Microparticulated whey protein can be used as a fat substitute in foods to enhance their nutritional value and improve their sensory and rheological characteristics. Microparticulation denatures whey proteins through heat treatment, followed by shear rate control to determine the aggregate sizes formed. The processing method used during microparticulation can modify the protein denaturation rate, which may enable the application of new proteins. This study sought to develop a methodology for quantifying whey protein fractions in commercial samples before and after microparticulation at different concentrations and physical states. The developed methodology proved to be applicable because of its selectivity, quantification, detection limits, and precision. The analyzed samples exhibited a higher denaturation rate for β-lactoglobulin than α-lactoalbumin; the denaturation rate varied among samples due to differences in the microparticulation process. characteristics treatment formed states selectivity quantification limits precision βlactoglobulin β lactoglobulin αlactoalbumin α lactoalbumin α-lactoalbumin process

The coronavirus disease (COVID-19) pandemic, caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has had a profound impact on global health and socio-economic conditions. To date, various vaccines have been administered worldwide in an effort to curb the spread of the virus. Despite vaccination efforts, there have been complications. Existing antiviral drugs have shown limited effectiveness, prompting the use of computational methods to understand the dynamics of the virus and develop suitable treatments. The current study focuses on using biocompatible para-sulfonato-calix[4]arenes to dock against two key proteins of SARS-CoV-2, namely ribonucleic acid (RNA)-dependent RNA polymerase, and helicase. Docking results indicate a strong binding affinity of these compounds to the target proteins, with higher scores compared to commonly used medications. Molecular dynamics (MD) simulation validates the docking results, showing stable protein-ligand complexes over time. The compounds are also screened for absorption, distribution, metabolism, and excretion properties and toxicity, suggesting their potential as lead candidates for inhibiting the virus’s key proteins. However, further in vivo and in vitro studies are recommended to confirm these findings. COVID19 COVID 19 (COVID-19 pandemic SARSCoV2, SARSCoV2 SARSCoV SARS CoV , (SARS-CoV-2) socioeconomic socio economic conditions date efforts complications effectiveness treatments parasulfonatocalix4arenes parasulfonatocalixarenes para sulfonato calix 4 arenes 2, SARS-CoV-2 RNAdependent dependent polymerase helicase medications MD (MD proteinligand protein ligand time absorption distribution metabolism toxicity viruss s However findings COVID1 1 (COVID-1 (SARS-CoV-2 parasulfonatocalix SARS-CoV- (COVID- (SARS-CoV- SARS-CoV (COVID (SARS-CoV

ABSTRACT Purpose: To characterize the extracellular vesicle protein cargo in the aqueous humor and plasma of patients with ocular toxoplasmosis. Methods: Aqueous humor and plasma were collected from six patients with active ocular toxoplasmosis and six patients with cataract. Extracellular vesicles were isolated, and western blotting and mass spectrometry were performed for protein analysis. Results: All plasma samples from patients with ocular toxoplasmosis and cataract were positive for the tetraspanins CD63 and TSG101. However, the aqueous humor from patients with ocular toxoplasmosis was positive only for CD63. Sixty-seven new unreported proteins were identified in the aqueous humor and plasma of patients with the ocular toxoplasmosis and cataract. Of the 67 proteins, 10 and 7 were found only in the cataract and ocular toxoplasmosis groups, respectively. In general, these proteins were involved in immune system activation and retina homeostasis and were related to infections and retina-associated diseases. Conclusion: The distinct protein signatures between ocular toxoplasmosis and cataract may be helpful in the differential diagnosis of ocular toxoplasmosis. However, more studies are needed to better understand the role of these proteins in the pathogenesis of ocular toxoplasmosis. Purpose Methods isolated analysis Results CD CD6 TSG101 TSG However Sixtyseven Sixty seven 6 1 groups respectively general retinaassociated associated diseases Conclusion TSG10 TSG1

ABSTRACT Purpose: The epithelial–mesenchymal transition of human lens epithelial cells plays a role in posterior capsule opacification, a fibrotic process that leads to a common type of cataract. Hyaluronic acid has been implicated in this fibrosis. Studies have investigated the role of transforming growth factor (TGF)-β2 in epithelial–mesenchymal transition. However, the role of TGF-β2 in hyaluronic acid-mediated fibrosis of lens epithelial cell remains unknown. We here examined the role of TGF-β2 in the hyaluronic acid-mediated epithelial–mesenchymal transition of lens epithelial cells. Methods: Cultured human lens epithelial cells (HLEB3) were infected with CD44-siRNA by using the Lipofectamine 3000 transfection reagent. The CCK-8 kit was used to measure cell viability, and the scratch assay was used to determine cell migration. Cell oxidative stress was analyzed in a dichloro-dihydro-fluorescein diacetate assay and by using a flow cytometer. The TGF-β2 level in HLEB3 cells was examined through immunohistochemical staining. The TGF-β2 protein level was determined through western blotting. mRNA expression levels were determined through quantitative real-time polymerase chain reaction. Results: Treatment with hyaluronic acid (1.0 μM, 24 h) increased the epithelial–mesenchymal transition of HLEB3 cells. The increase in TGF-β2 levels corresponded to an increase in CD44 levels in the culture medium. However, blocking the CD44 function significantly reduced the TGF-β2-mediated epithelial–mesenchymal transition response of HLEB3 cells. Conclusions: Our study showed that both CD44 and TGF-β2 are critical contributors to the hyaluronic acid-mediated epithelial-mesenchymal transition of lens epithelial cells, and that TGF-β2 in epithelial-mesenchymal transition is regulated by CD44. These results suggest that CD44 could be used as a target for preventing hyaluronic acid-induced posterior capsule opacification. Our findings suggest that CD44/TGF-β2 is crucial for the hyaluronic acid-induced epithelial-mesenchymal transition of lens epithelial cells. Purpose epithelialmesenchymal mesenchymal opacification cataract TGFβ2 TGFβ TGF β2 β (TGF)-β However TGF-β acidmediated mediated unknown Methods HLEB (HLEB3 CD44siRNA CDsiRNA siRNA CD 300 reagent CCK8 CCK 8 CCK- viability migration dichlorodihydrofluorescein dichloro dihydro fluorescein cytometer staining blotting realtime real time reaction Results 1.0 10 1 0 (1. μM 2 h CD4 medium TGFβ2mediated TGFβmediated Conclusions acidinduced induced CD44/TGFβ2 CD44TGFβ2 CDTGFβ CD44/TGF CD44/TGF-β (HLEB 30 1. (1 CD44/TGFβ CD44TGFβ CD44TGF CDTGF 3 (

The in silico study aimed to suggest phytochemicals from Commiphora leptophloeos leaves essential oil as therapies against coronavirus disease (COVID-19) (or severe acute respiratory syndrome coronavirus 2, SARS-CoV-2). For that, we used molecular docking (MD), absorption, distribution, metabolism, excretion, and toxicity (ADMET) parameters, and pharmacokinetic analysis. MD showed the presence of hydrophobic and hydrogen bonds for nine compounds. β-Selinene and bicyclogermacrene showed the best interaction values with the protein Mpro (ID: 6Y2F) (-5.9) and chimeric receptor complexed with its receptor human ACE2 (ID: 6VW1) (-6.1 and -5.5) proteins, as well as low gastrointestinal and high epithelial absorptions, medium permeability for Caco-2 cells and high epithelial permeability, low oral excretion, and were inhibitors of CYP2C19 and CYP2C9 enzymes. On the other hand, both were not permeable to the blood-brain barrier and inhibitors or substrates for glycoprotein-D, and showed carcinogenic potential, but only β-selinene was considered mutagenic. Although little bioavailable, both presented aspects structurally adequate, being compounds with moderate synthesis difficulty. Therefore, more analyses are necessary to evaluate the mechanism of action and unsatisfactory parameters foreseen in bioinformatic assays. COVID19 COVID 19 (COVID-19 2 SARSCoV2. SARSCoV2 SARSCoV SARS CoV . SARS-CoV-2) that MD, , (MD) absorption distribution metabolism excretion ADMET (ADMET analysis βSelinene β Selinene ID (ID 6Y2F YF Y F 5.9 59 5 9 (-5.9 ACE 6VW1 VW 6.1 61 6 1 (-6. 5.5 55 -5.5 proteins absorptions Caco2 Caco Caco- CYPC CYP C CYP2C1 CYP2C enzymes hand bloodbrain blood brain glycoproteinD, glycoproteinD glycoprotein D, D glycoprotein-D potential βselinene selinene mutagenic bioavailable adequate difficulty Therefore assays COVID1 (COVID-1 SARS-CoV-2 (MD 5. (-5. 6VW 6. (-6 -5. (COVID- SARS-CoV- (-5 (- -5 (COVID SARS-CoV ( -

Seaweeds have been explored by humans for thousands of years as a source of chemical compounds. This study describes the content of minerals, ash, carbohydrates, protein, lipids, and main metabolites of dichloromethane / methanol extracts of the seaweed Ulva lactuca, Padina gymnospora, Palisada perforata and Gelidiella acerosa from sandstone reefs on the Brazilian tropical coast (Pernambuco, Northeastern of Brazil). The content (% dry weight) of carbohydrates ranged from 14.35-48.52, proteins 7.49-14.98, total lipids 0.40-8.92, and ash 18.51-37.02. The concentration (mg kg dry algae−1) of Ca (900-3468), Mg (1655-4902), K (810-1707), Na (1062-4580), Mn (19-4462), and Cu (3.6-6.4) were maximum in Palisada and minimum in Padina. In turn, the lowest and highest contents (mg kg dry algae−1) of Fe (100-2312), Zn (18-43), and Cr (0.08-0.93) were recorded in Gelidiella and Ulva, respectively. Neophytadiene was the major compound. Phytol and palmitic acid were found in all seaweeds, although in low quantities. Palisada had the highest contents (% dry weight) of metabolites (neophytadiene: 23.89, phytol: 8.29; palmitic acid: 8.32), while Ulva had the lowest, except phytone, which was present only in this species. Our findings highlight the potential of these macroalgae from the coastal reefs as a source of chemical compounds. compounds minerals protein lactuca gymnospora Pernambuco, Pernambuco (Pernambuco Brazil. Brazil . Brazil) % ( weight 14.3548.52, 14354852 14.35 48.52, 14 35 48 52 14.35-48.52 7.4914.98, 7491498 7.49 14.98, 7 49 98 7.49-14.98 0.408.92, 040892 0.40 8.92, 0 40 8 92 0.40-8.92 18.5137.02. 18513702 18.51 37.02. 18 51 37 02 18.51-37.02 mg algae−1 algae1 algae 1 9003468, 9003468 900 3468 , (900-3468) 16554902, 16554902 1655 4902 (1655-4902) 8101707, 8101707 810 1707 (810-1707) 10624580, 10624580 1062 4580 (1062-4580) 194462, 194462 19 4462 (19-4462) 3.66.4 3664 3.6 6.4 3 6 4 (3.6-6.4 turn 1002312, 1002312 100 2312 (100-2312) 1843, 1843 43 (18-43) 0.080.93 008093 0.08 0.93 08 93 (0.08-0.93 respectively compound seaweeds quantities neophytadiene (neophytadiene 2389 23 89 23.89 phytol 8.29 829 29 8.32, 832 8.32 32 8.32) phytone species 3548 14.3548.52 1435485 1435 14.3 4852 48.52 5 14.35-48.5 4914 7.4914.98 749149 749 7.4 1498 14.98 9 7.49-14.9 408 0.408.92 04089 040 0.4 892 8.92 0.40-8.9 5137 18.5137.02 1851370 1851 18.5 3702 37.02 18.51-37.0 algae− 900346 90 346 (900-3468 1655490 165 490 (1655-4902 810170 81 170 (810-1707 1062458 106 458 (1062-4580 19446 446 (19-4462 66 3.66. 366 36 3. 64 6. (3.6-6. 100231 10 231 (100-2312 184 (18-43 080 0.080.9 00809 008 0.0 093 0.9 (0.08-0.9 238 2 23.8 8.2 82 83 8.3 354 14.3548.5 143548 143 14. 485 48.5 14.35-48. 491 7.4914.9 74914 74 7. 149 14.9 7.49-14. 0.408.9 0408 04 0. 8.9 0.40-8. 513 18.5137.0 185137 185 18. 370 37.0 18.51-37. 90034 34 (900-346 165549 16 (1655-490 81017 17 (810-170 106245 45 (1062-458 1944 44 (19-446 3.66 (3.6-6 10023 (100-231 (18-4 0.080. 0080 00 09 (0.08-0. 23. 8. 14.3548. 14354 48. 14.35-48 7.4914. 7491 7.49-14 0.408. 0.40-8 18.5137. 18513 37. 18.51-37 9003 (900-34 16554 (1655-49 8101 (810-17 10624 (1062-45 194 (19-44 (3.6- 1002 (100-23 (18- 0.080 (0.08-0 14.3548 14.35-4 7.4914 7.49-1 0.408 0.40- 18.5137 18.51-3 (900-3 (1655-4 (810-1 (1062-4 (19-4 (3.6 (100-2 (18 (0.08- 14.354 14.35- 7.491 7.49- 18.513 18.51- (900- (1655- (810- (1062- (19- (3. (100- (1 (0.08 (900 (1655 (810 (1062 (19 (3 (100 (0.0 (90 (165 (81 (106 (10 (0. (9 (16 (8 (0

Brazilian Savanna biodiversity is abundant in native fruits with nutritional and sensory properties with potential technological applications. However, the information available regarding the proprieties of Brazilian native fruits is still limited. This study aimed to analyze the metabolic profile of Brosimum gaudichaudii fruits (BGF) using metabolomics approaches, besides its physicochemical and nutritional proprieties. BGF exhibited protein content at least two times higher than other native fruits. Fifty-five metabolites were identified by the metabolomics approaches, mainly fatty acids (n = 22), carbohydrates (n = 13), organic acids (n = 6), and polyphenols (n = 5). The most abundant constituents were D-fructose (42.97%) for sugars, L-asparagine (6.47%) for amino acids, succinic acid (44.11%) for organic acids, and linolenic acid (3.04%) for fatty acids. Highlighted phytochemicals included chlorogenic acid, α-amyrin, and γ-tocopherol. Additionally, this study revealed that BGF is a good source of vitamin C and dietary fiber and an excellent source of vitamin A. Future studies providing nutritional information and suggesting consumption methods and potential industrial applications are essential to familiarize consumers with these foods. Additionally, clarifying the appropriate planting and harvesting seasons can revive traditional practices, especially for native fruits like BGF. However limited (BGF approaches Fiftyfive Fifty five n 22, 22 , 22) 13, 13 13) 6, 6 6) 5. 5 . 5) Dfructose D fructose 42.97% 4297 42 97 (42.97% sugars Lasparagine L asparagine 6.47% 647 47 (6.47% 44.11% 4411 44 11 (44.11% 3.04% 304 3 04 (3.04% αamyrin, αamyrin α amyrin, amyrin α-amyrin γtocopherol. γtocopherol γ tocopherol. tocopherol γ-tocopherol Additionally A foods practices 2 1 42.97 429 4 9 (42.97 6.47 64 (6.47 44.11 441 (44.11 3.04 30 0 (3.04 42.9 (42.9 6.4 (6.4 44.1 (44.1 3.0 (3.0 42. (42. 6. (6. 44. (44. 3. (3. (42 (6 (44 (3 (4 (

The emergence of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) stands for being the most serious epidemic (so far) of the 21st century. However, only a few computational studies have investigated the molecular mechanisms underlying the neutralization of the spike protein by antibodies of different classes. Hence, bioinformatic methods were employed to unravel the factors contributing to the remarkable neutralization capacity exhibited by specific antibodies. Initially, crystallographic structures of IgA monomeric / dimeric, IgG, and IgM antibodies binding with the receptor-binding domain region of the SARS-CoV-2 spike protein were retrieved. Subsequently, rigid molecular docking and molecular dynamic simulations were performed over 100 ns with explicit water solvation. Lastly, an energy decomposition was conducted to estimate the binding affinity using the last frames from molecular dynamics. The results revealed a higher binding affinity for both monomeric and dimeric forms of IgA antibodies against the spike protein. Additionally, a greater number of hydrogen bonds were observed during their interaction with the spike protein, as well as greater structural instability along the time and especially a more thermodynamically favorable interaction affinity. In this way, the research contributes a small piece to the complex puzzle of understanding the humoral immune response induced by the SARS-CoV-2 virus. SARSCoV2 SARSCoV SARS CoV (SARS-CoV-2 so far st century However classes Hence Initially IgG receptorbinding receptor SARS-CoV- retrieved Subsequently 10 solvation Lastly dynamics Additionally way virus (SARS-CoV- SARS-CoV 1 (SARS-CoV

RESUMO Este estudo foca no desempenho de vacas para produção de carne, criadas nos pântanos do Paraguai, examinando cinco genótipos bovinos; Brahman, Brangus, Nelore, bem como duas raças locais em risco de extinção. O principal objetivo é identificar e classificar variáveis fenotípicas que incluem variáveis sanguíneas, clínicas, de pelagem e saúde, demonstrando ligação causal com o peso vivo das vacas analisadas. Inicialmente, foram identificadas correlações elevadas entre diferentes variáveis incluídas neste estudo, e, então, utilizando técnicas avançadas de aprendizado de máquina e a aplicação de explicações aditivas de Shapley (SHAP), foi proporcionado um entendimento mais profundo dos fatores fortemente associados à adaptabilidade nestes ambientes, e, portanto, o respectivo desempenho zootécnico. A associação entre o componente genotípico bovino ligado à estação do ano mostrou ser o fator influente mais predominante sobre o peso vivo bovino. Variáveis como comprimento do pelo, hematócrito, fosfatase, fósforo, creatina phosphokinase, creatinina, proteína, cortisol, cálcio e a presença de endoparasitas foram destacadas, demonstrando sua importância hierárquica para a seleção animal. Os modelos de ML são ferramentas eficazes para estabelecer hierarquias de relevância em multivariáveis fenotípicas complexas, o que é crucial em programas de melhoramento genético em diferentes espécies zootécnicas, bem como em ambientes especiais e específicos, como os pântanos. carne Paraguai bovinos Brahman Brangus Nelore extinção sanguíneas clínicas saúde analisadas Inicialmente então SHAP, SHAP , (SHAP) portanto zootécnico pelo hematócrito fosfatase fósforo phosphokinase creatinina proteína cortisol destacadas animal complexas zootécnicas específicos (SHAP

ABSTRACT This study focuses on the performance of cows for meat production raised in the wetlands of Paraguay, examining five cattle genotypes: Brahman, Brangus, and Nelore, as well as two local breeds at risk of extinction. The main objective is to identify and rank phenotypic variables, including blood, clinical, hair, and health variables, demonstrating causal linkage with the live weight of the cows analyzed. Initially, high correlations were identified between different variables included in this study; then, using advanced Machine learning (ML) techniques and the application of Shapley additive explanations (SHAP), a deeper understanding was provided of the factors strongly associated with adaptability in these environments, and, therefore, the respective zootechnical performance. The association between cattle genotypic components linked with the season of the year proved to be the most influential factor on cattle live weight. Variables such as hair length, hematocrit, phosphatase, phosphorus, creatine phosphokinase, creatinine, protein, cortisol, calcium, and the presence of endoparasites were highlighted, demonstrating their hierarchical importance for animal selection. ML models are effective tools for establishing hierarchies of relevance in complex phenotypic multivariable, which is crucial in breeding programs for different zootechnical species and in special and specific environments like wetlands. Paraguay genotypes Brahman Brangus Nelore extinction blood clinical analyzed Initially then (ML SHAP, SHAP , (SHAP) therefore length hematocrit phosphatase phosphorus phosphokinase creatinine protein cortisol calcium highlighted selection multivariable (SHAP

ABSTRACT The development of aquaculture through the cultivation of what are considered unconventional species is an option with the potential to solve the problem of food scarcity. The aim of the present study was to evaluate the zootechnical performance of juvenile Pomacea haustrum fed different commercial diets, and subjected to different levels of salinity. After seven P. haustrum spawn had hatched in the laboratory, the juveniles were randomly selected and transferred to the rearing containers. Two different commercial diets were offered: rabbit feed with a high proportion of ingredients rich in crude vegetable protein (VP), and dog food containing balanced levels of ingredients rich in crude vegetable and animal protein (MP). The selected juveniles were randomly stocked at different salinities (0, 2 and 4 parts per thousand - ppt). The experiment lasted 65 days. Twenty-one hours after the start of the experiment, 100% of the individuals reared at a salinity of 4 ppt had died. There was a significant difference (p ≤ 0.05) between the results for zootechnical performance (average final weight, weight gain, average final growth, specific growth rate, protein efficiency and apparent feed conversion) for animals reared in fresh water or at a salinity of 2 ppt, regardless of the type of diet. The origin of the protein in the diet had no significant effect on the zootechnical performance of the individuals; however, the salinity of the water impaired development of the animals in direct proportion to its concentration. scarcity P laboratory containers offered VP, VP , (VP) MP. MP . (MP) 0, 0 (0 ppt. ppt) 6 days Twentyone Twenty one 100 died p 0.05 005 05 gain rate conversion however concentration (VP (MP ( 10 0.0 00 1 0.

Computational predictive approaches present themselves as an attractive alternative in the search for new compounds that may present toxic or growth regulatory activity in Aedes aegypti. Aspergillus species is a producer of secondary metabolites with diverse biological activities. In this study, it was possible to evaluate the pharmacokinetic and toxicological properties in silico, as well as the prediction of the insecticidal biological activities of the juvenile hormone of natural substances produced by the genus Aspergillus. Initially, 187 molecules were cataloged from various species of Aspergillus spp., and only seven molecules of indole alkaloids were found to exhibit a toxic dose 50% (TD50) after conducting toxicity predictions. The molecular dynamic simulation trajectories were utilized to study parameters such as root mean square deviation, root mean square fluctuation, radius of gyration and intermolecular hydrogen bond to understand the behavior and stability of proposed compounds in the binding pocket of the target protein. The N-β-acetyltryptamine molecule showed a binding affinity value of –8.100 ± 0.200 kcal mol-1, which classifies the compound as a potential insecticidal agent against the Aedes aegypti vector. silico Initially 18 spp spp. 50 TD50 TD (TD50 predictions deviation fluctuation protein Nβacetyltryptamine N β acetyltryptamine 8100 8 100 –8.10 0200 0 200 0.20 mol1, mol1 mol 1, 1 mol-1 vector 5 TD5 (TD5 810 10 –8.1 020 20 0.2 mol- (TD 81 –8. 02 2 0. –8 –

Resumen Introducción. La contaminación por metales pesados es un problema que ha ido en aumento, debido a las actividades antrópicas. Las plantas pueden eliminar, destruir o transformar contaminantes del suelo, agua y aire. Con el empleo de reguladores del crecimiento vegetal y agentes quelantes se puede reducir la toxicidad por metales pesados. Objetivo. Evaluar el efecto de la aplicación de una mezcla de oligogalacturónidos sobre la actividad peroxidasa y los contenidos de malondialdehido, proteinas totales y clorofilas, en plantas de tomate (Solanum lycopersicum L.) var. Amalia sometidas a altos niveles de metales pesados. Materiales y métodos. Se colectó suelo contaminado con Cu, Cd y el Fe, en la Empresa cerámica blanca Adalberto Vidal de Mayabeque, Cuba en el año 2020. El experimento se desarrolló en bolsas de 7 kg de capacidad y se utilizaron plantas de tomate, en condiciones semi controladas en áreas del Instituto Nacional de Ciencias Agrícolas (INCA), con un diseño experimental completamente aleatorizado con siete tratamientos y diez réplicas. Los tratamientos del producto a base de oligogalacturónidos (Ogal) fueron: aplicación al suelo (30 mg kg-1); aspersión foliar (20 mg L-1), la combinación de estas formas de aplicación y un testigo. A los 35 y 56 días de emergidas las plantas, se determinó la actividad peroxidasa y los contenidos de malondialdehido, proteinas totales y clorofilas. Resultados. El efecto que provocan el Cu, Cd y el Fe en las plántulas, fue atenuado por todas las variantes de uso de la mezcla Ogal, por lo que las mismas han desarrollado mecanismos de adaptación ante las altas concentraciones metálicas que le permiten subsistir al estrés abiótico. Conclusiones. La mezcla de Ogal atenuó el efecto que provocan los metales pesados en la actividad peroxidasa, el contenido malondialdehido, proteínas totales y clorofilas en plantas de tomate var. Amalia.

Abstract Introduction. Heavy metal pollution is a growing problem due to human activities. Plants can remove, destroy, or transform contaminants from soil, water, and air. By using plant growth regulators and chelating agents, heavy metal toxicity can be reduced. Objective. To evaluate the effect of applying a mixture of oligogalacturonides on peroxidase activity and the contents of malondialdehyde, total protein, and chlorophylls in tomato plants (Solanum lycopersicum L.) var. Amalia subjected to high levels of heavy metals. Materials and methods. Soil contaminated with Cu, Cd, and Fe was collected from the Adalberto Vidal White Ceramic Company in Mayabeque, Cuba, in 2020. The experiment was conducted in 7 kg capacity bags using tomato plants, under semi-controlled conditions at the Instituto Nacional de Ciencias Agrícolas (INCA), with a completely randomized experimental design with each treatment having ten replicates. The treatments with the oligogalacturonides (Ogal) were soil application (30 mg kg-1); foliar spray (20 mg L-1), the combination of these application forms, and a control. Peroxidase activity and the contents of malondialdehyde, total protein, and chlorophylls were determined at 35 and 56 days after the plant’s emergence. Results. The effect of Cu, Cd, and Fe on the seedlings was attenuated by all the use of the Ogal mixture variants use, indicating that they have developed adaptation mechanisms to high metal concentrations which allows to withstand abiotic stress. Conclusions. The Ogal mixture attenuated the effect of heavy metals on peroxidase activity, malondialdehyde content, total proteins, and chlorophylls in Amalia tomato plants.

Summary: Biofortification is a process that increases the nutrient content in plants’ edible parts. The objective was to determine the effect of iron chelate and iron sulfathe applied through foliar and edaphic methods on the mineral, nutritional, and bioactive compound content of the Verdin bean grain. Foliar doses of 0, 25, 50 and 100 mM of iron chelate and edaphic doses of 0, 0.25 and 0.50 g per plant of iron sulfate were applied, which generated 12 treatments that were evaluated in a randomized block design in 4×3 factorial arrangement where the first factor was the foliar doses, and the second factor was the edaphic doses. The content of iron, zinc, manganese, nickel, calcium and potassium, ash, proteins, fats, fiber, total phenols, flavonoid, anthocyanins, and antioxidant capacity were determined in the grain. The individual applications of foliar and edaphic iron had significant effects (P ≤ 0.05), with the edaphic dose of 0.50 g presenting the greatest increases in iron (17.38%), nickel (20.69%), protein (10.93%), crude fiber (5.82%) and antioxidant capacity (2.84%) regarding doses without edaphic iron. The simultaneous application of edaphic and foliar iron showed statistical differences (P ≤ 0.05), presenting the greatest increase in iron (75.91%), nickel (30.61%), ash (114.69%), protein (18.14%) and crude fiber (15.34%) with the 100 mM foliar - 0.50 g edaphic combination regarding to the combination without iron. For individual and simultaneous iron applications, it was observed that increasing the foliar and edaphic doses had antagonistic effects on the zinc and fat content. The application of iron can be beneficial to increase the mineral and nutritional content, and some bioactive compounds, but care must be taken to minimize the negative effects on other nutrients and properties of the grain.

Resumen: La biofortificación es el proceso por medio del cual se puede incrementar el contenido de nutrientes en la parte comestible de las plantas cultivadas. El objetivo fue determinar el efecto del quelato y sulfato de hierro aplicado de forma foliar y edáfica en el contenido mineral, nutricional, y compuestos bioactivos del grano de frijol variedad Verdín. Se aplicaron las dosis de 0, 25, 50 y 100 mM de quelato de hierro foliar y dosis de 0, 0.25 y 0.50 g por planta de sulfato de hierro edáfico, lo que origino 12 tratamientos que se evaluaron bajo un diseño de bloques al azar en arreglo factorial 4×3 donde el primer factor fueron las dosis foliares y el segundo factor las dosis edáficas. En el grano se determinó el contenido de hierro, zinc, manganeso, níquel, calcio, potasio, cenizas, proteínas, grasas, fibra, fenoles totales, flavonoides, antocianinas y capacidad antioxidante. Las aplicaciones individuales de hierro edáfico y foliar tuvieron efectos significativos (P < 0.0001), presentando la aplicación edáfica de 0.50 g de hierro los mayores incrementos de 17.38%, 20.69%, 10.93%, 5.82% y 2.84% del contenido de hierro, níquel, proteína, fibra cruda y capacidad antioxidante con respecto a la dosis sin hierro edáfico. Pero se observaron efectos antagónicos en el contenido de zinc y grasas al incrementar la dosis de hierro. La aplicación simultánea de hierro edáfico y foliar presentó diferencias estadísticas (P ≤ 0.05), presentando el mayor incremento de hierro (75.91%), níquel (30.61%), cenizas (114.69%), proteína (18.14%) y fibra cruda (15.34%) la aplicación de 100 mM foliar y 0.50 g edáfico con respecto a la combinación sin hierro. La aplicación combinada de hierro puede ser benéfica para aumentar el contenido mineral, nutricional y algunos compuestos bioactivos; pero se debe tener cuidado para minimizar los efectos negativos en otros nutrientes y propiedades del grano.

Resumen (Objetivo) Tryapanosoma cruzi, un agente causal de la enfermedad de Chagas, es un parásito que presenta una alternancia en su ciclo de vida entre un hospedador invertebrado (triatomino) y uno vertebrado (mamífero). Varios estudios han demostrado que en T. cruzi, HSP83 (homólogo de HSP90) es esencial para la división celular y el control de la respuesta al estrés térmico. Esta investigación se concentró en el estudio de la clonación, la caracterización bioinformática y la expresión del gen de la proteína de choque térmico HSP83 de T. cruzi para estudios posteriores sobre señalización. (Metodología) El ARN fue extraído de epimagostigotes T. cruci (clon EPm6/MOHM/VE/2007/ 6c), utilizando un kit comercial. El ADNc que codifica HSP83 se obtuvo utilizando RT-PCR, a partir del ARNm extraído, para lo cual se diseñaron los cebadores con base en la secuencia HSP83 de la cepa T. cruzi CL Brener. La clonación se realizó usando pGEM®T-Easy y se subclonó en el vector de expresión pQE30. Se efectuó la caracterización bioinformática y de secuencias. El gen se expresó y la proteína recombinante se purificó por cromatografía de afinidad, igual que se identificó por inmunotransferencia. (Resultados) El análisis de secuencia mostró similitud con el gen HSP83 de Trypanosoma cruzi y se observaron dominios HSP, así como epítopos B en la secuencia. Después de 3 horas de inducción con IPTG, se obtuvo una proteína recombinante con un peso aproximado de 83 kDa. La reacción de inmunotransferencia con suero hiperinmune anti-epimastigote de T. cruzi permitió la detección de una sola banda con un peso molecular de aproximadamente 83 kDa. (Conclusiones) Todos los resultados indican que se logró la clonación y caracterización de HSP83 de Trypanosoma cruzi.

Resumo (Objetivo) O Tryapanosoma cruzi, agente causador da doença de Chagas, é um parasita que apresenta uma alternância em seu ciclo de vida entre um hospedeiro invertebrado triatomíneo) e um vertebrado(mamífero). Vários estudos demonstraram que, no T. cruzi, a HSP83 (homóloga da HSP90) é essencial para a divisão celular e o controle da resposta ao estresse térmico. Esta pesquisa concentrou-se no estudo da clonagem, caracterização bioinformática e expressão do gene da proteína de choque térmico HSP83 do T. cruzi para estudos adicionais de sinalização. (Metodologia) O RNA foi extraído de epimastigotas de T. cruzi (clone EPm6/MOHM/VE/2007/ 6c) usando um kit comercial. O cDNA que codifica a HSP83 foi obtido por meio de RT-PCR a partir do mRNA extraído, para o qual os primers foram projetados com base na sequência da HSP83 da cepa T. cruzi CL Brener. A clonagem foi realizada com o pGEM®T-Easy e subclonada no vetor de expressão pQE30. Foi realizada a caracterização bioinformática e de sequência. O gene foi expresso e a proteína recombinante foi purificada por cromatografia de afinidade e identificada por imunotransferência. (Resultados) A análise da sequência mostrou similaridade com o gene HSP83 do Trypanosoma cruzi e os domínios HSP, bem como os epítopos B, foram observados na sequência. Após 3 horas de indução com IPTG, foi obtida uma proteína recombinante com peso aproximado de 83 kDa. A imunotransferência com soro hiperimune antiepimastigota de T. cruzi permitiu a detecção de uma única banda com peso molecular de aproximadamente 83 kDa. (Conclusões) Todos os resultados indicam que a clonagem e a caracterização da HSP83 do Trypanosoma cruzi foram alcançadas.

Abstract (Objective) Trypanosoma cruzi, a causal agent of Chagas’ disease, is a parasite whose life cycle alternates between an invertebrate (triatomine) and a vertebrate (mammal) host. Various studies have shown that in T. cruzi, HSP83 (a homolog of HSP90) is essential for cell division and control of the response to thermal stress. This investigation focused on studying the cloning, bioinformatic characterization, and expression of the heat shock protein HSP83 T. cruzi gene for further cell signaling studies. (Methodology) RNA was extracted from T. cruzi epimastigotes (EPm6 clone, MHOM/VE/2007/ 6c) using a commercial kit. The cDNA encoding HSP83 was determined using RT-PCR on extracted mRNA, for which the primers were designed based on the HSP83 sequence of T. cruzi strain CL Brener. Cloning was performed using pGEM®T-Easy and subcloned into the expression vector pQE30. Sequence and bioinformatic characterization were performed. The gene was expressed, and the recombinant protein was purified using affinity chromatography and identified through immunoblotting. (Results) Sequence analysis showed similarity to the gene encoding HSP83 from Trypanosoma cruzi, and HSP domains and B epitopes in the sequence were also observed. After 3 hours of induction with IPTG, a recombinant protein with an approximate weight of 83 kDa was obtained. The immunoblotting reaction with hyperimmune anti-T. cruzi epimastigote serum helped detect a single band with a molecular weight of nearly 83 kDa. (Conclusions) All results indicate that the cloning and characterization of HSP83 from Trypanosoma cruzi was achieved.

Resumen (Objetivo) Plasmodium vivax infecta ampliamente a muchas personas en algunas regiones. La singularidad de formar una etapa latente hace que la malaria vivax pueda ser inducida por una recaída por una infección adicional. En el presente estudio, utilizamos un enfoque de simulación dinámica y acoplamiento molecular ara predecir la posibilidad de que compuestos bioactivos de la familia de plantas Zingiberaceae sean candidatos a fármaco contra la malaria dirigiéndose a la proteína de unión a Duffy de Plasmodium vivax (PvDBP). Se requiere la interacción molecular PvDBP-DARC para mediar el proceso de invasión de merozoitos en los glóbulos rojos. La inhibición de este proceso posiblemente pueda controlar el crecimiento y desarrollo del parásito. (Metodología) El análisis de acoplamiento molecular dio como resultado los dos compuestos principales con el valor de energía de unión más bajo, incluida la 5,7-dihidroxiflavanona (-9,3 kcal/mol) y la pinostrobina (-9,2 kcal/mol). (Resultados) Estos compuestos previstos tienen una interacción más fuerte que la cloroquina como control. Además, los compue potenciales también interactúan con los residuos del sitio de unión de DARC y lo mantienen durante el proceso de simulación de dinámica molecular. De lo contrario, la cloroquina como control no puede retener el 75% de los residuos de unión hacia PvDBP. El estudio de dinámica molecular reveló que los tres complejos tienen una estabilidad relativamente similar. (Conclusiones) Predijimos que los dos compuestos bioactivos tienen potencial como inhibidores de la invasión de merozoitos.

Resumo (Objetivo) O Plasmodium vivax infecta amplamente muitas pessoas em algumas regiões. A singularidade de formar um estágio latente significa que a malária vivax pode ser induzida por recaída devido à infecção adicional. No presente estudo, utilizamos uma abordagem de simulação dinâmica e de acoplamento molecular para prever a possibilidade de compostos bioativos da família de plantas Zingiberaceae como candidatos a medicamentos antimaláricos, visand o aproteína de ligação Duffy do Plasmodium vivax (PvDBP). A interação molecular PvDBP-DARC é necessária para mediar o processo de invasão de merozoítos nos glóbulos vermelhos. A inibição deste processo pode possivelmente controlar o crescimento e desenvolvimento do parasita. (Metodologia) A análise de acoplamento molecular resultou nos dois principais compostos com o menor valor de energia de ligação, incluindo ,7-dihidroxiflavanona (-9,3 kcal/mol) e pinostrobina (-9,2 kcal/mol). (Resultados) Esses compostos previstos têm uma interação mais forte do que a cloroquina como controle. Além disso, os compostos potenciais também interagem e mantêm resíduos do sítio de ligação do DARC durante o processo de simulação de dinâmica molecular. Caso contrário, a cloroquina como controle não pode reter 75% dos resíduos de ligação ao três complexos apresentam estabilidade relativa compostos bioativos têm potencial como inibidor Palavras chave: Medicamentos antimalária; ePvmeesdmsilDBP. ntesa invício;O estudo de dinâmica molecular revelou que os emelhante. (Conclusões) Previmos que os dois asão de merozoítos. Plasmodium vivax; PvDBP; Zingiberáceas.

Abstract (Objective) Plasmodium vivax predominantly infects many people in numerous tropical areas, including Southeast Asia, the Western Pacific, the Americas, and the Eastern Mediterranean. The uniqueness of forming dormant stages can lead to relapse in vivax malaria upon further infection. This study used molecular docking and dynamic simulation to predict potential bioactive compounds from the Zingiberaceae plant family as inhibitors by targeting Plasmodium vivax Duffy Binding Protein (PvDBP). PvDBP-DARC molecular interaction is required to mediate the merozoite invasion process into red blood cells. Inhibiting this process can possibly control parasite growth and development. (Methodology) Molecular docking screening was conducted by using 138 natural compounds from the Zingiberaceae plant family targeting Plasmodium vivax Duffy binding protein (PvDBP). The top two compounds with the lowest binding energy were selected to be analyzed by pharmacokinetics prediction and molecular dynamic (MD) simulation. (Results) Molecular docking screening resulted in the top two compounds with the lowest binding energy value, including 5,7-dihydroxyflavanone (-9.3 kcal/mol) and pinostrobin (-9.2 kcal/mol). These compounds are predicted to have stronger interaction than chloroquine as a control. Furthermore, the potential compounds also interact with DARC binding site residues and maintain them during the molecular dynamic simulation process. Otherwise, chloroquine as a control cannot retain 75% binding residues towards PvDBP. A molecular dynamic study revealed that all three complexes have relatively similar stability. (Conclusions) We predicted that the two bioactive compounds (5,7-dihydroxyflavanone and pinostrobin) have the potential as merozoite invasion inhibitors.