Non-natural amino acids into LfcinB-derived peptides: effect in their (i) proteolytic degradation and (ii) cytotoxic activity against cancer cells.

The dimeric peptide 26[F]: (RRWQWRFKKLG)2-K-Ahx has exhibited a potent cytotoxic effect against breast cancer cell lines, with position 26 (F) being the most relevant for anti-cancer activity. In this investigation, six analogues of the 26[F] peptide were synthesized in which the 26th position was replaced by non-natural hydrophobic amino acids, finding that some modifications increased the resistance to proteolytic degradation exerted by trypsin or pepsin. Additionally, these modifications increased the cytotoxic effect against breast cancer cells and generated cell death mediated by apoptosis pathways, activating caspases 8 and 9, and did not compromise the integrity of the cytoplasmic membrane. Finally, it was found that the modified peptides have a broad spectrum of action, since they also have a cytotoxic effect against the HeLa human cervical cancer cell line. Peptide 26[F] was inoculated in mice by ip administration and the lethal dose 50 (LD50) was between 70 and 140 mg kg-1. While for the 26[1-Nal]: (RRWQWR-1-Nal-KKLG)2-K-Ahx peptide, a dose-response test was performed, and the survival rate was 100%. These results suggested that these peptides are safe in this animal model and could be considered as promissory to develop a treatment against breast cancer.

Extract of Calyces from Physalis peruviana Reduces Insulin Resistance and Oxidative Stress in Streptozotocin-Induced Diabetic Mice.

Diabetes mellitus is a metabolic disorder mainly characterized by obesity, hyperglycemia, altered lipid profile, oxidative stress, and vascular compromise. Physalis peruviana is a plant used in traditional Colombian medicine for its known activities of glucose regulation. This study aimed to evaluate the anti-diabetic activity of the butanol fraction from an extract of Physalis peruviana calyces in two doses (50 mg/kg and 100 mg/kg) in induced type 2 diabetic mice. Blood glucose levels were evaluated once a week, demonstrating that a dose of 100 mg/kg resulted in greater regulation of blood glucose levels in mice throughout the experiment. The same overall result was found for the oral glucose tolerance test (OGTT) and the homeostatic model assessment for insulin resistance (HOMA- IR). The lipid profile exhibited improvement compared to the non-treated group, a dose of 100 mg/kg having greater protection against oxidative stress (catalase, superoxide dismutase, and malondialdehyde levels). Histopathological findings in several tissues showed structure preservation in most of the animals treated. The butanol fraction from Physalis peruviana at 100 mg/kg showed beneficial results in improving hyperglycemia, lipidemia, and oxidative stress status, and can therefore be considered a beneficial coadjuvant in the therapy of diabetes mellitus.

Lipid nanoparticles with improved biopharmaceutical attributes for tuberculosis treatment.

Tuberculosis is a topic of relevance worldwide because of the social and biological factors that triggered the disease and the economic burden on the health-care systems that imply its therapeutic treatment. Challenges to handle these issues include, among others, research on technological breakthroughs modifying the drug regimens to facilitate therapy adherence, avoid mycobacterium drug resistance, and minimize toxic side-effects. Lipid nanoparticles arise as a promising strategy in this respect as deduced from the reported scientific data. They are prepared from biodegradable and biocompatible starting materials and compared to the use of the free drugs, the entrapment of active molecules into the carriers might lead to both dose reduction and controlled delivery. Moreover, the target to the lung, the organ mainly affected by the disease, could be possible if the particle surface is modified. Although conclusive statements cannot be made considering the limited number of available research works, looking into what has been achieved up to now definitively encourages to continue investigations in this regard.

Nicotinamide mononucleotide adenylyltransferase of Trypanosoma cruzi (TcNMNAT): a cytosol protein target for serine kinases

Nicotinamide/nicotinate adenine dinucleotide (NAD+/NaAD) performs essential functions in cell metabolism and energy production due to its redox properties. The nicotinamide/nicotinate mononucleotide adenylyltransferase (NMNAT, EC 2.7.7.1/18) enzyme catalyses the key step in the biosynthesis of NAD+. Previously, the enzyme NMNAT was identified in Trypanosoma cruzi (TcNMNAT), a pathogenic agent with epidemiological importance in Latin America. To continue with the functional characterisation of this enzyme, its subcellular location and its possible post-translational modifications were examined in this study. For this, polyclonal antibodies were generated in mice, with soluble and denatured recombinant protein being used to detect the parasite’s NMNAT. Immunodetection assays were performed on whole extracts of T. cruzi, and an approximation of its intracellular location was determined using confocal microscopy on wild and transgenic parasites, which revealed the cytosol distribution patterns. This localisation occurs according to the needs of the dinucleotides that exist in this compartment. Additionally, a bioinformatics study was performed as a first approach to establish the post-translational modifications of the enzyme. Possible phosphorylation events were experimentally analysed by western blot, highlighting TcNMNAT as a potential target for serine kinases.

Nicotinamide mononucleotide adenylyltransferase of Trypanosoma cruzi (TcNMNAT): a cytosol protein target for serine kinases.

Nicotinamide/nicotinate adenine dinucleotide (NAD+/NaAD) performs essential functions in cell metabolism and energy production due to its redox properties. The nicotinamide/nicotinate mononucleotide adenylyltransferase (NMNAT, EC 2.7.7.1/18) enzyme catalyses the key step in the biosynthesis of NAD+. Previously, the enzyme NMNAT was identified in Trypanosoma cruzi (TcNMNAT), a pathogenic agent with epidemiological importance in Latin America. To continue with the functional characterisation of this enzyme, its subcellular location and its possible post-translational modifications were examined in this study. For this, polyclonal antibodies were generated in mice, with soluble and denatured recombinant protein being used to detect the parasite's NMNAT. Immunodetection assays were performed on whole extracts of T. cruzi, and an approximation of its intracellular location was determined using confocal microscopy on wild and transgenic parasites, which revealed the cytosol distribution patterns. This localisation occurs according to the needs of the dinucleotides that exist in this compartment. Additionally, a bioinformatics study was performed as a first approach to establish the post-translational modifications of the enzyme. Possible phosphorylation events were experimentally analysed by western blot, highlighting TcNMNAT as a potential target for serine kinases.

Estudio fitoquímico y actividad antiinflamatoria de hojas, flores y frutos de Bejaria resinosa Mutis ex L. (Pegamosco) / Phytochemical study and anti-inflammatory activity of leaves, flowers and fruits of Bejaria resinosa Mutis ex L. (pegamosco)

Introducción: Bejaria resinosa Mutis ex L. es una especie vegetal conocida en Colombia como pegamosco; es empleada por diferentes comunidades para atrapar insectos y para el tratamiento de dolencias respiratorias; además, es una especie que cuenta con escasos estudios desde el punto de vista químico y biológico. Objetivos: contribuir al estudio fitoquímico de las hojas, flores y frutos de B. resinosa (Ericaceae) y evaluar su actividad antiinflamatoria. Métodos: hojas, flores y frutos por separado fueron extraídos por maceración en frío con éter de petróleo y etanol 96 por ciento; estos extractos se fraccionaron por partición líquido/líquido y métodos cromatográficos; su actividad antiinflamatoria se evaluó utilizando el modelo murino de edema auricular inducido por 13-acetato de 12-tetradecanoilforbol (TPA). La elucidación estructural de los compuestos aislados se llevó a cabo mediante las técnicas de CG-EM y RMN (experimentos 1H, 13C, COSY, J-MOD, HSQC y HMBC). Resultados: la separación de los extractos y fracciones por cromatografías en columna, en capa delgada y preparativa, permitieron obtener una mezcla de compuestos tipo triterpeno compuesta por germanicol, -amirina y -amirina y el aislamiento de lupeol, salicilato de metilo, 3,5,7,3´,4´ pentahidroxiflavona (quercetina), 3,5-dihidroxi-6,7,8-trimetoxiflavona y 3,5,7,3´,4´ pentahidroxiflavanol. Se encontró que la fracción que contiene la mezcla de triterpenos y la quercetina fueron las que presentaron un efecto antiinflamatorio mayor al 65 por ciento. Conclusiones: el estudio fitoquímico de la especie vegetal B. resinosa permitió establecer similitud en cuanto a la composición química de los diferentes órganos ya que se encontraron metabolitos secundarios comunes para hojas flores y frutos como la mezcla de triterpenos, lupeol y quercetina; además se logró establecer que la mezcla de triterpenos y la quercetina son fuertes agentes antiinflamatorios, pues redujeron significativamente el edema causado por el TPA en la oreja del ratón con porcentajes de inhibición cercanos a los presentados por el fármaco de referencia, indometacina(AU)

Introduction: Bejaria resinosa Mutis ex L. is a plant species known as pegamosco in Colombia. It is used by several communities to catch insects and to treat respiratory disorders. Few chemical and biological studies have been conducted about this species. Objectives: Contribute to the phytochemical study of leaves, flowers and fruits of B. resinosa (Ericaceae) and evaluate its anti-inflammatory activity. Methods: Leaves, flowers and fruits were extracted separately by cold maceration with petroleum ether and 96 percent ethanol. The extracts obtained were fractioned using liquid / liquid partition and chromatographic methods. Their anti-inflammatory activity was evaluated with the mouse model of ear edema induced by 12-tetradecanoyl phorbol 13-acetate (TPA). Structural characterization of the compounds isolated was based on GC-MS and NMR techniques (experiments 1H, 13C, COSY, J-MOD, HSQC and HMBC). Results: Separation of extracts and fractions by thin-layer and preparative column chromatography made it possible to obtain a mixture of triterpene compounds made up of germanicol, α-amyrin and ß-amyrin, and isolate lupeol, methyl salicylate, 3,5,7,3´,4´ pentahydroxyflavone (quercetin), 3,5-dihydroxy-6,7,8-trimetoxyflavona and 3,5,7,3´,4´-pentahydroxyflavanol. It was found that the fraction containing the mixture of triterpenes and quercetin had an anti-inflammatory effect above 65 percent. Conclusions: The phytochemical study of the plant species B. resinosa revealed similarities between the chemical composition of the different organs, since common secondary metabolites were found in leaves, flowers and fruits, such as the mixture of triterpenes, lupeol and quercetin. It was also established that the mixture of triterpenes and quercetin is a strong anti-inflammatory agent, for it significantly reduced the mouse ear edema caused by TPA with inhibition percentages close to those of the drug of reference indometacin(AU)

ACTIVIDAD ANTIINFLAMATORIA DE EXTRACTOS Y FRACCIONES DE Myrcianthes leucoxila, Calea prunifolia, Curatella americana Y Physalis peruviana EN LOS MODELOS EDEMA AURICULAR POR TPA, EDEMA PLANTAR POR CARRAGENINA Y ARTRITIS INDUCIDA POR COLÁGENO / ANTIINFLAMMATORY ACTIVITY OF EXTRACTS AND FRACTIONS OF Myrcianthes leucoxila, Calea prunifolia, Curatella americana Y Physalis peruviana OBTAINED FROM ON TPA-INDUCED EAR OEDEMA, CARRAGEENANINDUCED PAW OEDEMA AND COLLAGEN-INDUCED ARTHRITIS

Introducción. El empleo etnofarmacológico de plantas en el manejo de procesos inflamatorios crónicos y la necesidad su caracterización farmacológica, promueven la evaluación de actividad antiinflamatoria de sustancias en modelos in vivo. Materiales y métodos. Evaluación de extractos y fracciones de Calea prunifolia (CP), Curatella americana (CA), Myrcianthes leucoxila (ML) y Physalis peruviana (PP) sobre los modelos edema auricular por acetato de tetradecanoilforbol (TPA) en ratón albino ICR y edema plantar por carragenina en ratas Wistar, seleccionando un extracto para valorar su actividad antiartrítica en el modelo artritis inducida por colágeno en ratones DBA. Resultados. Las fracciones con mayor porcentaje de inhibición del edema en el modelo edema auricular por TPA fueron ML etanólica total (82±6%), CP rica en terpenos (81±6%) y CA rica en terpenos (81±7%) (P<0,05). No se obtuvo actividad antiinflamatoria significativa sobre el modelo edema plantar por carragenina. Se evaluó la actividad antiartrítica de la fracción rica en terpenos de ML sobre el modelo artritis inducida por colágeno, sin encontrarse efecto significativo sobre edema de patas traseras, peso corporal, escala histopatológica de severidad de artritis ni inmunohistoquímica para factor de necrosis tumoral alfa (P>0,05). Discusión. La actividad antiinflamatoria en el modelo de inflamación aguda edema auricular por TPA para los extractos y fracciones de CP, CA y ML se puede relacionar con la afectación de mediadores relacionados con fosfolipasa A2 dado el nivel de efecto similar a indometacina encontrado. La fracción terpénica de ML no mostró actividad antiartrítica ni modificó la expresión de TNF-a en el modelo de artritis crónica autoinmune empleado, por lo cual no posee actividad inmunomoduladora ni antiinflamatoria en la dosis evaluada. Conclusión: Las fracciones terpénicas de los extractos de CA y CP y los extractos metanólicos de ML mostraron una actividad antiinflamatoria significativa en el edema auricular inducida por TPA. Estos extractos tuvieron poca actividad sobre el edema inducido por carragenina en la pata. La fracción terpénica del extracto ML no presentó actividad antiartrítica en el modelo de artritis inducido por el colágeno.

Introduction. Ethnopharmacological use of plants in management of chronic inflammatory diseases and the need to have their pharmacological characterization promote the evaluation of anti-inflammatory activity over in vivo models. Materials and methods. Evaluation of extracts and fractions of Calea prunifolia (CP), Curatella americana (CA), Myrcianthes leucoxila(ML) and Physalis peruviana (PP) on auricular edema by tetradecanoylphorbol acetate (TPA)-in ICR albino mice and carrageenan-induced leg edema in Wistar rats selecting an extract to evaluate its anti-arthritic activity in collagen-induced arthritis model in DBA mice. Results. The fractions with greater edema inhibition percentage on TPA-induced ear edema included whole ethanolic fraction of ML (82±6%), CP terpenes rich fraction (81±6%) and CA terpenes rich fraction (81±7%) (P<0.05). Significant antiinflamatory activity was not obtained on the carrageenan-induced leg edema. Evaluation of antiarthritic activityof the ML terpenes rich fraction was carried out on collagen induced arthritis. Without finding any significant effect on back leg edema, corporal weight, arthritis histopathology severity scale or immunohistochemical tumoral necrosis factor alfa immunohistochemical evaluation (P>0.05). Discussion. Anti-inflammatory activity in TPAinduced acute ear edema model for CP, CA, and ML extracts and fractions can be interfered by phospholipase A2 related mediators due to similar effect with indomethacin was found. The ML terpens rich fraction neither show any anti-arthritic activity nor affected TNF-a expression on the autoimmune chronic arthritis model used, reason why it does not have immunomodulatory/anti-inflammatory effect on the evaluated dose. Conclusion. CA and CP terpenic rich fractions and ML ethanol extract showed significant anti-inflammatory activity in TPA-induced ear edema. They did have some activity in carrageenan-induced leg edema. The ML terpenic rich fraction did not have anti-arthritic activity in the collagen-induced arthritis model.