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Plants are a source of valuable organic chemical compounds with complex structures rich in therapeutic activities. The encapsulation of compounds in nanostructured systems is an alternative to avoid limitations, such as instability and low solubility, and to promote therapeutic use. The objective of the present review was to summarize the data in the literature on the physicochemical characteristics, biomedical efficacy, and toxicity of nanostructures containing extracts and oils obtained from annatto (Bixa orellana). For this, searches were conducted in the CINAHL, LILACS, Embase, FSTA, MEDLINE, ProQuest, PubMed, ScienceDirect, Scopus, and Web of Science databases. Studies that carried out the development, physical-chemical characterization, and evaluation of therapeutic efficacy and/or in vitro, in vivo, or clinical toxicity of nanostructures containing extracts and oils derived from annatto were included in the review. Of the 708 articles found, nine met the inclusion criteria. The included studies developed different nanostructures (nanofibers, nanocochleates, chitosan, lipid, polymeric, and metallic nanoparticles). These nanostructures showed leishmanicidal, photoprotective, antioxidant, antimicrobial, and immunomodulatory efficacy, and tissue regeneration potential with no or low toxic effects in the tested models. Thus, the present work supports the nanostructuring of annatto extracts and oils as a relevant approach to the development of new technologies for biomedical applications. [ABSTRACT FROM AUTHOR]

Annatto (Bixa orellana L.) is an important dye-yielding species harbouring huge variations in qualitative as well as quantitative traits. Identification of factors influencing bixin content and seed yield is an important breeding aspect, considering the economic importance of annatto for selection of elite genotypes based on morphological traits. Thirtyone trees plus were selected for the study, and sixteen quantitative traits were recorded for two years (2021 and 2022) to understand their association with bixin content and seed yield. Annatto has bivalve pods; however, trivalve pods were also observed during the study. A positive and significant association (r = 0.65) between bixin content and seed yield was observed from the correlation analysis. The study revealed that bixin content was highly correlated and influenced directly by the number of primary and secondary branches and hundred seed weight, while tree height, basal diameter, and the number of primary and secondary branches exhibit a significantly positive direct effect on seed yield, suggesting these traits as a reliable morphological marker for selecting highly productive genotypes. The thirtyone plus trees were grouped through hierarchical clustering into six clusters, with maximum bixin content and seed yield in Cluster I and Cluster III. The genotypes TNBi 0020, TNBi 0021, TNBi 0022, TNBi 0023, TNBi 0024 and TNBi 0037 can be utilized for mass multiplication and popularization for commercial cultivation of Bixa orellana. [ABSTRACT FROM AUTHOR]

This research aims to optimize the silk and wool dyeing process using natural dyes from Bixa orellana (annatto) through response surface methodology. Central composite design experiments highlight the significant enhancement of color outcomes achieved through microwave treatment. For silk, the optimal conditions (80 °C for 40 min) with annatto extract yield a color strength (K/S) of 17.8588, while wool achieves a K/S of 7.5329. Introducing eco-friendly bio-mordants, such as pomegranate peel and red sumac tannins, enhances color strength. Pre-dyeing treatments with 2% red sumac, 1.5% pomegranate peel, and weld flower extracts for silk produce high color strength, with K/S values of 16.4063, 16.3784, and 12.1658, respectively. Post-dyeing, the K/S values increase to 40.1178, 17.4779, and 21.6494. Wool yarn exhibits similar improvements, with pre-dyeing K/S values of 13.1353, 13.5060, and 16.3232, escalating to 10.5892, 15.3141, and 23.4850 post-dyeing. Furthermore, this research underscores improved colorfastness properties, including notable enhancements in light, wash, and rubbing fastness for both silk fabric and wool yarn. These findings underscore the efficacy of the proposed sustainable dyeing methods, offering valuable insights for eco-friendly textile production. [ABSTRACT FROM AUTHOR]

Aging is a complex process resulting from internal and external changes, culminating in a decreased efficacy of cognitive and other biological functions. Studies report that oxidative stress, immunosenescence, inflammaging, and cell senescence are involved in aging. Continuous research is performed seeking treatments delaying such processes to achieve healthy aging, including nutrition and nutraceuticals. In this article, we review the potential of some plants from the Amazon rainforest that have been gaining attention in healthspan, namely Euterpe oleraceae (Açaí), Myrciaria dubia (Camu-camu), and Bixa orellana (Annatto). [ABSTRACT FROM AUTHOR]

Bixin is a commercially valuable apocarotenoid pigment found in the seed aril of Bixa orellana. The dynamics and regulation of its biosynthesis and accumulation during seed development remain largely unknown. Here, we combined chemical, anatomical, and transcriptomic data to provide stage-specific resolution of the cellular and molecular events occurring during B. orellana seed development. Seeds at five developmental stages (S1–S5) were used for analysis of bixin content and seed anatomy, and three of them (S1, S3, and S4) were selected for Illumina HiSeq sequencing. Bixin accumulated in large quantities in seeds compared with other tissues analyzed, particularly during the S2 stage, peaking at the S4 stage, and then decreasing slightly in the S5 stage. Anatomical analysis revealed that bixin accumulated in the large central vacuole of specialized cells, which were scattered throughout the developing mesotesta at the S2 stage, but enlarged progressively at later stages, until they occupied most of the parenchyma in the aril. A total of 13 million reads were generated and assembled into 73,381 protein-encoding contigs, from which 312 were identified as containing 1-deoxy-D-xylulose-5-phosphate/2-C-methyl-D-erythritol-4-phosphate (DOXP/MEP), carotenoid, and bixin pathways genes. Differential transcriptome expression analysis of these genes revealed that 50 of them were sequentially and differentially expressed through the seed developmental stages analyzed, including seven carotenoid cleavage dioxygenases, eight aldehyde dehydrogenases, and 22 methyltransferases. Taken together, these results show that bixin synthesis and accumulation in seeds of B. orellana are a developmentally regulated process involving the coordinated expression of DOXP/MEP, carotenoid, and bixin biosynthesis genes. [ABSTRACT FROM AUTHOR]

Bixa orellana L. is a plant popularly known as "ucurum", "annatto", and "achiote". It is native to South America, and its seeds are an abundant source of geranylgeraniol and tocotrienols. Nanoencapsulation is a valuable technique that can decrease the drug needed to achieve an effect, decreasing potential toxicity, side effects and potentiate the anti-inflammatory effect. This study aimed to evaluate the acute toxicity of an intramuscular application of a nanodispersion containing a standardized extract from the seeds of Bixa orellana (NBO) in Wistar rats. The chemical evaluation showed δ-tocotrienol at 0.725 ± 0.062 mg/mL (72.6 ± 0.9%). The stability study showed the nanoparticles had an average size from 53.15 ± 0.64 to 59.9 ± 3.63 nm, with a polydispersity index ranging from 0.574 ± 0.032 to 0.574 ± 0.32, Zeta potential from 18.26 ± 0.59 to 19.66 ± 1.45 mV. After testing the intramuscular application of NBO with doses from 1 to 5 mg/kg in animals, it was observed that the acute treatment did not elicit any toxic effects within this range. The dose of 10 mg/kg, although not affecting hematological and biochemical parameters (CPK, LDH, myoglobin, AST, ALT, TC, TG, glucose levels, creatinine, and urea), could induce some muscle tissue changes, including leukocyte infiltration, morphological chances, and potentially necrosis. In conclusion, the results showed that the treatments devoided toxicity between 1 and 5 mg/kg. [ABSTRACT FROM AUTHOR]

Background: Alternatively to conventional treatments, chemo-mechanical caries removal agents can be used. A modality of treatment that has been increasing in dentistry is antimicrobial photodynamic therapy (aPDT). Bixa orellana is being researched for application in aPDT. This protocol aims to determine the effectiveness of aPDT with Bixa orellana extract in deep caries lesions. Methods: A total of 160 teeth with deep occlusal dental caries will be selected and divided into 4 groups: G1 - control group (Caries removal with a low-speed drill); G2 - Partial Caries Removal with Papacarie™ (Fórmula e Ação, São Paulo, SP, Brazil); G3 - Partial Caries Removal with Papacarie™ and application Bixa orellana extract (20%) (Fórmula e Ação, São Paulo, SP, Brazil); G4 - Partial Caries Removal with Papacarie™ and application Bixa orellana extract (20%) with LED (Valo Cordless Ultradent®, South Jordan, UT, USA) (aPDT). After treatment, all the teeth will be restored with glass ionomer cement and followed up clinically and radiographically, with evaluations at immediately, 1 week, and 1, 3, 6, and 12 months. Dentin samples before and after treatment will be analyzed microbiologically. The efficacy of treatments will be assessed with microbiological (colony-forming units, before and after carious tissue removal), radiographic (integrity of the periapical area and eventual changes in the radiolucent zones), and clinical examinations (retention of the restorative material in the cavity and occurrence of secondary caries), as well as with the time required for the procedures and the need for anesthesia during the procedures. In case data distribution is normal, analysis of variance (ANOVA) will be used for both the dependent and independent variables. In case the data distribution is not normal, the Friedman test will be used for the dependent variables. For independent variables, the Kruskal-Wallis test will be used. Discussion: Procedures using aPDT have been developed for the treatment of dental caries, but there are few controlled clinical trials in the literature confirming its efficacy. Trial registration: This protocol is registered at ClinicalTrials.gov under the number NCT05236205 and it was first posted on 01/21/2022 and last updated on 05/10/2022. [ABSTRACT FROM AUTHOR]

Objective. To describe the in vitro antimicrobial activity of the methanolic extract of Bixa orellana L. leaves against anaerobic bacteria associated to bacterial vaginosis and Lactobacillus spp. Materials and methods. Eight ATCC reference strains; Gardnerella vaginalis, Prevotella bivia, Peptococcus niger, Peptostreptococcus anaerobius, Mobiluncus curtisii, Atopobium vaginae, Veillonella parvula, and Lactobacillus crispatus, and twenty-two clinical isolates; eleven Gardnerella vaginalis and eleven Lactobacillus strains, were included in the study. The antimicrobial susceptibility was determined by the agar diffusion method. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined by using agar dilution and a modified dilution plating method, respectively. Results. All ATCC reference strains showed high levels of susceptibility to the extract, except P. vibia, V. parvula and L. crispatus. Interestingly, all G. vaginalis clinical isolates and the G. vaginalis ATTC strain were the most susceptible to the extract, given their low MIC (1.0 -- 2.0 mg/mL) and MBC (1.0 -- 4.0 mg/mL) values, whereas, the Lactobacillus spp. clinical isolates and the L. crispatus ATCC strain were the least susceptible bacteria given their high MIC (32.0 mg/mL) and MBC (≥ 32.0 mg/mL) values. Conclusions. In vitro experiments suggest that the extract possesses selective antimicrobial properties given its high activity against bacterial vaginosis-associated anaerobic bacteria and low activity against Lactobacillus species. [ABSTRACT FROM AUTHOR]

This work developd nanomaterials formulated from annatto seed oily extract (ASE), myristic acid (tetradecanoic acid), and their fatty acid esters. The annatto seed oily extract was obtained using only soybean oil (ASE + SO) and Brazil nut oil (ASE + BNO). The UV/VIS analysis of the oily extracts showed three characteristic peaks of the bixin molecule at 430, 456 and 486 nm. The lipid nanoparticles obtained using myristic acid and ASE + BNO or only BNO showed better results than the oil soybean extract, i.e., the particle size was <200 nm, PDI value was in the range of 0.2–0.3, and had no visual physical instability as they kept stable for 28 days at 4 °C. Lipid nanoemulsions were also produced with esters of myristic acid and ASE + BNO. These fatty acid esters significantly influenced the particle size of nanoemulsions. For instance, methyl tetradecanoate led to the smallest particle size nanoemulsions (124 nm), homogeneous size distribution, and high physical stability under 4 and 32 °C for 28 days. This work demonstrates that the chemical composition of vegetable oils and myristic acid esters, the storage temperature, the chain length of fatty acid esters (FAE), and their use as co-lipids improve the physical stability of lipid nanoemulsions and nanoparticles from annatto seed oily extract. [ABSTRACT FROM AUTHOR]

[Display omitted] • Ultrasound acoustic power (4.6 to 20 W) and extraction time (0.5 to 5 min) were evaluated. • The more intense ultrasound conditions allowed redder colorants consistent with higher bixin content. • Specific energies (0.02 to 0.20 kJ/g) affected the microstructure and X-ray diffractograms. • The extraction condition did not impact the colorant solubility. The interest in natural colorants derived from sustainable processes has prompted research into obtaining bixin from defatted annatto (Bixa orellana L.) seeds. Bixin is a compound that imparts yellow-orange-red coloration, known for its high biodegradability, low toxicity, and wide industrial applicability. Meanwhile, high-intensity ultrasound (HIUS) technology has emerged as a promising method for extracting natural colorants, offering higher yields through shorter processes and minimizing thermal degradation. Although some studies have demonstrated the efficiency of HIUS technology in bixin extraction, research on the effects of acoustic cavitation on the properties of the colorant remains limited. Therefore, this study aimed to investigate the influence of HIUS-specific energy levels (0.02, 0.04, 0.12, and 0.20 kJ/g) on the chemical, physical, and morphological characteristics of annatto extracts containing bixin and geranylgeraniol. Single-step extractions of bixin using ethanol as a solvent were evaluated at various acoustic powers (4.6, 8.5, 14.5, and 20 W) and extraction times (0.5, 1, 3, and 5 min) to determine their impact on the yield of natural colorant extraction. Increasing the acoustic power from 4.6 to 20 W and extending the extraction time from 0.5 to 5 min resulted in higher yields of natural colorant, likely due to the effects of acoustic cavitation and increased heat under more intense conditions. However, elevated levels of mechanical and thermal energy did not affect the chemical properties of the colorant, as indicated by UV–Vis and FTIR spectra. Conversely, higher specific energies yielded colorants with a more intense red hue, consistent with increased bixin content, and altered the microstructure and physical state, as observed in X-ray diffractograms. Nevertheless, these alterations did not impact the solubility of the colorant. Therefore, employing a cleaner extraction procedure aided by one-step ultrasound facilitated the recovery of natural colorants and contributed to the biorefining of annatto seeds, enabling the production of a rich geranylgeraniol colorant through a sustainable approach. [ABSTRACT FROM AUTHOR]

Tissue repair is a systemic process divided into different molecular, biochemical, and cellular steps. Alternatives that aid tissue repair are constantly being studied, as well as ways to avoid complications to the patient. Thus, it is important to highlight the therapeutic use of annatto, which has compounds with excellent therapeutic properties that allow a positive evolution in the repair of these lesions. The aim of this study was to conduct a literature review on the therapeutic potential of annatto compounds (Bixa orellana L.) in tissue repair. The research was carried out by consulting the online databases: Pubmed, Lilacs, BVS, EBSCO and Capes Journals. Works published in the last 10 years in Portuguese, English or Spanish were selected. The results were organized in a descriptive way, pointing out in each study the relevant data regarding the theme. The results showed that bioactive compounds, such as trochotrienols and carotenoids (bixin and norbixin), can be obtained from annatto extract. These substances have anti-inflammatory, antioxidant, antimicrobial, osteogenic and antiosteoporotic properties and are being used in the treatment of health problems. Therefore, it was concluded that compounds found in annatto (Bixa orellana L.) have therapeutic potential in the process of repairing skin and bone injuries. [ABSTRACT FROM AUTHOR]

This study describes the purification of Bixa orellana L. leaf lectin (BoLL) and the evaluation of its antimicrobial activity and in vivo toxicity. BoLL was isolated from saline extract of leaves through protein precipitation with ammonium sulfate (60% saturation) followed by ion exchange (CM-Sephadex) and size exclusion (Sephadex G-75) chromatographies. The lectin was characterized by polyacrylamide gel electrophoresis under denaturing conditions (SDS-PAGE), stability towards heating and pH variation, as well as influence of ions on its hemagglutinating activity (HA). HA inhibition assay by different carbohydrates and glycoproteins was also performed. The antimicrobial activity of BoLL was investigated against pathogens for humans and plants. In vivo acute toxicity in mice was assessed using a dose of 100 mg/kg per os or i.p. BoLL was isolated with high specific HA (purification fold: 53.89) and showed a single 19-kDa polypeptide band in SDS-PAGE. BoLL SHA increased in the presence of 20 mM Mg2+ ions, was resistant to heating at 100 °C, and stable at acidic pH range. HA inhibition was detected especially by glycoproteins, but also by monosaccharides. BoLL showed bacteriostatic and bactericidal effects against Escherichia coli [minimum inhibitory (MIC) and bactericidal (MBC) concentrations of 400 and 800 μg/mL, respectively] and Staphylococcus aureus (MIC and MBC: 400 μg/mL), as well as bacteriostatic effect against Bacillus megaterium and Micrococcus luteus (MIC: 18.2 and 800 μg/mL, respectively). No changes were observed in hematological, biochemical, and behavioral parameters in the acute toxicity test. However, animals treated with BoLL at 100 mg/kg per os or i.p. showed histopathological alterations in the liver, spleen, and kidneys. In conclusion, our data add new biotechnological value for B. orellana leaves, since they contain a thermostable lectin with antibacterial activity, which stimulate future studies aiming at its biomedical application. However, caution should be considered in future studies due to some signs of toxicity in mice. • A protocol for purification of a lectin from Bixa orellana leaf (BoLL) was defined. • BoLL showed a single 19-kDa polypeptide band in SDS-PAGE. • Hemagglutinating activity of BoLL increased in the presence of Mg2+ ions, was resistant to heating at 100 °C, and stable at acidic pH range. • BoLL showed bacteriostatic and bactericidal effects • BoLL (100 mg/kg per os or i.p.) did not cause the death of Swiss mice. [ABSTRACT FROM AUTHOR]

Background: Full chloroplast genomes provide high resolution taxonomic discrimination between closely related plant species and are quickly replacing single and multi-locus barcoding regions as reference materials of choice for DNA based taxonomic annotation of plants. Bixa orellana, commonly known as “achiote” and “annatto” is a plant used for both human and animal foods and was thus identified for full chloroplast sequencing for the Center for Veterinary Medicine (CVM) Complete Chloroplast Animal Feed database. This work was conducted in collaboration with the Instituto de Medicina Tradicional (IMET) in Iquitos, Peru. There is a wide range of color variation in pods of Bixa orellana for which genetic loci that distinguish phenotypes have not yet been identified. Here we apply whole chloroplast genome sequencing of “red” and “yellow” individuals of Bixa orellana to provide high quality reference genomes to support kmer database development for use identifying this plant from complex mixtures using shotgun data. Additionally, we describe chloroplast gene content, synteny and phylogeny, and identify an indel and snp that may be associated with seed pod color. Results: Fully assembled chloroplast genomes were produced for both red and yellow Bixa orellana accessions (158,918 and 158,823 bp respectively). Synteny and gene content was identical to the only other previously reported full chloroplast genome of Bixa orellana (NC_041550). We observed a 17 base pair deletion at position 58,399–58,415 in both accessions, relative to NC_041550 and a 6 bp deletion at position 75,531–75,526 and a snp at position 86,493 in red Bixa orellana. Conclusions: Our data provide high quality reference genomes of individuals of red and yellow Bixa orellana to support kmer based identity markers for use with shotgun sequencing approaches for rapid, precise identification of Bixa orellana from complex mixtures. Kmer based phylogeny of full chloroplast genomes supports monophylly of Bixaceae consistent with alignment based approaches. A potentially discriminatory indel and snp were identified that may be correlated with the red phenotype. [ABSTRACT FROM AUTHOR]

Objective: Bixa orellana is a plant from the Bixaceae family, for which, limited information is available on hepatoprotective properties. This study aimed at evaluating the protective effects of this plant on sub-acute acetaminophen (APAP)-induced liver injury in mice. Materials and Methods: Various aqueous extracts were prepared from roots, leaves, and barks. Albino mice were divided into six groups: a control group, an APAP group; a silymarin group (positive control) and three test groups. Mice were treated orally with APAP (250 mg/kg) followed 3 hr later by plant extracts, silymarin (50 mg/kg) or distilled water (10 ml/kg) administration once daily, for seven days. After treatment, animals were sacrificed, the liver was collected and different biochemical parameters were measured. Histological analyses were performed using hematoxylin/eosin staining and the qualitative phytochemical content of plant extracts was evaluated using conventional methods. Results: Administration of B. orellana barks decoction (250 mg/kg) significantly reduced alanine aminotransferase levels (p<0.001), unlike leaves and roots extracts. Moreover, the bark infusion had the highest activity compared to macerate and decoction. It significantly reduced malondialdehyde levels (p<0.001) and increased the levels of glutathione, superoxide dismutase and catalase, at doses of 250 and 500 mg/kg compared to the APAP group. A significant (p<0.001) decrease of tumor necrosis factor-a levels and leukocyte infiltration was found following treatment with bark infusion. The infusion content evaluation revealed the presence of polyphenols, saponins, tannins, sterols, anthraquinones, and coumarins and the absence of alkaloids. Conclusion: These results show that infusion from B. orellana barks is hepatoprotective against APAP-induced toxicity via antioxidant and anti-inflammatory mechanisms. [ABSTRACT FROM AUTHOR]

Background: Residual annatto seed meal (RASM) is a byproduct after extraction of bixin in the cosmetic industry and may be used in animal diets. Objective: Two experiments were carried out to determine the nutritive value and effect of RASM inclusion levels in the diets of Japanese quails. Methods: For the digestibility experiment, 160 Japanese quails were distributed in a randomized study with two treatments and 10 replicates with eight birds each. The treatments consisted of a reference diet and a test diet containing 75% of the reference diet and 25% of RASM. For the performance assay, 192 Japanese quails were used in a randomized study with six treatments and four replicates with eight birds each. Treatments consisted of a control corn-based diet (Contr1) and another diet based on sorghum + canthaxanthin (Contr2), and four diets based on sorghum with increasing inclusion levels of RASM (0, 3, 6, and 9%). Results: Digestibility of dry matter and crude protein, metabolization coefficient of gross energy, and retention of calcium and phosphorus were 41.05, 43.12, 47.97, 25.49, and 30.35%, respectively. RASM inclusion did not influence bird productivity; however, feed conversion worsened in treatments with 0-6% RASM inclusion relative to that in Contr1. Yolk color increased due to RASM treatment. Conclusion: RASM may be included in sorghum-based diets for quails at 9%, with improvement in yolk color and no negative effects on productivity and egg quality. [ABSTRACT FROM AUTHOR]

• Green synthesis of TiO 2 nanoparticles using natural Bixa orellana seed extract as capping and stabilizing agent. • Extensive enhancement in efficiency of DSSC with the use of as synthesized G-TNP compared to conventionally synthesized TiO 2. • Current conversion efficiency 3% achieved with use of G-TNP photoanode. Mesoporous anatase TiO 2 nanoparticles are the best photoanode materials for high performance DSSC. Chemical and physical methods used for the preparation of TiO 2 nanoparticles suffer with environment related issues e.g. use of toxic solvents, hazardous by-products and consumptions of high energy. We have synthesized mesoporous anatase TiO 2 nanoparticles from Titanium (IV) butoxide solution using Bixa orellana seed extract and further used it for making DSSC photoanode. The method offers environment friendly, cheaper and efficient DSSC. Structure, morphology, size and porosity of TiO 2 nanoparticles were characterized using XRD, SEM, TEM and N 2 sorption measurements. The amount of dye loading by TiO 2 was obtained using desorption technique which revealed much higher dye loading for the plant seed grown nanoparticles (G-TNP). TNP and G-TNP demonstrated photovoltaic conversion efficiencies 1.03% and 2.97% respectively. This study therefore demonstrates that the synthesized mesoporous anatase TiO 2 naoparticles can serve as promising photoanode materials for DSSC applications. [ABSTRACT FROM AUTHOR]

Background: Epilepsy and psychosis have a complex and interesting relationship. The drugs used in the treatment of epilepsy have the chances of reducing the symptoms and the risk of psychosis. Tardionally, seeds of Bixa Orellana L. were used to treat epilepsy in Brazilian tribes. Objective: To explore the anti-epileptic and anti-psychotic activity of bixin on experimental animals. Methodology: The anti-epileptic activity was assessed by pentylenetetrazol (PTZ) and maximal electro shock (MES)-induced convulsions. The effect of bixin against psychosis was assessed by an apomorphine-induced stereotyped behaviour in rats along with the climbing activities in mice. Bixin (2.5 and 5.0 mg/kg, p.o.) was pre-treated for a period of fifteen days. Bixin was administered one hour prior to the respective induction (MES/PTZ/apomorphine). After the induction, the assigned rodents were monitored for the different stages of convulsions as well as behavioural changes with respect to stereotyped and climbing behaviours in experimental animals. Result: Pre-treatment of bixin exhibited dose dependent prevention of seizures in the MES and PTZ models at P<0.001 significant level. The high dose of bixin showed complete inhibition of tonic-extension phase seizure and improved the mean time latency in the onset of tonic-clonic convulsions. The bixin pre-treatment also showed a significant (P<0.01) deterrence in stereotyped and climbing behaviour dose dependently. Conclusion: The present study provided the scientific footage to the traditional usage of the herb in the manifestation of convulsion and psychosis. [ABSTRACT FROM AUTHOR]

The effect of salt stress on pigment synthesis and antioxidant enzyme activity as well as in the genes involved in the biosynthetic pathway of bixin was studied. The 14-day germinated seedlings of Bixa orellana were induced into the various NaCl concentration (0, 25, 50, 75, 100 mM). After 45 days, leaves were taken for pigment analysis, antioxidant assays, and gene expression analysis to study the response of salt stress. The pigment content such as chlorophyll level was increased upon salt stress with a reduction in total carotenoid clearly indicating the adaptability of plants towards the stressed state. The level of β-carotene was increased in the highest concentration of salt stress treatment. The secondary metabolites such as bixin and abscisic acid (ABA) content were also high in elevated concentration of salt-treated seedling than control. The antioxidant enzyme activity was increased with the highest dose of salt stress suggesting the antioxidant enzymes to protect the plant from the deleterious effects. The mRNA transcript gene of the carotenoid biosynthetic pathway such as phytoene synthase (PSY), 1-deoxyxylulose-5-phosphate synthase (DXS), phytoene desaturase (PDS), beta-lycopene cyclase (LCY-β), epsilon lycopene cyclase (LCY-ε), carboxyl methyl transferase (CMT), aldehyde dehydrogenase (ADH), lycopene cleavage dioxygenase (LCD), and carotenoid cleavage dioxygenase (CCD) showed differential expression pattern under salt stress. In addendum, we studied the co-expression network analysis of gene to assess the co-related genes associated in the biosynthesis pathway of carotenoid. From the co-expression analysis result showed, the LCY, PDS, and PSY genes were closely correlated with other genes. These finding may provide insight to the plants to exist in the stress condition and to improve the industrially important pigment production. [ABSTRACT FROM AUTHOR]

Plant-parasitic nematodes are known as one of the most important pests attacking various plants in the world, and investigating the nematode component is very essential for management of this pest and prevent damage to plants in general. Our survey of plant-parasitic nematodes on medicinal plants in Me Linh Station for Biodiversity, a place for conservation of precious plants and animals in Vietnam, identified ten species that belong to nine genera, five families, and two orders of plant-parasitic nematodes parasitizing six medicinal plants. Excoecaria cochinchinensis was parasitized by the highest number of nematode genera (5 genera, including Xiphinema, Discocriconemella, Meloidogyne, Helicotylenchus, and Hemicriconemoides), while Hymenocallis littoralis was associated with the highest number of plant-parasitic nematodes (2060 nematodes/250g soil). The results also showed that Discocriconemella limitanea was found to be a dominant species with the highest number of individuals on 6 medicinal plants, and the genus Helicotylenchus had the highest frequency of appearance (5/6 plants or 83.3%). These nematodes caused symptoms such as yellowing leaves, root galls, and root lesions, which directly affect the quality and yield of medicinal plants. Based on the results, this study showed that plant-parasitic nematodes are a potential threat to the cultivation of medicinal plants in Me Linh Station for Biodiversity, and thus, control measures should be applied to ensure sustainable cultivation of medicinal plants in this place. [ABSTRACT FROM AUTHOR]

Background: Bixa orellana extracts are used to food coloring and it is important in pharmaceutical industry as a potential source of antioxidant activity. The application may be limited because it is unstable, but the process of inclusion in β-cyclodextrin (β-CD) by supercritical fluid CO2 (FSC-CO2) is an alternative to mediate this disadvantage. Objectives: we compared different conditions of temperature and pressure of FSC-CO2 extraction, to obtain a Bixa orellana extract with excellent antioxidant activity, and we evaluated the effect of inclusion in β-CD in the antioxidant capacity. Methods: extracts were obtained by FSC-CO2 with different conditions of pressure and temperature: (I) 3583 psi, 35°C, (II) 1413 psi, 35°C, (III) 2184 psi, 45°C, (IV) 5076 psi, 45°C, and (V) 2300 psi, 40°C. The antioxidant activity by DPPH assay was determined and the IC50 was evaluated. We performed the inclusion complex in β-CD with the highest antioxidant activity extract. The extract/β-CD complex was characterized by IR, DSC and NMR. This complexation was verified by comparative analysis of the spectra of the β-CD, free extract and β-CD/extract complex. The antioxidant capacity of this inclusion complex was evaluated. Results: the extract with highest antioxidant activity was obtained under the extraction condition IV, with an IC50 of 23.55 μg/mL, followed extract II (28.76 μg/mL), extract III (37.23 μg/mL), extract V (81.09 μg/mL) and extract I (193.82 μg/mL). Analysis showed significant differences (P<0.01) between these extracts. The spectra obtained by IR, DSC and NMR evidence the encapsulation process. The IC50 value of the extract/β-CD complex (104.84 μg/mL) was significantly higher than the value obtained for the pure extract (23.55 µg/mL). Conclusions: the highest antioxidant activity of Bixa orellana extracts was obtained by supercritical fluid pressure at 5076 psi and temperature of 45°C. Variations in IR, DSC and NMR spectra showed the inclusion of the β-CD/extract, and the IC50 values indicated the protective effect of β-CD to the reaction with DPPH radical. [ABSTRACT FROM AUTHOR]

Supercritical fluid extraction (SFE) coupled to low-pressure solvent extraction (LPSE) was studied for producing tocotrienols-rich oil and bixin-rich extract from annatto seeds. In the SFE step using CO2, 2.90 ± 0.01 g oil/100 g seeds was obtained, which was composed of 14.85 wt.% δ-tocotrienol and 2.06 wt.% γ-tocotrienol. In the LPSE step using ethanol, 11.3 ± 0.6 g extract/100 g defatted seeds was obtained, which was composed of 30 wt.% bixin. The process integration was also evaluated in the economic viewpoint with the objective to show its feasibility in different scenarios, which included different purchasing costs of seeds, costs of plants, processing times in the SFE step, bed height to internal diameter ratios in the SFE step, solvent type in the LPSE step, and selling prices of oil and extract fractions. Among 64 scenarios assessed, the more promising one provided values of cost of manufacturing (COM) of US$ 336.96/kg oil and US$ 12.05/kg bixin extract, payback time of 0.26 year, return on investment of 389.7% and gross margin of 66.5%. For a 100 L scale plant, 4968 kg oil and 7643 kg bixin can be recovered per year using approximately 220 tons of seeds. [ABSTRACT FROM AUTHOR]

The objective of this study was to correlate fruit epicarp colorimetry with bixin content in urucum seeds (Bixa orellana L.). Bixin is used in several industrial segments, such as pharmaceuticals, textiles, cosmetics and food. The fruits were harvested from adult trees and were classified visually in four maturation stages based on the external color of the epicarp. After visual segregation, the color of the fruit was measured with a digital color analyzer. The initial characterization was through seed morphometry and determination of water content. The experiment was completely randomized, with four replicates. Green fruits and light brown seeds showed larger seeds and bixin content. To obtain bixin the seeds can be harvested when the fruit is closed, with green and / or light brown or with reflectance values in the range of 93-162; 61-144; Or 35-62 nm on the red, green or blue scales, respectively, of the color spectrum. [ABSTRACT FROM AUTHOR]

The current study was undertaken to analyse the effect of short-term UV-B and UV-C radiations in provoking carotenoid biosynthesis in Bixa orellana . Seeds of B . orellana were germinated and exposed to the short term UV pre-treatment under controlled environmental condition for 5 days. The UV treated young seedlings response in pigment contents; antioxidant enzyme activity and mRNA gene expression level were analysed. The pigment content such as chlorophyll was increased in both UV-B and UV-C treated seedlings, but the total carotenoid level was decreased when compared to the control seedlings this can be attributed to the plant adaptability to survive in a stressed condition. The β-carotene level was increased in UV-B, and UV-C treated young seedlings. No significant changes have occurred in the secondary pigment such as bixin and ABA. The activity of the antioxidant enzymes such as catalase, peroxidase, and superoxide dismutase was significantly increased in UV-B treated seedlings when compared to the UV-C treated seedlings and control. The mRNA expression of the genes involved in bixin biosynthesis pathways such as DXS , PSY , PDS , LCY - β , LCY - ε , CMT , LCD , ADH and CCD genes showed different expression pattern in UV-B and UV-C treated young seedlings. Further we analysed the gene co-expression network to identify the genes which are mainly involved in carotenoid/bixin biosynthesis pathway. Form our findings the CCD , LCY , PDS , ZDS and PSY showed a close interaction. The result of our study shows that the short term UV-B and UV-C radiations induce pigment content, antioxidant enzyme activity and different gene expression pattern allowing the plant to survive in the oxidative stress condition. [ABSTRACT FROM AUTHOR]

The arils of Bixa orellana L. seeds contain carotenoid storage cells (CSCs). The main compounds in these cells include bixin and norbixin, which are important pigments in the food and pharmaceutical industries. Although many studies have been conducted on these chemical constituents, the cellular events that occur during the development of the carotenoid-accumulating cells in the arils and their relationship with the final carotenoid accumulation in the vacuoles remain unknown. In this study, the development of the CSCs in B. orellana arils was analyzed by light and transmission electron microscopy. Carotenoids formed in specialized cells, whose number and size increased during aril development. At various stages of development, the cytoplasm of the CSCs contained chromoplasts that held an extensive network of tubules and plastoglobules. Next to the chromoplasts, lipid droplets may fuse one another to form osmiophilic bodies. In addition, vesicles were observed next to the tonoplast. At the final stages of development, both the osmiophilic bodies and vesicles, which became quadrangular or rectangular, were stored in the vacuoles of the CSCs. This study reported for the first time the occurrence of different storage unit types within the vacuole of carotenoid storage cells. [ABSTRACT FROM AUTHOR]

Background: Bixin or annatto is a commercially important natural orange-red pigment derived from lycopene that is produced and stored in seeds of Bixa orellana L. An enzymatic pathway for bixin biosynthesis was inferred from homology of putative proteins encoded by differentially expressed seed cDNAs. Some activities were later validated in a heterologous system. Nevertheless, much of the pathway remains to be clarified. For example, it is essential to identify the methylerythritol phosphate (MEP) and carotenoid pathways genes. Results: In order to investigate the MEP, carotenoid, and bixin pathways genes, total RNA from young leaves and two different developmental stages of seeds from B. orellana were used for the construction of indexed mRNA libraries, sequenced on the Illumina HiSeq 2500 platform and assembled de novo using Velvet, CLC Genomics Workbench and CAP3 software. A total of 52,549 contigs were obtained with average length of 1,924 bp. Two phylogenetic analyses of inferred proteins, in one case encoded by thirteen general, single-copy cDNAs, in the other from carotenoid and MEP cDNAs, indicated that B. orellana is closely related to sister Malvales species cacao and cotton. Using homology, we identified 7 and 14 core gene products from the MEP and carotenoid pathways, respectively. Surprisingly, previously defined bixin pathway cDNAs were not present in our transcriptome. Here we propose a new set of gene products involved in bixin pathway. Conclusion: The identification and qRT-PCR quantification of cDNAs involved in annatto production suggest a hypothetical model for bixin biosynthesis that involve coordinated activation of some MEP, carotenoid and bixin pathway genes. These findings provide a better understanding of the mechanisms regulating these pathways and will facilitate the genetic improvement of B. orellana. [ABSTRACT FROM AUTHOR]

Natural dyes with highly delocalized π-electron systems are considered as promising organic materials for nonlinear optical applications. Among these dyes, Bixa Orellana dye with extended π-electron delocalization is one of the most attractive dyes. Bixa Orellana dye-doped Poly(methyl methacrylate) (PMMA) thin films were prepared through spin coating process for linear and nonlinear optical properties investigation. Atomic force microscopy (AFM) was used to evaluate the roughness of the thin films. The optical constants n and k were evaluated by ellipsometric spectroscopy. The refractive index had a maximum of about 1.456 at 508.5, 523.79 and 511.9 nm, while the maximum of k varies from 0.070 to 0.080 with the thickness. The third order nonlinear optical properties of the hybrid Bixa Orellana dye-PMMA polymer were investigated under 30 ps laser irradiation at 1064 nm with a repetition rate of 10 Hz. In particular the third-order nonlinear susceptibility has been determined by means of the Maker Fringes technique. The nonlinear third order susceptibility was found to be 1.00 × 10 −21 m 2 V −2 or 0.72 × 10 −13 esu. Our studies provide concrete evidence that the hybrid-PMMA composites of Bixa dye are prospective candidates for nonlinear material applications. [ABSTRACT FROM AUTHOR]

Natural dyes are receiving increasing attention from researchers and manufacturers, given its perceived eco-friendly nature. Yet, adjunct agents known as mordants that help bond the molecules of the fabric being dyed and the colorant used to dye it are often toxic. There is scant published information describing the dyeing potential and toxicity of colorant-bearing plants and the dye uptake with respect to the mordant treatments. A preliminary survey based on a range of sources of information showed that of the Yucatan peninsula harbors 23 plants showing dyeing properties. Four of them ( Justicia spicigera , Bixa orellana , Bougainvillea glabra and Rhoe discolor ) were selected to extract their natural dyes, which were chemical and toxically characterized. B. orellana was successfully employed to dye the fabric, viz. ordinary cotton cloth, manta. The dye baths without mordant only present low toxicity, the J . spicigera and B . glabra dyes being the most toxic. The B . orellana dye was less toxic, although its toxicity is increased when mordants are used. Regarding color performances of the dyed fabrics, mordanting influenced the depth of the shades, improving dyeing and light fastness. Nevertheless, mordants had no significant effects on color values. [ABSTRACT FROM AUTHOR]

A process to obtain tocotrienols from annatto seeds was studied in a supercritical fluid extraction (SFE) system containing two 1 L extractor vessels with height ( H B ) to internal diameter ( D B ) ratios of 7.1 (E-1) and 2.7 (E-2). Pseudo continuous operation of the SFE system was validated by producing clove volatile oil at 40 °C and 15 MPa. To obtain tocotrienols, the system was operated in pseudo continuous mode at 40 °C, 20 MPa and CO 2 flow rate of 20.7 g/min. The extraction time, from a technical perspective, was set to 60 min, according to kinetic parameters obtained from an overall extraction curve (OEC) with processing time of 360 min. OECs were initially measured in batch mode (BM) and the kinetic parameters for the constant extraction rate (CER) period were determined to be lengths of the CER period ( t CER ) equal to 43 ± 1 and 42 ± 2 min for E-1 and E-2, respectively; mass transfer rates ( M CER ) of 0.22 ± 0.01 g/min for both E-1 and E-2; and total yields ( R CER ) of 50 ± 1 and 49 ± 1 g of extract/100 g of extractable mass for E-1 and E-2, respectively. No differences were observed between each group of kinetic parameters ( t CER ; M CER ; R CER ) obtained in BM. Following the measurement of OECs, the SFE system was operated in pseudo continuous mode for four extraction stages of 60 min each with the extractors organized in parallel. This organization improved the productivity of the process. We also proposed the integration of SFE with low-pressure solvent extraction (LPSE) using water and ethanol as solvents to remove bixin from the defatted annatto seeds. LPSE conditions were: 60 °C and solvent mass to feed mass ratio ( S/F ) of 8 g solvent/g annatto for ethanol and 50 °C and S/F of 8 g solvent/g annatto for water. LPSE assays were performed in an agitated bed over a 95 min time period. Using partially defatted annatto seeds as the feedstock for LPSE resulted in increased bixin extraction. LPSE with ethanol was found to increase the bixin content in the extract. Thus, the proposed process integration appears to be useful, mainly due to three aspects: (1) the initial removal of the oil layer via supercritical CO 2 favors bixin extraction, (2) LPSE is a low-cost methodology for extracting bixin and (3) the cost of raw material in the second step is null. [ABSTRACT FROM AUTHOR]

[Display omitted] • Norbixin (NBIX) inhibits human LDL oxidation in vitro. • NBIX reduces triglycerides and increases HDL levels of atherogenic rabbits (AR) • NBIX reduces oxidized LDL and autoantibodies levels in AR. • NBIX inhibits lipid and protein oxidation in the aortic tissue of AR. • NBIX counteracts changes in endogenous antioxidant defense systems of AR. We hypothesized that norbixin, which is a carotenoid used as an orange/red natural food coloring additive, has anti-atherogenic properties. An in vitro oxidation assay with human LDL and a rabbit model of atherosclerosis were used to test this hypothesis. Norbixin inhibited the oxidation of isolated human LDL in a concentration-dependent manner. In the in vivo assay, rabbits were fed with a regular chow (control) or an atherogenic diet (0.5% cholesterol) alone or supplemented with norbixin (10, 30 or 100 mg/kg b.w.) for 60 days. Norbixin supplementation (30 and 100 mg/kg b.w.) increased HDL levels and reduced triglyceride levels and the atherogenic index of rabbits. This effect was associated with the decrease of serum levels of oxidized LDL, oxidized LDL antibodies and aortic tissue levels of lipid and protein oxidation in the atherogenic rabbits supplemented with norbixin. Atherogenic diet increased enzymatic (superoxide dismutase, catalase, glutathione reductase, and thioredoxin reductase-1) and non-enzymatic (non-protein thiol groups content) antioxidant defense systems in the aortic tissue but reduced the activity of paraoxonase-1 in the serum. All these changes were prevented by norbixin supplementation (10, 30 and 100 mg/kg b.w.). These results suggest that norbixin has atheroprotective potential by improving serum lipid profile and preventing oxidative modifications of circulating LDL and aortic tissue. Norbixin may, therefore, be beneficial in the control of atherosclerosis risk factors and can be further investigated as a candidate to be used not only as a functional food ingredient but also for therapeutic applications and in the nutraceutical industry. [ABSTRACT FROM AUTHOR]

Abstract: Bixin is the major carotenoid in the seed of the Annatto plant (Bixa orellana L.). The aim of this study was to obtain extracts containing bixin from seeds that had been partially defatted by supercritical fluid extraction. Pressurized liquid extraction (PLE) and low-pressure solvent extraction (LPSE) methods were used, and the effects of the solvent, temperature, pressure, solvent mass to feed mass (S/F) ratio and ultrasonication were evaluated for the global yield (X 0(%)) and the bixin yield (BY(%)). Extraction conditions producing high yields of bixin were established for both the PLE and LPSE methods. Analysis of variance was used to examine the influence of the individual extraction variables in LPSE and PLE. For LPSE; significant effects were found for solvent, temperature, and the interactions of temperature with solvent and temperature with S/F. Solvent was the only variable that significantly affected X 0(%) and BY(%), for PLE. While ultrasonication did not significantly affect X 0(%) or BY(%), scanning electron microscopy analysis revealed structural changes in the vegetal matrix following this treatment. [Copyright &y& Elsevier]

Highlights: [•] Bixin has no genotoxic effect in liver and colon cells by comet assay. [•] Bixin significantly reduced the DNA damage in hepatocytes. [•] Bixin not reduce the frequency of DNA damage in colon cells. [•] Bixin not suppress the formation of pre-neoplastic lesions in colon. [ABSTRACT FROM AUTHOR]

Ulcerative colitis is an idiopathic inflammatory bowel disease characterized by tissue damage, diarrhea, anemia, and loss of body weight. Tissue damage occurs as a result of uncontrolled activation of the immune response and an increase in free radicals, which have a strong effect on the pathogenesis of inflammatory bowel disease. The incidence and prevalence of this inflammatory disease continue to increase worldwide. Maceration of Bixa orellana leaves in palm wine is used in traditional medicine to treat diarrhea, dysentery, and hemorrhoids in the Adamaoua region of Cameroon. The present work evaluated the preclinical effects (ie, antioxidant, hematological, and histological activities) of the hydroethanolic extract of Bixa orellana leaves in an in vivo, rat acetic acid-induced ulcerative colitis model. Thirty-six female rats weighing between 165 and 180 g were fasted for 18 hours and then anesthetized with ether. A dose of 1 mL acetic acid (5%) was administered rectally through a catheter in all rats except the normal control group, which received distilled water (1 mL) instead. Treatments began 48 hours after rectal administrations of acetic acid or water, and all animals were treated twice daily for 7 days. The normal control group and the colitis control group received PO distilled water (10 mL/kg), the positive control received orally loperamide (5 mg/kg, and the 3 test groups received orally the hydroethanolic extract of Bixa orellana at 100, 200, and 400 mg/kg, respectively. During treatment, the number of diarrheal stools and weight change were assessed. At the end of the treatment, the animals were put to death under ether anesthesia. Blood was collected postmortem for evaluation of hematological and antioxidant parameters. The abdomen was opened via a midline incision and the colon was removed and emptied of all contents to assess histological and antioxidant parameters. During treatment, the number of diarrheal stools was significantly decreased from day 3 in animals treated with 100 (P < 0.05), 200 (P < 0.05), and 400 (P < 0.01) mg/kg extract compared with the colitis control group. The change in body weight of all extract-treated rats decreased significantly from day 3 (–5.55%; P < 0.05) to day 8 (–13.80%; P < 0.01) compared with the normal control. In the colitis control, this change ranges from –6.15% on day 2 to –15.13% on day 8. Extract treatment with 100, 200, and 400 mg/kg significantly reduced (P < 0.05) the number of lesions and the relative weight of the colon. The levels of red blood cells, neutrophils, and total white blood cells decreased in the colitis control group, whereas treatment with the extract at doses of 100, 200, and 400 mg/kg was associated with a significant increase in these hematological parameters. Catalase and superoxide dismutase activity and glutathione concentrations all increased significantly (P < 0.01) in blood and colon in all extract-treated animals, whereas levels of malondialdehyde and nitric oxide were significantly decreased (P < 0.01) compared with the colitis control animals. The hydroethanolic extract of Bixa orellana leaves had protective effects against acetic acid-induced ulcerative colitis in rats that was associated with inhibited production of free radicals believed to be responsible for oxidative stress, hematological disorders, and tissue damage in this animal model. [ABSTRACT FROM AUTHOR]

Bixa orellana (annatto or lipstick tree) is a perennial tropical plant that stores and produces considerable quantities of the apocarotenoid bixin, a culturally and economically important pigment used worldwide. However, the mechanisms underlying the gene regulation and pigment accumulation of bixin and carotenoids in annatto remain unknown. Bixin is present in the different tissues of the plant, although this pigment is primarily accumulated in the seed coat. Thus, the leaves are useful organs for understanding carotenoid and bixin production, thereby facilitating the study of this pigment, which would otherwise be difficult in ligneous adult plants. To study the regulation of bixin synthesis and to determine which genes are important regulatory molecules at the transcription level, the herbicide norflurazon (NF) was used to block carotenoid synthesis and bixin concentrations in B. orellana leaves. The genes activated in the early stages of the carotenoid pathway are involved in lycopene production ( dxs, psy and pds), and those induced in the later stages of the carotenoid pathway, such as β and ε-lycopene cyclases and boccd1, were differentially expressed compared with the control. The expression of some genes was more susceptible to certain concentrations of NF, potentially reflecting the roles of these genes in carotenoid synthesis in B. orellana. These results suggest that apocarotenoids, such as bixin, are synthesized using alternative precursors through the actions of genes that have not yet been identified. [ABSTRACT FROM AUTHOR]

The pigment extraction process using annatto (Bixa orellana L.) seed produces a large amount of seed waste. Although most of these seeds are discarded, a number of studies report promising results with their use in animal feed. The good fiber content also suggests human nutrition applications, with possible incorporation in dietary foods. In the present study, annatto seeds derived from color extraction were dried, with and without the layer of oil left over from the process. Seeds were dried at 40, 50, 60 and 70 °C. Drying data were fitted to the Diffusion Approximation, Two Term, Midilli, Page and Thompson models. Drying was carried out up to a moisture content of approximately 5% wet basis. All the models studied exhibited adequate fit to the drying kinetics data of the annatto seeds, with coefficients of determination above 0.98 and root mean squared error (RMSE) below 1.0. Seeds with oil had longer drying times at 40 and 50 °C and shorter times at 60 and 70 °C. The coefficients of diffusion showed values between 2.67 x 10-11and 9.50 x 10-11m2 s-1 and between 2.7 x 10-11 and 6.21 x 10-11 m² s-1, while activation energies for liquid diffusions were 38.04 and 23.52 kJ mol-1, for residual seed drying with and without oil, respectively. [ABSTRACT FROM AUTHOR]

Annatto ( Bixa orellana L.) is an important dye-yielding medium-sized tree occurring in the tropics. Investigations aimed at elucidating the reproductive characters as well as establishing a set of breeding parameters for genetic improvement of B. orellana was carried out. Records on pollen/ovule ratios of annatto showed that this species is xenogamous. The acetolyzed pollen of B. orellana was tricolporate. The fluorochromatic test showed 95% pollen viability. The viability of the pollen grains decreased sharply after 4 h from anther dehiscence. The optimum time range for artificial pollination was standardized between 12:00 and 13:00 h. A high in vitro germination rate (92%) of freshly collected pollen was recorded in modified Brewbaker's medium with 15% sucrose. Resource allocation studies showed that the construction of each flower cost 1.162 g. The floral vegetative cost was 75%. Of the total allocation, 21% was spent by the stamens. Based on identified reproductive characters, controlled pollination experiments were conducted by selecting four morphologically distinct parent plants: (1) green-fruited with reddish spines, (2) red-fruited, (3) brown large-fruited, and (4) with brown distorted fruits. In artificial cross-pollination experiments, wide variation ranges in fruit set (0-95%) and fruit maturation (0-85%) were observed in different crosses. The number of seeds per capsule also showed variation (0-40 per capsule). Germination studies of seeds raised from different crosses showed a maximum 93% germination. The present study proved that improvement in B. orellana could be achieved by adopting breeding techniques using specifically selected parents. [ABSTRACT FROM AUTHOR]

Carotenoid cleavage dioxygenases (CCDs) are a class of enzymes involved in the biosynthesis of a broad diversity of secondary metabolites known as apocarotenoids. In plants, CCDs are part of a genetic family with members which cleave specific double bonds of carotenoid molecules. CCDs are involved in the production of diverse and important metabolites such as vitamin A and abscisic acid (ABA). Bixa orellana L. is the main source of the natural pigment annatto or bixin, an apocarotenoid accumulated in large quantities in its seeds. Bixin biosynthesis has been studied and the involvement of a CCD has been confirmed in vitro. However, the CCD genes involved in the biosynthesis of the wide variety of apocarotenoids found in this plant have not been well documented. In this study, a new CCD1 gene member (BoCCD1) was identified and its expression was charaterized in different plant tissues of B. orellana plantlets and adult plants. The BoCCD1 sequence showed high homology with plant CCD1s involved mainly in the cleavage of carotenoids in several sites to generate multiple apocarotenoid products. Here, the expression profiles of the BoCCD1 gene were analysed and discussed in relation to total carotenoids and other important apocarotenoids such as bixin. [ABSTRACT FROM PUBLISHER]

Abstract: Bixa orellana L. (annatto), from Bixaceae family, is a native plant of tropical America, which accumulates several carotenoids (including bixin and norbixin), terpenoids, tocotrienols and flavonoids with potential antioxidant activity. In the present study, the in vitro scavenging capacity of annatto seed extracts against reactive oxygen species (ROS) and reactive nitrogen species (RNS) was evaluated and compared to the bixin standard. Annatto extracts were obtained using solvents with different polarities and their phenolic compounds and bixin levels were determined by high performance liquid chromatography coupled to diode array detector. All annatto extracts were able to scavenge all the reactive species tested at the low μg/mL range, with the exception of superoxide radical. The ethanol:ethyl acetate and ethyl acetate extracts of annatto seeds, which presented the highest levels of hypolaetin and bixin, respectively, were the extracts with the highest antioxidant capacity, although bixin standard presented the lowest IC50 values. [Copyright &y& Elsevier]

nnatto ( Bixa orellana) is the sole source of the commercially important pigment bixin. The broad diversity of annatto phenotypes and the wide variation in pigment contents have restricted its use. Therefore, genetic improvement focusing on this variation is important in order to achieve optimal exploitation of annatto. To characterize and understand some of the genetic basis governing carotenoid production, two annatto cultivars with either pink or white flowers were selected on the basis of their contrasting pigment levels. Several complementary DNA (cDNA)-probes corresponding to genes involved in carotenoid biosynthesis ( dxs, psy, pds, β-lcy, and ε-lcy) were cloned and analyzed by reverse transcriptase-polymerase chain reaction (RT-PCR) in different tissues of these two annatto cultivars during flower and fruit development. In general, expression of most selected genes could be associated with pigment overaccumulation in these cultivars. The pink cultivar exhibited higher carotenoid contents and higher pigment-related gene expression, and the two variants analyzed showed notable differences in gene expression in the highest pigment producing tissues. [ABSTRACT FROM AUTHOR]

Objective: The present study was conducted to assess the anti-inflammatory effect of a crude aqueous extract of Bixa orellana leaves (AEBO) and to examine the possible involvement of nitric oxide (NO) in its anti-inflammatory mechanism. Materials and Methods: The air-dried, powdered leaves were soaked in distilled water (1:20 w/v) at 50°C for 24 h and the supernatant obtained was freeze-dried (yield 8.5% w/w). The dosage was recorded as the mass of extract per kg b.w. of rats in all inflammatory assays (bradykinin-induced paw edema, peritoneal vascular permeability and NO assay). Results: Pretreatment with AEBO for 4 consecutive days exhibited significant inhibitory activity against inflammatory models, the bradykinin-induced hind paw edema model and bradykinin-induced increased peritoneal vascular permeability at both doses in dose-dependent manner. In addition, AEBO was also found to significantly suppress the production of NO at doses of 50 and 150 mg/kg. Conclusion: This study provides scientific data to support the traditional use of B. orellana leaves in treating inflammation. Results from this study suggest that AEBO exerts anti-inflammatory effects. Part of this anti-inflammatory effect may be associated with its antibradykinin activity and may be related to a reduction of the NO production. Copyright © 2011 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Abstract: Bixin is commercially important and increasingly in demand as an apocarotenoid pigment derived from the seed coats of the tropical shrub Bixa orellana. It has been widely used in food and cosmetics for a very longer time. Recently, the biosynthetic pathway of bixin has been elucidated and three genes from B. orellana govern bixin synthesis from the precursor lycopene. These genes code for lycopene cleavage dioxygenase, bixin aldehyde dehydrogenase, and norbixin carboxyl methyltransferase, which catalyze the sequential conversion of lycopene into bixin. Based on available chemical data and by analogy to biosynthetic pathway for bixin, comparative genome sequence analysis has been carried out. We have identified and functionally characterized that bixin coding genes are not only present in Bixa, but also in Crocus and Vitis. Our chromatographic study based on TLC, FT-IR and GC–MS shows evidence of presence of bixin in the other two organisms. This is the first report and Crocus and Vitis could be an alternative and competitive source for natural bixin production. [Copyright &y& Elsevier]

Various traditional systems of medicine enlightened the importance of the leaves of Bixa orellana (Bixaceae) to have a great medicinal value. The present study was aimed at pharmacognostic and preliminary phytochemical evaluations of B. orellana leaves. The pharmacognostic investigations were carried out in terms of organoleptic, microscopic and physical parameters. The dried leaves were subjected to successive Soxhlet extraction using petroleum ether, chloroform, ethyl acetate, methanol and water. These solvent extracts were subjected to a preliminary phytochemical screening to detect the different chemical principles present viz., carbohydrates, proteins, amino acids, steroids, glycosides, alkaloids, tannins and phenolic compounds. The phytochemical evaluation revealed the presence of carbohydrates, steroids, alkaloids, proteins, flavonoids, terpenoids, phenolics, tannins and glycosides. [ABSTRACT FROM AUTHOR]

Some significant compounds present in annatto are geranylgeraniol and tocotrienols. These compounds have beneficial effects against hyperlipidemia and chronic diseases, where oxidative stress and inflammation are present, but the exact mechanism of action of such activities is still a subject of research. This study aimed to evaluate possible mechanisms of action that could be underlying the activities of these molecules. For this, in silico approaches such as ligand topology (PASS and SEA servers) and molecular docking with the software GOLD were used. Additionally, we screened some pharmacokinetic and toxicological parameters using the servers PreADMET, SwissADME, and ProTox-II. The results corroborate the antidyslipidemia and anti-inflammatory activities of geranylgeraniol and tocotrienols. Notably, some new mechanisms of action were predicted to be potentially underlying the activities of these compounds, including inhibition of squalene monooxygenase, lanosterol synthase, and phospholipase A2. These results give new insight into new mechanisms of action involved in these molecules from annatto and Chronic®. [ABSTRACT FROM AUTHOR]

The plants with insecticide activities constitute a main compound of integrated pest management. Under this premise, the aim of the current research was to evaluate mortality and repellence of Eupalamides cyparissias Fab. (Lepidoptera: Castniidae) larvae, pest of oil palm Elaeis guineensis Jacquin, employing ten plants with insecticide potential: Indian heliotrope (Heliotropium indicum L., Boraginaceae), Angel's trumpets (Brugmansia x candida Pers., Solanaceae), Wandering Jew (Tradescantia zebrina Hort ex Bosse, Commelinaceae), Nettles-purge (Jathropa curcas L., Euphorbiaceae), Soapberry (Paullinia clavigera Schltdl., Sapindaceae), Red spurge (Euphorbia cotinifolia L., Euphorbiaceae), Annato (Bixa orellana L., Bixaceae), Golden shower (Cassia fistula L., Fabaceae), Birthwort fruit (Aristolochia pilosa Kunth, Aristolochiaceae) and Pareira (Chondrodendron tomentosum Ruiz & Pavon, Menispermaceae). Bioessays with E. cyparissias were performed at 1 h and 24 h under standardized laboratory conditions. At 24 h exposure, the highest percentage of mortality of E. cyparissias was observed with Soapberry (63.3%: bark and leaves in decoction), Annato (63.3%: liquefied seeds) and Red spurge (48.3%: liquefied leaves). In the case of repellence, the highest effects were found in Annato (83.3%), Soapberry (75%) and Angels' trumpets (66.7%: liquefied leaves). [ABSTRACT FROM AUTHOR]

The plants with insecticide activities constitute a main compound of integrated pest management. Under this premise, the aim of the current research was to evaluate mortality and repellence of Eupalamides cyparissias Fab. (Lepidoptera: Castniidae) larvae, pest of oil palm Elaeis guineensis Jacquin, employing ten plants with insecticide potential: Indian heliotrope (Heliotropium indicum L., Boraginaceae), Angel's trumpets (Brugmansia x candida Pers., Solanaceae), Wandering Jew (Tradescantia zebrina Hort ex Bosse, Commelinaceae), Nettles-purge (Jathropa curcas L., Euphorbiaceae), Soapberry (Paullinia clavigera Schltdl., Sapindaceae), Red spurge (Euphorbia cotinifolia L., Euphorbiaceae), Annato (Bixa orellana L., Bixaceae), Golden shower (Cassia fistula L., Fabaceae), Birthwort fruit (Aristolochia pilosa Kunth, Aristolochiaceae) and Pareira (Chondrodendron tomentosum Ruiz & Pavon, Menispermaceae). Bioessays with E. cyparissias were performed at 1 h and 24 h under standardized laboratory conditions. At 24 h exposure, the highest percentage of mortality of E. cyparissias was observed with Soapberry (63.3 %: bark and leaves in decoction), Annato (63.3 %: liquefied seeds) and Red spurge (48.3 %: liquefied leaves). In the case of repellence, the highest effects were found in Annato (83.3 %), Soapberry (75 %) and Angels' trumpets (66.7 %: liquefied leaves). [ABSTRACT FROM AUTHOR]

A protocol for direct organogenesis in Bixa orellana (pink flowers variety) has been developed with significant organogenic response from rooted hypocotyls, hypocotyl segments, and cotyledonary leaf explants on Murashige and Skoog (MS) medium supplemented with 2.0 mg L-1 thidiazuron and 0.25% coconut water or 7.0 mg L-1 N6-Benzyladenine and 0.1 mg L-1 α-naphthalene acetic acid. Thidiazuron in combination with coconut water promoted higher organogenic response in rooted hypocotyls. Similarly direct organogenesis was noticed from hypocotyls, cotyledonary leaf explants and shoot tip explants on half MS medium with 0.25 mg L-1 N6-Benzyladenine (BA) and 0.5 mg L-1 indole-3-acetic acid (IAA). Best shoot elongation of shoot buds was achieved in the presence of 1.5 mg L-1 (BA) + 1.0 mg L-1indole-3-butyric acid (IBA). The in vitro rooting of microshoots was good in the presence of 3.0 mg L-1 (IBA). Seventy percent of rooted plants survived after acclimatization. [ABSTRACT FROM AUTHOR]

Abstract: Preliminary pharmacological studies were performed on the methanol extract of Bixa orellana L. (Bixaceae) leaves to investigate neuropharmacological, anticonvulsant, analgesic, antidiarrhoeal activity and effect on gastrointestinal motility. All studies were conducted in mice using doses of 125, 250 and 500mg/kg of body weight. In the pentobarbitone-induced hypnosis test, the extract statistically reduced the time for the onset of sleep at 500mg/kg dose and (dose-dependently) increased the total sleeping time at 250 and 500mg/kg dose. A statistically significant decrease in locomotor activity was observed at all doses in the open-field and hole-cross tests. In the strychnine-induced anticonvulsant test, the extract increased the average survival time of the test animals (statistically significant at 250 and 500mg/kg). The extract significantly and dose-dependently reduced the writhing reflex in the acetic acid-induced writhing test. Antidiarrhoeal activity was supported by a statistically significant decrease in the total number of stools (including wet stools) in castor oil-induced diarrhoea model. A statistically significant delay in the passage of charcoal meal was observed at 500mg/kg in the gastrointestinal motility test. The extract was further evaluated in vitro for antioxidant and antibacterial activity. It revealed radical scavenging properties in the DPPH assay (IC50 =22.36μg/ml) and antibacterial activity against selected causative agents of diarrhoea and dysentery, including Shigella dysenteriae. [Copyright &y& Elsevier]

Abstract: Annatto (Bixa orellana L.) is a native species of the Neotropics. It is commercially grown for the high apocarotenoid pigment content in its seed coat, particularly bixin (BXN) (9′Z-6,6′-diapocarotene-6,6′-dioate). Bixin is a natural orange-red pigment used primarily as a coloring agent in dairy products, baked goods, vegetable oils, and drinks. Large-scale propagation of annatto is problematic because it is cross-pollinated, highly heterozygous, and has heterogeneous bixin content. Another drawback is dehiscence in dry capsules in high bixin content varieties. Previous research on B. orellana does not address the biochemical or molecular aspects of apocarotenoid content or procedures for artificial pollination of Mexican variants. In an initial effort toward genetic improvement, preliminary studies were done to begin developing procedures for artificial pollination of annatto, understand its reproductive biology, and investigate the bixin and carotenoid content in the seeds of variants selected based on flower and fruit morphological traits. In the state of Yucatan, Mexico; annatto begins flowering in September, but the optimum period for artificial pollination and fruit set is October and November. Under natural conditions an average of 32±5.3 seeds per fruit are produced. Pistil receptivity lasts approximately 4h and peaks between 08:00 and 12:00h. Based on in vitro germination, average pollen viability is about 86%, and pollen longevity declines by about 50% 24h after release from anther. Significant differences in carotenoids and bixin contents in mature seeds were observed between the studied annatto variants. The pink flower variant had the highest contents of both pigments, and one of the white flower variants (‘peruana verde’) had the lowest. [Copyright &y& Elsevier]