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5. The International development of PROQOL-HCV: An instrument to assess the health-related quality of life of patients treated for Hepatitis C virus

Author

Armstrong, A.R., Herrmann, S.E., Chassany, O., Lalanne, C., Da Silva, M.H., Galano, E., Carrieri, P.M., Estellon, V., Sogni, P., Duracinsky, M., Armstrong, A.R., Herrmann, S.E., Chassany, O., Lalanne, C., Da Silva, M.H., Galano, E., Carrieri, P.M., Estellon, V., Sogni, P., and Duracinsky, M.

Abstract

Background Hepatitis C virus (HCV) compromises Health-related Quality of Life (HRQL) with detriments to Physical, Mental and Social health domains. Treatment with interferon and ribavirin is associated with side effects which further impair HRQL. New treatments appear potent, effective and tolerable. However, Patient Reported Outcomes instruments that capture the impact on HRQL for people with hepatitis C are largely non-specific and will be needed in the new treatment era. Therefore, we developed a conceptually valid multidimensional model of HCV-specific quality of life and pilot survey instrument, the Patient Reported Outcome Quality of Life survey for HCV (PROQOL-HCV). Methods HCV patients from France (n = 30), Brazil (n = 20) and Australia (n = 20) were interviewed to investigate HCV-HRQL issues raised in the scientific literature and by treatment specialists. Interviews were recorded, transcribed and translated into English and French. Results Fifteen content dimensions were derived from the qualitative analysis, refined and fitted to four domains: (1) Physical Health included: fatigue, pain, sleep, sexual impairment and physical activity; (2) Mental Health: psychological distress, psychosocial impact, and cognition; (3) Social Health: support, stigma, social activity, substance use; (4) Treatment: management, side effects, and fear of treatment failure. The impact of some dimensions extended beyond their primary domain including: physical activity, cognition, sleep, sexual impairment, and the three treatment dimensions. A bank of 300 items was constructed to reflect patient reports and, following expert review, reduced to a 72-item pilot questionnaire. Conclusion We present a conceptually valid multidimensional model of HCV-specific quality of life and the pilot survey instrument, PROQOL-HCV. The model is widely inclusive of the experience of hepatitis C and the first to include the treatment dimension.

Published
2016

7. Psychometric Validation of the New International Questionnaire to Assess Health-Related Quality of Life (Hrql) Specific to Viral Hepatitis C: Proqol-Hcv

Author

Duracinsky, M., Armstrong, A., Herrmann, S., Lalanne, C., Galano, E., Da Silva, M.H., Carrieri, P.M., Chassany, O., Duracinsky, M., Armstrong, A., Herrmann, S., Lalanne, C., Galano, E., Da Silva, M.H., Carrieri, P.M., and Chassany, O.

Abstract

Objectives To validate a new international questionnaire to assess Health-Related Quality of Life (HRQL) in patients living with HCV. Existing tools used in HCV were mostly generic and missing important concepts such as impact of the treatment. Methods 660 HCV mono-infected or HIV co-infected patients included in a cross-sectional survey in 3 countries (France [59%], Brazil [30%], and Australia [11%]) completed paper- or internet-based version of 3 HRQL questionnaires and symptom, sociodemographic and biomedical checklist. Psychometric item analysis and factor analysis were used to reduce the original 72-item PROQOL-HVC questionnaire. Reliability (Cronbach’s alpha), convergent validity (EQ-5D, SF-12, symptoms), and clinical validity (biological markers) were assessed on the final version using correlation and two-group comparison at a fixed 5% level. Results Sample characteristics: 57% men, 52 years, 80% vocational level and higher, 35% living alone. Regarding comorbidities and treatment status: 35% co-infected with VIH (95% under ART), 15% not treated, 25% undergoing interferon, 35% ribavirine, 43% Direct-Acting Antivirals (35% without associated interferon); 27% depressive (71% treated). The PROQOL-HCV questionnaire was reduced to 38 items spanning 7 dimensions scored on a 100-point scale: Physical Health (PH, 6 items, α=0.90), Emotional Health (EH, 9 items, α=0.93), Future Uncertainty (FU, 6 items, α=0.79), Intimate Relationships (IR, 3 items, α=0.86), Social Health (SH, 3 items, α=0.47), Cognitive Functioning (CF, 3 items, α=0.80), and Treatment Impact (TI, 8 items, α=0.61). Pearson correlations of PROQOL-HCV dimension with the EQ5-D (general health) and SF-12 mental and physical composites were in the acceptable range. The PH dimension was negatively correlated to the number of bothersome symptoms (p<0.001). Lower HRQL (PH, EH, CF) was reported in women (p<0.01), depressive patients (p<0.001), patients under interferon (p<0.01, except CF), patients without trea

Published
2015

8. A biochemical and cellular approach to explore the antiproliferative and prodifferentiative activity of Aloe Arborescens leaf extract

Author

Di Luccia B, Manzo N, Vivo M, Galano E, Amoresano A, Crescenzi E, Pollice A, Tudisco R, Infascelli F, Calabrò V., DI LUCCIA, Blanda, Manzo, Nicola, Vivo, Maria, Galano, Eugenio, Amoresano, Angela, Crescenzi, E., Pollice, Alessandra, Tudisco, Raffaella, Infascelli, Federico, and Calabro', Viola

Subjects
Keratinocytes, Proteomics, Aloe arborescence, differentiation, proliferation, Aloe, Anti-Bacterial Agents, Antineoplastic Agents, Phytogenic, Cell Differentiation, Cell Line, Cell Proliferation, Gas Chromatography-Mass Spectrometry, Humans, Microbial Sensitivity Tests, Plant Extracts, Plant Leaves, Aloe arborescens, Antineoplastic Agents, cell proliferation, Phytogenic, Cell differentiation
Abstract

Aloe arborescens Miller, belonging to the Aloe genus (Liliaceae family), is one of the main varieties of Aloe used worldwide. Although less characterized than the commonest Aloe vera, Aloe arborescens is known to be richer in beneficial phytotherapeutic, anticancer, and radio-protective properties. It is commonly used as a pharmaceutical ingredient for its effect in burn treatment and ability to increase skin wound healing properties. However, very few studies have addressed the biological effects of Aloe at molecular level. The aim of the research is to provide evidences for the antiproliferative properties of Aloe arborescens crude leaf extract using an integrated proteomic and cellular biological approach. We analysed the composition of an Aloe arborescens leaf extract by gas chromatography-mass spectrometry analysis. We found it rich in Aloe-emodin, a hydroxylanthraquinone with known antitumoral activity and in several compounds with anti-oxidant properties. Accordingly, we show that the Aloe extract has antiproliferative effects on several human transformed cell lines and exhibits prodifferentiative effects on both primary and immortalized human keratinocyte. Proteomic analysis of whole cell extracts revealed the presence of proteins with a strong antiproliferative and antimicrobial activity specifically induced in human keratinocytes by Aloe treatment supporting its application as a therapeutical agent. Copyright © 2013 John Wiley & Sons, Ltd.

Published
2013

9. The constituents of the ink from a Qumran inkwell: new prospects for provenancing the ink on the Dead Sea Scrolls

Author

Rasmussen, Kl, Tenorio, Al, Bonaduce, Ilaria, Colombini, MARIA PERLA, Birolo, L, Galano, E, Amoresano, A, Doudna, G, Bond, Ad, Palleschi, V, Lorenzetti, G, Legnaioli, S, van der Plicht, J, Gunneweg, J., LLUVERAS TENORIO, Anna, Isotope Research, Kaare Lund, Rasmussen, Anna Lluveras, Tenorio, Ilaria, Bonaduce, Maria Perla, Colombini, Birolo, Leila, Galano, Eugenio, Amoresano, Angela, Greg, Doudna, Andrew D., Bond, Vincenzo, Palleschi, Giulia, Lorenzetti, Stefano, Legnaioli, Johannes van der, Plicht, and Jan, Gunneweg

Subjects
Radiocarbon dating, Archeology, LIBS, SPECTROSCOPY, Inkwell, STRATEGIES, media_common.quotation_subject, Dead Sea Scrolls, Art, Qumran, Archaeology, AMORPHOUS CALCIUM-CARBONATE, MONOHYDROCALCITE, FT-IR, MEDIA, PXRD, Proteomic analyses, ICP-MS, Inorganic materials, Ink, Second Temple period, GC-MS, NESQUEHONITE, Raman, media_common
Abstract

A unique sample of ink from an inkwell in the Schoyen Collection allegedly found at Qumran has been subjected to analyses by several analytical techniques: GC-MS, proteomic analysis, PXRD, Raman, (ATR) FT-IR, LIBS, ICP-MS and MS. The results reveal to an unexpected level of detail how the ink was manufactured, which gives insight into, the industrial processes and craftsmanship that were practiced at the Qumran settlement during the Second Temple period (100 BCE-CE 70). The identified minerals and other organic and inorganic materials are sufficiently multiple and diverse that it is probable that this specific ink can be recognized if analyses. of inks are performed on manuscripts from Qumran and other locations in Israel and the Middle East. The present work exposes a distinct and unique possibility to shed light on early Jewish manuscript controversies, including their provenance. (C) 2012 Elsevier Ltd. All rights reserved.

Published
2012
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10. Living Lab MACVIA Handicap

Author

Laffont, I., primary, Jourdan, C., additional, Coroian, F., additional, Blain, H., additional, Carre, V., additional, Viollet, E., additional, Tavares, I., additional, Fattal, C., additional, Gelis, A., additional, Nouvel, F., additional, Bakhti, K., additional, Cros, V., additional, Patte, K., additional, Schifano, L., additional, Porte, M., additional, Galano, E., additional, Dray, G., additional, Fouletier, M., additional, Rivier, F., additional, Morales, R., additional, Labauge, P., additional, Camu, W., additional, Combe, B., additional, Morel, J., additional, Froger, J., additional, Coulet, B., additional, Cottalorda, J., additional, Kouyoumdjian, P., additional, Jonquet, O., additional, Landreau, L., additional, Bonnin, H.-Y., additional, Hantkié, O., additional, Nicolas, P., additional, Enjalbert, M., additional, Leblond, C., additional, Soua, B., additional, Coignard, P., additional, Guiraud, D., additional, Azevedo, C., additional, Mottet, D., additional, Fraisse, P., additional, Pastor, E., additional, Mercier, J., additional, Bourret, R., additional, Bousquet, J., additional, Pélissier, J., additional, Bardy, B., additional, Herisson, C., additional, and Dupeyron, A., additional

Published
2015
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17. A simple and reliable methodology to detect egg white in art samples

Author

Gambino, M., Cappitelli, F., Cattò, C., Carpen, A., Principi, P., Ghezzi, L., Bonaduce, I., Galano, E., Pucci, P., Birolo, L., Villa, F., Forlani, F., Gambino, M., Cappitelli, F., Cattò, C., Carpen, A., Principi, P., Ghezzi, L., Bonaduce, I., Galano, E., Pucci, P., Birolo, L., Villa, F., and Forlani, F.

Published
2013

34. Monitoring of shaft voltage and shaft grounding current in rotating machinery

Author

Nippes, P.I. and Galano, E.

Subjects
Ships -- Design and construction -- Usage, Vibration -- Analysis -- Usage, Shafting -- Usage, Engineering and manufacturing industries, Design and construction, Analysis, Usage
Abstract

06-1524 Nippes, P.I., and E. Galano. Monitoring of shaft voltage and shaft grounding current in rotating machinery. Proceedings of the 60th Meeting of the Society for Machinery Failure Prevention Technology [...]

Published
2006

36. Introduction

Author

Bousquet, J., Bourret, R., Camuzat, T., Augé, P., Domy, P., Bringer, J., Best, N., Jonquet, O., de la Coussaye, J.-E., Noguès, M., Robine, J.-M., Avignon, A., Blain, H., Combe, B., Dray, G., Dufour, V., Fouletier, M., Giraudeau, N., Hève, D., Jeandel, C., Laffont, I., Larrey, D., Laune, D., Laurent, C., Mares, P., Marion, C., Pastor, E., Pélissier, J.-Y., Radier-Pontal, F., Reynes, J., Royère, E., Ychou, M., Bedbrook, A., Granier, S., Abecassis, F., Albert, S., Adnet, P.-A., Alomène, B., Amouyal, M., Arnavielhe, S., Asteriou, T., Attalin, V., Aubas, P., Azevedo, C., Badin, M., Bakhti, Baptista, G., Bardy, B., Battesti, M.-P., Bénézet, O., Bernard, P.-L., Berr, C., Berthe, J., Bobia, X., Bockaert, J., Boegner, C., Boichot, S., Bonnin, H.-Y., Boulet, P., Bouly, S., Boubakri, C., Bourdin, A., Bourrain, J.-L., Bourrel, G., Bouix, V., Breuker, C., Bruguière, V., Burille, J., Cade, S., Caimmi, D., Calmels, M.-V., Camu, W., Canovas, G., Carre, V., Cavalli, G., Cayla, G., Chiron, R., Claret, P.-G., Coignard, P., Coroian, F., Costa, D.-J., Costa, P., Cottalorda, Coulet, B., Coupet, A.-L., Courrouy-Michel, M.-C., Courtet, P., Cristol, J.-P., Cros, V., Cuisinier, F., Daien, C., Danko, M., Dauenhauer, P., Dauzat, M., David, M., Davy, J.-M., Delignières, D., Demoly, P., Desplan, J., Dhivert-Donnadieu, H., Dujols, P., Dupeyron, A., Dupeyron, G., Engberink, O., Enjalbert, M., Fattal, C., Fernandes, J., Fesler, P., Fraisse, P., Froger, J., Gabrion, P., Galano, E., Gellerat-Rogier, M., Gellis, A., Goucham, A.-Y., Gouzi, F., Gressard, F., Gris, J.-C., Guillot, B., Guiraud, D., Handweiler, V., Hantkié, H., Hayot, M., Hérisson, C., Heroum, C., Hoa, D., Jacquemin, S., Jaber, S., Jakovenko, D., Jorgensen, C., Journot, L., Kaczorek, M., Kouyoudjian, P., Labauge, P., Landreau, L., Lapierre, M., Leblond, C., Léglise, M.-S., Lemaitre, J.-M., Le Moing, V., Le Quellec, A., Leclercq, F., Lehmann, S., Lognos, B., Lussert, J.-M., Makinson, A., Mandrick, K., Marmelat, V., Martin-Gousset, P., Matheron, A., Mathieu, G., Meissonnier, M., Mercier, G., Messner, P., Meunier, C., Mondain, M., Morales, R., Morel, J., Morquin, D., Mottet, D., Nérin, P., Nicolas, P., Ninot, G., Nouvel, F., Ortiz, J.-P., Paccard, D., Pandraud, G., Pasdelou, M.-P., Pasquié, J.-L., Patte, K., Perrey, S., Pers, Y.-M., Picot, M.-C., Pin, J.-P., Pinto, N., Porte, E., Portejoie, F., Pujol, J.-L., Quantin, X., Quéré, I., Raffort, N., Ramdani, S., Ribstein, J., Rédini-Martinez, I., Richard, S., Ritchie, K., Riso, J.-P., Rivier, F., Rolland, C., Roubille, F., Sablot, D., Savy, J.-L., Schifano, L., Senesse, P., Sicard, R., Soua, B., Stephan, Y., Strubel, D., Sultan, A., Taddei-Ologeanu, Tallon, G., Tanfin, M., Tassery, H., Tavares, I., Torre, K., Touchon, J., Tribout, V., Uziel, A., Van de Perre, P., Vasquez, X., Verdier, J.-M., Vergne-Richard, C., Vergotte, G., Vian, L., Viarouge-Reunier, C., Vialla, F., Viart, F., Villain, M., Villiet, M., Viollet, E., Wojtusciszyn, A., Aoustin, M., Bourquin, C., and Mercier, J.

Published
2015
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37. Living LabMACVIA Handicap

Author

Laffont, I., Jourdan, C., Coroian, F., Blain, H., Carre, V., Viollet, E., Tavares, I., Fattal, C., Gelis, A., Nouvel, F., Bakhti, K., Cros, V., Patte, K., Schifano, L., Porte, M., Galano, E., Dray, G., Fouletier, M., Rivier, F., Morales, R., Labauge, P., Camu, W., Combe, B., Morel, J., Froger, J., Coulet, B., Cottalorda, J., Kouyoumdjian, P., Jonquet, O., Landreau, L., Bonnin, H.-Y., Hantkié, O., Nicolas, P., Enjalbert, M., Leblond, C., Soua, B., Coignard, P., Guiraud, D., Azevedo, C., Mottet, D., Fraisse, P., Pastor, E., Mercier, J., Bourret, R., Bousquet, J., Pélissier, J., Bardy, B., Herisson, C., and Dupeyron, A.

Published
2015
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38. Family structure and adolescent sexual behavior in a poor area of Sao Paulo, Brazil.

Author

Peres CA, Rutherford G, Borges G, Galano E, Hudes ES, and Hearst N

Abstract

PURPOSE: In Brazil, as elsewhere, behavior during adolescence can place young people at risk for serious medical and social problems, including sexually transmitted infections, unintended pregnancy, drugs, crime, and violence. Few studies internationally have examined the influence of family structure on risk behavior among low-income youths. METHODS: This cross-sectional study included 296 young people in one of the poorest areas of Sao Paulo who were recruited through a vocational school and completed an anonymous, self-administered questionnaire. We examined associations between family structure and various risk behaviors. RESULTS: Ages ranged from 13-24 years (82%, 15-18); 67% were of Afro-Brazilian ancestry, and 56% were female. Median family monthly income was about US$200. Less than half lived with both parents, and 14% lived with neither parent. Rates of many risk behaviors, including involvement in crime and violence, drug and alcohol use, and sexual risk, were lowest among those living with both parents, higher among those living with one parent, and highest among those living with neither parent. For example, 26% of females living with both parents, 37% with one parent, and 71% with neither parent were sexually active (p = .003). Family structure and a personal or parental history of drug or alcohol problems were significant independent predictors of sexual activity. CONCLUSIONS: The presence of both parents is an important protective factor for Brazilian youth vulnerable to multiple risks. Prevention programs should explore ways to support parents to be present and involved in the lives of their adolescent children. [ABSTRACT FROM AUTHOR]

Published
2008
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39. Multisectorial responses to HIV/AIDS. Vulnerability, human rights, and comprehensive health care needs of young people living with HIV/AIDS.

Author

Ayres JR, Paiva V, França I Jr, Gravato N, Lacerda R, Della Negra M, Marques HH, Galano E, Lecussan P, Segurado AC, and Silva MH

Abstract

We sought to identify and understand the health care needs of young people living with HIV/AIDS, particularly in terms of their psychosocial well-being. We conducted a qualitative analysis of HIV-positive young people and their caregivers, focusing on the implications of an HIV diagnosis for health care needs. Stigma was a recurrent issue that arose in the interviews conducted with the respondents, and it was evident that youths had been denied many rights related to health. We concluded that young people living with HIV need comprehensive care based on a human rights approach. In this regard, we offer some practical recommendations for health programs. [ABSTRACT FROM AUTHOR]

Published
2006
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44. Molecular fingerprint of the alcoholic Grappa beverage by mass spectrometry techniques

Author

Eugenio Galano, Matteo Imbelloni, Antonio Malorni, Angela Amoresano, Angela Chambery, Galano, E, Imbelloni, M, Chambery, Angela, Malorni, A, Amoresano, A., Galano, Eugenio, Imbelloni, Matteo, Antonio, Malorni, and Amoresano, Angela

Subjects
Chromatography, Aromatic compound, MALDI fingerprint, Mass spectrometry, Chemistry, Analytical chemistry, Grappa, Time-of-flight mass spectrometry, Molecular Fingerprint, Food Science
Abstract

Grappa is the traditional Italian spirit derived from marc. The molecular characterization of volatile compounds in the liquid Grappa, responsible of its bouquet, has been achieved by headspace solid-phase microextraction (HS-SPME) followed by gas chromatography–mass spectrometry (GC-MS). Of these, 12 compounds represent more than 1% of Grappa volatile fraction. In addition, a molecular fingerprint of Grappa samples was performed by MALDI-TOF and MALDI-TOF/TOF MS, allowing the identification of molecular structures of major compounds within the non-volatile fraction of Grappa. The − 82 Da shifts on precursor ion masses due to the loss of 5-methylfuran ion also allowed the identification of some heterocyclic compounds, such as furan derivatives, formed during the Grappa aging within barrels. Overall, by integrating different mass spec techniques, we obtained a comprehensive molecular fingerprint of selected Grappa samples.

Published
2015

45. Qualitative screening in doping control by MALDI-TOF/TOF mass spectrometry: A proof-of-evidence

Author

L. Palomba, M. Fidani, Gennaro Marino, F. Baia, Angela Amoresano, Eugenio Galano, Galano, E, Fidani, M, Baia, F, Palomba, L, Marino, G, and Amoresano, Angela

Subjects
Chromatography, Chemistry, Illicit Drugs, Clinical Biochemistry, Forensic toxicology, Pharmaceutical Science, Reproducibility of Results, Mass spectrometry, Analytical Chemistry, Substance Abuse Detection, Cocaine, Tandem Mass Spectrometry, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Drug Discovery, Analytical Toxicology, Screening method, Biological fluids, Animals, Humans, Horses, Spectroscopy, Cocaine urine
Abstract

The analysis of doping agents in biological fluids is of top significance in clinical and forensic toxicology. Herein we describe the study of a screening method for the detection of a mixture of drugs of potential abuse including cocaine and its metabolites. By using matrix-assisted laser desorption/ionization MALDI-TOF/TOF mass spectrometry. This screening procedure to detect the presence of different drugs, avoiding time consuming procedures could be useful in different fields of forensic analytical toxicology, including antidoping analysis.

Published
2012

46. Advantages and limitations of laser desorption/ionization mass spectrometric techniques in the chemical characterization of complex carbonaceous materials

Author

Eugenio Galano, Michela Alfè, Angela Amoresano, Barbara Apicella, Anna Ciajolo, Apicella, B., Alfe, M., Amoresano, Angela, Galano, E, and Ciajolo, A.

Subjects
Chemical ionization, Mass spectrometry, Chemistry, Analytical chemistry, Combustion, Condensed Matter Physics, Laser, Characterization (materials science), law.invention, Time of flight, Carbonaceous samples, law, Desorption, Ionization, Physics::Atomic and Molecular Clusters, Laser power scaling, Physics::Atomic Physics, Physical and Theoretical Chemistry, Instrumentation, Spectroscopy
Abstract

Laser desorption/ionization techniques coupled with mass spectrometry analyzers have evolved rapidly in the recent years and are currently capable of providing valuable information about the chemical composition and structure of very high molecular weight species, mainly biopolymers or synthetic polymers. In view of this rapidly increasing interest a thorough understanding of the desorption/ionization process is not only of scientific interest, but also important for a correct spectra interpretation and for further improvements of the technique. In the present paper, the effect of main experimental parameters on mass range detectable by laser desorption/ionization techniques has been investigated for standard aromatic molecules, like polycyclic aromatic hydrocarbons (PAH), fullerenes, polyacenaphthylene (PACE) and for complex carbonaceous materials like heavy fractions of fuel oils and combustion-formed particulate. In particular, it has been shown that laser power (or more specifically, the surface power density of the laser spot, named laser fluence) as well as the surface concentration of samples are crucial parameters controlling the highest detected molecular weight range. However, neither of these parameters is easily well-controlled and therefore more work is necessary for the standardization of laser/desorption techniques in the analysis of complex samples.

Published
2010
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47. Glycosylation Pattern and in vitro Bioactivity of Reference Follitropin alfa and Biosimilars.

Author

Riccetti L, Sperduti S, Lazzaretti C, Klett D, De Pascali F, Paradiso E, Limoncella S, Potì F, Tagliavini S, Trenti T, Galano E, Palmese A, Satwekar A, Daolio J, Nicoli A, Villani MT, Aguzzoli L, Reiter E, Simoni M, and Casarini L

Abstract

Recombinant follicle-stimulating hormone (FSH) (follitropin alfa) and biosimilar preparations are available for clinical use. They have specific FSH activity and a unique glycosylation profile dependent on source cells. The aim of the study is to compare the originator (reference) follitropin alfa (Gonal-f®)- with biosimilar preparations (Bemfola® and Ovaleap®)-induced cellular responses in vitro . Gonadotropin N-glycosylation profiles were analyzed by ELISA lectin assay, revealing preparation specific-patterns of glycan species (Kruskal-Wallis test; p < 0.05, n = 6) and by glycotope mapping. Increasing concentrations of Gonal-f® or biosimilar (1 × 10 -3 -1 × 10 3 ng/ml) were used for treating human primary granulosa lutein cells (hGLC) and FSH receptor (FSHR)-transfected HEK293 cells in vitro . Intracellular cAMP production, Ca 2+ increase and β-arrestin 2 recruitment were evaluated by BRET, CREB, and ERK1/2 phosphorylation by Western blotting. 12-h gene expression, and 8- and 24-h progesterone and estradiol synthesis were measured by real-time PCR and immunoassay, respectively. We found preparation-specific glycosylation patterns by lectin assay (Kruskal-Wallis test; p < 0.001; n = 6), and similar cAMP production and β-arrestin 2 recruitment in FSHR-transfected HEK293 cells (cAMP EC 50 range = 12 ± 0.9-24 ± 1.7 ng/ml; β-arrestin 2 EC 50 range = 140 ± 14.1-313 ± 18.7 ng/ml; Kruskal-Wallis test; p ≥ 0.05; n = 4). Kinetics analysis revealed that intracellular Ca 2+ increased upon cell treatment by 4 μg/ml Gonal-f®, while equal concentrations of biosimilars failed to induced a response (Kruskal-Wallis test; p < 0.05; n = 3). All preparations induced both 8 and 24 h-progesterone and estradiol synthesis in hGLC, while no different EC 50 s were demonstrated (Kruskal-Wallis test; p > 0.05; n = 5). Apart from preparation-specific intracellular Ca 2+ increases achieved at supra-physiological hormone doses, all compounds induced similar intracellular responses and steroidogenesis, reflecting similar bioactivity, and overall structural homogeneity.

Published
2019
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48. Generation of site-distinct N-glycan variants for in vitro bioactivity testing.

Author

Brühlmann D, Vuillemin T, Satwekar A, Galano E, Palmese A, D'Angelo A, Manco Z, Souquet J, Broly H, Sauer M, Hemberger J, and Jordan M

Subjects
Animals, Antibodies, Monoclonal genetics, CHO Cells, Cricetulus, Glycosylation, Immunoglobulin Fc Fragments genetics, Mannose genetics, Polysaccharides genetics, Receptors, IgG genetics, Antibodies, Monoclonal biosynthesis, Immunoglobulin Fc Fragments biosynthesis, Mannose metabolism, Polysaccharides metabolism, Receptors, IgG metabolism
Abstract

Glycosylation, a critical product quality attribute, may affect the efficacy and safety of therapeutic proteins in vivo. Chinese hamster ovary fed-batch cell culture batches yielded consistent glycoprofiles of a Fc-fusion antibody comprizing three different N-glycosylation sites. By adding media supplements at specific concentrations in cell culture and applying enzymatic glycoengineering, a diverse N-glycan variant population was generated, including high mannose, afucosylated, fucosylated, agalactosylated, galactosylated, asialylated, and sialylated forms. Site-specific glycosylation profiles were elucidated by glycopeptide mapping and the effect of the glycosylation variants on the FcγRIIIa receptor binding affinity and the biological activity (cell-based and surface plasmon resonance) was assessed. The two fusion body glycosylation sites were characterized by a high degree of sialic acid, more complex N-glycan structures, a higher degree of antennarity, and a site-specific behavior in the presence of a media supplement. On the other hand, the media supplements affected the Fc-site glycosylation heterogeneity similarly to the various studies described in the literature with classical monoclonal antibodies. Enzymatic glycoengineering solely managed to generate high levels of galactosylation at the fusion body sites. Variants with low core fucosylation, and to a lower extent, high mannose glycans exhibited increased FcγRIIIa receptor binding affinity. All N-glycan variants exhibited weak effects on the biological activity of the fusion body. Both media supplementation and enzymatic glycoengineering are suitable to generate sufficient diversity to assess the effect of glycostructures on the biological activity., (© 2019 Wiley Periodicals, Inc.)

Published
2019
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50. GC/MS and proteomics to unravel the painting history of the lost Giant Buddhas of Bāmiyān (Afghanistan).

Author

Lluveras-Tenorio A, Vinciguerra R, Galano E, Blaensdorf C, Emmerling E, Perla Colombini M, Birolo L, and Bonaduce I

Subjects
Afghanistan, Animals, Asia, Eastern, Gas Chromatography-Mass Spectrometry methods, Milk chemistry, Milk Proteins chemistry, Paintings, Proteins chemistry, Proteomics methods, Paint analysis
Abstract

A chemical investigation of the organic paint binders of the Giant Buddhas of Bāmiyān was performed using an analytical approach based on mass spectrometry, combining traditional gas chromatography/mass spectrometry protocols with advanced proteomics methodologies. The research was carried out on a selection of rescued fragments. The data revealed the use of egg proteins as the paint binders of the original layers, in accordance with the traditional use of this proteinaceous medium in antiquity, spanning from the Mediterranean basin to the Far East, and already in the Bronze Age. Egg tempera was thus known to artists of the region in the first centuries AD, probably also due to the position of the Bāmiyān valley, which was connected to the Silk Road. Milk was found in the first historical overpaintings. A new proteomics approach was used, which was able to identify the source of the milk proteins present in the restoration layers, despite their age and degradation. In particular cow's and goat's milk were both found, in agreement with the documented presence of rich pastures in the Bāmiyān valley when the historical restorations were carried out. Investigating the materials of the Giant Buddhas not only enabled us to obtain isolated data on these invaluable works of art, which are now lost, but contributes to understanding the big "puzzle" of our past and the development of our culture, by implementing and supporting written sources, stylistic and anthropological studies with molecular data.

Published
2017
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