ABSTRACT
Urinary tract infection caused by Klebsiella, Proteus and Streptococcus is a urease dependent process, hence treatment of these infections by antibacterial compounds lies in inhibition of their virulence factors. The crude methanolic extracts derived from sumac fruit, pomegranate peel and Indian almond leaves were separated into anthocyanin and non-anthocyanin fractions using solid phase cartridges. The inhibitory effect of these fractions was determined on the growth of urease producing species and jack bean urease activity. Known compounds in the fractions were also docked with ureases of different biological origins viz. K. pneumoniae (PDB ID: 8HCN), K. aerogenes (PDB ID: 2KAU), Helicobacter pylori (PDB ID:8HC1)and Canavalia ensiformis (jack bean) (PDB ID: 3LA4) to determine their binding affinities and interaction with the enzyme. All the fractions showed significant inhibition growth for P. mirabilis, S. epidermidis and K.pneumoniae. Among the samples, sumac showed greatest inhibition against all (MIC 6-25 mg.mL-1) while among the fractions, anthocyanin was found to be most active (MIC 6-12 mg/mL). Likewise, all fractions inhibited urease with lowest ICs50 shown by sumac fractions (21-116 µg.mL-1). Out of 39 compounds docked, 27 showed interaction with movable flaps and/or active site of ureases which explains their mode of inhibition.
Subject(s)
Polyphenols , Pomegranate , Prunus dulcis , Urease , Urease/antagonists & inhibitors , Urease/metabolism , Polyphenols/pharmacology , Polyphenols/chemistry , Pomegranate/chemistry , Prunus dulcis/chemistry , Canavalia/enzymology , Plant Extracts/pharmacology , Plant Extracts/chemistry , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/chemistry , Enzyme Inhibitors/pharmacology , Enzyme Inhibitors/chemistry , Microbial Sensitivity Tests , Molecular Docking Simulation , Bacteria/drug effects , Bacteria/enzymologyABSTRACT
Leptospirosis is a global public health problem caused by Gram-negative pathogenic bacteria belonging to the genus Leptospira. The disease is transmitted through the urine of infected animals, which contaminates water and soil, leading to the infection of other animals and humans. Currently, several approaches exist to detect these bacteria; however, a new sensitive method for the live-cell imaging of Leptospira is required. In this study, we report the green synthesis of cadmium telluride quantum dots (CdTe QDs) which are unique fluorescent nanocrystals with a high fluorescence quantum yield capable of modifying cell surfaces and are biocompatible with cells. The fabrication of QDs with concanavalin A (ConA), a carbohydrate-binding lectin and known biological probe for Gram-negative bacteria, produced ConA-QDs which can effectively bind on Leptospira and exhibit strong fluorescence under simple fluorescence microscopy, allowing the live-cell imaging of the bacteria. Overall, we performed the simple synthesis of ConA-QDs and demonstrated their potential use as versatile fluorescent probes for the live-cell imaging of Leptospira. This technique could be further applied to track leptospiral cells and study the infection mechanism, contributing to a more thorough understanding of leptospirosis and how to control it in the future.
Subject(s)
Leptospira , Quantum Dots , Quantum Dots/chemistry , Fluorescent Dyes/chemistry , Cadmium Compounds/chemistry , Tellurium/chemistry , Concanavalin A/chemistry , Canavalia/chemistry , Biocompatible Materials/chemistry , Microscopy, FluorescenceABSTRACT
The poor extractability and digestibility of jack beans restrict their application in food systems. Thermal treatment could be a processing tool to disrupt the compact conformation of the plant matrix and inactivate inherent antinutrients. Therefore, this research investigated the impact of conventional heat-aided (HA-) and microwave-aided (MA-) extraction treatments on the structure, functional properties, and digestibility of jack bean protein concentrate (JBPC) under varying extraction pH. The novelty brought by the present study is establishing the thermal treatment/extraction pH combinations for improving techno-functionalities and digestibility of JBPC. Heat (50 °C for 1 h) and sequential microwave power (400 W, 600 W, and 800 W for 5 min) at three extraction pH (9.0, 10.0, and 11.0) were studied. Upon increasing extraction pH, a significant decrease in the protein content, and ß-Sheet structure was observed, in the order of pH 11.0 > 10.0 > 9.0. JBPC extracted using HA treatments displayed the highest contents of surface hydrophobicity (90.02) and sulfhydryl groups. In functional properties, MA-extracted JBPC under 400 W showed significantly improved solubility (93.45 %), emulsifying activity index (45.23 m2/g), and foaming capacity (141.70 %) when compared to other thermal treatments. The degree of hydrolysis result revealed that MA treatment improved the JBPC in vitro digestibility at a low power level of 400 W. These findings suggest that MA extraction treatment can improve the functional and nutritional properties of JBPC regardless of the extraction pH, and thus, expand the potential application in food systems.
Subject(s)
Digestion , Hot Temperature , Hydrophobic and Hydrophilic Interactions , Microwaves , Plant Proteins , Hydrogen-Ion Concentration , Plant Proteins/chemistry , Plant Proteins/isolation & purification , Food Handling/methods , Canavalia/chemistryABSTRACT
Leishmaniases, caused by Leishmania parasites, are widespread and pose significant health risks globally. Visceral leishmaniasis (VL) is particularly prevalent in Brazil, with high morbidity and mortality rates. Traditional treatments, such as pentavalent antimonials, have limitations due to toxicity and resistance. Therefore, exploring new compounds like lectins is crucial. Concanavalin A (ConA) has shown promise in inhibiting Leishmania growth. This study aimed to evaluate its leishmanicidal effect on L. infantum promastigotes and understand its mechanism of action. In vitro tests demonstrated inhibition of promastigote growth when treated with ConA, with IC50 values ranging from 3 to 5 µM over 24-72 h. This study suggests that ConA interacts with L. infantum glycans. Additionally, ConA caused damage to the membrane integrity of parasites and induced ROS production, contributing to parasite death. Scanning electron microscopy confirmed morphological alterations in treated promastigotes. ConA combined with the amphotericin B (AmB) showed synergistic effects, reducing the required dose of AmB, and potentially mitigating its toxicity. ConA demonstrated no cytotoxic effects on macrophages, instead stimulating their proliferation. These findings reinforce that lectin exhibits promising leishmanicidal activity against L. infantum promastigotes, making ConA a potential candidate for leishmaniasis treatment.
Subject(s)
Antiprotozoal Agents , Canavalia , Concanavalin A , Leishmania infantum , Leishmania infantum/drug effects , Concanavalin A/pharmacology , Animals , Antiprotozoal Agents/pharmacology , Antiprotozoal Agents/chemistry , Seeds/chemistry , Reactive Oxygen Species/metabolism , Mice , Amphotericin B/pharmacology , Lectins/pharmacology , Lectins/chemistry , Lectins/metabolism , Plant Lectins/pharmacology , Plant Lectins/chemistry , Macrophages/drug effects , Macrophages/metabolism , Macrophages/parasitologyABSTRACT
Thirteen novel hydrazone-Schiff bases (3-15) of fexofenadine were succesfully synthesized, structurally deduced and finally assessed their capability to inhibit urease enzyme (inâ vitro). In the series, six compounds 12 (IC50=10.19±0.16â µM), 11 (IC50=15.05±1.11â µM), 10 (IC50=17.01±1.23â µM), 9 (IC50=17.22±0.81â µM), 13 (IC50=19.31±0.18â µM), and 14 (IC50=19.62±0.21â µM) displayed strong inhibitory action better than the standard thiourea (IC50=21.14±0.24â µM), while the remaining compounds displayed significant to less inhibition. LUMO and HOMO showed the transferring of charges from molecules to biological transfer and MEP map showed the chemically reactive zone appropriate for drug action are calculated using DFT. AIM charges, non-bonding orbitals, and ELF are also computed. The urease protein binding analysis benefited from the docking studies.
Subject(s)
Drug Design , Enzyme Inhibitors , Hydrazones , Molecular Docking Simulation , Schiff Bases , Terfenadine , Urease , Urease/antagonists & inhibitors , Urease/metabolism , Hydrazones/chemistry , Hydrazones/pharmacology , Hydrazones/chemical synthesis , Enzyme Inhibitors/chemistry , Enzyme Inhibitors/pharmacology , Enzyme Inhibitors/chemical synthesis , Schiff Bases/chemistry , Schiff Bases/pharmacology , Schiff Bases/chemical synthesis , Terfenadine/analogs & derivatives , Terfenadine/chemistry , Terfenadine/metabolism , Terfenadine/pharmacology , Terfenadine/chemical synthesis , Density Functional Theory , Molecular Structure , Structure-Activity Relationship , Canavalia/enzymologyABSTRACT
The effect of two processing methods of Jack beans (i.e. cooked bean (CB) and cooked tempeh (CT)) on the in vitro digestibility of protein and starch, as well as the production of short chain fatty acids (SCFAs), γ-aminobutyric acid (GABA), and tryptophan (Trp) metabolites after in vitro colonic fermentation, was investigated. CT was obtained by fungal fermentation after cooking under acidic conditions. CT had significantly higher protein, lower digestible starch, lower total fiber, higher free phenolic compounds, and higher ash content compared to CB. CT exhibited better in vitro protein digestibility than CB and less glucose release during in vitro digestion than CB. A comparable concentration of total SCFAs and GABA was produced after in vitro fermentation of CB and CT, but CB produced more indole than CT, resulting in higher amounts of total Trp metabolites. In summary, our findings show that tempeh fermentation improves the nutritional quality of Jack beans and describe the impact of fermentation on the digestibility of nutrients and the formation of metabolites during colonic fermentation.
Subject(s)
Fabaceae , Soy Foods , Canavalia/metabolism , Fermentation , Fabaceae/metabolism , Starch/metabolism , Fatty Acids, Volatile/metabolism , gamma-Aminobutyric Acid/metabolism , DigestionABSTRACT
Cells use glycans to encode information that modulates processes ranging from cell-cell recognition to programmed cell death. This information is encoded within a glycocode, and its decoding is performed by carbohydrate-binding proteins. Among these, lectins stand out due to their specific and reversible interaction with carbohydrates. Changes in glycosylation patterns are observed in several pathologies, including cancer, where abnormal glycans are found on the surfaces of affected tissues. Given the importance of the bioprospection of promising biomolecules, the current work aimed to determine the structural properties and anticancer potential of the mannose-specific lectin from seeds of Canavalia villosa (Cvill). Experimental elucidation of the primary and 3D structures of the lectin, along with glycan array and molecular docking, facilitated the determination of its fine carbohydrate-binding specificity. These structural insights, coupled with the lectin's specificity, have been combined to explain the antiproliferative effect of Cvill against cancer cell lines. This effect is dependent on the carbohydrate-binding activity of Cvill and its uptake in the cells, with concomitant activation of autophagic and apoptotic pathways.
Subject(s)
Canavalia , Lectins , Lectins/pharmacology , Lectins/analysis , Canavalia/metabolism , Molecular Docking Simulation , Plant Lectins/metabolism , Seeds/metabolism , Carbohydrates/analysis , Polysaccharides/analysisABSTRACT
Sword bean (SB) contains various phytochemicals, such as flavonoids, tannins, saponins, and terpenoids. Although the evaluation of its potential functions, including antioxidant, anti-obesity, anti-inflammatory, liver protection, and antiangiogenic activities, has been widely reported, research on their use in osteoporosis prevention is insufficient. Furthermore, while various studies are conducted on SB, research on sword bean pods (SBP) is not yet active, and little is known about it. Therefore, this study investigated the effects of promoting osteoblast differentiation of MC3T3-E1 cells using SB and SBP extracts and their mechanisms. We show that SBP extracts increase osteoblast proliferation, mineralization-activated alkaline phosphatase (ALP), and collagen synthesis activities. Additionally, treatment with SBP extract increased the expression of markers related to osteoblast differentiation, such as ALP, SPARC, RUNX2, COL-I, BMP2, OCN, and OPN. It was confirmed that SBP induces differentiation by activating the BMP2/SMAD/RUNX2 pathway. We also show that SBP is more effective than SB, and SBP may be useful in assimilating bone minerals and preventing osteoporosis.
Subject(s)
Canavalia , Osteoporosis , Humans , Canavalia/metabolism , Core Binding Factor Alpha 1 Subunit/metabolism , Cell Differentiation , Bone Morphogenetic Protein 2/metabolism , Osteoblasts , Osteogenesis , Osteoporosis/prevention & control , Osteoporosis/metabolismABSTRACT
BACKGROUND: Sword bean (Canavalia gladiata) is an underutilized legume that has the potential to become an important food source owing to its wide range of nutritional and medicinal properties. In May 2023, symptoms induced by a possible virus infection such as mosaic, mottling and vein banding were observed on the leaves of about 20% of the Sword bean plants growing at the experimental research farm of the Indian Agricultural Research Institute in Pune, Maharashtra, India. METHODS AND RESULTS: Symptomatic and asymptomatic samples were screened by ELISA for the presence of Potyvirus, Cucumber mosaic virus and Tobacco mosaic virus. All symptomatic samples tested positive for Potyvirus in ELISA as well as in RT-PCR assay using the universal potyvirus primer pair (CPUP /P9502) which amplify c. 700 bp of the partial coat protein region and 3'UTR. Asymptomatic samples tested negative for all tested viruses in both serological and molecular assays. BLASTn sequence analysis of the amplicons revealed that the sequence shares more than 98% identity with an Indian isolate of Bean common mosaic virus (BCMV). Sequence analysis enabled the identification of the Potyvirus as BCMV. Furthermore, the present Sword bean isolate clustered with other BCMV isolates in the phylogenetic analysis. CONCLUSION: In the present study, BCMV was found to be naturally infecting Sword bean for the first time in the world. This is of epidemiological importance, as BCMV is known to cause significant yield losses in legumes and could severely hamper Sword bean production.
Subject(s)
Fabaceae , Potyvirus , Canavalia , Phylogeny , India , Potyvirus/geneticsABSTRACT
The development of novel scaffolds that can increase the effectiveness, safety, and convenience of medication therapy using drug conjugates is a promising strategy. As a result, drug conjugates are an active area of research and development in medicinal chemistry. This research demonstrates acetamide-sulfonamide scaffold preparation after conjugation of ibuprofen and flurbiprofen with sulfa drugs, and these scaffolds were then screened for urease inhibition. The newly designed conjugates were confirmed by spectroscopic techniques such as IR, 1HNMR, 13CNMR, and elemental analysis. Ibuprofen conjugated with sulfathiazole, flurbiprofen conjugated with sulfadiazine, and sulfamethoxazole were found to be potent and demonstrated a competitive mode of urease inhibition, with IC50 (µM) values of 9.95 ± 0.14, 16.74 ± 0.23, and 13.39 ± 0.11, respectively, and urease inhibition of 90.6, 84.1, and 86.1% respectively. Ibuprofen conjugated with sulfanilamide, sulfamerazine, and sulfacetamide, whereas flurbiprofen conjugated with sulfamerazine, and sulfacetamide exhibited a mixed mode of urease inhibition. Moreover, through molecular docking experiments, the urease receptor-binding mechanisms of competitive inhibitors were anticipated, and stability analysis through MD simulations showed that these compounds made stable complexes with the respective targets and that no conformational changes occurred during the simulation. The findings demonstrate that conjugates of approved therapeutic molecules may result in the development of novel classes of pharmacological agents for the treatment of various pathological conditions involving the urease enzyme.
Subject(s)
Flurbiprofen , Molecular Docking Simulation , Flurbiprofen/pharmacology , Ibuprofen/pharmacology , Enzyme Inhibitors/pharmacology , Sulfacetamide , Kinetics , Urease , Sulfamerazine , Canavalia , Structure-Activity Relationship , Sulfanilamide , Sulfonamides/pharmacology , Molecular StructureABSTRACT
Different degrees in the biological activities of Canavalia rosea had been previously reported . In this study, our group assessed the cardioprotective effects of the ethyl acetate fraction (EAcF) of the Canavalia rosea leaves. Firstly, it was confirmed, by in vitro approach, that the EAcF has high antioxidant properties due to the presence of important secondary metabolites, as flavonoids. In order to explore their potential protector against cardiovascular disorders, hearts were previously perfused with EAcF (300 µg.mL-1) and submitted to the global ischemia followed by reperfusion in Langendorff system. The present findings have demonstrated that EAcF restored the left ventricular developed pressure and decreased the arrhythmias severity index. Furthermore, EAcF significantly increased the glutathiones peroxidase activity with decreased malondialdehyde and creatine kinase levels. EAcF was effective upon neither the superoxide dismutase, glutationes reductase nor the catalase activities. In addition, the Western blot analysis revealed that ischemia-reperfusion injury significantly upregulates caspase 3 protein expression, while EAcF abolishes this effect. These results provide evidence that the EAcF reestablishes the cardiac contractility and prevents arrhythmias; it is suggested that EAcF could be used to reduce injury caused by cardiac reperfusion. However more clinical studies should be performed, before applying it in the clinic.
Subject(s)
Antioxidants , Myocardial Reperfusion Injury , Rats , Animals , Humans , Antioxidants/pharmacology , Antioxidants/metabolism , Myocardial Reperfusion Injury/metabolism , Canavalia/metabolism , Rats, Sprague-Dawley , Superoxide Dismutase/metabolism , Plant Leaves/metabolism , Myocardium/metabolismABSTRACT
Mannich bases consisting of 1,3,4-oxadiazole-2-thione (3 a-3 l) bearing various substituents were synthesized and found potent jack bean urease inhibitors. The prepared compounds showed significantly good inhibitory activities with IC50 values from 9.45±0.05 to 267.42±0.23â µM. The compound 3 k containing 4-chlorophenyl (-R) and 4-hydroxyphenyl (-R') was most active with IC50 9.45±0.05â µM followed by 3 e (IC50 22.52±0.15â µM) in which -R was phenyl and -R' was isopropyl group. However, when both -R and -R' were either 4-chlorophenyl groups (3 l) or only -R' was 4-nitrophenyl (3 i), both compounds were found inactive. The detailed binding affinities of the produced compounds with protein were explored through molecular docking and data-supported in-vitro enzyme inhibition profiles. Drug likeness was confirmed by in silico ADME investigations and molecular orbital analysis (HOMO-LUMO) and electrostatic potential maps were got from DFT calculations. ESP maps exposed that there are two potential binding sites with the most positive and most negative parts.
Subject(s)
Enzyme Inhibitors , Urease , Molecular Docking Simulation , Enzyme Inhibitors/chemistry , Structure-Activity Relationship , Oxadiazoles/pharmacology , Oxadiazoles/chemistry , Mannich Bases/pharmacology , Canavalia , Molecular StructureABSTRACT
Because of its high protein content, Jack bean (Canavalia ensiformis) is a promising alternative protein source. However, the utilization of Jack bean is limited due to the long cooking time to achieve palatable softness. We hypothesize that the cooking time may influence protein and starch digestibility. In this study, we characterized seven Jack bean collections with different optimal cooking times in terms of their proximate composition, microstructure and protein and starch digestibility. Kidney bean was included as a reference for microstructure and protein and starch digestibility. Proximate composition showed that Jack bean collections have a protein content ranging from 28.8 to 39.3%, a starch content ranging from 31 to 41%, a fiber content from 15.4 to 24.6%, and a concanavalin A content in the range 35-51 mg/g dry cotyledon. Particle sizes ranging between 125 and 250 µm were chosen as a representative sample of the whole bean to characterize microstructure and digestibility of the seven collections. Confocal laser microscopy (CLSM) revealed that Jack bean cells have an oval shape and contain starch granules embedded in a protein matrix similar to kidney bean cells. The diameter of Jack bean cells was measured by image analysis of CLSM micrographs and ranged from 103 to 123 µm, while the diameter of starch granules was 31-38 µm, comparatively larger than that of the kidney bean starch granules. Isolated intact cells were used to determine the starch and protein digestibility in the Jack beans collections. The digestion kinetics of starch followed a logistic model, whereas the digestion kinetics of protein followed a fractional conversion model. We found no correlation between optimal cooking time and kinetic parameters of protein and starch digestibility, implying that optimal cooking time is not predictive of protein and starch digestibility. In addition, we tested the effect of reduced cooking times on protein and starch digestibility on one Jack bean collection. The result showed that reducing cooking time significantly reduces starch digestibility, but not protein digestibility. The present study contributes to our understanding of the effect of food processing on protein and starch digestibility in legumes.
Subject(s)
Canavalia , Phaseolus , Cooking , Microscopy, Confocal , StarchABSTRACT
Cysteine-rich transmembrane module (CYSTM) proteins constitute small molecular protein families and have been identified across eukaryotes, including yeast, humans, and several plant species. Plant CYSTMs play vital roles in growth regulation, development, phytohormone signal transduction, pathogen defense, environmental stress response, and even heavy metal binding and detoxification. Canavalia rosea (Sw.) DC is a perennial halophyte with great semi-arid and saline-alkali tolerance. In this study, the CrCYSTM family including 10 members were identified in the C. rosea genome, with the purpose of clarifying the possible roles of CrCYSTMs in C. rosea plants development and stress resistance. The phylogenetic relationships, exon-intron structure, domain structure, chromosomal localization, and putative cis-acting elements in promoter regions were predicted and analyzed. Transcriptome analysis combined with quantitative reverse transcription PCR showed that different CrCYSTM members exhibited varied expression patterns in different tissues and under different abiotic stress challenges. In addition, several CrCYSTMs were cloned and functionally characterized for their roles in abiotic stress tolerance with yeast expression system. Overall, these findings provide a foundation for functionally characterizing plant CYSTMs to unravel their possible roles in the adaptation of C. rosea to tropical coral reefs. Our results also lay the foundation for further research on the roles of plant CYSTM genes in abiotic stress signaling, especially for heavy metal detoxification.
Subject(s)
Canavalia , Cysteine , Humans , Cysteine/metabolism , Canavalia/genetics , Canavalia/metabolism , Saccharomyces cerevisiae/metabolism , Phylogeny , Stress, Physiological/genetics , Plant Proteins/genetics , Plant Proteins/metabolism , Gene Expression Regulation, Plant , Multigene FamilyABSTRACT
Organic fertilization and mycorrhization can increase the phytoremediation of copper-contaminated soils. The time of vermicomposting alters the properties of vermicompost, which can affect copper's availability and uptake. Therefore, this study sought to evaluate the effect of different organic fertilizers and mycorrhization on copper-contaminated soil phytoremediation. The soil was contaminated with 100 mg Cu kg-1 dry soil and received mineral fertilizer (MIN), bovine manure (CM), and vermicompost produced in 45 days (V45) or 120 days (V120), all in doses equivalent to 40 mg kg-1 dry soil of phosphorus. Half of the jack bean (Canavalia ensiformis) plants were inoculated with the arbuscular mycorrhizal fungus Rhizophagus clarus. At plant flowering, the dry mass and concentrations of Cu, Zn, Mn, Ca, Mg, P, and K in the soil, solution, and plant tissue were determined, in addition to mycorrhizal colonization, nodulation, photosynthetic pigments, and oxidative stress enzyme activity. Organic fertilization increased plant growth and copper accumulation in aerial tissues. These effects were more evident with the V120, making it suitable for use in copper phytoextraction. Mycorrhization increased root and nodule dry mass, making it recommended for phytostabilization. C. ensiformis nodulation in Cu-contaminated soils depends on vermicompost fertilization and mycorrhization. Hence, the copper phytoremediation by C. ensiformis is increased by using organic fertilization and mycorrhization.
Subject(s)
Mycorrhizae , Soil Pollutants , Animals , Cattle , Copper/analysis , Soil , Biodegradation, Environmental , Sand , Canavalia , Soil Pollutants/analysis , FertilizationABSTRACT
Helicobacter pylori and Cryptococcus ssp. are pathogenic ureolytic microorganisms that cause several disorders in the host organism and, in severe cases, lead to death. Both infections have the urease enzyme as a key virulence factor since they use its ability to produce ammonia to soften the inhospitable pH to which they are subjected. In this review, we describe two ureases as possible molecular targets for drug discovery and provide insights for developing potent inhibitors against ureases from these pathogenic microorganisms through computer-aided drug discovery approaches, such as structure-based drug design (SBDD) and structure-activity relationship (SAR). The SAR studies have indicated several essential subunits and groups to be present in urease inhibitors that are critical for inhibitory activity against H. pylori or Cryptococcus spp. Since the threedimensional structure of C. neoformans urease has yet to be determined experimentally, the plant urease of Canavalia ensiformis was used in this study due to its structural similarity. Therefore, in the SBDD context, FTMap and FTSite analyses were performed to reveal characteristics of the urease active sites in two protein data bank files (4H9M, Canavalia ensiformis, and 6ZJA, H. pylori). Finally, a docking-based analysis was performed to explore the best inhibitors described in the literature to understand the role of the ligand interactions with the key residues in complex ligand-urease stabilization, which can be applied in the design of novel bioactive compounds.
Subject(s)
Cryptococcus neoformans , Helicobacter pylori , Humans , Urease/chemistry , Urease/metabolism , Cryptococcus neoformans/metabolism , Ligands , Canavalia/metabolismABSTRACT
The aim of this study was to investigate the effect of Canavalia gladiata extract (CGE) on the regulation of AMP-activated protein kinase (AMPK) in 3T3-L1 preadipocytes and evaluate the adipogenesis and lipogenesis mechanisms. In 3T3-L1 preadipocytes, lipid accumulation and differentiation were suppressed by 1.1, 1.3, and 1.4 times under the CGE treatment at 0.25, 0.5, and 1.0 mg/mL, respectively. The expression of the main genes involved in the inhibition of adipogenesis was evaluated at the mRNA level via a transcription-polymerase chain reaction. The extract at 1.0 mg/mL increased the mRNA expressions of AMPK and carnitine palmitoyl transferase-1 (CPT-1) by 1.9 and 1.2 times, respectively, while it decreased the expression of sterol regulatory element binding proteins-1c (SREBP-1c), peroxisome proliferator activated receptor-γ (PPAR-γ), CCAAT enhancer binding protein-α (C/EBP-α), and fatty acid synthase (FAS) by 1.1, 1.2, 1.8, and 1.5 times, respectively, indicating inhibition of the adipogenesis and lipogenesis potential of CGE. Gallic acid (4.02 mg/g) was identified as the main component of the CGE via LC-MS/MS and HPLC analysis. The results of this study suggested that CGE can be utilized as an anti-obesity food additive or medication by activating the AMPK-induced regulation and suppressing adipogenesis transcription factors.
Subject(s)
Adipogenesis , Lipogenesis , Mice , Animals , Adipogenesis/genetics , AMP-Activated Protein Kinases/metabolism , Canavalia/genetics , Chromatography, Liquid , Adipocytes/metabolism , Tandem Mass Spectrometry , RNA, Messenger/metabolism , 3T3-L1 Cells , PPAR gamma/genetics , PPAR gamma/metabolism , Cell Differentiation , Lipid Metabolism , CCAAT-Enhancer-Binding Protein-alpha/metabolismABSTRACT
In plants, the Gibberellic Acid-Stimulated Arabidopsis (GASA) gene family is unique and responds to ubiquitous stress and hormones, playing important regulatory roles in the growth and development of plants, as well as in the resistance mechanisms to biotic and abiotic stress. In this study, a total of 23 CrGASAs were characterized in C. rosea using a genome-wide approach, and their phylogenetic relationships, gene structures, conserved motifs, chromosomal locations, gene duplications, and promoter regions were systematically analyzed. Expression profile analysis derived from transcriptome data showed that CrGASAs are expressed at higher levels in the flowers or fruit than in the leaves, vines, and roots. The expression of CrGASAs also showed habitat- and environmental-stress-regulated patterns in C. rosea analyzed by transcriptome and quantitative reverse transcription PCR (qRT-PCR). The heterologous induced expression of some CrGASAs in yeast enhanced the tolerance to H2O2, and some CrGASAs showed elevated heat tolerance and heavy metal (HM) Cd/Cu tolerance. These findings will provide an important foundation to elucidate the biological functions of CrGASA genes, especially their role in the ecological adaptation of specific plant species to tropical islands and reefs in C. rosea.
Subject(s)
Arabidopsis , Arabidopsis/genetics , Arabidopsis/metabolism , Canavalia/genetics , Canavalia/metabolism , Gene Expression Regulation, Plant , Multigene Family , Phylogeny , Plant Proteins/metabolism , Hydrogen Peroxide/metabolismABSTRACT
Asthma is one of the most common inflammatory diseases of the lung worldwide. There has been considerable progress in recent studies to treat and prevent allergic asthma, however, various side effects are still observed in clinical practice. Six-week-old male BALB/c mice were orally administered with either sword bean pod extracts (SBP; 100 or 300 mg/kg) or dexamethasone (DEX; 5 mg/kg) once daily over 3 weeks, followed by ovalbumin sensitization (OVA/Alum.; intraperitoneal administration, 50 µg/2 mg/per mouse). Scoring of lung inflammation was performed to observe pathological changes in response to SBP treatment compared to OVA/Alum.-induced lung injury. Additionally, inflammatory cytokines were quantified in serum, bronchoalveolar lavage fluid (BALF), and lung tissue using ELISA and Western blot analyses. SBP treatment significantly reduced the infiltration of inflammatory cells, and release of histamine, immunoglobulin E, and leukotriene in serum and BALF. Moreover, the therapeutic effect of SBP was also assessed to analyze the inflammatory changes in the lung tissues. SBP markedly suppressed the activation of the MAPK signaling pathway and the expression of key inflammatory proteins (e.g., TNF-α) and Th2 type cytokines (IL-5 and IL-13). SBP was effective in ameliorating the allergic inflammation against OVA/Alum.-induced asthma by suppressing pulmonary inflammation.
Subject(s)
Asthma , Pneumonia , Alum Compounds , Animals , Asthma/chemically induced , Asthma/drug therapy , Asthma/metabolism , Bronchoalveolar Lavage Fluid , Canavalia , Cytokines/metabolism , Dexamethasone/pharmacology , Disease Models, Animal , Histamine/pharmacology , Immunoglobulin E , Inflammation/drug therapy , Interleukin-13 , Interleukin-5/adverse effects , Lung , Male , Mice , Mice, Inbred BALB C , Ovalbumin/adverse effects , Plant Extracts/therapeutic use , Pneumonia/drug therapy , Tumor Necrosis Factor-alpha/pharmacologyABSTRACT
Heat shock transcription factors (Hsfs) are key regulators in plant heat stress response, and therefore, they play vital roles in signal transduction pathways in response to environmental stresses, as well as in plant growth and development. Canavalia rosea (Sw.) DC. is an extremophile halophyte with good adaptability to high temperature and salt-drought tolerance, and it can be used as a pioneer species for ecological reconstruction on tropical coral islands. To date, very little is known regarding the functions of Hsfs in the adaptation mechanisms of plant species with specialized habitats, especially in tropical leguminous halophytes. In this study, a genome-wide analysis was performed to identify all the Hsfs in C. rosea based on whole-genome sequencing information. The chromosomal location, protein domain or motif organization, and phylogenetic relationships of 28 CrHsfs were analyzed. Promoter analyses indicated that the expression levels of different CrHsfs were precisely regulated. The expression patterns also revealed clear transcriptional changes among different C. rosea tissues, indicating that the regulation of CrHsf expression varied among organs in a developmental or tissue-specific manner. Furthermore, the expression levels of most CrHsfs in response to environmental conditions or abiotic stresses also implied a possible positive regulatory role of this gene family under abiotic stresses, and suggested roles in adaptation to specialized habitats such as tropical coral islands. In addition, some CrHsfAs were cloned and their possible roles in abiotic stress tolerance were functionally characterized using a yeast expression system. The CrHsfAs significantly enhanced yeast survival under thermal and oxidative stress challenges. Our results contribute to a better understanding of the plant Hsf gene family and provide a basis for further study of CrHsf functions in environmental thermotolerance. Our results also provide valuable information on the evolutionary relationships among CrHsf genes and the functional characteristics of the gene family. These findings are beneficial for further research on the natural ecological adaptability of C. rosea to tropical environments.