Anti-inflammatory effect of Trichospira verticillata via suppression of the NLRP3 inflammasome in neutrophilic asthma.

Trichospira verticillata is an annual herb that belongs to the family Asteraceae. Trichospira verticillata extract (TVE) elicits anti-plasmodial activity; however, there has been no detailed report about its anti-inflammatory effects and molecular mechanisms. In addition, herbal plants exhibit anti-inflammatory effects by suppressing the NLRP3 inflammasome. Therefore, the primary goal of this study was to examine the effects of TVE on NLRP3 inflammasome activation by measuring interleukin-1ß (IL-1ß) secretion. We treated lipopolysaccharides (LPS)-primed J774A.1 and THP-1 cells with TVE, which attenuated NLRP3 inflammasome activation. Notably, TVE did not affect nuclear factor-kappa B (NF-κB) signalling or intracellular reactive oxygen species (ROS) production and potassium efflux, suggesting that it inactivates the NLRP3 inflammasome via other mechanisms. Moreover, TVE suppressed the formation of apoptosis-associated speck-like protein (ASC) speck and oligomerization. Immunoprecipitation data revealed that TVE reduced the binding of NLRP3 to NIMA-related kinase 7 (NEK7), resulting in reduced ASC oligomerization and speck formation. Moreover, TVE alleviated neutrophilic asthma (NA) symptoms in mice. This study demonstrates that TVE modulates the binding of NLPR3 to NEK7, thereby reporting novel insights into the mechanism by which TVE inhibits NLRP3 inflammasome. These findings suggest TVE as a potential therapeutic of NLRP3 inflammasome-mediated diseases, particularly NA.

Guarea microcarpa C. DC. extract inhibits NLRP3 inflammasome by suppressing its ATPase activity.

ETHNOPHARMACOLOGICAL RELEVANCE: Guarea genus comprises tropical and subtropical terrestrial herbs inhabiting Central and South America. These plants, including Guarea guidonia (L.) Sleumer, have anti-inflammatory, analgesic, antibacterial, antiviral, and immune-enhancing properties. AIM OF THE STUDY: Although various species of the Guarea genus are known for their medicinal properties, comprehensive data on their anti-inflammatory effects remain limited. Therefore, we investigated the NLRP3 inflammasome-inhibiting effects of the Guarea genus in this study. MATERIALS AND METHODS: To evaluate the anti-inflammatory activities of 18 members of the Guarea genus, we treated NLRP3 inflammasome activators with their extracts in LPS-primed J774A.1 and THP-1 cells. Cell viability was determined by water soluble tetrazolium salt (WST) and cytokine production, protein expression, and nuclear fractionation were determined by western blotting. Reactive oxygen species (ROS) production and apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC) oligomerization were measured using confocal microscopic analysis. Inflammation-induced zebrafish was used in the in vivo experiments. RESULTS: Among the 18 Guarea members tested, Guarea microcarpa C. DC. extract (GM) exhibited no cytotoxicity and specifically suppressed the activation of the NLRP3 inflammasome, but not of the AIM2 or NLRC4 inflammasomes, by inhibiting the ATPase activity of NLRP3. This was achieved without affecting NF-κB signaling, potassium efflux, or intracellular ROS production, all of which are involved in NLRP3 activation. The reduced ATPase activity of NLRP3 led to decreased ASC oligomerization. Furthermore, GM exhibited anti-inflammatory effects in vivo. Additionally, GM treatment alleviated inflammation at the organismal level in an LPS-induced inflammation model using zebrafish embryos. CONCLUSION: Our results demonstrate the anti-inflammatory effects of GM via suppressing the NLRP3 inflammasome. Therefore, GM can be a potential therapeutic candidate for various inflammatory diseases caused by aberrant NLRP3 inflammasome activation.

Ameliorative Effects of Daphnopsis costaricensis Extract against Oxazolone-Induced Atopic Dermatitis-like Lesions in BALB/c Mice.

The genus Daphnopsis has been traditionally used as a purgative, diuretic, stimulant, and psoriasis treatment. In this study, the anti-AD (atopic dermatitis) activities of the Daphnopsis costaricensis EtOH extract (DCE) were investigated in an oxazolone (OX)-induced mouse model of AD, and the anti-inflammatory effects of its active compounds were confirmed in PI-sensitized or IgE/DNP-BSA-sensitized RBL-2H3 cells. DCE improved the symptoms of OX-induced inflammatory dermatitis (swelling, erythema, and increased ear thickening) in OX-induced BALB/c mice ears and reduced epidermal thickness and mast cell infiltration. Eleven flavonoid compounds were isolated from DCE, and two compounds (7,8-dimethoxyflavone and 7,2'-dimethoxyflavone) significantly inhibited IL-4 overexpression in PI-induced RBL-2H3 cells and mast cell degranulation in IgE + DNP-BSA-induced RBL-2H3 cells. Our study indicates that DCE and two compounds (7,8-dimethoxyflavone and 7,2'-dimethoxyflavone) might effectively improve inflammatory and atopic skin symptoms.

Phylotranscriptomics of Theaceae: generic-level relationships, reticulation and whole-genome duplication.

BACKGROUND AND AIMS: Theaceae, with three tribes, nine genera and more than 200 species, are of great economic and ecological importance. Recent phylogenetic analyses based on plastomic data resolved the relationships among the three tribes and the intergeneric relationships within two of those tribes. However, generic-level relationships within the largest tribe, Theeae, were not fully resolved. The role of putative whole-genome duplication (WGD) events in the family and possible hybridization events among genera within Theeae also remain to be tested further. METHODS: Transcriptomes or low-depth whole-genome sequencing of 57 species of Theaceae, as well as additional plastome sequence data, were generated. Using a dataset of low-copy nuclear genes, we reconstructed phylogenetic relationships using concatenated, species tree and phylogenetic network approaches. We further conducted molecular dating analyses and inferred possible WGD events by examining the distribution of the number of synonymous substitutions per synonymous site (Ks) for paralogues in each species. For plastid protein-coding sequences , phylogenies were reconstructed for comparison with the results obtained from analysis of the nuclear dataset. RESULTS: Based on the 610 low-copy nuclear genes (858 606 bp in length) investigated, Stewartieae was resolved as sister to the other two tribes. Within Theeae, the Apterosperma-Laplacea clade grouped with Pyrenaria, leaving Camellia and Polyspora as sister. The estimated ages within Theaceae were largely consistent with previous studies based mainly on plastome data. Two reticulation events within Camellia and one between the common ancestor of Gordonia and Schima were found. All members of the tea family shared two WGD events, an older At-γ and a recent Ad-ß; both events were also shared with the outgroups (Diapensiaceae, Pentaphylacaceae, Styracaceae and Symplocaceae). CONCLUSIONS: Our analyses using low-copy nuclear genes improved understanding of phylogenetic relationships at the tribal and generic levels previously proposed based on plastome data, but the phylogenetic position of the Apterosperma-Laplacea clade needs more attention. There is no evidence for extensive intergeneric hybridization within Theeae or for a Theaceae-specific WGD event. Land bridges (e.g. the Bering land bridge) during the Late Oligocene may have permitted the intercontinental plant movements that facilitated the putative ancient introgression between the common ancestor of Gordonia and Schima.

The Anti-Inflammatory Effect of Trichilia martiana C. DC. in the Lipopolysaccharide-Stimulated Inflammatory Response in Macrophages and Airway Epithelial Cells and in LPS-Challenged Mice.

A number of species of the genus Trichilia (Meliaceae) exhibit anti-inflammatory effects. However, the effect of Trichilia martiana C. DC. (TM) on lipopolysaccharide (LPS)-induced inflammation has not, to the best of our knowledge, yet been determined. Therefore, in the present study, the antiinflammatory effect of TM on LPS-stimulated RAW264.7 macrophages was evaluated. The ethanol extract of TM (TMEE) significantly inhibited LPS-induced nitric oxide (NO), prostaglandin 2 (PGE2), inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). TMEE also reduced the levels of inflammatory cytokines, including tumor necrosis factor-alpha (TNF-α), interleukin (IL)-1ß and IL-6. The upregulation of mitogen-activated protein kinases (MAPKs) and NF-κB activation was revealed to be downregulated following TMEE pretreatment. Furthermore, TMEE was indicated to lead to the nucleus translocation of nuclear factor erythroid-derived 2-related factor 2 (Nrf2) and the expression of heme oxygenase-1 (HO-1). In H292 airway epithelial cells, the pretreatment of TMEE significantly downregulated the production of LPS-stimulated IL-1ß, and TMEE was indicated to increase the expression of HO-1. In animal models exhibiting LPS-induced acute lung injury (ALI), treatment with TMEE reduced the levels of macrophages influx and TNF-α production in the bronchoalveolar lavage fluid (BALF) of ALI mice. Additionally, TMEE significantly downregulated the activation of ERK, JNK and IκB, and upregulated the expression of HO-1 in the lungs of ALI mice. In conclusion, the results of the current study demonstrated that TMEE could exert a regulatory role in the prevention or treatment of the endotoxin-mediated inflammatory response.

Anti-inflammatory effects of ethanol extract from the leaves and shoots of Cedrela odorata L. in cytokine-stimulated keratinocytes.

Cedrela odorata L. is a native plant of the Amazon region. The bark is used in folk remedies for the treatment of diarrhea, vomiting, fever and inflammation. Atopic dermatitis (AD) is a chronic, relapsing inflammatory skin disease accompanied by itching. It is a complex disease involving environmental factors and genetic factors. In the present study, the anti-inflammatory and anti-allergic effects of C. odorata L. methanol extract (COEE) on tumor necrosis factor (TNF)-α and interferon (IFN)-γ-stimulated HaCaT keratinocyte cells were investigated. ELISA and RT-PCR analysis revealed that the extract had anti-inflammatory effects, and reduced the interleukin (IL)-6 and IL-8 levels of the HaCaT cells. In addition, COEE exhibited anti-allergic effects, comprising a reduction in the thymus and activation-regulated chemokine and macrophage-derived chemokine levels. In addition, pathway analysis and comparison with Bay11-7082 indicated that these effects are due to the inhibition of nuclear factor (NF)-κB in TNF-α/IFN-γ-induced HaCaT cells. Therefore, the results of the present study suggest that COEE has anti-inflammatory and anti-allergic properties in TNF-α and IFN-γ-stimulated HaCaT cells, which are associated with the inhibition of pro-inflammatory cytokines and chemokines via the NF-κB pathway.

Twelve new and exciting Annonaceae from the Neotropics.

As a result of concerted efforts of the community of Annonaceae taxonomists, increasingly detailed knowledge of the diversity of the Neotropical genera has been documented. With the exception of just two large genera, Annona and Xylopia, all Neotropical Annonaceae have been revised within the last 25 years. Subsequent to these publications, many new specimens have been collected and sent to us in Leiden for identification. These included a number that, despite the advanced state of taxonomic knowledge, proved to represent rarely collected, undescribed species. Here we describe 12 new species of Annona, Guatteria, Klarobelia, Tetrameranthus, and Xylopia. These species serve to illustrate the still underestimated diversity of the Neotropical flora, even in well studied plant groups like Annonaceae.

Drivers of tropical rainforest composition and alpha diversity patterns over a 2,520 m altitudinal gradient.

AIM: We sought to determine the relationship of forest composition and alpha diversity (the species diversity of a local assemblage) to altitude, soil, and spatial factors over a 440-2,950 m a.s.l gradient. LOCATION: Altitudinal gradient on the Caribbean slope of the Talamanca Cordillera, Costa Rica. TAXON: Angiosperm and gymnosperm trees, palms, and tree ferns. METHODS: We measured and identified all stems ≥10 cm dbh in 32 0.25-ha undisturbed rain forest plots over the gradient. We determined compositional patterns using Non-Metric Multidimensional Scaling (NMS) ordination, and used linear regressions to explore the relationship between four alpha diversity metrics and altitude. With variation partitioning (VARPART), we determined the compositional variation explained by altitude, soil, and spatial variables quantified using Principle Components of Neighbor matrices. RESULTS: We identified 425 species. NMS axis 1 separated a lowland zone (440-1,120 m asl) from a transitional one dominated by holarctic Oreomunnea mexicana (1,400-1,600 m asl) and Quercus-dominated forests at altitudes >2,100 m asl. The lowland zone was separated into two clusters of plots on NMS axis 2, the first in the 430-620 m asl range and the second at 1,000-1,120 masl. Regressions showed that all alpha diversity metrics were strongly negatively related to altitude (R 2 > 0.78). Overall, adjusted R 2 from VARPART was 0.43, with 0.30, 0.21, and 0.17 for altitude, soil, and space respectively. The respective adjusted R 2 of individual matrices, on controlling for the other two, was 0.06, 0.05 and 0.09 (p < 0.001). MAIN CONCLUSIONS: There are two well-defined forest compositional zones on this gradient-lowlands 430-1,120 m asl and montane forests >2,150 m asl-with a transitional zone at 1,400-1,600 m asl, where lowland tropical and montane holarctic species are found together. Montane forests are very distinct in their composition and low alpha diversity. Vegetation and soil respond to altitude, and therefore temperature, as an integrated system, a model that goes beyond niche assembly as shown by the significant effect of space in the VARPART.

Dimensionamiento del sistema de microfiltración para la producción de la Vacuna Pertussis elaborada en el Instituto Nacional de Higiene “Rafael Rangel” / Microfiltration sistem dimension for using in Pertussis Vaccine toxin production in the Instituto Nacional de Higiene "Rafael Rangel"

Se evaluó el uso de la tecnología de Flujo de Filtración Tangencial (FFT), para obtener la Vacuna Pertussis Celular a partir de cultivos de la bacteria Bordetella pertussis, usando el proceso de Microfiltración (MF) a objeto de recuperar el paquete celular. Se determinaron las características de los filtros, condiciones de trabajo y el dimensionamiento del equipo a adquirir para la nueva producción in dustrial de Vacuna Pertussis Celular. Se evaluaron el flujo y tiempo de proceso, rendimiento y las características del producto obtenido. Utilizando cultivos con Vacuna Pertus sis en un equipo de filtración de laboratorio, diseñado para producir el efecto de FFT. Se seleccionó las membranas tipo cassettes, formato Suspended Screen, porosidad 0,2 μm, como las adecuadas para el proceso de MF, ya que mostraron un 100% de recuperación del paquete celular sin transmisión de células al filtrado y con un flujo promedio de filtrado de 54.00 L/m2h. Estos resultaron permitieron dimensionar, considerando las variables a utilizar en la nueva producción industrial (Volumen 650 Litros, Tiempo de Procesos, 3 a 4 horas), el área de filtración del equipo de MF a adquirir, estimado en 20 m² .

Tangential Flow Filtration (TFF) technology was evaluated to process Whole Cell Pertussis Vaccine which is produced by Bordetella pertussis bacterium. Microfiltration (MF) is used to recovery cells to produ ce the vaccine. MF pro - cesses was evaluated to specify the filters and corresponding critical process parameters to scale-up the application. As part of the evaluation, flow rate, processing time, yield and product attributes were characterized. The cell harvest con taining the Whole Cell Pertussis was processed using a laboratory scale TFF system designed to pro duct the TFF effect. The evaluation demonstrated that a cassette in suspended screen format and membrane with 0.2μm pore is the right selection for the MF step. It showed 100% of cell recovery without cell transmission to the filtrate and average process flux of 54.00 L/m2h. These results were used to scale-up the application to process the industrial volume of 650 liters in 3 hours of processing time. Membrane area sizing of MF to be acquired is estimated in 20 m².

Evolución histórica y logros de la gerencia sectorial de producción del Instituto Nacional de Higiene Rafael Rangel / Historical evolution and achievements of the sector's production management of the Instituto Nacional de Higiene Rafael Rangel

La Gerencia Sectorial de Producción elabora vacunas bacterianas y virales de uso humano y veterinario, para satisfacer necesidades específicas de nuestra población. Igualmente responde a los requerimientos del resto de las áreas técnicas de la institución, a las que provee de insumos, reactivos químicos y biológicos como cultivos celulares y animales de experimentación, entre otros. También presta servicios con fines diagnósticos, productivos, de control e investigación. Un ejemplo relevante de proyección de la Gerencia Sectorial de Producción, y del Instituto, lo constituye la nueva Planta de Vacunas, instalación de alta tecnología que representa un contundente aporte para consolidar la autosuficiencia nacional en la producción de tan importantes rubros para la salud.

The Sectorial Productión Management produces bacterial and viral vaccines for human and veterinary use to satisfy specific needs of our population. It also responds, to the requirements of the rest of the technical units of the Institute, to whom it provides with chemical and biological reagents like cellular cultures and laboratory animals, among others. It also carry out activities for diag nostic purposes, production, control and research. An excellent example of projection of The Sectorial Productión Management and the Institute constitutes the new Vaccine Plant, installation of high technology that will allow to consolidate the national self-sufficiency in the production of such important headings for the health.

Diterpenoid, steroid, and triterpenoid agonists of liver X receptors from diversified terrestrial plants and marine sources.

It has been demonstrated that liver X receptors (LXR) play a significant role in cholesterol homeostasis. Agonists of LXR are expected to increase cellular cholesterol efflux, lower LDL, and raise HDL levels. Screening of a natural product library of plant extracts using a LXR-SPA binding assay and bioassay-guided fractionation of a number of plant and marine gorgonian extracts led to the isolation of a number of active compounds. These included acanthoic acid (1) and alcohol (2), viperidone (3), polycarpol (4), rosacea acid (5), a cycloartane derivative (6), a new cycloartane analogue (7), betulinic acid (8), and gorgostane derivatives (9, 10, and 11). Of these compounds, 1, 4, and 11 exhibited potent binding affinity for alpha-receptor with IC(50) values of 0.25, 0.12, and 0.07 microM, respectively. Functionally they also showed strong coactivator association stimulation for LXRalpha receptor with EC50 values of 0.18, 0.03, and 0.05 microM, respectively. They also exhibited 15-, 8-, and 13-fold induction of the alpha-receptor in a transactivation assay in HEK-293 cells, respectively. In general these compounds were selective for the LXR alpha-receptor over the beta-receptor in all assays and were much better stimulators of the alpha-receptor than the endogenous steroid ligands.

Sobrevivencia de Sitophilus oryzae (L. ) en harina cruda y extruida de granos de Canavalia ensiformis / Survival of Sitophilus oryzae (L. ) in raw flour and extruded of grains of Canavalia ensiformis

El objetivo de este trabajo fue evaluar la sobrevivencia de gorgojos adultos del arroz (Sitophilus oryzae) en harina cruda y extruida de granos de Canavalia ensiformis. Se determinó el contenido de los factores antinutricionales (título hemaglutinante y canavanina). La actividad hemaglutinante de la harina cruda fue eliminada, como resultado de la alta temperatura (155°C) empleada durante el proceso de extrusión. El contenido de canavanina en la harina cruda y extruida no fue afectado por la extrusión. Se midió la sobrevivencia de gorgojos adultos del arroz como respuesta a dietas con harinas y almidón de granos de Canavalia. La sobrevivencia de los gorgojos alimentados con harina cruda y extruida fue menor que la de los insectos mantenidos en ayunas (control negativo, es decir, sin alimento). La diálisis de las harinas mejoró la sobrevivencia de los gorgojos en relación al control negativo, pero no se alcanzó la sobrevivencia de los gorgojos alimentados con harina de cotiledones de arvejas (control positivo). La suplementación de las harinas con arginina (0,1 por ciento) no produjo mejora en la sobrevivencia de los insectos. La mortalidad de los gorgojos en almidón de harina extruida fue similar al control negativo. En general, los resultados indican que S. oryzae es susceptible a los factores antinutritivos tales como canavanina

[Effect of extrusion on the activity of anti-nutritional factors and in vitro digestibility of protein and starch in flours of Canavalia ensiformis]. / Efecto de la extrusión sobre la actividad de factores antinutricionales y digestibilidad in vitro de proteínas y almidón en harinas de Canavalia ensiformis.

The effect of the extrusion (155 degrees C, 20% moisture, screw speed 75 rpm, feed speed 205 g min-1) on antinutritional factors of Canavalia ensiformis was studied. In vitro protein and starch digestibilities were assessed. The extrusion not affect protein content (23%) in the flours, but significantly (P < 0.01) decrease moisture content. The protein digestibility values were improved from 57.5 to 89.5%, these values were lower than casein (98.19%). The digestibility of starch values were improved from 37.7 to 53%. The protease inhibitors activities (trypsin and chymotrypsin) and alpha-amylase inhibitor activity were reduced by 95%. The haemagglutinating activity was eliminated as result of the high temperature employed during the extrusion process. The canavanine content in the flours were not affect by the treatment of extrusion.

Evaluación y mejoramiento del aspecto físico de las vacunas pertussis y DPT elaboradas en el Instituto Nacional de Higiene "Rafael Rangel" / Improvement and evaluation of physical appearance of the pertussis vaccines and DPT elaborates in the Instituto Nacional de Higiene \"Rafael Rangel\"

Se evaluaron los procesos de recuperación de la vacuna pertussis y formulación de la vacuna DPT, elaborados en el Instituto Nacional de Higiene "Rafael Rangel", con el fin de mejorar el aspecto físico u homogeneidad de la vacuna pertussis antes de ser incorporada como fracción en la vacuna DPT. Se evaluaron la calidad del agua, concentración del adyuvante y la concentración y tiempo de almacenamiento de la vacuna pertussis. Se demostró que cuando fueron mezclados lotes de vacunas pertussis de baja concentración (menor de 200 Uop/ml), los cuales habían sido almacenados durante un período menor de dos años y habían sido previamente agitados, con calidad inyectable y a una concentración del adyuvante de 0,52 mg/dosis, generaron una vacuna DPT homogénea, comparable con otras vacunas similares producidas por laboratorios de reconocida trayectoria. Vacunas pertussis concentradas a valores superiores, generaron una vacuna DPT con presencia de aglomerados consistentes de la bacteria Bordetella pertussis. El incremento de la concentración del adyuvante en la vacuna DPT, hasta valores permisibles por la Organización Mundial de la Salud (1,25 mg/dosis) y el uso de agua de calidad inyectable mejoran el aspecto físico de la vacuna DPT. La visualización macroscópica y microscópica de las vacunas

Optimacion del proceso de production de la vacina pertussis / Optimization of pertussis vaccine production process

The production of Pertussis Vaccine was reevaluated at the Instituto Nacional de Higiene(r)Rafael Rangel(r) in order to optimise it in terms of vaccine yield, potency, specific toxicity and efficiency (cost per doses). Four different processes, using two culture media (Cohen-Wheeler and Fermentacion Glutamato Prolina-1) and two types of bioreactors (25 L Fermentador Caracas and a 450 L industrial fermentor) were compared. Runs were started from freeze-dried strains (134 or 509) andcontinued until the obtention of the maximal yield. It was found that the combination Fermentacion Glutamato Prolina-1/industrial fermentor, shortened the process to 40 hours while consistently yielding a vaccine of higher potency (7.91 +/- 2.56 IU/human dose) and lower specific toxicity in a mice bioassay. In addition, the physical aspect of the preparation was rather homogeneous and free of dark aggregates. Most importantly, the biomass yield more than doubled those of the Fermentador Caracas using the two different media and that in the industrial fermentor with the Cohen-Wheeler medium. Therefore, the cost per doses was substantially decreased