OR 1.08 [95% CI 1.02–1.15], P value = 0.012), small size for gestational age (OR 2.15 [95% CI 2.07–2.23], P value < 0.001), and childhood overweight (OR 1.42 [95% CI 1.35–1.48], P value < 0.001). Mothers who reduced the number of cigarettes between the first and third trimester, without quitting, still had a higher risk of small size for gestational age. However, the corresponding risk estimates were smaller than for women who continued the same amount of cigarettes throughout pregnancy (OR 1.89 [95% CI 1.52–2.34] instead of OR 2.20 [95% CI 2.02–2.42] when reducing from 5–9 to ≤4 cigarettes/day; OR 2.79 [95% CI 2.39–3.25] and OR 1.93 [95% CI 1.46–2.57] instead of OR 2.95 [95% CI 2.75–3.15] when reducing from ≥10 to 5–9 and ≤4 cigarettes/day, respectively [P values < 0.001]). Reducing the number of cigarettes during pregnancy did not affect the risks of preterm birth and childhood overweight. Among nonsmoking mothers, paternal smoking was associated with childhood overweight (OR 1.21 [95% CI 1.16–1.27], P value < 0.001) but not with adverse birth outcomes. Limitations of this study include the self-report of parental smoking information and the possibility of residual confounding. As this study only included participants from Europe and North America, results need to be carefully interpreted regarding other populations. Conclusions. We observed that as compared to nonsmoking during pregnancy, quitting smoking in the first trimester is associated with the same risk of preterm birth and small size for gestational age, but with a higher risk of childhood overweight. Reducing the number of cigarettes, without quitting, has limited beneficial effects. Paternal smoking seems to be associated, independently of maternal smoking, with the risk of childhood overweight. Population strategies should focus on parental smoking prevention before or at the start, rather than during, pregnancy.This collaborative project received funding from the European Union’s Horizon 2020 research and innovation programme (Grant Agreement No. 733206 LifeCycle). EMP and LT were supported by grant R01ES022972 from the National Institutes of Health, USA. LC was supported by the National Institute for Environmental Health Sciences: P30ES007048, R21ES029681, R01ES029944, R01ES030364, R21ES028903. DAL works in a unit that receives UK MRC funding (MC_UU_12013/5) and is an NIHR senior investigator (NF-SI-0611-10196). ACS holds an FCT Investigator contract IF/01060/2015. RG received funding from the Dutch Heart Foundation (grant number 2017T013), the Dutch Diabetes Foundation (grant number 2017.81.002) and the Netherlands Organization for Health Research and Development (ZonMW, 543003109). VWVJ received grant from the European Research Council (Consolidator Grant, ERC-2014-CoG-648916)

Hypothalamic AMPK-ER Stress-JNK1 Axis Mediates the Central Actions of Thyroid Hormones on Energy Balance

Thyroid hormones (THs) act in the brain to modulate energy balance. We show that central triiodothyronine (T3) regulates de novo lipogenesis in liver and lipid oxidation in brown adipose tissue (BAT) through the parasympathetic (PSNS) and sympathetic nervous system (SNS), respectively. Central T3 promotes hepatic lipogenesis with parallel stimulation of the thermogenic program in BAT. The action of T3 depends on AMP-activated protein kinase (AMPK)-induced regulation of two signaling pathways in the ventromedial nucleus of the hypothalamus (VMH): decreased ceramide-induced endoplasmic reticulum(ER) stress, which promotes BAT thermogenesis, and increased c-Jun N-terminal kinase (JNK) activation, which controls hepatic lipid metabolism. Of note, ablation of AMPK alpha 1 in steroidogenic factor 1 (SF1) neurons of the VMH fully recapitulated the effect of central T3, pointing to this population in mediating the effect of central THs on metabolism. Overall, these findings uncover the underlying pathways through which central T3 modulates peripheral metabolism

Ultra-high Pressure LC Method to Determine Astaxanthin in Shrimp By-Products and Migration Evaluation from an Active Plastic Film Produced with Shrimp Waste to Fatty Food Simulants

Carotenoids have antioxidant properties allowing protection of tissues from oxidative damages and they are also beneficial in cardiovascular, immune, inflammatory and neurodegenerative diseases. Astaxanthin (3,3’-dihydroxy-β-β´-carotene-4-4´-dione) is a carotenoid classified as xanthophyl and it is one of the major carotenoids in crustaceans. The project ‘Preparation of active packaging with antioxidant and antimicrobial activity based on astaxanthin and chitosan’ aims to develop a methodology for the incorporation of compounds obtained from shrimp waste in plastic matrices for the development of an active packaging with antimicrobial and antioxidant properties. In the frame of this project, shrimp by-products were fermented and the silage was centrifuged. Three fractions were obtained and the upper phase, corresponding to the lipid fraction, was further analysed to determine astaxanthin content. The aim of the present work was to optimize a method to determine astaxanthin as well as seven other carotenoids and two vitamins (A and E) by ultra-high pressure liquid chromatography (Ultra Performance Liquid Chromatography, UPLC) with diode array detection (DAD) method in shrimp by-products. The chromatographic separation is achieved using a vanguard pre-column (UPLCÒ BEH, 1.7 µm particle size) and a column (UPLCÒ BEH, 2.1 x 50 mm, 1.7 µm particle size) at 20 °C. The mobile phase is a gradient of A (dichoromethane/methanol with ammonium acetate/acetonitrile 5:20:75 (v/v)) and B (ultrapure water) with a flow rate of 0.5 mL/min. The optimized UPLC method allows excellent peaks separation. Shrimp by-products (lipid fraction of shrimp waste, head and shell of cooked and raw shrimp) have been analysed. Moreover, low density polyethylene plastic films produced by extrusion with different amount of the lipid fraction of shrimp waste were prepared and tested regarding migration into fatty food simulants. Migration tests were carried out with isooctane and ethanol 95% (v/v), both alternative fatty food simulants. No migration was detected at the conditions tested, which are conventionally considered the most severe when the food contact material is intended for use at room temperature.This work was funded under Project ‘Preparation of active packaging with antioxidant and antimicrobial activity based on astaxanthin and chitosan’ (PAPAAABAC, in Spanish PEACAABAQ) no. 95935 from FONCICYT C002-2008-1/ALA – 127 249

Migration of chitosan films prepared by solvent evaporation and extrusion

Introduction: Chitosan has multiple applications and as inhibitor of microbial growth, there is interest in developing new methodologies to its incorporation into plastics, which would avoid microbial growth in foods. In the frame of the project “Preparation of active packaging with antioxidant and antimicrobial activity based on astaxanthin and chitosan” (PAPAAABAC), chitosan was extracted from shrimp by-products and used to prepare plastic films by solvent evaporation (casting) and extrusion. Material and Methods: Chitosan films were prepared by extrusion of polyamide and by casting. By casting, films were prepared at different concentrations (1%, 2%, and 3%) by dissolving chitosan in acetic acid aqueous solution 1% (w/v), with and without plasticizer (1% glycerol). By extrusion, chitosan pellets were made of polyamide 6 at 2%, 5%, 6%, 8%, and 10% with two different particle sizes (180 and 300 µm). Then, they were rolled in an extruder using a specific screw for polyamide. In order to determine chitosan in food simulants (ultrapure water and ethanol 95% (v/v)), it was degraded into the glucosamine units by hydrolysis and quantified by ultra high pressure liquid chromatography coupled with diode array detection after its derivatization with 9-fluorenylmethyl chloroformate. Results and conclusion: From the plastic films prepared by casting and by extrusion, only those prepared by casting without plasticizer presented chitosan migration that increases with the amount of chitosan added. These films could not be used to pack aqueous foodstuffs. However, the addition of a plasticizer (glycerol) has avoided the migration of chitosan. Therefore, the use of casting films with chitosan shall include a plasticizer in the formulation. Films prepared by extrusion presented no migration into both simulants indicating suitability to pack both aqueous and fatty foodstuffs.This work was funded under the Project no. 95935 from FONCICYT C002-2008-1/ ALA – 127 249. The authors are grateful to the postdoctoral contract of Ana Sanches Silva in the frame of the Program “Science 2007” funded by “Fundação para a Ciência e para a Tecnologia”

Active Packaging Produced by Extrusion with Shrimp Waste: Migration of Astaxanthin into Food Simulants

Introduction: Astaxanthin (3,3’-dihydroxy-β-β´-carotene-4-4´-dione), a potent antioxidant, is one of the major carotenoids in crustaceans. In the frame of the project ‘Preparation of active packaging with antioxidant and antimicrobial activity based on astaxanthin and chitosan’, a methodology for the incorporation of compounds obtained from shrimp waste in plastic matrices was developed to produce an active packaging with antioxidant properties. The aim of the present work was to develop and optimize a method to determine astaxanthin by ultra-high pressure liquid chromatography in fermented shrimp waste. Moreover, the method was also applied to determine the migration of astaxanthin from plastic films containing different amounts of shrimp waste to food simulants. Material and Methods: The method was optimized to determine astaxanthin by ultra-high pressure liquid chromatography (UHPLC) with diode array detection (DAD). The chromatographic separation was achieved using a vanguard pre-column (UPLCÒ BEH, 1.7 µm particle size) and a column (UPLCÒ BEH, 2.1 x 50 mm, 1.7 µm particle size) at 20 °C. The mobile phase was a gradient of A (dichloromethane/methanol with ammonium acetate/acetonitrile 5:20:75 (v/v)) and B (ultrapure water) with a flow rate of 0.5 mL/min. The optimized UPLC method allowed an excellent resolution of astaxanthin. The method was also evaluated in what concerns to validation parameters such as linearity, precision, limit of detection, limit of quantification and recovery. Low density polyethylene plastic films produced by extrusion with different amounts of the lipid fraction of shrimp waste were prepared and tested regarding migration into fatty food stimulants (isooctane and ethanol 95%, v/v). Results and conclusion: The proposed method to determine astaxanthin in shrimp waste is simple and has a low detection level (0.054 μg/mL). The concentration of astaxanthin found in the lipid fraction of fermented shrimp waste was 453.8 μg/g. The films produced by extrusion with the lipid fraction of the fermented shrimp waste did not originate astaxanthin migration into the tested fatty food simulants. Further studies could be made in order to evaluate the capacity of these films in protecting packed food from oxidation.This work was funded under the Project no. 95935 from FONCICYT C002-2008-1/ ALA – 127 249. The authors are grateful to the postdoctoral contract of Ana Sanches Silva in the frame of the Program “Science 2007” funded by “Fundação para a Ciência e para a Tecnologia”