ABSTRACT
A novel MADS-box transcription factor from Pinus radiata D. Don was characterized. PrMADS11 encodes a protein of 165 amino acids for a MADS-box transcription factor belonging to group II, related to the MIKC protein structure. PrMADS11 was differentially expressed in the stems of pine trees in response to 45° inclination at early times (1 h). Arabidopsis thaliana was stably transformed with a 35S::PrMADS11 construct in an effort to identify the putative targets of PrMADS11. A massive transcriptome analysis revealed 947 differentially expressed genes: 498 genes were up-regulated, and 449 genes were down-regulated due to the over-expression of PrMADS11. The gene ontology analysis highlighted a cell wall remodeling function among the differentially expressed genes, suggesting the active participation of cell wall modification required during the response to vertical stem loss. In addition, the phenylpropanoid pathway was also indicated as a PrMADS11 target, displaying a marked increment in the expression of the genes driven to the biosynthesis of monolignols. The EMSA assays confirmed that PrMADS11 interacts with CArG-box sequences. This TF modulates the gene expression of several molecular pathways, including other TFs, as well as the genes involved in cell wall remodeling. The increment in the lignin content and the genes involved in cell wall dynamics could be an indication of the key role of PrMADS11 in the response to trunk inclination.
Subject(s)
Gene Expression Regulation, Plant , Pinus , Plant Proteins , Pinus/genetics , Pinus/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Arabidopsis/genetics , Arabidopsis/metabolism , Plant Stems/metabolism , Plant Stems/genetics , Cell Wall/metabolism , Cell Wall/genetics , Gene Expression Profiling , Transcription Factors/metabolism , Transcription Factors/genetics , Lignin/metabolism , Lignin/biosynthesis , MADS Domain Proteins/genetics , MADS Domain Proteins/metabolism , Plants, Genetically Modified/geneticsABSTRACT
Fragaria chiloensis is a Chilean native species that softens intensively during its ripening. Its softening is related to cell wall disassembly due to the participation of cell wall degrading enzymes. Softening of F. chiloensis fruit can be accelerated by ABA treatment which is accompanied by the increment in the expression of key cell wall degrading genes, however the molecular machinery involved in the transcriptional regulation has not been studied until now. Therefore, the participation of two MADS-box transcription factors belonging to different subfamilies, FchAGL9 and FchSHP, was addressed. Both TFs are members of type-II MADS-box family (MIKC-type) and localized in the nucleus. FchAGL9 and FchSHP are expressed only in flower and fruit tissues, rising as the fruit softens with the highest expression level at C3-C4 stages. EMSA assays demonstrated that FchAGL9 binds to CArG sequences of RIN and SQM, meanwhile FchSHP interacts only with RIN. Bimolecular fluorescence complementation and yeast two-hybrid assays confirmed FchAGL9-FchAGL9 and FchAGL9-FchSHP interactions. Hetero-dimer structure was built through homology modeling concluding that FchSHP monomer binds to DNA. Functional validation by Luciferase-dual assays indicated that FchAGL9 transactivates FchRGL and FchPG's promoters, meanwhile FchSHP transactivates those of FchEXP2, FchRGL and FchPG. Over-expression of FchAGL9 in C2 F. chiloensis fruit rises FchEXP2 and FchEXP5 transcripts, meanwhile the over-expression of FchSHP also increments FchXTH1 and FchPL; in both cases there is a down-regulation of FchRGL and FchPG. In summary, we provided evidence of FchAGL9 and FchSHP participating in the transcription regulation associated to F. chiloensis's softening.
ABSTRACT
Chilean papaya, also known as mountain papaya (Vasconcellea pubescens), is a fruit valued for its nutritional value and pleasant fragrance. The oblong fruit, featuring five ridges and a seed-filled mucilage cavity, is typically consumed cooked due to its high protease content. The mucilage and the seeds are usually discarded as byproducts. This study analyzed the biochemical composition of mountain papaya seed mucilage using methods such as HPAEC and immunolabeling. Results revealed that papaya seeds yield nearly 20% of their weight in mucilage polysaccharides, which can be separated into soluble and adherent layers. The mucilage exhibited a high proportion of acidic sugars, indicating that homogalacturonan (HG) is the predominant domain. It also contained other domains like rhamnogalacturonan-I (RG-I) and hemicelluloses, predominantly xyloglucan. The HG-rich mucilage, currently considered waste, emerges as a promising source of polysaccharides, indicating its multifaceted utility in various industrial applications.
ABSTRACT
Environmental pollution caused by petroleum-derived plastics continues to increase annually. Consequently, current research is interested in the search for eco-friendly bacterial polymers. The importance of Bacillus bacteria as producers of polyhydroxyalkanoates (PHAs) has been recognized because of their physiological and genetic qualities. In this study, twenty strains of Bacillus genus PHA producers were isolated. Production was initially evaluated qualitatively to screen the strains, and subsequently, the strain B12 or Bacillus sp. 12GS, with the highest production, was selected through liquid fermentation. Biochemical and molecular identification revealed it as a novel isolate of Bacillus cereus. Production optimization was carried out using the Taguchi methodology, determining the optimal parameters as 30 °C, pH 8, 150 rpm, and 4% inoculum, resulting in 87% and 1.91 g/L of polyhydroxybutyrate (PHB). Kinetic studies demonstrated a higher production within 48 h. The produced biopolymer was analyzed using Fourier-transform infrared spectroscopy (FTIR), confirming the production of short-chain-length (scl) polyhydroxyalkanoate, named PHB, and differential scanning calorimetry (DSC) analysis revealed thermal properties, making it a promising material for various applications. The novel B. cereus isolate exhibited a high %PHB, emphasizing the importance of bioprospecting, study, and characterization for strains with biotechnological potential.
ABSTRACT
INTRODUCTION: Pectin-oligosaccharides (POS) serve diverse purposes as a food ingredient, antimicrobial and biostimulant in plants, and their functionality is linked to the degree of esterification. Grape and broccoli wastes emerge as environmentally friendly alternatives to obtaining pectin, serving as a sustainable source to producing POS. For example, microwaves have proven to be an effective and sustainable method to extract polysaccharides from plant matrices. OBJECTIVE: This work aims to use grape and broccoli wastes as alternative sources for obtaining pectin by microwave-assisted extraction and biotransformation into POS, which possess biological properties. MATERIAL AND METHODS: The extraction conditions were identified at a power of 400 W, 300 s for the extraction of pectin from grape pomace and broccoli waste. Biotransformation of pectins into POS, using commercial enzyme preparations (Viscozyme L and Pectinase). Characterisation was carried out by Fourier-transform infrared spectroscopy, thermogravimetric analysis, and scanning electron microscopy. RESULTS: Physicochemical analysis indicated grape pomace and broccoli waste pectins had galacturonic acid content of 63.81 ± 1.67 and 40.83 ± 2.85 mg 100 mg-1, low degree of esterification of 34.89% and 16.22%, respectively. Biotransformation of pectins into POS resulted in a 20% hydrolysis rate. The main enzymatic activity was polygalacturonase for the degradation of the main structure of the pectin. CONCLUSION: Production of POS from agro-industrial wastes by emerging technologies, such as the combined use of microwave-assisted extraction and enzymatic processes, represents an alternative method for the generation of bioactive compounds with distinctive properties suitable for different applications of interest.
ABSTRACT
Grape pomace (GP), a by-product of wine production, contains bioactive polyphenols with potential health benefits. This study investigates the anti-inflammatory properties of a polyphenolic fraction derived from GP, obtained by ultrasound-microwave hybrid extraction and purified using ion-exchange chromatography. In the inflammation model, mice were divided into six groups: intact, carrageenan, indomethacin, and three GP polyphenols treatment groups. Paw edema was induced by subplantar injection of carrageenan, and the GP polyphenols were administered intraperitoneally at doses of 10, 20, and 40â mg/kg. The anti-inflammatory effect was evaluated by measuring paw volume, and expression of inflammatory markers: cyclooxygenase-2 (COX-2), myeloperoxidase (MPO), and cytokines (IL-1ß and IL-6), along with lipid peroxidation levels. The GP polyphenols significantly reduced paw edema and expression levels of COX-2, MPO, and cytokines in a dose-dependent manner effect, with the highest dose showing the greatest reduction. Additionally, lipid peroxidation levels were also decreased by GP polyphenols treatment at doses of 10 and 20â mg/kg. These findings suggest that ultrasound-microwave extraction combined with amberlite purification proved to be effective in obtaining a polyphenolic-rich fraction from GP. Thus, GP polyphenols may serve as a natural anti-inflammatory and antioxidant agent for treating inflammation and oxidative stress-related diseases.
Subject(s)
Carrageenan , Disease Models, Animal , Edema , Inflammation , Polyphenols , Vitis , Animals , Polyphenols/pharmacology , Polyphenols/isolation & purification , Polyphenols/chemistry , Mice , Vitis/chemistry , Inflammation/drug therapy , Inflammation/chemically induced , Inflammation/metabolism , Male , Edema/drug therapy , Edema/chemically induced , Lipid Peroxidation/drug effects , Cyclooxygenase 2/metabolism , Anti-Inflammatory Agents/pharmacology , Anti-Inflammatory Agents/chemistry , Anti-Inflammatory Agents/isolation & purification , Anti-Inflammatory Agents/therapeutic use , Dose-Response Relationship, Drug , Peroxidase/metabolism , Cytokines/metabolismABSTRACT
The Pacific whiteleg shrimp Penaeus (Litopenaeus) vannamei is a highly relevant species for the world's aquaculture development, for which an incomplete genome is available in public databases. In this work, PacBio long-reads from 14 publicly available genomic libraries (131.2 Gb) were mined to improve the reference genome assembly. The libraries were assembled, polished using Illumina short-reads, and scaffolded with P. vannamei, Feneropenaeus chinensis, and Penaeus monodon genomes. The reference-guided assembly, organized into 44 pseudo-chromosomes and 15,682 scaffolds, showed an improvement from previous reference genomes with a genome size of 2.055 Gb, N50 of 40.14 Mb, L50 of 21, and the longest scaffold of 65.79 Mb. Most orthologous genes (92.6%) of the Arthropoda_odb10 database were detected as "complete," and BRAKER predicted 21,816 gene models; from these, we detected 1,814 single-copy orthologues conserved across the genomic references for Marsupenaeus japonicus, F. chinensis, and P. monodon. Transcriptomic-assembly data aligned in more than 99% to the new reference-guided assembly. The collinearity analysis of the assembled pseudo-chromosomes against the P. vannamei and P. monodon reference genomes showed high conservation in different sets of pseudo-chromosomes. In addition, more than 21,000 publicly available genetic marker sequences were mapped to single-site positions. This new assembly represents a step forward to previously reported P. vannamei assemblies. It will be helpful as a reference genome for future studies on the evolutionary history of the species, the genetic architecture of physiological and sex-determination traits, and the analysis of the changes in genetic diversity and composition of cultivated stocks.
Subject(s)
Genome , Penaeidae , Penaeidae/genetics , Animals , Databases, Genetic , Genomics/methods , Molecular Sequence AnnotationABSTRACT
Pomegranate peel (PP) and coffee pulp (CP) are by-products of the food industry that can cause environmental pollution if not handled adequately. These by-products contain a significant amount of polyphenolic compounds which have antioxidant and possibly anticancer properties. We investigated the antiproliferative and cytotoxic activities of polyphenols from PP, CP and a 50-50% mixture of both against HeLa, A549, MDA-MB and Hek-293 cells. The total phenolic content of the PP and CP extracts was determined by high performance liquid chromatography/electrospray ionisation/mass spectrometry, and the antiproliferative and cytotoxic potentials were evaluated by MTT (3- (4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide) and Lactate Dehydogenase assays, respectively. Results showed antiproliferative and cytotoxic effects of polyphenols from PP and CP when administered at different concentrations or mixtures on HeLa, A549 and MDA-MB cells. No significant antiproliferative effects were observed on Hek-293 cells treated under similar conditions. These results suggest the potential of PP and CP polyphenols, individually or in combination, to modulate biological mechanisms involved in cervical, breast and lung cancer.
ABSTRACT
The present study aims to obtain manganese ferrite nanoparticles functionalized with chitosan (C-MNP) or ethylenediamine (E-MNP) by coprecipitation and polyol one-step methods, characterize their interaction with S. griseus demonstrating cell immobilization, and evaluate the antimicrobial activity of the free cell extracts obtained from immobilized S. griseus fermentation in the presence of different concentrations of MNP. The adsorption isotherms were analyzed mathematically using Langmuir and Freundlich models. The highest coefficient of determination (R2) for the S. griseus cell adsorption isotherm with C-MNP was observed with a linear function of the Langmuir model. The adsorption isotherm of S. griseus cells with E-MNP was better fitted to the Freundlich model. Cell immobilization by adsorption on magnetic nanoparticles was demonstrated in both cases. Different concentrations of C-MNP and E-MNP were used in fermentations to prepare cell-free extracts with antifungal activity. The best results were obtained with E-MNP, with a 91.5% inhibition of radial fungal growth. Magnetic nanoparticles offer potential applications in different fields and easy biomass separation.
ABSTRACT
Over the past few decades, efforts to eradicate hunger in the world have led to the generation of sustainable development goals to reduce poverty and inequality. It is estimated that the current coronavirus pandemic could add between 83 and 132 million to the total number of undernourished people in the world by 2021. Food insecurity is a contributing factor to the increase in malnutrition, overweight and obesity due to the quality of diets to which people have access. It is therefore necessary to develop functional foods that meet the needs of the population, such as the incorporation of sprouts in their formulation to enhance nutritional quality. Germination of grains and seeds can be used as a low-cost bioprocessing technique that provides higher nutritional value and better bioavailability of nutrients. Consequently, the manuscript describes relevant information about the germination process in different seeds, the changes caused in their nutritional value and the use of techniques within the imbibition phase to modify the metabolic profiles within the sprouts such as inoculation with lactic acid bacteria and yeasts, to generate a functional symbiotic food.
ABSTRACT
Porophyllum ruderale (Jacq.) Cass. (Asteraceae) has antiprotozoal properties and contains extractable phenolic compounds by the maceration method (M). However, new extraction proposals such as ultrasound (U), microwaves (MW), and ultrasound/microwaves (U/MW) have emerged to optimise yields, but it is unknown if these methods modify effectiveness. Therefore, the study consisted of extracting the aerial part of P. ruderale with ethanol using the M, U, MW and U/MW methods to study its composition by RP-HPLC-ESI-MS, its total polyphenol content and its effect against Entamoeba histolytica. The study showed that U, MW and U/MW did not modify the extraction yield compared to M, but they did change the composition and the total polyphenol content. All extracts contain phloretin, caffeic acid 4-O-glucoside, todolactol A, quercetin 3-O-glucoside, quercetin 3-O-rhamnoside, luteolin and 3,7-dimethylquercetin, and affected the growth of E. histolytica. However, M and U extracts were the most effective at 5 mg/mL.
ABSTRACT
Giardia lamblia is a globally distributed protozoan parasite that causes intestinal disease. Recently, there is an increase in refractory cases of giardiasis to chemotherapeutic agents, and drugs available cause side effects that may limit its use or cause therapeutic non-compliance. Therefore, search for alternative and less harmful drugs to treat giardiasis is an important task. In this sense, resveratrol (RSV) is a polyphenol with a wide range of pharmacological effects such as antimicrobial, anticarcinogenic and antioxidant. The aim of this study was to evaluate the effects of RSV on Giardia lamblia trophozoites in vitro and in silico, focusing on tubulin affectation, a major protein of the Giardia cytoskeleton which participates in relevant processes for cell survival. In vitro determinations showed that RSV inhibits parasite growth and adherence, causes morphological changes, and induces apoptosis-like cell death through tubulin alterations demonstrated by immunolocalization and Western blot assays. Bioinformatic analysis by molecular docking suggested that RSV binds to Giardia tubulin interface heterodimer, sharing binding residues to those reported with depolymerization inhibitors. These findings suggest that RSV affects microtubular dynamics and make it an interesting compound to study for its safety and antigiardiasic potential.
Subject(s)
Giardia lamblia , Giardiasis , Animals , Humans , Giardiasis/drug therapy , Giardiasis/parasitology , Tubulin/metabolism , Tubulin/pharmacology , Tubulin/therapeutic use , Resveratrol/pharmacology , Trophozoites , Molecular Docking SimulationABSTRACT
Resumen Introducción: La obesidad mórbida se asocia a alteraciones de la capacidad de caminar, sin embargo se desconoce cómo es el comportamiento de la prueba de caminata de 6 minutos en sujetos con incrementos del índice de masa corporal (IMC). Objetivo: Describir el comportamiento de la prueba de caminata de 6 minutos en sujetos con IMC normal hasta la obesidad mórbida. Métodos: Mediante un diseño transversal analítico se estudiaron sujetos de ambos sexos de 18 a 60 años con IMC: normal (18.5-24.9 kg/m2), sobrepeso (25-29.9 kg/m2), obesidad (30-39.9 kg/m2), obesidad mórbida (> 40 kg/m2). Se les realizó prueba de caminata de 6 minutos, se acotaron variables demográficas y antecedentes personales patológicos. Se analizaron las categorías de IMC con ANOVA de una vía y ajuste de Bonferroni y los sexos con prueba t, ambos para grupos independientes y correlaciones de Pearson para las diversas variables. Una p < 0.05 fue considerada estadísticamente significativa. Resultados: Se estudiaron 480 sujetos de ambos sexos en cuatro grupos. Edad: hombres 43 ± 11 y mujeres 45 ± 10 años. Porcentaje diabetes mellitus (6.7%), hipertensión arterial (18.3%). Metros caminados hombres vs. mujeres por IMC (normal: 483 ± 56 vs. 449 ± 61; sobrepeso: 471 ± 55 vs. 441 ± 44; obesidad: 455 ± 70 vs. 421 ± 47; obesidad mórbida: 443 ± 49 vs. 403 ± 54; p < 0.05). Correlación IMC-metros caminados r: -0.446 (p < 0.0001). Conclusiones: Los metros caminados en la prueba de caminata de 6 minutos disminuyeron conforme incrementó el IMC. El sexo masculino caminó más metros en todas las categorías.
Abstract Introduction: Morbid obesity is associated with alterations in the ability to walk, however, the behavior of the 6-minute walk test in subjects with increases in body mass index is unknown. Objective: To describe the behavior of the 6-minute walk test in subjects with normal body mass index to morbid obesity. Methods: Through an analytical cross-sectional design, subjects of both genders from 18 to 60 years old with body mass index were studied: Normal (BMI: 18.5-24.9); overweight (BMI: 25-29.9); obesity (BMI: 30-39.9); morbid obesity (BMI: > 40) kg/m2. A 6-minute walk test was performed, demographic variables and pathological personal history were delimited. BMI categories were analyzed with one-way ANOVA and Bonferroni adjustment, and gender with t-test, both for independent groups, and Pearson's correlations for the various variables. Results: 480 subjects of both genders were studied in four groups. Age: men 43 ± 11 and women 45 ± 10 years old. Percentage diabetes mellitus (6.7%), arterial hypertension (18.3%). Meters walked men vs. women by body mass index (normal: 483 ± 56 vs. 449 ± 61; overweight: 471 ± 55 vs. 441 ± 44; obesity: 455 ± 70 vs. 421 ± 47; morbid obesity: 443 ± 49 vs. 403 ± 54, p < 0.05). Correlation body mass index-meters walked: r: -0.446 (p < 0.0001). Conclusions: Meters walked in the 6-minute walk test decreased as body mass index increased. The male gender walked more meters in all categories.
ABSTRACT
In fishes, the availability of taurine is regulated during ontogenetic development, where its endogenous synthesis capacity is species dependent. Thus, different pathways and involved enzymes have been described: pathway I (cysteine sulfinate-dependent pathway), cysteine dioxygenase type 1 (cdo1) and cysteine sulfinic acid decarboxylase (csad); pathway II (cysteic acid pathway), cdo1 and glutamic acid decarboxylase (gad); and pathway III (cysteamine pathway), 2-aminoethanethiol dioxygenase (ado); whereas taurine transporter (taut) is responsible for taurine entry into cells on the cell membrane and the mitochondria. This study determined if the tropical gar (Atractosteus tropicus), an ancient holostean fish model, has the molecular mechanism to synthesize taurine through the identification and analysis expression of transcripts coding for proteins involved in its biosynthesis and transportation, at different embryo-larvae stages and in different organs of juveniles (31 dah). We observed a fluctuating expression of all transcripts involved in the three pathways at all analyzed stages. All transcripts are expressed during the beginning of larval development; however, ado and taut show a peak expression at 9 dah, and all transcripts but csad decreased at 23 dah, when the organism ended the larval period. Furthermore, at 31 dah, we observed taut expression in all examined organs. The transcripts involved in pathways I and III are expressed differently across all organs, whereas pathway II was only observed in the brain, eye, and skin. The results suggested that taurine biosynthesis in tropical gar is regulated during its early development before first feeding, and the pathway might also be organ-type dependent.
Subject(s)
Carboxy-Lyases , Fishes , Animals , Fishes/metabolism , Larva/genetics , Larva/metabolism , Taurine/metabolism , Carboxy-Lyases/genetics , Carboxy-Lyases/metabolismABSTRACT
Living in environments whose ecologies vary in periods as short as 24 h is a challenge for animals as Drosophila species that inhabit pear and apple orchards. These orchards have sunny and shady sections. The size and shape of these habitats change daily according to the position of the sun in the sky. Sunny areas are related to dryness and water loss, and shady places have lower temperatures and higher humidity. The presence of heterospecific flies may lead to competition for space and food. In sunny habitats we did not find adult Drosophila. In shady sections we found conspecific groups D. melanogaster, D. simulans, D. immigrans, D. subobscura, and the Chilean endemic D. pavani perched on grasses and herbs at 8-10 cm from fruits that had fallen on the ground. In the fruits, 99% of the adults were females and they were not grouped. The way in which daily changes in the size and shape of shady habitats together with the presence of heterospecific adults influence the selection of places to live is poorly understood in Drosophila. Our experiments show that adults of the five species prefer dark areas. The experimental results show that the odors of each species: i) influence conspecifics to select similar perch sites and decrease mobility, and ii) increase mobility in heterospecific adults and modify their perch site preferences. Attractions between conspecifics, the repulsions between species, and preferences for shaded areas matter in choosing a place to live in the five Drosophila species. These behaviors seem to have evolved as coordinated routines, contributing to the coexistence of the five Drosophila species in the apple and pear orchards examined.
Subject(s)
Drosophila melanogaster , Drosophila , Animals , Female , Male , Sexual Behavior, Animal , Ecosystem , OdorantsABSTRACT
Hormones act as master ripening regulators. In non-climacteric fruit, ABA plays a key role in ripening. Recently, we confirmed in Fragaria chiloensis fruit that in response to ABA treatment the fruit induces ripening-associated changes such as softening and color development. In consequence of these phenotypic changes, transcriptional variations associated with cell wall disassembly and anthocyanins biosynthesis were reported. As ABA stimulates the ripening of F. chiloensis fruit, the molecular network involved in ABA metabolism was analyzed. Therefore, the expression level of genes involved in ABA biosynthesis and ABA perception was quantified during the development of the fruit. Four NCED/CCDs and six PYR/PYLs family members were identified in F. chiloensis. Bioinformatics analyses confirmed the existence of key domains related to functional properties. Through RT-qPCR analyses, the level of transcripts was quantified. FcNCED1 codifies a protein that displays crucial functional domains, and the level of transcripts increases as the fruit develops and ripens, in parallel with the increment in ABA. In addition, FcPYL4 codifies for a functional ABA receptor, and its expression follows an incremental pattern during ripening. The study concludes that FcNCED1 is involved in ABA biosynthesis; meanwhile, FcPYL4 participates in ABA perception during the ripening of F. chiloensis fruit.
Subject(s)
Fragaria , Fragaria/metabolism , Fruit/metabolism , Chile , Anthocyanins/metabolism , Perception , Plant Proteins/genetics , Plant Proteins/metabolism , Gene Expression Regulation, Plant , Abscisic Acid/metabolismABSTRACT
Recent work demonstrating that asymptomatic carriers of P. falciparum parasites make up a large part of the infectious reservoir highlights the need for an effective malaria vaccine. Given the historical challenges of vaccine development, multiple parasite stages have been targeted, including the sexual stages required for transmission. Using flow cytometry to efficiently screen for P. falciparum gamete/zygote surface reactivity, we identified 82 antibodies that bound live P. falciparum gametes/zygotes. Ten antibodies had significant transmission-reducing activity (TRA) in a standard membrane feeding assay and were subcloned along with 9 nonTRA antibodies as comparators. After subcloning, only eight of the monoclonals obtained have significant TRA. These eight TRA mAbs do not recognize epitopes present in any of the current recombinant transmission-blocking vaccine candidates, Pfs230D1M, Pfs48/45.6C, Pf47 D2 and rPfs25. One TRA mAb immunoprecipitates two surface antigens, Pfs47 and Pfs230, that are expressed by both gametocytes and gametes/zygotes. These two proteins have not previously been reported to associate and the recognition of both by a single TRA mAb suggests the Pfs47/Pfs230 complex is a new vaccine target. In total, Pfs230 was the dominant target antigen, with five of the eight TRA mAbs and 8 of 11 nonTRA gamete/zygote surface reactive mAbs interacting with Pfs230. Of the three remaining TRA mAbs, two recognized non-reduced, parasite-produced Pfs25 and one bound non-reduced, parasite-produced Pfs48/45. None of the TRA mAbs bound protein on an immunoblot of reduced gamete/zygote extract and two TRA mAbs were immunoblot negative, indicating none of the new TRA epitopes are linear. The identification of eight new TRA mAbs that bind epitopes not included in any of the constructs currently under advancement as transmission-blocking vaccine candidates may provide new targets worthy of further study.
Subject(s)
Malaria Vaccines , Malaria, Falciparum , Humans , Plasmodium falciparum , Antibodies, Blocking , Epitopes , Antibodies, Protozoan , Antibodies, Monoclonal , Protozoan Proteins , Antigens, ProtozoanABSTRACT
Development of a malaria vaccine that blocks transmission of different parasite stages to humans and mosquitoes is considered critical for elimination efforts. A vaccine using Pfs25, a protein on the surface of zygotes and ookinetes, is under investigation as a transmission-blocking vaccine (TBV) that would interrupt parasite passage from mosquitoes to humans. The most extensively studied Pfs25 TBVs use Pichia pastoris-produced recombinant forms of Pfs25, chemically conjugated to a recombinant carrier protein, ExoProtein A (EPA). The recombinant form of Pfs25 first used in humans was identified as Pfs25H, which contained a total of 14 heterologous amino acid residues located at the amino- and carboxyl-termini including a His6 affinity tag. A second recombinant Pfs25, identified as Pfs25M, was produced to remove the heterologous amino acid residues and conjugated to EPA (Pfs25M-EPA). Here, monomeric Pfs25M was characterized biochemically and biophysically for identity, purity, and integrity including protein structure to assess its comparability with Pfs25H. Although the biological activities of Pfs25H and Pfs25M, whether generated by monomeric forms or conjugated nanoparticles, appeared similar, fine-mapping studies with two transmission-blocking monoclonal antibodies detected structural and immunological differences. In addition, evaluation of antisera generated against conjugated Pfs25H or Pfs25M nanoparticles in nonhuman primates identified polyclonal IgG that recognized these structural differences.
ABSTRACT
Five samples of agricultural soil and five samples of Aloe barbadensis (P. Mill., 1768) plants with symptoms of wilt and root necrosis were collected in five localities of the state of Tamaulipas, México. The aims of this study were the morphological identification, molecular identification and in vitro evaluation of the antagonistic activity of Trichoderma spp. on Fusarium spp. Four strains of Trichoderma asperellum, one strain of Trichoderma harzianum and five strains of Fusarium oxysporum were identified by morphological and molecular methods. The evaluation of the antagonistic activity of T. harzianum isolate (TP) showed the highest inhibition in Fusarium spp. (78.80%). The evaluation of the antagonistic activity of Trichoderma spp. extracts in Fusarium spp. did not show significant differences between treatments (P ≤ 0.05), with Trichoderma growth percentages that oscillated between 81.08 and 94.38%. The native isolate of T. harzianum (TP) showed significant competitive capability against the mycelial growth of F. oxysporum. Trichoderma species are promising agents of biological control in the central area of the State Tamaulipas, Mexico.
Subject(s)
Fusarium , Trichoderma , Soil , Soil Microbiology , Mexico , Plant Diseases/prevention & controlABSTRACT
Fructans are a polydisperse mixture of fructose polymers generally bound to a glucose molecule, in recent years, interest in their use has grown, either as a potential ingredient in functional foods or for their technological properties. The diversity of its applications lies in its structure and origin. Until now, the scientific approach has been more focused on inulin-type fructans and not so much on the effect of those of mixed branched structure as agave fructans. These have a complex structure with the presence of ß (2 - 1) and ß (2 - 6) bonds that give it prebiotic properties. In this context, a review is made of the general processes of extraction of agave fructans, as well as their technological functionality in the obtaining of base structures for the development of food products.