Antileishmanial Activity and In Silico Molecular Docking Studies of Malachra alceifolia Jacq. Fractions against Leishmania mexicana Amastigotes.

Malachra alceifolia Jacq. (family Malvaceae), known as "malva," is a medicinal plant used as a traditional therapy in many regions of America, Africa and Asia. Traditionally, this plant is used in the form of extracts, powder and paste by populations for treating fever, stomachache, inflammation, and parasites. However, the ethnopharmacological validation of M. alceifolia has been scarcely researched. This study showed that the chloroform fraction (MA-IC) and subfraction (MA-24F) of the leaves of M. alceifolia exhibited a potential antileishmanial activity against axenic amastigotes of Leishmania mexicana pifanoi (MHOM/VE/60/Ltrod) and had high and moderate cytotoxic effects on the viability and morphology of macrophages RAW 264.7. This study reports, for the first time, possible terpenoid metabolites and derivatives present in M. alceifolia with activity against some biosynthetic pathways in L. mexicana amastigotes. The compounds from the subfractions MA-24F were highly active and were analyzed by gas chromatography-mass spectrometry (GC-MS) and by a molecular docking study in L. mexicana target protein. This study demonstrates the potential modes of interaction and the theoretical affinity energy of the metabolites episwertenol, α-amyrin and methyl commate A, which are present in the active fraction MA-24F, at allosteric sites of the pyruvate kinase, glyceraldehyde-3-phosphate dehydrogenase, triose phosphate isomerase, aldolase, phosphoglucose isomerase, transketolase, arginase and cysteine peptidases A, target proteins in some vital biosynthetic pathways were responsible for the survival of L. mexicana. Some phytoconstituents of M. alceifolia can be used for the search for potential new drugs and molecular targets for treating leishmaniases and infectious diseases. Furthermore, contributions to research and the validation and conservation of traditional knowledge of medicinal plants are needed globally.

An Overview of Genus Malachra L.-Ethnobotany, Phytochemistry, and Pharmacological Activity.

The genus Malachra L. belongs to the family Malvaceae. It includes herbs or subshrubs of nine accepted species with approximately thirty synonyms, and it has been widely used in community folk medicine to treat health problems including inflammation, nasal obstruction, leishmaniasis, malaria, childbirth, kidney disorders, fever, respiratory tract diseases, among others. From the genus Malachra L., flavonoids, steroids, triterpenes, anthocyanins, leucoanthocyanins, saponins, carbohydrates, phenols, glycosides, and alkaloids have been isolated and identified. Some pharmacological reports have indicated that the genus has antidiarrheal, antiepileptic, antiulcerogenic, antioxidant, anticonvulsant, antiviral, anticancer, antibacterial, anthelmintic, and hepatoprotective properties. However, there have been limited studies of bioactive molecules with pharmacological and biological activities associated with Malachra alceifolia Jacq., Malachra capitata (L.) L., Malachra fasciata Jacq., Malachra radiata (L.) L., Malachra ruderalis Gürke., Malachra rudis Benth., Malachra helodes Mart., Malachra urens Poit. ex Ledeb. & Alderstam., and Malachra officinalis Klotzsch. In this review, we consider the conservation of these species to save the ancestral knowledge of their traditional use in populations, and their pharmacological potential for future studies in search of alternatives for solutions to diseases in humans and animals and tools for the design and search of potential bioactive compounds against infectious and non-infectious agents.

Virtual screening using docking and molecular dynamics of cannabinoid analogs against CB1 and CB2 receptors.

BACKGROUND: Cannabis sativa has been attributed to different pharmacological properties. A number of secondary metabolites such as tetrahydrocannabinol (THC), cannabinol (CBD), and different analogs, with highly promising biological activity on CB1 and CB2 receptors, have been identified. METHODS: Thus, this study aimed was to evaluate the activity of THC, CBD, and their analogs using molecular docking and molecular dynamics simulations (MD) methods. Initially, the molecules (ligands) were selected by bioinformatics searches in databases. Subsequently, CB1 and CB2 receptors were retrieved from the protein data bank database. Afterward, each receptor and its ligands were optimized to perform molecular docking. Then, MD Simulation was performed with the most stable ligand-receptor complexes. Finally, the Molecular Mechanics-Generalized Born Surface Area (MM-PBSA) method was applied to analyze the binding free energy between ligands and cannabinoid receptors. RESULTS: The results obtained showed that ligand LS-61176 presented the best affinity in the molecular docking analysis. Also, this analog could be a CB1 negative allosteric modulator like CBD and probably an agonist in CB2 like THC and CBD according to their dynamic behavior in silico. The possibility of having a THC and a CBD analog (LS-61176) as a promising molecule for experimental evaluation since it could have no central side-effects on CB1 and have effects of CB2 useful in pain, inflammation, and some immunological disorders. Docking results were validate using ROC curve for both cannabinoids receptor where AUC for CB1 receptor was 0.894±0.024, and for CB2 receptor AUC was 0.832±0032, indicating good affinity prediction.

In Silico Analysis to Identify Molecular Targets for Chemicals of Concern: The Case Study of Flocoumafen, an Anticoagulant Pesticide.

Rodenticides are pesticides used worldwide, with little information available regarding health consequences in wildlife and humans. The aim of the present study was to use virtual screening to identify potential targets for flocoumafen, a superwarfarin rodenticide. Blind docking of more than 841 human proteins was carried out employing AutoDock Vina. The strength of the ligand interaction with the proteins was quantified based on the binding affinity score (kcal/mol). Results indicate that flocoumafen could be a promiscuous ligand for diversity of cellular protein targets. The best complexes were obtained for prostaglandin F synthase (-14.2 kcal/mol) and serum albumin (-14.0 kcal/mol) followed by glucocorticoid receptor 2, matrix metalloproteinase-9, nuclear receptor ROR-alpha, and activin receptor type-1, all with values equal or better than -13.5 kcal/mol. Docking method validation based on the root-mean-square deviation showed that flocoumafen had good capability to predict corresponding co-crystallized poses; and molecular dynamics simulations suggested the complex with greater binding affinity was thermodynamically stable. Protein-protein interaction networks built with main protein targets revealed that protein kinase B (AKT1), ribosomal protein S6 kinase B1 (RPS6KB1), phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA), retinoid X receptor alpha (RXRA), and protein phosphatase 2 catalytic subunit alpha (PPP2CA) were major hub proteins, whereas the gene ontology analysis reported that cellular response to endogenous stimulus, protein binding, and the TOR complex were the biological processes, molecular function, and cell component enrichments, respectively. These results should motivate more ecotoxicity testing for flocoumafen and other superwarfarins, as well as precautionary legislation to minimize exposure to these highly toxic chemicals. Environ Toxicol Chem 2021;40:2034-2043. © 2021 SETAC.

Homology Modeling of Leishmanolysin (gp63) from Leishmania panamensis and Molecular Docking of Flavonoids.

Leishmaniasis is a chronic disease caused by protozoa of the distinct Leishmania genus transmitted by sandflies of the genus Phlebotomus (old world) and Lutzomyia (new world). Among the molecular factors that contribute to the virulence and pathogenesis of Leishmania are metalloproteases, e.g., glycoprotein 63 (gp63), also known as leishmanolysin or major surface protease (MSP). This protease is a zinc-dependent metalloprotease that is found on the surface of the parasite, abundant in Leishmania promastigote and amastigote. This study describes the prediction of three-dimensional (3D) structures of leishmanolysin (UniProt ID A0A088RJX7) of Leishmania panamensis employing a homology modeling approach. The 3D structure prediction was performed using the SWISS-MODEL web server. The tools PROCHECK, Molprobyty, and Verify3D were used to check the quality of the model, indicating that they are reliable. Best docking configurations were identified applying AutoDock Vina in PyRx 0.8 to obtain a potential antileishmanial activity. Biflavonoids such as lanaroflavone, podocarpusflavone A, amentoflavone, and podocarpusflavone B showed good scores among these molecules. Lanaroflavone appears to be the most suitable compound from binding affinity calculations.

Quantification of microplastics along the Caribbean Coastline of Colombia: Pollution profile and biological effects on Caenorhabditis elegans.

The Caribbean Coast of Colombia has a flourishing plastic industry with weak and insufficient waste management policies and practices, leading to plastic pollution along its touristic beaches. In this work, primary and secondary microplastics (MPs) were surveyed at four different locations along the Colombian Caribbean Coast. Primary microplastics, specifically white new plastic pellets, represented the largest amount of MPs found, with densities decreasing in the order Cartagena > Coveñas > Puerto Colombia > Riohacha. This distribution was connected to the vicinity of MPs sources, marine currents and wind direction. The presence of secondary MPs was associated with urban centers and proximity to river mouths. The FTIR characterization showed polyethylene as the predominant resin type, with different degrees of surface oxidation. Aqueous extracts from sampled MPs were tested on Caenorhabditis elegans. Secondary MPs elicited greater toxicological responses than pellets, especially those from Cartagena Bay, suggesting MPs act as carriers for biologically-active pollutants.

Validación de una metodología analítica USP por HPLC para la cuantificación de warfarina sódica en tabletas / Validation of an analytical HPLC method for the quantification of warfarin sodium in tablets

La metodología analítica para la cuantificación de warfarina sódica en tabletas de 5 mg fue estandarizada y validada mediante cromatografía líquida de alta eficiencia acoplada a detector de arreglo de diodo (HPLC-DAD). Se usó como fase móvil una mezcla compuesta por MeOH-AcOH-H2O (68:1:32), una columna cromatográfica C8e Phenomenex® de 150 x 4,6 mm y tamaño de partícula de 5 μm. Los resultados analíticos muestran que el método es preciso, exacto y selectivo para este fármaco. La curva de calibración se realizó en un intervalo del 80 al 120%, en una concentración nominal de 0,1mg/mL, demostrando ser lineal con un coeficiente de correlación a r²> 0,990; el porcentaje de recuperación fue de 98,1% ± 1,3, la precisión medida a través de la repetibilidad y la precisión intermedia fue adecuada (%RSD<2). En la prueba de disolución se encontró un porcentaje de 92,3% ± 3.0. Por lo tanto, la metodología desarrollada cumple con las especificaciones establecida por la USP 38/ NF33.

The analytic methodology for the quantification of sodium warfarin tablets of 5 mg was standardized and validated by High Efficiency Liquid Chromatography coupled to Diode Array Detector (HPLC-DAD). A mixture of MeOH-AcOH-H2O (68:1:32) was used like mobil phase, a chromatographic column C8e Phenomenex® of 150x4.6 mm and a particle size of 5 μm. The analytic results show that the method is precise, accurate and selective for this drug. The calibration curve was performed in the range of 80 to 120% at a nominal concentration of 0.1 mg/mL, demonstrating to be linear with a correlation coefficient at r²> 0.990; recovery rate was 98.1% ± 1.3 and the precision measured through repeatability and intermediate accuracy was adequate (% RSD <2). A percent of 92.3% ± 3.0 was found in dissolution test, thus, the methodology developed complies with the specifications established by USP 38/NF33.

Estudio biofarmacéutico comparativo de tabletas de ácido acetilsalicílico disponibles en el mercado colombiano / A comparative biopharmaceutical study of brands of acetylsalicylic acid tablets available on the colombian market

Objetivo: determinar la equivalencia biofarmacéutica de cinco marcas comerciales de tabletas de ácido acetilsalicílico 100 mg disponibles en el mercado colombiano. Métodos: se tomaron cinco productos comerciales de ácido acetilsalicílico tabletas 100 mg, adquiridos en droguerías y farmacias de las ciudades de Bogotá, Cali, Medellín, Barranquilla y Cartagena, a los cuales se les evaluaron las carac­terísticas físicas, químicas y biofarmacéuticas de las tabletas, tales como variación de peso, dureza, desintegración, test y perfil de disolución, eficiencia de la disolución y valoración del ingrediente farmacéuticamente activo, los ensayos farmacopeicos se evaluaron según lo establecido en la USP-37. Los resultados se analizaron a fin de establecer diferencias estadísticamente significativas y posible intercambiabilidad entre los productos evaluados. Resultados: el análisis comparativo de los productos, permitió evidenciar marcadas diferencias en cuanto a la liberación in vitro del ingrediente farmacéuticamente activo, con uno de los productos evaluados, incumpliendo este importante parámetro de calidad. Cabe resaltar que cuatro de los productos evaluados cumplen con las especificaciones oficiales de identificación y valoración del ingrediente activo farmacéuticamente, uniformidad de dosificación y tiempo de desintegración. En cuanto a la cinética de disolución se encontraron diferencias entre las formulaciones, con un producto (marca E) de deficiente eficiencia de disolución (ED). Conclusiones: cuatro productos cumplen con todas las especificaciones establecidas en la USP-37. Los resultados de este trabajo constituyen una valiosa información para las autoridades sanitarias y para los pacientes que consumen este tipo de productos ya que genera confianza en términos de efectividad del medicamento, sobre si se considera que el ácido acetilsalicílico está exento de realizar estudios de bioequivalencia por pertenecer a la Clase I del Sistema de Clasificación Biofarmacéutica y muestra una alta solubilidad y absorción en humanos(AU)

Objective: to determine the biopharmaceutical equivalency of five brands if 100 mg acetylsalicylic acid tablets available on the Colombian market. Methods: five brands of 100 mg acetylsalicylic acid tablets obtained from drugstores and pharmacies in Bogota, Cali, Medellin, Barranquilla and Cartagena, were evaluated in terms of their physical, chemical and biopharmaceutical characteristics such as variation in weight, hardness, disintegration, dissolution test, dissolution profile and efficiency and quantitation of pharmaceutically active ingredient; the pharmacopeial assays were evaluated according to the USP 37. The results analysis was used to determine the statistically significant differences and the possible interchangeability among the evaluated products. Results: the comparative analysis of products allowed showing marked differences in terms of the in vitro release of pharmaceutically active ingredient and the non-compliance by one of the evaluated products with this important quality parameter. It is worth noting that four of the tested products comply with the official specifications of identification and measurement of the pharmaceutically active ingredient, dosage uniformity and disintegration time. In terms of the dissolution kinetics, differences were found among formulations, with a product (Mark E) having poor dissolution efficiency. Conclusions: four products meet all the the USP-37 specifications. The results of this study provide valuable information for health authorities and patients who take these products because they bring confidence on the drug effectiveness, considering that aspirin is exempted from bioequivalence studies since it belong to Class I Biopharmaceutics Classification System showing high solubility and absorption in human consumers(AU)

Níquel en cavidad oral de individuos con agrandamiento gingival inducido por tratamiento ortodóncico / Nickel in oral cavity of individuals with gingival overgrowth induced by orthodontic treatment

El agrandamiento gingival es una de las alteraciones orales generadas por el uso de aparatología ortodóncica fija. Inicialmente fue descrito como producto de la respuesta alérgica del huésped al níquel, y estudios recientes demostraron que este metal induce proliferación fibroblástica. Sin embargo, se desconoce el grado de bioacumulación de este metal en la cavidad oral. El objetivo del presente estudio consistió en cuantificar la concentración de níquel en muestras de saliva, placa dental y encía de individuos con aparatología ortodóncica fija con y sin agrandamiento gingival. El tamaño de la muestra se estableció según tendencia histórica, evaluando un total de 24 individuos con tratamiento ortodóncico activo, los cuales fueron clasificados en 2 grupos: 12 con agrandamiento gingival (grupo A) y 12 sin agrandamiento gingival (grupo B). De cada participante se obtuvieron muestras de: encía, saliva estimulada y placa dental. Una vez procesadas las muestras se midieron las concentraciones de níquel en mg/l, mediante espectrofotometría de absorción atómica (Thermo Scientific. Atomic absorption spectrometer iCE 3000, Reino Unido). El análisis estadístico se realizó mediante el software SPSS v20. Al comparar los niveles de níquel en muestras de saliva estimulada, placa dental y encía entre los individuos con y sin agrandamiento gingival, se observaron niveles significativamente mayores de níquel en las muestras de encía del grupo A (medias = 0,61 mg/l vs 0,36 mg/l; p = 0,001). Sin embargo, no se observó diferencia estadísticamente significativa al comparar las concentraciones de Ni en saliva entre el grupo A y B (medias = 0,502 mg/l vs 0,473 mg/l; p = 0,178, respectivamente). A partir de estos resultados se concluye que el uso de aparatología ortodóncica fija conlleva distintos niveles de bioacumulación de iones níquel en la encía al comparar ambos grupos. Es probable que altas concentraciones de este metal estén relacionadas con la etiología del agrandamiento gingival, por lo que se requieren nuevos estudios para establecer causalidad.

Gingival overgrowth is an oral condition generated by the use of fixed orthodontic appliances. It was initially described as a product of the allergic response of the host to Nickel, and recent studies showed that this metal induces fibroblast proliferation. However, the degree of bioaccumulation of this metal in the oral cavity is unknown. The aim of this study was to quantify the concentrations of nickel in samples of saliva, dental plaque, and gingiva of individuals with fixed orthodontic appliances with and without gingival overgrowth. The sample size was established according to historical trends, evaluating 24 individuals with active orthodontic treatment, which were classified in two groups: 12 with gingival overgrowth (Group A) and 12 without gingival overgrowth (Group B). Samples of gingival, stimulated saliva and dental plaque were obtained from each participant. Once the sample were processed, nickel concentrations were measured in [mg/L] by atomic absorption spectrophotometry (Thermo Scientific. Atomic absorption spectrometer iCE 3000, UK). Statistical analysis was performed using SPSS v20. The levels of Nickel in samples of stimulated saliva, dental plaque and gingiva were compared between individuals in group A and group B. There were significantly higher levels of nickel in gingiva samples in group A (mean = 0.61 mg/L vs. 0.36 mg/L, P = .001). However, no statistical difference was observed when comparing the concentrations of nickel in saliva between groups A and B (mean = 0.502 mg/L vs. 0.473 mg/L, P = .178, respectively). The use of fixed orthodontic appliances leads to different levels of bioaccumulation of nickel in gingiva when comparing both groups. High concentrations of this metal are probably associated with the origin of gingival overgrowth. Further studies are required to establish causation.

Folk medicine in the northern coast of Colombia: an overview.

BACKGROUND: Traditional remedies are an integral part of Colombian culture. Here we present the results of a three-year study of ethnopharmacology and folk-medicine use among the population of the Atlantic Coast of Colombia, specifically in department of Bolívar. We collected information related to different herbal medicinal uses of the local flora in the treatment of the most common human diseases and health disorders in the area, and determined the relative importance of the species surveyed. METHODS: Data on the use of medicinal plants were collected using structured interviews and through observations and conversations with local communities. A total of 1225 participants were interviewed. RESULTS: Approximately 30 uses were reported for plants in traditional medicine. The plant species with the highest fidelity level (Fl) were Crescentia cujete L. (flu), Eucalyptus globulus Labill. (flu and cough), Euphorbia tithymaloides L. (inflammation), Gliricidia_sepium_(Jacq.) Kunth (pruritic ailments), Heliotropium indicum L. (intestinal parasites) Malachra alceifolia Jacq. (inflammation), Matricaria chamomilla L. (colic) Mentha sativa L. (nervousness), Momordica charantia L. (intestinal parasites), Origanum vulgare L. (earache), Plantago major L. (inflammation) and Terminalia catappa L. (inflammation). The most frequent ailments reported were skin affections, inflammation of the respiratory tract, and gastro-intestinal disorders. The majority of the remedies were prepared from freshly collected plant material from the wild and from a single species only. The preparation of remedies included boiling infusions, extraction of fresh or dry whole plants, leaves, flowers, roots, fruits, and seeds. The parts of the plants most frequently used were the leaves. In this study were identified 39 plant species, which belong to 26 families. There was a high degree of consensus from informants on the medical indications of the different species. CONCLUSIONS: This study presents new research efforts and perspectives on the search for new drugs based on local uses of medicinal plants. It also sheds light on the dependence of rural communities in Colombia on medicinal plants.

Contracaecum sp. infection in Hoplias malabaricus (moncholo) from rivers and marshes of Colombia.

Nematode infection indices were recorded in Hoplias malabaricus captured in six different rivers and a marsh belonging to the North Coast Basin of Colombia, and from the Amazon River, during February 2003-December 2004. Preliminary morphological analysis of nematodes indicated the presence of Contracaecum sp. Parasites were mostly found in the intestinal mesenteries and a very low percentage in muscle. Parasite prevalence in all sampling locations at the north of Colombia was 100%, whereas in the Amazon River it was 6.12%. The mean intensity in the different stations were as follows: Magdalena River at the City of Magangué (58.92+/-7.59), Magdalena river at the city of Zambrano (128.9+/-7.08), Sinú River (53.88+/-4.92), Dique Channel (207.3+/-59.52), Cauca River (77.26+/-9.35), Atrato River (21.11+/-2.6), San Jorge River (39.5+/-7.13), and Totumo Marsh (62.5+/-6.38). In average, all specimens of Hoplias malabaricus from the north coast basin of Colombia were infected with a mean intensity of 77.82+/-4.81 (1-466 parasites per host) whereas in fish from the Amazon River this value was significantly lower (intensity 1.0+/-0.0). Size and weight correlated significantly with parasite intensity in fish collected from sampling locations at the north of Colombia (R=0.240, P<0.001 and R=0.199, P=0.008, respectively). Moreover, a significant, but low and negative correlation was found between condition factor and parasite intensity (R=-0.159, P=0.034), suggesting a possible impact of parasites on fish health. These results suggest, for the first time, that the parasitism in Moncholo is a widespread phenomenon in Colombian rivers and could represent a risk factor for human consumers.