Journal of Pharmaceutical and Biomedical Analysis, Jun 1, 2010
A new photoisomer of the promising &a... more A new photoisomer of the promising "anti-Alzheimer" drug candidate (+/-) huperzine A is described. The new substance was formed via a photoisomerization reaction and was found to be 1-amino-13-ethylidene-11-methyl-6-aza-tetracyclo-[7.3.1.0(2.7).0(4.7)]-trideca-2,10-diene-5-one using NMR analysis. The kinetics of its formation was studied and proven to be of first-order. The described photoisomer showed a significant loss in activity, being more than 100 times less active than (-) huperzine A itself. The new substance was named photohuperzine A, referring to its photopyridone substructure.
A new photoisomer of the promising “anti-Alzheimer” drug candidate (±) huperzine A is described. ... more A new photoisomer of the promising “anti-Alzheimer” drug candidate (±) huperzine A is described. The new substance was formed via a photoisomerization reaction and was found to be 1-amino-13-ethylidene-11-methyl-6-aza-tetracyclo-[7.3.1.02.7.04.7]-trideca-2,10-diene5-one using NMR analysis. The kinetics of its formation was studied and proven to be of first-order. The described photoisomer showed a significant loss in activity, being more than 100 times less active than (−) huperzine A itself. The new substance was named photohuperzine A, referring to its photopyridone substructure.
Journal of Pharmaceutical and Biomedical Analysis, 2010
A new photoisomer of the promising &a... more A new photoisomer of the promising "anti-Alzheimer" drug candidate (+/-) huperzine A is described. The new substance was formed via a photoisomerization reaction and was found to be 1-amino-13-ethylidene-11-methyl-6-aza-tetracyclo-[7.3.1.0(2.7).0(4.7)]-trideca-2,10-diene-5-one using NMR analysis. The kinetics of its formation was studied and proven to be of first-order. The described photoisomer showed a significant loss in activity, being more than 100 times less active than (-) huperzine A itself. The new substance was named photohuperzine A, referring to its photopyridone substructure.
Alzheimer's is a neurodegenerative disease. Its symptoms are attributed to a deficiency of ch... more Alzheimer's is a neurodegenerative disease. Its symptoms are attributed to a deficiency of cholinergic neurotransmission. The drugs of choice for the treatment of Alzheimer's disease are acetylcholinesterase (AChE) inhibitors. Starting in the 1980's from non‐specific AChE inhibitors, the first‐generation drugs such as physostigmine, a second generation of more selective and better tolerated products has been developed. Methods to detect and quantify these drugs and their metabolites in biological samples have been developed for analysis in plasma, blood, urine and cerebrospinal fluid. Diverse detection techniques have been used, such as ultraviolet, fluorescence, electrochemical and mass spectrometry. In this review, the methods applied to the analysis of these drugs and their metabolites in different biological matrices are reviewed and discussed. The stability of these drugs in biological matrices and under stress‐conditions is also included in the discussion. Copyrigh...
Journal of Pharmaceutical and Biomedical Analysis, 2011
Galantamine hydrobromide was subjected to different stress conditions (acidic, alkaline, thermal,... more Galantamine hydrobromide was subjected to different stress conditions (acidic, alkaline, thermal, photolytic and oxidative). Degradation was found to occur under acidic, photolytic and oxidative conditions, while the drug was stable under alkaline and elevated temperature conditions. A stability-indicating reversed-phase liquid chromatographic method was developed for the determination of the drug in the presence of its degradation products. The method was validated for linearity, precision, accuracy, specificity, selectivity and intermediate precision. Additionally, the degradation kinetics of the drug was assessed in relevant cases. The kinetics followed a first order behavior in the case of acidic and photolytic degradation, while a two-phase kinetics behavior was found for the oxidative degradation. The degradation products were characterized by mass spectrometry and nuclear magnetic resonance spectroscopy. Dehydration, epimerization and N-oxidation were the main processes observed during the degradation of galantamine. Moreover, if sufficient material could be isolated the inhibitory activity against the target enzyme acetylcholinesterase was also assessed.
Cure of severe infections, sepsis, and septic shock with antimicrobial drugs is a challenge becau... more Cure of severe infections, sepsis, and septic shock with antimicrobial drugs is a challenge because morbidity and mortality in these conditions are essentially caused by improper immune response. We have tested the hypothesis that repeated reactivation of established memory to pathogens may reset unfavorable immune responses. We have chosen for this purpose a highly stringent mouse model of polymicrobial sepsis by cecum ligation and puncture. Five weeks after priming with a diverse Ag pool, high-grade sepsis was induced in C57BL/6j mice that was lethal in 24 h if left untreated. Antimicrobial drug (imipenem) alone rescued 9.7% of the animals from death, but >5-fold higher cure rate could be achieved by combining imipenem and two rechallenges with the Ag pool (p < 0.0001). Antigenic stimulation fine-tuned the immune response in sepsis by contracting the total CD3+ T cell compartment in the spleen and disengaging the hyperactivation state in the memory T subsets, most notably CD...
Dental materials : official publication of the Academy of Dental Materials, 2017
To evaluate the cytotoxic effects of exposing odontoblast cells to a variety of commercial self-a... more To evaluate the cytotoxic effects of exposing odontoblast cells to a variety of commercial self-adhesive cements polymerized using different activation modes. Five cements: MaxCem Elite (MAX), Bifix SE (BSE), G-Cem LinkAce (GCE), Clearfil SA Luting (CAS), and RelyX U200 (U200) were mixed, dispensed into molds, and distributed in groups, according to polymerization protocols: immediate photoactivation; delayed photoactivation (10min self-curing plus light-activation); and chemical activation (no light exposure). Immortalized rat odontoblast cells (MDPC-23) were cultured. Cell viability was assessed by Trypan Blue staining and total cell death was assessed by annexin V-APC/7-AAD double staining and flow cytometry. Volatilized compounds from polymerized specimens of cements were evaluated by gas chromatography/mass spectrometry (GC-MS). Data was analyzed with 2-way ANOVA/Tukey tests (α=0.05). Exposure to all of the cements tested significantly reduced the cell viability, irrespective o...
The influence of in vitro maturation (IVM) in oocytes is still not totally understood. The aim of... more The influence of in vitro maturation (IVM) in oocytes is still not totally understood. The aim of this study was to determine the influence of IVM on the metabolism and homeostasis of bovine cumulus-oocyte complexes. In the present study, we demonstrated that IVM leads to accumulation of neutral lipids associated with differential levels of the mono-, di-, and tri-acylglycerols in both cumulus cells and oocytes. We observed that in vitro-matured oocytes exhibited decreased glutathione and reactive oxygen species levels and a lower ATP/ADP ratio when compared to in vivo-matured oocytes, with no significant differences in metabolism and stress related mRNA or miRNA levels. Moreover, in addition to an increase in lipids in in vitro-matured cumulus cells, fatty acid synthesis and accumulation as well as glycolysis pathway genes were upregulated, whereas those affiliated with the -oxidation pathway were decreased. Our gene expression data in cumulus cells suggest the disruption of endopl...
Lysine is catabolized in mammals through the saccharopine and pipecolate pathways - the former is... more Lysine is catabolized in mammals through the saccharopine and pipecolate pathways - the former is mainly hepatic and renal, and the latter is believed to play a role in the cerebral lysine oxidation. Both pathways lead to the formation of aminoadipic semialdehyde (AASA) that is then oxidized to aminoadipate (AAA) by antiquitin (ALDH7A1). Mutations in the ALDH7A1 gene result in the accumulation of AASA and its cyclic form, piperideine-6-carboxylate (P6C), which causes pyridoxine-dependent epilepsy (PDE). P6C reacts with pyridoxal 5'-phosphate (PLP) causing its inactivation. Here, we used liquid chromatography-mass spectrometry to investigate lysine catabolism in mice injected with lysine labelled at either its nitrogen epsilon (ε-(15)N) or nitrogen alpha (α-(15)N). Analysis of ε-(15)N and α-(15)N lysine catabolites in plasma, liver and brain suggested the saccharopine as the main pathway for AAA biosynthesis. Although there was evidence for upstream cerebral pipecolate pathway ac...
The present paper describes the influence of the chemical structure of two aminoalkoxysilanes: 3-... more The present paper describes the influence of the chemical structure of two aminoalkoxysilanes: 3-aminopropyltriethoxysilane (APTS) and N-(3-(trimethoxysilyl)-propyl)-ethylenediamine (TSPEN) on the morphology of thin layer hybrid films with ...
European Journal of Lipid Science and Technology, 2010
Easy ambient sonic spray ionization (EASI), a novel desorption/ionization technique for ambient m... more Easy ambient sonic spray ionization (EASI), a novel desorption/ionization technique for ambient mass spectrometry analysis, is shown to permit the characterization of different types of vegetable and animal fats. The taxonomic markers of the oils, mainly fatty acids and phenols, are extracted with methanol/water (1:1) solution and made alkaline with NH4OH, then placed and dried on the surface of a glass tip and directly analyzed by EASI‐MS in the negative‐ion mode. EASI provides a supersonic cloud of charged droplets that causes efficient desorption and ionization of the oil markers directly from the surface of the glass tip. As proof‐of‐principle cases, EASI(−)‐MS was applied to genuine samples of olive oil, hazelnut oil, soybean oil, grape seed oil, canola oil, butter, and lard. Characteristic metabolomics EASI(−)‐MS profiles of fatty acids and eventually phenols were obtained.
Journal of Pharmaceutical and Biomedical Analysis, Jun 1, 2010
A new photoisomer of the promising &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;a... more A new photoisomer of the promising &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;quot;anti-Alzheimer&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;quot; drug candidate (+/-) huperzine A is described. The new substance was formed via a photoisomerization reaction and was found to be 1-amino-13-ethylidene-11-methyl-6-aza-tetracyclo-[7.3.1.0(2.7).0(4.7)]-trideca-2,10-diene-5-one using NMR analysis. The kinetics of its formation was studied and proven to be of first-order. The described photoisomer showed a significant loss in activity, being more than 100 times less active than (-) huperzine A itself. The new substance was named photohuperzine A, referring to its photopyridone substructure.
A new photoisomer of the promising “anti-Alzheimer” drug candidate (±) huperzine A is described. ... more A new photoisomer of the promising “anti-Alzheimer” drug candidate (±) huperzine A is described. The new substance was formed via a photoisomerization reaction and was found to be 1-amino-13-ethylidene-11-methyl-6-aza-tetracyclo-[7.3.1.02.7.04.7]-trideca-2,10-diene5-one using NMR analysis. The kinetics of its formation was studied and proven to be of first-order. The described photoisomer showed a significant loss in activity, being more than 100 times less active than (−) huperzine A itself. The new substance was named photohuperzine A, referring to its photopyridone substructure.
Journal of Pharmaceutical and Biomedical Analysis, 2010
A new photoisomer of the promising &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;a... more A new photoisomer of the promising &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;quot;anti-Alzheimer&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;quot; drug candidate (+/-) huperzine A is described. The new substance was formed via a photoisomerization reaction and was found to be 1-amino-13-ethylidene-11-methyl-6-aza-tetracyclo-[7.3.1.0(2.7).0(4.7)]-trideca-2,10-diene-5-one using NMR analysis. The kinetics of its formation was studied and proven to be of first-order. The described photoisomer showed a significant loss in activity, being more than 100 times less active than (-) huperzine A itself. The new substance was named photohuperzine A, referring to its photopyridone substructure.
Alzheimer's is a neurodegenerative disease. Its symptoms are attributed to a deficiency of ch... more Alzheimer's is a neurodegenerative disease. Its symptoms are attributed to a deficiency of cholinergic neurotransmission. The drugs of choice for the treatment of Alzheimer's disease are acetylcholinesterase (AChE) inhibitors. Starting in the 1980's from non‐specific AChE inhibitors, the first‐generation drugs such as physostigmine, a second generation of more selective and better tolerated products has been developed. Methods to detect and quantify these drugs and their metabolites in biological samples have been developed for analysis in plasma, blood, urine and cerebrospinal fluid. Diverse detection techniques have been used, such as ultraviolet, fluorescence, electrochemical and mass spectrometry. In this review, the methods applied to the analysis of these drugs and their metabolites in different biological matrices are reviewed and discussed. The stability of these drugs in biological matrices and under stress‐conditions is also included in the discussion. Copyrigh...
Journal of Pharmaceutical and Biomedical Analysis, 2011
Galantamine hydrobromide was subjected to different stress conditions (acidic, alkaline, thermal,... more Galantamine hydrobromide was subjected to different stress conditions (acidic, alkaline, thermal, photolytic and oxidative). Degradation was found to occur under acidic, photolytic and oxidative conditions, while the drug was stable under alkaline and elevated temperature conditions. A stability-indicating reversed-phase liquid chromatographic method was developed for the determination of the drug in the presence of its degradation products. The method was validated for linearity, precision, accuracy, specificity, selectivity and intermediate precision. Additionally, the degradation kinetics of the drug was assessed in relevant cases. The kinetics followed a first order behavior in the case of acidic and photolytic degradation, while a two-phase kinetics behavior was found for the oxidative degradation. The degradation products were characterized by mass spectrometry and nuclear magnetic resonance spectroscopy. Dehydration, epimerization and N-oxidation were the main processes observed during the degradation of galantamine. Moreover, if sufficient material could be isolated the inhibitory activity against the target enzyme acetylcholinesterase was also assessed.
Cure of severe infections, sepsis, and septic shock with antimicrobial drugs is a challenge becau... more Cure of severe infections, sepsis, and septic shock with antimicrobial drugs is a challenge because morbidity and mortality in these conditions are essentially caused by improper immune response. We have tested the hypothesis that repeated reactivation of established memory to pathogens may reset unfavorable immune responses. We have chosen for this purpose a highly stringent mouse model of polymicrobial sepsis by cecum ligation and puncture. Five weeks after priming with a diverse Ag pool, high-grade sepsis was induced in C57BL/6j mice that was lethal in 24 h if left untreated. Antimicrobial drug (imipenem) alone rescued 9.7% of the animals from death, but >5-fold higher cure rate could be achieved by combining imipenem and two rechallenges with the Ag pool (p < 0.0001). Antigenic stimulation fine-tuned the immune response in sepsis by contracting the total CD3+ T cell compartment in the spleen and disengaging the hyperactivation state in the memory T subsets, most notably CD...
Dental materials : official publication of the Academy of Dental Materials, 2017
To evaluate the cytotoxic effects of exposing odontoblast cells to a variety of commercial self-a... more To evaluate the cytotoxic effects of exposing odontoblast cells to a variety of commercial self-adhesive cements polymerized using different activation modes. Five cements: MaxCem Elite (MAX), Bifix SE (BSE), G-Cem LinkAce (GCE), Clearfil SA Luting (CAS), and RelyX U200 (U200) were mixed, dispensed into molds, and distributed in groups, according to polymerization protocols: immediate photoactivation; delayed photoactivation (10min self-curing plus light-activation); and chemical activation (no light exposure). Immortalized rat odontoblast cells (MDPC-23) were cultured. Cell viability was assessed by Trypan Blue staining and total cell death was assessed by annexin V-APC/7-AAD double staining and flow cytometry. Volatilized compounds from polymerized specimens of cements were evaluated by gas chromatography/mass spectrometry (GC-MS). Data was analyzed with 2-way ANOVA/Tukey tests (α=0.05). Exposure to all of the cements tested significantly reduced the cell viability, irrespective o...
The influence of in vitro maturation (IVM) in oocytes is still not totally understood. The aim of... more The influence of in vitro maturation (IVM) in oocytes is still not totally understood. The aim of this study was to determine the influence of IVM on the metabolism and homeostasis of bovine cumulus-oocyte complexes. In the present study, we demonstrated that IVM leads to accumulation of neutral lipids associated with differential levels of the mono-, di-, and tri-acylglycerols in both cumulus cells and oocytes. We observed that in vitro-matured oocytes exhibited decreased glutathione and reactive oxygen species levels and a lower ATP/ADP ratio when compared to in vivo-matured oocytes, with no significant differences in metabolism and stress related mRNA or miRNA levels. Moreover, in addition to an increase in lipids in in vitro-matured cumulus cells, fatty acid synthesis and accumulation as well as glycolysis pathway genes were upregulated, whereas those affiliated with the -oxidation pathway were decreased. Our gene expression data in cumulus cells suggest the disruption of endopl...
Lysine is catabolized in mammals through the saccharopine and pipecolate pathways - the former is... more Lysine is catabolized in mammals through the saccharopine and pipecolate pathways - the former is mainly hepatic and renal, and the latter is believed to play a role in the cerebral lysine oxidation. Both pathways lead to the formation of aminoadipic semialdehyde (AASA) that is then oxidized to aminoadipate (AAA) by antiquitin (ALDH7A1). Mutations in the ALDH7A1 gene result in the accumulation of AASA and its cyclic form, piperideine-6-carboxylate (P6C), which causes pyridoxine-dependent epilepsy (PDE). P6C reacts with pyridoxal 5'-phosphate (PLP) causing its inactivation. Here, we used liquid chromatography-mass spectrometry to investigate lysine catabolism in mice injected with lysine labelled at either its nitrogen epsilon (ε-(15)N) or nitrogen alpha (α-(15)N). Analysis of ε-(15)N and α-(15)N lysine catabolites in plasma, liver and brain suggested the saccharopine as the main pathway for AAA biosynthesis. Although there was evidence for upstream cerebral pipecolate pathway ac...
The present paper describes the influence of the chemical structure of two aminoalkoxysilanes: 3-... more The present paper describes the influence of the chemical structure of two aminoalkoxysilanes: 3-aminopropyltriethoxysilane (APTS) and N-(3-(trimethoxysilyl)-propyl)-ethylenediamine (TSPEN) on the morphology of thin layer hybrid films with ...
European Journal of Lipid Science and Technology, 2010
Easy ambient sonic spray ionization (EASI), a novel desorption/ionization technique for ambient m... more Easy ambient sonic spray ionization (EASI), a novel desorption/ionization technique for ambient mass spectrometry analysis, is shown to permit the characterization of different types of vegetable and animal fats. The taxonomic markers of the oils, mainly fatty acids and phenols, are extracted with methanol/water (1:1) solution and made alkaline with NH4OH, then placed and dried on the surface of a glass tip and directly analyzed by EASI‐MS in the negative‐ion mode. EASI provides a supersonic cloud of charged droplets that causes efficient desorption and ionization of the oil markers directly from the surface of the glass tip. As proof‐of‐principle cases, EASI(−)‐MS was applied to genuine samples of olive oil, hazelnut oil, soybean oil, grape seed oil, canola oil, butter, and lard. Characteristic metabolomics EASI(−)‐MS profiles of fatty acids and eventually phenols were obtained.
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Papers by Lygia Marques