Brazilin cream from Caesalpinia sappan inhibit periodontal disease: in vivo study.

Background: Gingivitis is an inflammation of the gums that is the initial cause of the development of periodontal disease by the activity of Nuclear Factor-kappa B (NF-κB), Interleukin-1ß (IL-1ß), Interleukin-6 (IL-6), p38, and Tumor Necrosis Factor-α (TNF-α). Unaddressed chronic inflammation can lead to persistent disturbances in other parts of the body. Brazilin is a naturally occurring plant chemical that may have antibacterial and anti-inflammatory effects. Treatment based on the natural plant compound, brazilin, is developed in the form of a topical cream for easy application. Objective: The aim is to develop the natural compound brazilin in the form of a topical cream as an anti-inflammatory agent to reduce NF-κB expression through Imunohistochemistry (IHC) methods, and the expression of pro-inflammatory genes IL-1ß, IL-6, p38, and TNF-α. Methods: Male Sprague-Dawley rats were induced with gingivitis using P. gingivalis bacteria. The observed groups included rats treated with a single application of brazilin cream and rats treated with two applications of brazilin cream. The treatment was administered for 15 days. On days 3, 6, 9, 12, and 15, anatomical wound observations and wound histology using hematoxylin-eosin and Masson's Trichrome staining were performed. NF-κB protein expression was analyzed using the IHC method. Gingival inflammation gene expression of NF-κB, IL-1ß, IL-6, p38, and TNF-α was measured using q-RTPCR. Results: Single and double applications of brazilin cream increased angiogenesis and decreased NF-κB protein expression, in addition to the IL-1ß, IL-6, p38, and TNF-α gene expressions. Conclusion: In a rat gingivitis model, Brazilin cream may function as an anti-inflammatory agent in the gingival tissue.

Strawberries treated with biodegradable film containing plant extracts.

Strawberry (Fragaria x ananassa Duch.) is a highly perishable fruit whose characteristics make it susceptible to developing microorganisms. Plant extracts have been studied as an alternative to pesticides to control spoilage microorganisms, responding to the expectation of the population seeking a healthier way of life. The fungus Botrytis cinerea is a facultative pathogen of vegetables, which can affect all stages of the development of several fruits, such as the strawberry, where it causes gray rot. Trichilia catigua (catuaba), Paullinia cupana (guarana), Stryphnodendron barbatiman (barbatimão), and Caesalpinia peltophoroides (sibipiruna) are planted in the Brazilian flora and have demonstrated pharmacological properties in their extracts. This work aimed to treat strawberries with a biodegradable film containing extracts of these species to evaluate strawberry conservation. There were notable distinctions in mass loss between the extract-treated and control samples. The pH, total acidity (TA), and soluble solids parameters exhibited consistently significant means across both sets of samples. Luminosity increased over the course of days in the color parameters, with the exception of strawberries coated with guarana. The red color showed greater intensity, except for those coated with barbatimão extract. Considering the results, it is possible to conclude that the coatings used can become an alternative to enhance the conservation of strawberries.

The potential of galactomannan from Caesalpinia ferrea on erosive dentin wear reduction in vitro.

Gels containing juca seed galactomannan (JSG) were evaluated for their potential to prevent the progression of dentin erosive wear in an in vitro study with four experimental groups (n = 9). The treatments included distilled water (DW), 0.05% stannous fluoride (121 ppm F), and 0.5% or 1% JSG. The specimens underwent a cycle (3 times/day) consisting of immersion in 1% citric acid (5 minutes), treatment (5 minutes), and artificial saliva exposure (2 hours/overnight) for 5 days. Surface changes were assessed using mechanical profilometry (wear), scanning electron microscopy (SEM), and energy-dispersive X-ray spectroscopy (EDS). The data were analyzed using ANOVA followed by Tukey's post-test (p < 0.05). The negative control group exhibited the highest wear (6.0 µm ± 3.5), significantly differing from the group treated with 0.05% stannous fluoride gel (p = 0.007), which showed less dentin loss. The groups treated with 0.5% and 1% JSG showed results similar to the negative control (p = 0.661; p = 0.212, respectively) and the stannous fluoride group (p = 0.103; p = 0.379, respectively). In the SEM images, the specimen treated with stannous fluoride showed obliterated tubules, while the JSG gels formed crystals on the dentin surface, as confirmed by the presence of oxygen and calcium in the EDS analysis. Although the JSG gels showed similar results to the stannous fluoride, did not exhibit superior efficacy at the tested concentrations.

Safety and efficacy of P2Et extract from Caesalpinia spinosa in breast cancer patients: study protocol for a randomized double blind phase II clinical trial (CS003-BC).

BACKGROUND: Chemotherapy in breast cancer is effective but can generate significant toxicity and lead to tumor resistance. Joint treatment with standardized plant extracts can be an alternative to improve the response and allow an effective activation of the antitumor immune response that favors recovery in the short and long term. The P2Et extract of Caesalpinia spinosa presents antitumor activity in cells and animal models of breast cancer, improves the tumor microenvironment, and induces activation of the specific immune response against the tumor and is synergistic when used together with anthracyclines, which makes it a good candidate for evaluation in patients. METHODS: Conducted at a single center, this phase II study is a randomized, double-blind, placebo-controlled trial aimed at assessing the safety and efficacy of P2Et extract in patients diagnosed with stage II and III breast cancer, who are eligible for neoadjuvant treatment. The study aims to determine the safety profile at the previously established optimal biological dose from phase I trial while investigating various efficacy outcomes. These outcomes include improvements in quality of life, immunomodulation, metabolic profile, microbiome, as well as clinical indicators such as tumor reduction, disease-free survival, and pathological response, assessed at different stages of the treatment regimen. DISCUSSION: Treatment with the P2Et extract in breast cancer patients is hypothesized to enhance overall well-being, positively influencing their quality of life, while also triggering an antitumor immune response and enhancing immune infiltration. These combined effects have the potential to contribute to improved long-term survival outcomes for patients receiving the phytomedicine alongside neoadjuvant chemotherapy treatment. TRIAL REGISTRATION: This trial was registered in the US National Library of Medicine with identifier NCT05007444. First Registered August 16th, 2021. Last Updated: August 9th, 2022.

Phylogenomic and morphological data reveal hidden patterns of diversity in the national tree of Brazil, Paubrasilia echinata.

PREMISE: Paubrasilia echinata (common names, pau brasil, brazilwood) is the national tree of Brazil and an endangered species endemic to the Brazilian Atlantic Forest. Over its wide distribution of 2000 km, its leaflets morphology exhibits extensive plasticity. Three morphotypes are commonly identified based on leaf size, but it is unclear if they represent distinct taxa or a single polymorphic species. This study aims to clarify the taxonomic position of the three morphotypes to inform conservation decisions. METHODS: A morphometric study of leaf characters of herbarium specimens was coupled with genetic analyses using genotype-by-sequencing data. We used maximum-likelihood and coalescent methods to evaluate the phylogenetic and population structure of the species. We compared these with a morphological dendrogram built from hierarchical clustering. RESULTS: Two of the three morphotypes formed separately evolving lineages, the third morphotype formed two geographically separate lineages, and northern trees with intermediate leaf morphology formed a separate fifth lineage. Leaflet size varied by over 35-fold, and although morphological clustering generally matched the genetic patterns, there were some overlaps, highlighting the cryptic diversity within this group. CONCLUSIONS: Our genetic and morphological results provide some evidence that cultivated trees from different states in Brazil seem to have a limited genetic origin and do not reflect the broader genetic and geographical diversity of the species. As a result, more care is likely needed to preserve the overall genomic diversity of this endangered and iconic species.

Investigation of antioxidant activity of ethanolic extract of the bark of Caesalpinia ferrea in Swiss mice exposed to paracetamol / Investigação da atividade antioxidante do extrato etanólico da casca de Caesalpinia ferrea em camundongos Swiss expostos ao paracetamol

Aims: it was evaluated the antioxidant effect of the ethanolic extract of Caesal-pinia ferrea bark in a model of oxidative stress induced by paracetamol (PCM). Methods: male Swiss mice were subdivided into four groups (control; PCM; PCM+extract; extract; n=8) in which a dose of paracetamol (250 mg.kg-1) was administered and after 3 hours the treatment with the extract (100 mg.kg-1/day) was administered for seven days, via gavage. Oxidative stress biomarkers were determined, such as catalase, glutathione-S-transferase, reduced gluta-thione, ascorbic acid, thiobarbituric acid reactive substances and carbonylated proteins of liver, kidneys and brain and plasma parameters through the dosage of glucose, cholesterol, triglycerides, aspartate aminotransferase and alanine aminotransferase. Results: the Caesalpinia ferrea extract was able to reverse the lipid and protein damage caused by the drug in the liver tissue and caused the same effect in the renal and brain tissues in the carbonylated proteins. The extract alone decreased liver glutathione-S-transferase and increased catalase and brain glutathione-S-transferase activity, in addition to lowering glucose and cholesterol, but without altering the triglycerides. Conclusions: it was possible to conclude that the ethanolic extract of the bark of Caesalpinia ferrea has a good antioxidant activity, probably due to dose of paracetamol in the samples investigated. However, more studies are needed for a better understanding of the effects of this extract compared to the effects found in this research

Objetivos: foi avaliado o efeito antioxidante do extrato etanólico da casca de Caesalpinia ferrea em modelo de estresse oxidativo induzido por paracetamol (acetaminofeno, PCM). Métodos: camundongos Swiss machos foram subdivididos em quatro grupos (controle; PCM; PCM+extrato; extrato; n=8) nos quais foi administrada uma dose de paracetamol (250 mg.kg-1) e após três horas foi administrado o tratamento com o extrato (100 mg.kg-1/ dia) por sete dias, via gavagem. Foram determinados biomarcadores de estresse oxidativo, como catalase, glutationa-S-transferase, glutationa reduzida, ácido ascórbico, substâncias reativas ao ácido tiobarbitúrico e proteínas carboniladas do fígado, rins e cérebro, além de parâmetros plasmáticos através da dosagem de glicose, colesterol, triglicerídeos, aspartato aminotransferase e alanina aminotransferase. Resultados: o extrato de Caesalpinia ferrea foi capaz de reverter os danos lipídicos e proteicos causados pela droga no tecido hepático, e também causou o mesmo efeito nos tecidos renal e cerebral nas proteínas carboniladas. O extrato sozinho diminuiu a atividade da glutationa-S-transferase hepática e aumentou a da catalase e glutationa-S-transferase cerebral, além de diminuir a glicose e o colesterol, mas sem alterar os triglicerídeos. Conclusões: foi possível concluir que o extrato etanólico da casca de Caesalpinia ferrea apresenta uma boa atividade antioxidante, provavelmente devido à presença de taninos, tendo em vista os danos causados pela alta dose de paracetamol nas amostras investigadas. Entretanto, mais estudos são necessários para um melhor entendimento dos efeitos deste extrato frente aos efeitos encontrados nesta pesquisa

Brazilin from Caesalpinia sappan inhibits viral infection against PRRSV via CD163ΔSRCR5 MARC-145 cells: an in silico and in vitro studies.

This research aimed to identify bioactive compounds from Caesalpinia sappan extract that function as novel porcine reproductive and respiratory syndrome virus (PRRSV) infection inhibitors by computational molecular screening. We obtained a set of small-molecule compounds predicted to target the scavenger receptor cysteine-rich domain 5 (SRCR5) of CD163. In addition, the functions of positive hits were assessed and verified utilizing an in vitro antiviral activity assay with PRRSV-infected MARC-145 cells. Combining molecular docking with the results of binding affinity and ligand conformation, it was found that brazilin had the highest binding energy with the SRCR5 receptor compared to catechin and epicatechin (- 5.8, - 5.5, and - 5.1 kcal/mol, respectively). In terms of molecular mechanics, the binding free energy between the SRCR5 receptor was - 15.71 kcal/mol based on the Poisson-Boltzmann surface area of brazilin. In addition, PRRSV infection in MARC-145 cells was significantly inhibited by brazilin compared to the control (virus titer, 4.10 vs. 9.25 TCID50/mL, respectively). Moreover, brazilin successfully limited the number of PRRSV RNA copies in MARC-145 cells as determined by RT-qPCR. By inhibiting the PRRSV-CD163 interaction with brazilin from Caesalpinia sappan, it may be possible to prevent PRRSV infection in pigs, as suggested by this research.

Protective role of Caesalpinia sappan extract and its main component brazilin against blue light-induced damage in human fibroblasts.

BACKGROUND: Ultraviolet (UV) radiation is a well-known factor that causes skin aging. Recently, with the development of technology, the skin has been exposed to not only the UV radiation but also the blue light from electronic devices. Blue light is a high-energy visible light that penetrates deep into the dermal layer, producing reactive oxygen species (ROS) and resulting in skin aging. In this study, we searched for candidate materials that can inhibit blue light-induced skin aging and found Caesalpinia sappan extract (CSE) to be effective. METHODS: Human dermal fibroblasts (HDFs) were treated with various concentrations of CSE and brazilin and exposed to blue light. We measured that antioxidant activity, MMP-1 levels using MMP-1 ELISA, changes in collagen type 1, collagen type 3, MMP-1, and MMP-3 mRNA expressions, and ROS generation. RESULTS: We confirmed that CSE has high absorption of blue light and antioxidant activity. Blue light irradiation at 30 J/cm2 decreased the expression of collagen types 1 and 3, increased the expression of matrix metalloproteinase (MMP)-1 and 3, and decreased the production of ROS in human dermal fibroblasts as compared to those of the nonirradiated group. However, pretreatment with CSE protected against the damage caused by the blue light. Brazilin, a major constituent of C. sappan, had high absorbance in the blue light region and antioxidant activities. Pretreatment with brazilin also inhibited the damage caused by the blue light in the cells. CONCLUSION: CSE and brazilin are potential agents for inhibiting skin aging caused by blue light-induced damage.

Polysaccharide extract of Caesalpinia ferrea (Mart) pods attenuates inflammation and enhances the proliferative phase of rat cutaneous wounds.

Pods of Caesalpinia ferrea, popularly used to treat inflammatory processes, were collected to obtain the polysaccharide-rich extract, presenting anti-inflammatory and antinociceptive effects in acute inflammation models. This study aimed to evaluate the anti-inflammatory, antinociceptive and healing activities of the polysaccharide-rich extract from Caesalpinia ferrea pods (PEp-Cf) in the rat model of cutaneous excisional wound. PEp-Cf (0.025-0.1%) or 0.9% NaCl was topically applied in the wounds at dorsal thoracic region (2×/day) during 21 days for measurement of clinical signs (hyperemia, inflammatory exudate, edema, nociception), wound size, histopathological/histomorphometric, oxidative/inflammatory markers and systemic toxicity. PEp-Cf at 0.1% reduced wound area and increased ulcer contraction [days 2 and 10 (21-78%)]. PEp-Cf reduced clinical signs [days 2 and 5 (2.2-2.8×)] and modulated the healing inflammatory phase via stimulation of epithelialization (days 10 and 14), and inhibition of polymorphonuclears [days 2 and 5 (71-74%)], protein leakage [days 2 and 5 (28-41%)], nitrate [days 2 and 5 (2.2-6×)] and malondialdehyde [days 2 and 5 (46-49%)]. PEp-Cf increased the number of blood vessels [days 5 and 7 (3.1-9.6×)], fibroblasts [days 5 and 7 (2.1-6.4×)] and collagen [days 5 to 14 (1.5-1.8×)]. In conclusion, the topical application of PEp-Cf at 0.1% accelerates the healing process of rat cutaneous wounds via modulation of the inflammatory and proliferative phases, being devoid of systemic alterations.

Molecular docking of brazilin and its analogs to barrier-to-autointegration factor 1 (BAF1).

The tetracyclic phenolic compound brazilin, derived from the wood of Caesalpinia sappan, has been shown to bind to the chromatin protein BAF1 (barrier-to-autointegration factor 1), a protein essential to maintain integrity of the nuclear envelope in cells. BAF1 plays a role in cancer development. Using molecular docking, we have located the binding site for brazilin on the surface of the BAF1 monomer and compared its binding to that of four analogs. The oxidized product brazilein (ΔE = -57.7 kcal/mol) exhibits a higher affinity for BAF1 compared to the reduced form brazilin (ΔE = -38.2 kcal/mol). Incorporation of a 4-hydroxyl substituent on the indenochromene unit affords hematoxylin and hematein. In silico analysis predicts that the oxidized form hematein (ΔE = -66.2 kcal/mol) displays a higher affinity for BAF1 than the reduced form hematoxylin (ΔE = -42.2 kcal/mol). In contrast, the atypical bis-lactone product brazilide A cannot form good complexes with BAF1. The analysis points to the formation of more stable BAF1 complexes with the oxidized molecules compared to the reduced ones, but the position of the binding site on the protein cavity is different for brazilin/hematoxylin compared to brazilein/hematein. Our study may be useful to guide the design of BAF1 ligands.

Changes in cell wall composition and ultrastructure related to desiccation during the seed maturation of Paubrasilia echinata (brazilwood).

Paubrasilia echinata (brazilwood) is an endangered native tree from the Brazilian Atlantic Forest whose seeds tolerate maturation drying, but, unlike classic orthodox seeds, they quickly lose viability after shedding. This work analyzed the biochemical and ultrastructural changes during the maturation of brazilwood seeds, with particular attention to the cell walls and organization of the cellular components. The physiological seed maturity was accompanied by increased starch content and decreased soluble sugars. Arabinose increased considerably and was the predominant cell-wall sugar during maturation, suggesting a rise in arabinans that contribute to greater cell wall flexibility. This increase was consistent with the cell wall infolding observed in the hypocotyl axis and cotyledons during the maturation of brazilwood seeds. Ultrastructural analyses showed changes in the number and distribution of protein bodies and amyloplasts and the reorganization of lipid droplets into large drops or masses during seed desiccation. Our findings demonstrate that brazilwood seeds behave like other orthodox seeds during maturation, performing the cell wall and metabolic changes before the major decline in the seed water content. However, the high vacuolization and reorganization of lipid bodies observed at 65 DAA suggest that cell deterioration occurs to some extent at the end of the maturation period and could be responsible for reducing the longevity of the brazilwood dried seeds.

Phytochemical characterisation and in vivo antilithiatic activity of the stems of Caesalpinia bahamensis (Brasilete).

The aim of this work was to identify the main chemical constituents and to evaluate the antilithiatic activity of the aqueous and hydroalcoholic extracts of stems of Caesalpinia bahamensis Lam. Fractionation and isolation of constituents from the hydroalcoholic extract was carried out by flash chromatography and semi-preparative liquid chromatography. The antilithiatic activity of the aqueous and hydroalcoholic extracts was evaluated in Wistar rats, where kidney stones were induced by ethylene glycol and ammonium chloride. Creatinine, calcium, and oxalate levels were evaluated and histological analysis was carried out. The homoisoflavonoids protosappanin B, 10-methyl-protosappanin B and brazilin were isolated and the antilithiatic activity of the aqueous and hydroalcoholic extracts was demonstrated by the reduction of the concentration of calcium and oxalate in urine compared to the lithiasis group. It was corroborated by histological analysis. Brazilin and protosappanin B were proposed as chemical markers for this plant species.

Efecto antimicrobiano in vitro de extractos de Caesalpinia Spinosa (algarrobo) sobre patógenos orales / In vitro antimicrobial effect of Caesalpinia Spinosa (carob tree) extracts on oral pathogens

La resistencia antimicrobiana es un problema de sa-lud pública mundial. Las infecciones por microorga-nismos resistentes pueden ser altamente transmisi-bles e incluso causar la muerte. Este hecho genera grandes costos para los pacientes y para los servi-cios de salud. El objetivo del presente trabajo fue de-terminar el efecto antimicrobiano in vitro de extractos etanólicos de Caesalpinia spinosa sobre el crecimien-to de Enterococcus faecalis, Staphylococcus aureus y Candida albicans. Se recolectaron y certificaron muestras de C. spinosa. Se obtuvieron extractos de hojas, vainas y semillas en concentraciones de 100%, 75%, 50% y 25%. Mediante Kirby - Bauer, se cargaron los discos con los extractos y se depositaron en el medio inoculado con cepas de E. faecalis, S. aureus y C. albicans; junto a un CP (antimicrobiano), y un CN (etanol). Las placas se incubaron a 370°C durante 24 horas, y posteriormente se midieron los halos de inhi-bición con un vernier digital. Destaca el valor del halo de extracto de vainas; superó al de Ampicilina 10mg, sobre el E. faecalis. El extracto de vainas presentó ma-yor diámetro de inhibición (19mm), el de semillas pre-sentó el más bajo (1mm). ANOVA arrojó diferencia es-tadísticamente significativa entre los datos obtenidos para todos los extractos. En conclusión, los extractos etanólicos de Caesalpinia spinosa tienen efecto anti-microbiano in vitro sobre Enterococcus faecalis, Sta-phylococcus aureus y Candida albicans. La actividad antimicrobiana del extracto es directamente propor-cional a su concentración. Los extractos de C. spinosa podrían ser utilizados como coadyuvantes en el trata-miento contra Enterococcus faecalis, Staphylococcus aureus, Candida albicans, que están relacionados con patologías orales (AU)

Antimicrobial resistance is a global public health problem. Infections with resistant microorganisms can be highly transmissible and even cause death. This fact generates great costs for patients and for health services. The objective of this work was to determine the in vitro antimicrobial effect of ethanolic extracts of Caesalpinia spinosa on the growth of Enterococcus faecalis, Staphylococcus aureus and Candida albicans. Samples of C. spinosa were collected and certified. Leaf, pod and seed extracts were obtained at concentrations of 100%, 75%, 50% and 25%. Using Kirby-Bauer, the disks were loaded with the extracts and deposited in the medium inoculated with strains of E. faecalis, S. aureus and C. albicans; together with a CP (antimicrobial), and a CN (ethanol). The plates were incubated at 370°C for 24 hours, then the inhibition halos were measured with a digital vernier. The value of the pod extract halo stands out, surpassing that of Ampicillin 10mg, over E. faecalis. The pod extract presented the greatest diameter of inhibition (19mm), the seed extract presented the lowest (1mm). ANOVA showed a statistically significant difference between the data obtained for all the extracts. In conclusion, the ethanolic extracts of Caesalpinia spinosa have an in vitro antimicrobial effect on Enterococcus faecalis, Staphylococcus aureus and Candida albicans. The antimicrobial activity of the extract is directly proportional to its concentration. C. spinosa extracts could be used as adjuvants in the treatment against Enterococcus faecalis, Staphylococcus aureus, Candida albicans, which are related to oral pathologies (AU)

Anthelmintic effect of four extracts obtained from Caesalpinia coriaria foliage against the eggs and larvae of Haemonchus contortus.

To investigate the in vitro anthelmintic efficacy of dividivi (Caesalpinia coriaria), a traditional medicinal plant used in Central America and the northern part of South America, extracts from the foliage of this plant were subjected to the egg hatching test (EHT) and larval exsheathment inhibition test (LEIT), against Haemonchus contortus. Four different extracts were evaluated: acetone-water (AW), methanol-water (MW), acetone-water-dichloromethane (AWD) and methanol-water-dichloromethane (MWD). The concentrations used for the EHT and LEIT tests ranged from 500 to 4000 µg mL-1 and six repetitions per concentration. The effective concentrations (EC50) were calculated using Probit analysis. The EC50 for EHT were 2947.0, 3347.0, 3959.6 and 4538.7 µg mL-1 for MWD, MW, AW and AWD, respectively. The EC50 for LEIT were 2883.4, 5927.4, 9876.3 and 9955.4 µg mL-1 for AWD, AW, MWD and MW, respectively. The methanol extracts were the most effective in inhibiting the hatching of eggs, while the acetone extracts showed efficacy in inhibiting larval exsheathment. This study explains the importance that C. coriaria has as a medicinal plant in Central and South American countries.

Plastome evolution in the Caesalpinia group (Leguminosae) and its application in phylogenomics and populations genetics.

MAIN CONCLUSION: The chloroplast genomes of Caesalpinia group species are structurally conserved, but sequence level variation is useful for both phylogenomic and population genetic analyses. Variation in chloroplast genomes (plastomes) has been an important source of information in plant biology. The Caesalpinia group has been used as a model in studies correlating ecological and genomic variables, yet its intergeneric and infrageneric relationships are not fully solved, despite densely sampled phylogenies including nuclear and plastid loci by Sanger sequencing. Here, we present the de novo assembly and characterization of plastomes from 13 species from the Caesalpinia group belonging to eight genera. A comparative analysis was carried out with 13 other plastomes previously available, totalizing 26 plastomes and representing 15 of the 26 known Caesalpinia group genera. All plastomes showed a conserved quadripartite structure and gene repertoire, except for the loss of four ndh genes in Erythrostemon gilliesii. Thirty polymorphic regions were identified for inter- or intrageneric analyses. The 26 aligned plastomes were used for phylogenetic reconstruction, revealing a well-resolved topology, and dividing the Caesalpinia group into two fully supported clades. Sixteen microsatellite (cpSSR) loci were selected from Cenostigma microphyllum for primer development and at least two were cross-amplified in different Leguminosae subfamilies by in vitro or in silico approaches. Four loci were used to assess the genetic diversity of C. microphyllum in the Brazilian Caatinga. Our results demonstrate the structural conservation of plastomes in the Caesalpinia group, offering insights into its systematics and evolution, and provides new genomic tools for future phylogenetic, population genetics, and phylogeographic studies.

Structural characterization of anticoagulant and antithrombotic polysaccharides isolated from Caesalpinia ferrea stem barks.

This study aimed to isolate, characterize chemical-structurally and evaluate the effects of polysaccharides from Caesalpinia (Libidibia) ferrea stem barks in the haemostatic system. The deproteinated-polysaccharide extract (PE-Cf) after being fractionated by ion exchange chromatography-DEAE-cellulose resulted in three fractions (FI, FII, FIII) containing total carbohydrates (14.3-38%), including uronic acid (5-16%), and polyphenols (0.94-1.7 mg/g GAE). The polysaccharide fractions presented polydisperse profile in polyacrylamide gel electrophoresis (detected by Stains-All) and molecular masses (9.5 × 104 Da-1.5 × 105 Da) identified by gel permeation chromatography. FT-IR showed absorption bands (1630 cm-1, 1396-1331 cm-1), indicative of uronic acid, and a band at 1071 cm-1, typical of COO- groups of galacturonic acid. The NMR spectra of C. ferrea polysaccharides revealed a central core composed mainly by 5-linked α-Araf and minority components as α-Rhap and α-GalAp. UV spectra of fractions revealed discrete shoulders at 269-275 nm, characteristic of polyphenolic compounds. In vitro, polysaccharides inhibited the intrinsic and/or common coagulation pathway (aPTT test) (2.0-3.7 fold) and the platelet aggregation induced by 3 µM adenosine diphosphate (25-48%) and 5 µg/mL collagen (24%), but not that induced by arachidonic acid. In vivo, the polysaccharides inhibited (36-69%) venous thrombosis induced by hypercoagulability and stasis, showing discrete hemorrhagic effect. In conclusion, the polysaccharides of C. ferrea barks, containing arabinose, galactose, rhamnose and uronic acid, possess anticoagulant, antiplatelet and antithrombotic properties of low hemorrhagic risk, suggesting potential applicability in thromboembolic disorders.

Successful adsorption of bright blue and methylene blue on modified pods of Caesalpinia echinata in discontinuous system.

Pods of the forest species Caesalpinia echinata were used as an alternative adsorbent to remove bright blue (BB) and methylene blue (MB) dyes. The raw and acid-treated samples were characterized by techniques like SEM, XRD, and FTIR. The acid-treated pod sample was characterized by an amorphous structure containing several cavities, bumps, and functional groups. The Elovich model was the most satisfactory to describe the adsorption kinetic data. The isothermal studies were better described by the Langmuir model for BB dye, with a maximum capacity of 261 mg g-1, and Tóth model for MB dye, giving a maximum capacity of 288 mg g-1. The thermodynamic study indicated a spontaneous and favorable process and endothermic nature for both dyes. In the treatment of two simulated effluents containing a mixture of different compounds such as dyes and salts, to simulate real wastewaters, the adsorbent was highly efficient, presenting around 80% of color removal for both effluents. Therefore, the acid-treated pods of Caesalpinia echinata have great potential to be applied as an alternative adsorbents in treating colored effluents in discontinuous systems.

Bioengineering of an elastase inhibitor from Caesalpinia echinata (Brazil wood) seeds.

Protease inhibitors have been widely used in several therapeutic applications such as in the treatment of bleeding disorders, hypertension, cancer and pulmonary diseases. In a previous work, we demonstrated that a Kunitz-type serine protease inhibitor isolated from the seeds of Caesalpinia echinata (CeEI) exhibits pharmacological potential in lung inflammatory diseases in which neutrophil elastase plays a crucial role. However, an important challenge in the use of natural products is to ensure a commercially viable production. In this work, we report the cloning, expression and purification of two recombinant CeEI isoinhibitors with 700 base pairs encoding two proteins with 181 amino acid residues (rCeEI-4 and rCeEI-5). After the expression, each yielding 22 mg/L of active protein, both isoinhibitors presented a molecular mass of about 23.0 kDa, evaluated by SDS-PAGE. The inhibition constants for human neutrophil elastase (HNE) were 0.67 nM (rCeEI-4) and 0.57 nM (rCeEI-5), i.e., similar to the native inhibitor (1.90 nM). Furthermore, rCeEI-4 was used as a template to design smaller functional peptides flanking the inhibitor reactive site: rCeEI-36, delimited between the amino acid residues N36 and S88 containing a disulfide bond in the reactive-site loop, and rCeEI-46, delimited between S46 and L75 without the disulfide bond. The yields were 18 mg/L (rCeEI-36) and 12 mg/L (rCeEI-46). Both peptides inhibit HNE in the nanomolar range (Ki 0.30 ± 0.01 and 8.80 ± 0.23, respectively). Considering their size and the inhibitory efficiency, these peptides may be considered in strategies for the development of drugs targeting pulmonary disorders where elastase is involved.

Polysaccaride-rich extract of Caesalpina ferrea stem barks attenuates mice acute inflammation induced by zymosan: Oxidative stress modulation.

ETHNOPHARMACOLOGICAL RELEVANCE: Stem barks of Caesalpinia ferrea Mart. Ex Tul. (Caesalpiniaceae), also known as pau-ferro jucá or jucaína, are popularly used to treat contusions, diabetes, rheumatism and other inflammatory conditions in the form of tea, lick or decoction. OBJECTIVE: To evaluate the effect of the polysaccharide-rich extract obtained from C. ferrea stem barks (PE-Cf) in mice models of acute inflammation induced by zymosan and the involvement of oxidative stress biomarkers. MATERIALS AND METHODS: Mice were treated with PE-Cf (0.001, 0.01, 0.1, 1 mg/kg) by endovenous route (i.v.) or per oral (p.o.) 30 or 60 min before injection of the inflammatory stimuli zymosan (0.5 mg; intraperitoneal or subcutaneous intraplantar). The inflammatory parameters (edema, nociception, leukocyte migration) and oxidative stress markers (myeloperoxidase-MPO, malondialdehyde-MDA, nitrite, reduced glutathione-GSH, glutathione peroxidase-GPx) were evaluated in the models of paw edema (hidropletysmometry/expressed as ml or area under curve-AUC) and peritonitis (optical microscopy/expressed as n° of cells/mm3 of peritoneal fluid). Statistical analysis was performed by ANOVA, followed by Bonferroni test. RESULTS: PE-Cf (0.1, 0.01 and 1 mg/kg) dose-dependently inhibited paw edema, showing maximal effect (74%) at 1 mg/kg in the 5th (52 ± 9.6 µl vs. zymosan: 204 ± 3.6 µl). PE-Cf (1 mg/kg) also inhibited by 43% MPO activity in the paw tissues (17 ± 1 vs. zymosan: 30 ± 2.6 U/mg). Besides, 4 h after peritonitis induction, PE-Cf (1 mg/kg) reduced neutrophil migration by 84% (432 ± 45 vs. zymosan: 2651 ± 643 cells/mm3); visceral nociception by 76% (3 ± 0.6 vs. zymosan: 16 ± 4 writhes); nitric oxide by 73% (0.131 ± 0.033 vs. zymosan: 0.578 ± 0.185 NO2-/NO3-ml); MDA (98 ± 10 vs. zymosan:156 ± 21 U/ml), and increased GSH by 65% (736 ± 65 vs. zymosan: 259 ± 58 µmol/ml) and GPx by 72% (0.037 ± 0.007 vs. zymosan: 0.010 ± 0.005 U/mg protein). CONCLUSION: The polysaccharide-rich extract of Caesalpinia ferrea stem barks present anti-inflammatory and antioxidant effects in mice models of acute inflammation induced by zymosan.

Antidepressant-like activity of ethanol extract of leaves of Caesalpinia pulcherrima in unstressed and stressed mice

In the present study, antidepressant-like activity of ethanol extract of leaves of Caesalpinia pulcherrima was evaluated in Swiss young male albino mice. Stress was induced in mice by subjecting them to unpredictable mild stress for 21 successive days. Ethanol extract of the leaves (100, 200 and 400 mg/ kg, p.o.) and fluoxetine (20 mg/kg, p.o.) were administered for 21 consecutive days to separate groups of unstressed and stressed mice. Ethanol extract (200 and 400 mg/kg) and fluoxetine significantly decreased immobility period of unstressed as well as stressed mice in tail suspension test (TST). However, the lowest dose (100 mg/kg) of the extract also significantly decreased immobility period of stressed mice in TST. The extract significantly restored reduced sucrose preference in stressed mice. There was no significant effect on locomotor activity of mice. Ethanol extract of the leaves significantly decreased plasma nitrite and corticosterone levels; brain MAO-A activity and MDA level; and increased brain reduced glutathione and catalase activity in unstressed as well as stressed mice as compared to their respective vehicle treated controls. Thus, ethanol extract of leaves of Caesalpinia pulcherrima showed significant antidepressant-like activity in unstressed and stressed mice probably through inhibition of brain MAO-Aactivity, reduction of oxidative stress and plasma corticosterone levels.