Production of Two Isomers of Sphaeralcic Acid in Hairy Roots from Sphaeralcea angustifolia.

The Sphaeralcea angustifolia plant is used as an anti-inflammatory and gastrointestinal protector in Mexican traditional medicine. The immunomodulatory and anti-inflammatory effects have been attributed to scopoletin (1), tomentin (2), and sphaeralcic acid (3) isolated from cells in suspension cultures and identified in the aerial tissues of the wild plant. The hairy roots from S. angustifolia established by infecting internodes with Agrobacterium rhizogenes were explored to produce active compounds based on biosynthetic stability and their capacity to produce new compounds. Chemical analysis was resumed after 3 years in these transformed roots, SaTRN12.2 (line 1) produced scopoletin (0.0022 mg g-1) and sphaeralcic acid (0.22 mg g-1); instead, the SaTRN7.1 (line 2) only produced sphaeralcic acid (3.07 mg g-1). The sphaeralcic acid content was 85-fold higher than that reported for the cells in the suspension cultivated into flakes, and it was similar when the cells in suspension were cultivated in a stirring tank under nitrate restriction. Moreover, both hairy root lines produced stigmasterol (4) and ß-sitosterol (5), as well as two new naphthoic derivates: iso-sphaeralcic acid (6) and 8-methyl-iso-sphaeralcic acid (7), which turned out to be isomers of sphaeralcic acid (3) and have not been reported. The dichloromethane-methanol extract from SaTRN7.1 hairy root line had a gastroprotective effect on an ulcer model in mice induced with ethanol.

Machine Learning Analysis of Essential Oils from Cuban Plants: Potential Activity against Protozoa Parasites.

Essential oils (EOs) are a mixture of chemical compounds with a long history of use in food, cosmetics, perfumes, agricultural and pharmaceuticals industries. The main object of this study was to find chemical patterns between 45 EOs and antiprotozoal activity (antiplasmodial, antileishmanial and antitrypanosomal), using different machine learning algorithms. In the analyses, 45 samples of EOs were included, using unsupervised Self-Organizing Maps (SOM) and supervised Random Forest (RF) methodologies. In the generated map, the hit rate was higher than 70% and the results demonstrate that it is possible find chemical patterns using a supervised and unsupervised machine learning approach. A total of 20 compounds were identified (19 are terpenes and one sulfur-containing compound), which was compared with literature reports. These models can be used to investigate and screen for bioactivity of EOs that have antiprotozoal activity more effectively and with less time and financial cost.

Early sensing of phosphate deprivation triggers the formation of extra root cap cell layers via SOMBRERO through a process antagonized by auxin signaling.

KEY MESSAGE: The role of the root cap in the plant response to phosphate deprivation has been scarcely investigated. Here we describe early structural, physiological and molecular changes prior to the determinate growth program of the primary roots under low Pi and unveil a critical function of the transcription factor SOMBRERO in low Pi sensing. Mineral nutrient distribution in the soil is uneven and roots efficiently adapt to improve uptake and assimilation of sparingly available resources. Phosphate (Pi) accumulates in the upper layers and thus short and branched root systems proliferate to better exploit organic and inorganic Pi patches. Here we report an early adaptive response of the Arabidopsis primary root that precedes the entrance of the meristem into the determinate developmental program that is a hallmark of the low Pi sensing mechanism. In wild-type seedlings transferred to low Pi medium, the quiescent center domain in primary root tips increases as an early response, as revealed by WOX5:GFP expression and this correlates with a thicker root tip with extra root cap cell layers. The halted primary root growth in WT seedlings could be reversed upon transfer to medium supplemented with 250 µM Pi. Mutant and gene expression analysis indicates that auxin signaling negatively affects the cellular re-specification at the root tip and enabled identification of the transcription factor SOMBRERO as a critical element that orchestrates both the formation of extra root cap layers and primary root growth under Pi scarcity. Moreover, we provide evidence that low Pi-induced root thickening or the loss-of-function of SOMBRERO is associated with expression of phosphate transporters at the root tip. Our data uncover a developmental window where the root tip senses deprivation of a critical macronutrient to improve adaptation and surveillance.

Objectivation of an Educational Model in Cranial Osteopathy Based on Experience.

Background and Objectives: The techniques directed to the cranial field in osteopathy are the most questioned due to the lack of scientific evidence. In osteopathic practice, manual palpation is essential and, therefore, measuring reliability is fundamental. The objective of this study is to assess the precision and objectification of an educational model in cranial osteopathy based on experience. Materials and Methods: A reliability study was conducted in a cadaver skull where a strain gauge was placed on the sphenobasilar synchondrosis (SBS) of the base of the skull. Three cranial osteopathic techniques (lateral compression, anteroposterior compression, and compression maneuver of the mastoids) were performed 25 times by osteopaths with different degrees of experience (5-10 years, 1-5 years, <1 year). Measurements were computed for each of the three techniques of each group in comparison with the osteopath with >15 years of experience. Data were analyzed to check for inter- and intra-observer reliability using intra-class correlation coefficients (ICC). Results: Reliability in osteopaths with 5-10 years' experience (observer 1 and observer 2) performing all three techniques was higher (p < 0.001) than the osteopath with >15 years' experience. Little or no reliability were observed in osteopaths with less experience. Conclusions: The experience of the osteopaths determines the reliability and effectiveness of the cranial techniques, a fundamental part in objectifying these techniques. This model can help implement objective training in cranial osteopathy formation.

Ligninolytic activity of the Penicillium chrysogenum and Pleurotus ostreatus fungi involved in the biotransformation of synthetic multi-walled carbon nanotubes modify its toxicity.

Multi-walled carbon nanotubes (MWCNTs) are of multidisciplinary scientific interest due to their exceptional physicochemical properties and a broad range of applications. However, they are considered potentially toxic nanoparticles when they accumulate in the environment. Given their ability to oxidize resistant polymers, mycorremediation with lignocellulolytic fungi are suggested as biological alternatives to the mineralization of MWCNTs. Hence, this study involves the ability of two fungi specie to MWCNTs biotransformation by laccase and peroxidases induction and evaluation in vivo of its toxicity using Caenorhabditis elegans worms as a model. Results showed that the fungi Penicillium chrysogenum and Pleurotus ostreatus were capable to grow on media with MWCNTs supplemented with glucose or lignin. Activities of lignin-peroxidase, manganese-peroxidase, and laccase in cultures of both fungi were induced by MWCNTs. Raman, FTIR spectroscopy, HR-TEM, and TGA analyses of the residue from the cultures of both fungi revealed structural modifications on the surface of MWCNTs and its amount diminished, correlating the MWCNTs structural modifications with the laccase-peroxidase activities in the fungal cultures. Results indicate that the degree of toxicity of MWCNTs on the C. elegans model was enhanced by the structure modification associated with the fungal ligninolytic activity. The toxic effect of MWCNTs on the in vivo model of worms reveals the increment of reactive oxygen species as a mechanism of toxicity. Findings indicate that the MWCNTs can be subject in nature to biotransformation processes such as the fungal metabolism, which contribute to modify their toxicity properties on susceptible organisms and contributing to environmental elimination.

Scientific Appearance and Homeopathy. Determinants of Trust in Complementary and Alternative Medicine.

Several studies have investigated the motivations driving the use of complementary and alternative medicines (CAM). Nevertheless, the general public view of these therapies remains relatively unexplored. Our study identifies the social factors that determine a person's trust in alternative therapies, like homeopathy or acupuncture, drawing conclusions from the results of the Spanish National Survey on the Social Perception of Science and Technology (N = 6,357). We show that trust in the effectiveness of CAM therapies is not mutually exclusive with a belief in science for the general public, pointing to a certain level of disinformation. The comparison with superstitions confirms a clear differentiation with the drivers of trust in analyzed CAM therapies. We argue that scientific appearance of these alternative therapies, in terms of prescription, communication and marketing, may play an important role in determining trust in them for a large part of the population. Furthermore, we confirm that women and those with higher socio-economic status are more likely to express trust in the effectiveness of CAM therapies. Additionally, distrust of the influence of big pharma on health policies seems to have an effect on viewing CAM therapies as more effective. Finally, we argue that media and pharmacies may have an effect on the scientific-like perception of CAM therapies, contributing to the social construction of trust in its effectiveness. Therefore, widespread confusion about the scientific validation of homeopathy may be among the main factors driving its successful extension as a practice.

Compuestos bioactivos, actividad antioxidante y perfil de ácidos grasos en aceite de semilla de Mezquite (Prosopis spp) / Bioactive Compounds, Antioxidant Activity and Profile of Fatty Acids in Mesquite Seed Oil (Prosopis spp)

La medicina tradicional y estudios realizados a diferentes especies del género Prosopis, del desierto sonorense, indican que es una fuente para la cualificación de compuestos bioactivos, con poder antioxidante y ácidos grasos (linoleico y linolénico) de la semilla. La actividad biológica, es atribuible a alcaloides, flavonoides, terpenos y compuestos fenólicos, para lo cual, se realizó el perfil fitoquímico en los extractos acuoso, etanólico, hexanico y clorofórmico (mediante técnicas colorimétricas), actividad antioxidante (método: 1,1-difenil-2-picrilhidrazil (DPPH)), fenoles totales (utilizando el reactivo de Folin-Ciocalteau) y perfil de ácidos grasos (cromatografía de gases) de la semilla de Prosopis spp. La extracción del aceite se realizó mediante Soxhlet. Se encontraron saponinas en todos los extractos, mientras que, en el etanólico, hexanico y clorofórmico, terpenos y esteroles. En el extracto etanólico se encontraron quinonas y en el acuoso aminoácidos libres. El valor más alto de la actividad antioxidante de EC50 fue de 3.272,41 ± 5,97, para el extracto etanólico, indicando su potencial como antioxidante. El contenido de fenoles totales, fue hexano> etanol > cloroformo> acuoso (81,95; 119,83; 125,18 y 127,57 mg equivalentes de ácido gálico/g de extracto seco). Los ácidos grasos en mayor proporción fueron los insaturados con 71,41 % (ácido linoleico: 42,68 %; oleico: 28.73 %) y ácidos grasos saturados: ácido palmítico (13.42 %) y estérico (4,73 %). Se concluye que este tipo de extractos presentan metabolitos importantes para la dieta, presentan actividad antioxidante y ácidos grasos esenciales para el organismo(AU)

Traditional medicine and studies with different species of the Prosopis genus, from the Sonoran Desert, is a source for the qualification of bioactive compounds, with antioxidant power and fatty acids (linoleic and linolenic) of the seed. The biological activity is attributable to alkaloids, flavonoids, terpenes and phenolic compounds, for which, the phytochemical profile was performed in the aqueous, ethanolic, hexane and chloroform extracts (using colorimetric techniques), antioxidant activity (method: 1,1-diphenyl-2-picrilhidrazil (DPPH)), total phenols (using the Folin-Ciocalteau reagent) and fatty acid profile (gas chromatography). The oil was extracted using Soxhlet. Saponins were found in all extracts, while, in ethanolic, hexanic and chloroform, terpenes and sterols. In the ethanolic extract quinones were found and in the aqueous free amino acids. The highest value of the antioxidant activity of EC50 was 3,272.41 ± 5.97, for the ethanolic extract, indicating its potential as an antioxidant. The total phenolic content was hexane> ethanol> chloroform> aqueous (81.95, 119.83, 125.18 and 127.57 mg equivalent of gallic acid / g of dry extract). The fatty acids in greater proportion were unsaturated with 71.41 % (linoleic acid: 42.68 %; oleic: 28.73 %) and saturated fatty acids: palmitic acid (13.42 %) and stearic (4.73 %). It is concluded that this type of extracts have important metabolites for the diet, have antioxidant activity and essential fatty acids for the body(AU)

Foliar persistence and residual activity of four insecticides of different mode of action on the predator Engytatus varians (Hemiptera: Miridae).

A greenhouse study was conducted to investigate the degradation kinetics of spinosad, flufenoxuron, dimethoate and imidacloprid in tomato (Solanum lycopersicum L.) foliage and their residual toxicity on Engytatus varians (Distant) (Hemiptera: Miridae), a predator of the tomato psyllid Bactericera cockerelli (Sulcer) (Hemiptera: Triozidae). Insecticides were sprayed at 100% and 50% of their maximum field-registered concentrations (MFRC). Starting 6 h after spraying, leaf samples were taken every 10 d for 40 d and analyzed while E. varians adults were exposed to treated leaves to evaluate residual toxicity. Immediately after application at 100% MFRC, the residue concentrations were 73.34 µg g-1 spinosyn A and 59.2 µg g-1 spinosyn D, 9.21 µg g-1 flufenoxuron, 71.49 µg g-1 dimethoate and 31.74 µg g-1 imidacloprid. At 50% MFRC, initial residue concentrations were between 75% and 90% those at 100% MFRC. The estimated half-life (DT50) of spinosyns A and D, flufenoxuron, and dimethoate was between 34 and 40 d, while that of imidacloprid was 112 d. Flufenoxuron caused no mortality, while mortality due to spinosad was less than 10%, and only during the first 10 d. Mortality caused by either imidacloprid or dimethoate was around 100% up to 10 d after application, then decreased to around 30% after 40 d. Dimethoate toxicity was approximately proportional to residue concentration, while for imidacloprid there was an apparent threshold around 15 µg g-1. These results can be used to establish periods harmless for release of E. varians in the control of B. cockerelli on tomato crops under greenhouse conditions.

Mitogen-activated protein kinase 6 integrates phosphate and iron responses for indeterminate root growth in Arabidopsis thaliana.

MAIN CONCLUSION: A MAPK module, of which MPK6 kinase is an important component, is involved in the coordination of the responses to Pi and Fe in the primary root meristem of Arabidopsis thaliana. Phosphate (Pi) deficiency induces determinate primary root growth in Arabidopsis through cessation of cell division in the meristem, which is linked to an increased iron (Fe) accumulation. Here, we show that Mitogen-Activated Protein Kinase6 (MPK6) has a role in Arabidopsis primary root growth under low Pi stress. MPK6 activity is induced in roots in response to low Pi, and such induction is enhanced by Fe supplementation, suggesting an MPK6 role in coordinating Pi/Fe balance in mediating root growth. The differentiation of the root meristem induced by low Pi levels correlates with altered expression of auxin-inducible genes and auxin transporter levels via MPK6. Our results indicate a critical role of the MPK6 kinase in coordinating meristem cell activity to Pi and Fe availability for proper primary root growth.

Side chain to main chain hydrogen bonds stabilize a polyglutamine helix in a transcription factor.

Polyglutamine (polyQ) tracts are regions of low sequence complexity frequently found in transcription factors. Tract length often correlates with transcriptional activity and expansion beyond specific thresholds in certain human proteins is the cause of polyQ disorders. To study the structural basis of the association between tract length, transcriptional activity and disease, we addressed how the conformation of the polyQ tract of the androgen receptor, associated with spinobulbar muscular atrophy (SBMA), depends on its length. Here we report that this sequence folds into a helical structure stabilized by unconventional hydrogen bonds between glutamine side chains and main chain carbonyl groups, and that its helicity directly correlates with tract length. These unusual hydrogen bonds are bifurcate with the conventional hydrogen bonds stabilizing α-helices. Our findings suggest a plausible rationale for the association between polyQ tract length and androgen receptor transcriptional activity and have implications for establishing the mechanistic basis of SBMA.

High accumulation of tilianin in in-vitro cultures of Agastache mexicana and its potential vasorelaxant action.

Agastache mexicana has gained importance during the last decade as a natural source of bioactive compounds, mainly due to the antidiabetic, antihyperlipidemic, and vasorelaxant effects derived from its flavonoids, particularly tilianin. The goal of this work was to evaluate the production of tilianin during the in-vitro process of morphogenesis leading to plant regeneration and to investigate the vasorelaxant activity of its methanolic extracts. The cultures were established from nodal segments and leaf explants, inoculated on Murashige and Skoog (MS) media supplemented with various concentrations of benzyl aminopurine (BAP) alone or in combination with 2,4-Dichlorophenoxyacetic acid (2,4-D). Callus inductions were obtained in all treatments from both types of explants, but the presence of auxin was essential. Maximal shoot multiplication and elongation was achieved with 0.1 mg/l 2,4-D and 1.0 mg/l BAP from nodal- segment explants. Shoots were rooted in 75% MS medium and the plantlets were transferred to a greenhouse with 33% average survival. Analysis of tilianin production in methanolic extracts from calli (0.15-2.01 ± 0.06 mg/g dry weight), shoots (4.45 ± 0.01 mg/g DW), and whole plants (9.77 ± 0.02 mg/g DW) derived from in-vitro cultured nodal segments reveals that tilianin accumulation is associated with high cell differentiation and morphogenetic response to the plant-growth regulators. All of the extracts showed strong vasorelaxant activity, as compared to those of wild plant extracts. These results indicate that plant-tissue cultures of A. mexicana possess vast potential as a source of tilianin and other bioactive compounds.

Long-chain omega-3 polyunsaturated fatty acid dietary intake is positively associated with bone mineral density in normal and osteopenic Spanish women.

The regular consumption of long-chain omega-3 polyunsaturated fatty acids (LCO3-PUFAs) results in general health benefits. The intake of LCO3-PUFAs has been reported to contribute to bone metabolism. We aimed to investigate the relationships between dietary intakes of LCO3-PUFAs and bone mineral density (BMD) in Spanish women aged 20-79 years old. A total of 1865 female subjects (20-79 years old) were enrolled, and lumbar (L2, L3, L3 and total spine), hip (femoral neck (FN), femoral trochanter (FT) and Ward's triangle (WT)) bone mineral density (BMD) were measured by dual energy X-ray absorptiometry (DXA). Dietary intakes of total energy, calcium, vitamin D, alpha-linolenic acid (ALA), eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and n-6 fatty acids (linoleic acid (LA) and arachidonic acid (AA)) were assessed by a self-administered food frequency questionnaire (FFQ). Spearman's rank correlations between LCO3-PUFAs and BMD were estimated. Partial correlations controlling for age, weight, height, dietary calcium, vitamin D, menopausal status and energy were calculated. A multiple regression analysis was computed to assess significant associations with BMD in this population. After adjustment for potential confounding factors, there were positive correlations between ALA, EPA and DHA intake and BMD. According to the WHO diagnosis criteria for osteoporosis, in this population of normal and osteopenic women, the dietary intake of ALA was also significantly associated with BMD at the hip. In normal women, the dietary intake of DHA was also significantly associated with BMD at the lumbar spine. No significant associations between LCO3-PUFAs and BMD were detected in the lumbar spine of osteopenic or osteoporotic women. The dietary intake of LCO3-PUFAs was positively associated with BMD in Spanish women at both the hips and the lumbar spine. We highlight that the intake of LCO3-PUFAs is not significantly associated with BMD in osteoporotic women; however, the intake of LCO3-PUFAs seems to be positively associated with BMD at both the hips and the lumbar spine in normal and osteopenic women.

Neuropharmacological and neuroprotective activities of some metabolites produced by cell suspension culture of Waltheria americana Linn.

Waltheria americana is a plant used in Mexican traditional medicine to treat some nervous system disorders. The aims of the present study were to isolate and determine the neuropharmacological and neurprotective activities of metabolites produced by a cell suspension culture of Waltheria americana. Submerged cultivation of W. americana cells provided biomass. A methanol-soluble extract (WAsc) was obtained from biomass. WAsc was fractionated yielding the chromatographic fractions 4WAsc-H2O and WAsc-CH2Cl2. For the determination of anticonvulsant activity in vivo, seizures were induced in mice by pentylenetetrazol (PTZ). Neuropharmacological activities (release of gamma amino butyric acid (GABA) and neuroprotection) of chromatographic fractions were determined by in vitro histological analysis of brain sections of mice post mortem. Fraction 4WAsc-H2O (containing saccharides) did not produce neuronal damage, neurodegeneration, interstitial tissue edema, astrocytic activation, nor cell death. Pretreatment of animals with 4WAsc-H2O and WAsc-CH2Cl2 from W. americana cell suspensions induced an increase in: GABA release, seizure latency, survival time, neuroprotection, and a decrease in the degree of severity of tonic/tonic-clonic convulsions, preventing PTZ-induced death of up to 100% of animals of study. Bioactive compounds produced in suspension cell culture of W. americana produce neuroprotective and neuropharmacological activities associated with the GABAergic neurotransmission system.

A Cytotoxic and Anti-inflammatory Campesterol Derivative from Genetically Transformed Hairy Roots of Lopezia racemosa Cav. (Onagraceae).

The genetically transformed hairy root line LRT 7.31 obtained by infecting leaf explants of Lopezia racemosa Cav with the Agrobacterium rhizogenes strain ATCC15834/pTDT, was evaluated to identify the anti-inflammatory and cytotoxic compounds reported previously for the wild plant. After several subcultures of the LRT 7.31 line, the bio-guided fractionation of the dichloromethane-methanol (1:1) extract obtained from dry biomass afforded a fraction that showed important in vivo anti-inflammatory, and in vitro cytotoxic activities. Chemical separation of the active fraction allowed us to identify the triterpenes ursolic (1) and oleanolic (2) acids, and (23R)-2α,3ß,23,28-tetrahydroxy-14,15-dehydrocampesterol (3) as the anti-inflammatory principles of the active fraction. A new molecule 3 was characterized by spectroscopic analysis of its tetraacetate derivative 3a. This compound was not described in previous reports of callus cultures, in vitro germinated seedlings and wild plant extracts of whole L. racemosa plants. The anti-inflammatory and cytotoxic activities displayed by the fraction are associated to the presence of compounds 1-3. The present study reports the obtaining of the transformed hairy roots, the bioguided isolation of the new molecule 3, and its structure characterization.

Cardiac activation-repolarization patterns and ion channel expression mapping in intact isolated normal human hearts.

BACKGROUND: The repolarization pattern of the human heart is unknown. OBJECTIVE: The purpose of this study was to perform a multisite analysis of the activation-repolarization patterns and mRNA expression patterns of ion channel subunits in isolated human hearts. METHODS: Hearts from 3 donors without reported cardiac disease were Langendorff perfused with the patient's own blood. A standard ECG was obtained before explantation. Up to 92 unipolar electrograms from 24 transmural needles were obtained during right atrial pacing. Local activation and repolarization times and activation-recovery intervals (ARI) were measured. The mRNA levels of subunits of the channels carrying the transient outward current and slow and rapid components of the delayed rectifier current were determined by quantitative reverse transcriptase polymerase chain reaction at up to 63 sites. RESULTS: The repolarization gradients in the 3 hearts were different and occurred along all axes without midmural late repolarization. A negative activation-repolarization relationship occurred along the epicardium, but this relationship was positive in the whole hearts. Coefficients of variation of mRNA levels (40%-80%) and of the Kv7.1 protein (alpha-subunit slow delayed rectifier channel) were larger than those of ARIs (7%-17%). The regional mRNA expression patterns were similar in the 3 hearts, unlike the ARI profiles. The expression level of individual mRNAs and of Kv7.1 did not correlate with local ARIs at the same sites. CONCLUSION: In the normal human heart, repolarization gradients encompass all axes, without late midmural repolarization. Last activated areas do not repolarize first as previously assumed. Gradients of mRNAs of single ion channel subunits and of ARIs do not correlate.

[Guide to clinical practice for the diagnosis, treatment and rehabilitation of non-syndromic craniosynostosis on 3 levels of care]. / Guía de práctica clínica para el diagnóstico, tratamiento y rehabilitación de craneosinostosis no sindrómica en los 3 niveles de atención.

INTRODUCTION: Craniosynostosis is a congenital anomaly resulting from the premature fusion of the cranial sutures changing growth patterns of the skull. METHODOLOGY: Focus, scope, target population and clinical questions to be solved were defined. A systematic search for evidence in different databases (Medline, Embase, KoreaMed, Cochrane Library and the website of the World Health Organization) in stages was performed: clinical practice guidelines; systematic reviews, and clinical trials and observational studies; using MeSH, Decs and corresponding free terms, unrestricted language or temporality. Risk of bias was evaluated using appropriate tools (AMSTAR, Risk of Bias or STROBE). The quality of evidence was graduated using the GRADE system. Modified Delphi Panel technique was used to assign the recommendation's strength and direction, as well as the degree of agreement with it, taking into account the quality of evidence, balance between risks and benefits of interventions, values and preferences of patients and availability of resources. RESULTS: There were 3,712 documents obtained by search algorithms; selecting 29 documents for inclusion in the qualitative synthesis. Due to heterogeneity between studies, it was not possible to perform meta-analysis. CONCLUSIONS: We issued 7 recommendations and 8 good practice points, which will serve as support for decision-making in the comprehensive care of patients with craniosynostosis.

Combined Use of Oligopeptides, Fragment Libraries, and Natural Compounds: A Comprehensive Approach To Sample the Druggability of Vascular Endothelial Growth Factor.

The modulation of protein-protein interactions (PPIs) is emerging as a highly promising tool to fight diseases. However, whereas an increasing number of compounds are able to disrupt peptide-mediated PPIs efficiently, the inhibition of domain-domain PPIs appears to be much more challenging. Herein, we report our results related to the interaction between vascular endothelial growth factor (VEGF) and its receptor (VEGFR). The VEGF-VEGFR interaction is a typical domain-domain PPI that is highly relevant for the treatment of cancer and some retinopathies. Our final goal was to identify ligands able to bind VEGF at the region used by the growth factor to interact with its receptor. We undertook an extensive study, combining a variety of experimental approaches, including NMR-spectroscopy-based screening of small organic fragments, peptide libraries, and medicinal plant extracts. The key feature of the successful ligands that emerged from this study was their capacity to expose hydrophobic functional groups able to interact with the hydrophobic hot spots at the interacting VEGF surface patch.

Protective effects of dietary avocado oil on impaired electron transport chain function and exacerbated oxidative stress in liver mitochondria from diabetic rats.

Electron transport chain (ETC) dysfunction, excessive ROS generation and lipid peroxidation are hallmarks of mitochondrial injury in the diabetic liver, with these alterations also playing a role in the development of non-alcoholic fatty liver disease (NAFLD). Enhanced mitochondrial sensitivity to lipid peroxidation during diabetes has been also associated to augmented content of C22:6 in membrane phospholipids. Thus, we aimed to test whether avocado oil, a rich source of C18:1 and antioxidants, attenuates the deleterious effects of diabetes on oxidative status of liver mitochondria by decreasing unsaturation of acyl chains of membrane lipids and/or by improving ETC functionality and decreasing ROS generation. Streptozocin-induced diabetes elicited a noticeable increase in the content of C22:6, leading to augmented mitochondrial peroxidizability index and higher levels of lipid peroxidation. Mitochondrial respiration and complex I activity were impaired in diabetic rats with a concomitant increase in ROS generation using a complex I substrate. This was associated to a more oxidized state of glutathione, All these alterations were prevented by avocado oil except by the changes in mitochondrial fatty acid composition. Avocado oil did not prevented hyperglycemia and polyphagia although did normalized hyperlipidemia. Neither diabetes nor avocado oil induced steatosis. These results suggest that avocado oil improves mitochondrial ETC function by attenuating the deleterious effects of oxidative stress in the liver of diabetic rats independently of a hypoglycemic effect or by modifying the fatty acid composition of mitochondrial membranes. These findings might have also significant implications in the progression of NAFLD in experimental models of steatosis.

Production of the anti-inflammatory compound 6-O-palmitoyl-3-O-ß-D-glucopyranosylcampesterol by Callus cultures of Lopezia racemosa Cav. (Onagraceae).

Lopezia racemosa Cav. is a plant used in Mexican traditional medicine to heal inflammatory diseases. From this plant we isolated the novel compound 6-O-palmitoyl- 3-O-ß-D-glucopyranosylcampesterol (1) and 6-O-palmitoyl-3-O-ß-D-glucopyranosyl-ß-sitosterol (2), previously reported to have cytotoxic activity on several cancer cell lines. We evaluated the anti-inflammatory activity of 1 in vivo by mouse ear edema induced with 12-O-tetradecanoylphorbol-13-acetate (TPA) and 57.14% inhibition was observed. The aim of our study was to obtain callus cultures derived from this plant species with the ability to produce the compounds of interest. Callus cultures were initiated on MS basal medium amended with variable amounts of naphthaleneacetic acid (NAA), or 2,4-dichlorophenoxyacetic acid (2,4-D), combined or not with 6-benzylaminopurine (BAP). Ten treatments with these growth regulators were carried out, using in vitro germinated seedlings as source of three different explants: hypocotyl, stem node, and leaf. Highest yield of 1 was observed on callus derived from leaf explants growing in medium containing 1.0 mg/L 2,4-D and 0.5 mg/L BAP. Selected callus lines produced less 1 than wild plants but the in vitro cultured seedlings showed higher production. So we conclude that it could be attractive to further investigate their metabolic potential.

Is there an optimal dose for dietary linoleic acid? Lessons from essential fatty acid deficiency supplementation and adipocyte functions in rats.

Differential effects of n-3 and n-6 polyunsaturated fatty acids (PUFAs) have been demonstrated on adipose tissue physiology. Facing to the widely recognized beneficial effects of n-3 PUFAs, the n-6 PUFA effects remain controversial. Thus, we further analyzed the linoleic acid (LA) influence on adipocyte functions. To this aim, we treated by LA supplementation at three distinct doses (1, 2.5, or 5% of energy intake) rats with essential fatty acids deficiency (EFAD). PUFA composition was determined in blood and white adipose tissue (WAT), while lipolytic and lipogenic activities were measured in isolated adipocytes. EFAD rats exhibited reduced WAT mass and increased EFAD biomarkers. WAT mass was completely recovered after supplementation, irrespective of LA dose. However, neither body mass nor EFAD biomarkers returned to control with 1% LA, while LA abundance doubled in adipocytes from rats supplemented with 5% LA. Regarding lipolysis, 2.5% LA normalized the EFAD-induced alterations. A trend to decrease the maximal stimulation of lipolysis was observed with 1 and 5% LA. Regarding lipogenesis, the lower and higher LA doses increased basal activity and hampered insulin to further stimulate glucose incorporation into lipids whereas 2.5% LA normalized the basal or insulin-stimulated levels. Our results show that dietary linoleate at 2.5% restored anatomical, biochemical, and functional disturbances induced by EFAD. At higher dose, LA tended to reduce triacylglycerol breakdown, to increase triacylglycerol assembly, and to provoke insulin resistance. Therefore, LA influence on adipocyte functions does not appear to follow a typical dose-response relationship, adding further complexity to the definition of its dietary requirement.