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    The Dimeric Peptide (KKYRYHLKPF)2K Shows Broad-Spectrum Antiviral Activity by Inhibiting Different Steps of Chikungunya and Zika Virus Infection
    (2023-05-01) Ayusso, Gabriela Miranda [UNESP]; Lima, Maria Letícia Duarte [UNESP]; da Silva Sanches, Paulo Ricardo [UNESP]; Santos, Igor Andrade; Martins, Daniel Oliveira Silva [UNESP]; da Conceição, Pâmela Jóyce Previdelli [UNESP]; Carvalho, Tamara [UNESP]; da Costa, Vivaldo Gomes [UNESP]; Bittar, Cíntia [UNESP]; Merits, Andres; Santos-Filho, Norival Alves [UNESP]; Cilli, Eduardo Maffud [UNESP]; Jardim, Ana Carolina Gomes [UNESP]; de Freitas Calmon, Marilia [UNESP]; Rahal, Paula [UNESP]; Universidade Estadual Paulista (UNESP); Universidade Federal de Uberlândia (UFU); The Rockefeller University; University of Tartu
    Chikungunya virus (CHIKV) and Zika virus (ZIKV) are important disease-causing agents worldwide. Currently, there are no antiviral drugs or vaccines approved to treat these viruses. However, peptides have shown great potential for new drug development. A recent study described (p-BthTX-I)2K [(KKYRYHLKPF)2K], a peptide derived from the Bothropstoxin-I toxin in the venom of the Bothrops jararacussu snake, showed antiviral activity against SARS-CoV-2. In this study, we assessed the activity of this peptide against CHIKV and ZIKV and its antiviral action in the different stages of the viral replication cycle in vitro. We observed that (p-BthTX-I)2K impaired CHIKV infection by interfering with the early steps of the viral replication cycle, reducing CHIKV entry into BHK-21 cells specifically by reducing both the attachment and internalization steps. (p-BthTX-I)2K also inhibited the ZIKV replicative cycle in Vero cells. The peptide protected the cells against ZIKV infection and decreased the levels of the viral RNA and the NS3 protein of this virus at viral post-entry steps. In conclusion, this study highlights the potential of the (p-BthTX-I)2K peptide to be a novel broad-spectrum antiviral candidate that targets different steps of the replication cycle of both CHIKV and ZIKV.
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    Adsorption of Cadmium(II) Ions from Aqueous Solutions Using Calcium Molybdate
    (2022-01-01) De Cássia Pereira, Sandra; Das Graças Barbosa, Amanda; De Figueiredo, Alberthmeiry Teixeira; Morita Barrado, Cristiano; Nunes Alves, Vanessa; Longo, Elson [UNESP]; Federal University of Catalão; Universidade Estadual Paulista (UNESP)
    The presence of toxic metals in aquatic environments poses serious problems for ecosystems and especially for human health. Numerous types of metal oxides have been used to remove these metals and other toxic organic compounds, using adsorption systems. In this work, CaMoO4 was synthesized via coprecipitation and processed for different periods of time using a microwave-assisted hydrothermal system. It was possible to synthesize CaMoO4 at room temperature without any heat treatment. In addition, small processing times in HTMW were able to produce CM with different morphologies. The effect of the reaction time on the morphology of the product and particle size was examined in SEM images. A plausible CaMO4 formation mechanism was proposed based on time and temperature parameters. The potential application of CaMO4 as an adsorbent in water treatment was also investigated and this material exhibited a favorable adsorption performance in the fast removal of cadmium(ii) ions from aqueous solution of 1 mg L-1 concentration. So, CM showed a promising potential for use in environmental remediation.
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    Efficacy of the combination of P5 peptide and photodynamic therapy mediated by bixin and chlorin-e6 against Cutibacterium acnes biofilm
    (2022-12-01) Pereira, Felipe [UNESP]; de Annunzio, Sarah Raquel [UNESP]; Lopes, Thais de Assis; de Oliveira, Kleber Thiago; Cilli, Eduardo Maffud [UNESP]; Barbugli, Paula Aboud [UNESP]; Fontana, Carla Raquel [UNESP]; Universidade Estadual Paulista (UNESP); Universidade Federal de São Carlos (UFSCar)
    In this study, the action of antimicrobial peptide (AMP) P5 and antimicrobial photodynamic therapy (aPDT) mediated by bixin and chlorin-e6 (Ce6) on Cutibacterium acnes (C. acnes) in planktonic phase and biofilm were evaluated both as monotherapies and combined therapies. Microbial viability after treatments were quantified by colony-forming units per milliliter of the sample (CFU/mL) and have demonstrated that all treatments employed exerted bactericidal activity, reducing the microbial load by more than 3 log10 CFU/mL, also demonstrating for the first time in the literature the antimicrobial photodynamic effect of bixin that occurs mostly through type I mechanism which was proved by the quantification of superoxide anion production. Bacterial biofilm was completely eliminated only after its exposure to aPDT mediated by this PS, however, Ce6 proved to be a more efficient PS, considering that most of the photodynamic effect of bixin- aPDT was exerted by excitation of the endogenous C porphyrins of C. acnes with blue light. The combination of P5 with Ce6-aPDT showed a synergistic effect on the bacterial biofilm with a reduction in microbial load by more than 10 log10 CFU/mL, in which the ability of P5 to permeabilize the polymeric extracellular matrix of the biofilm explains the obtained results, with greater internalization of the PS as shown by the Confocal Laser Scanning Microscopy. One-way ANOVA (Analysis of Variance) with Tukey's post-test and two-way ANOVA with Bonferroni's post-test were used to compare the values of continuous variables between the control group and the treatment groups.
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    Singular impasse points of planar constrained differential systems
    (2022-12-01) Perez, Otavio Henrique [UNESP]; da Silva, Paulo Ricardo [UNESP]; Universidade Estadual Paulista (UNESP); Instituto Federal de Educação Ciência e Tecnologia de São Paulo (IFSP)
    Planar analytic constrained differential systems (or impasse systems) are given by (Formular Presented) where F is a vector field and A is a matrix valued function. This class of systems differs from classical ODE’s due to existence of the so called impasse set ∆ = {x : det A(x) = 0}. The dynamics near smooth impasse points is well known in the literature (see for instance [17, 28, 31]). In this paper, we use Newton polygon and weighted blow ups in order to study its phase portrait near singular points of ∆. Moreover, we apply our results in electric circuit problems.
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    Adiponectin reverses β-Cell damage and impaired insulin secretion induced by obesity
    (2023-01-01) Munhoz, Ana Cláudia; Serna, Julian D. C.; Vilas-Boas, Eloisa Aparecida; Caldeira da Silva, Camille C.; Santos, Tiago G.; Mosele, Francielle C. [UNESP]; Felisbino, Sergio L. [UNESP]; Martins, Vilma Regina; Kowaltowski, Alicia J.; Universidade de São Paulo (USP); A. C. Camargo Cancer Center; Universidade Estadual Paulista (UNESP)
    Obesity significantly decreases life expectancy and increases the incidence of age-related dysfunctions, including β-cell dysregulation leading to inadequate insulin secretion. Here, we show that diluted plasma from obese human donors acutely impairs β-cell integrity and insulin secretion relative to plasma from lean subjects. Similar results were observed with diluted sera from obese rats fed ad libitum, when compared to sera from lean, calorically restricted, animals. The damaging effects of obese circulating factors on β-cells occurs in the absence of nutrient overload, and mechanistically involves mitochondrial dysfunction, limiting glucose-supported oxidative phosphorylation and ATP production. We demonstrate that increased levels of adiponectin, as found in lean plasma, are the protective characteristic preserving β-cell function; indeed, sera from adiponectin knockout mice limits β-cell metabolic fluxes relative to controls. Furthermore, oxidative phosphorylation and glucose-sensitive insulin secretion, which are completely abrogated in the absence of this hormone, are restored by the presence of adiponectin alone, surprisingly even in the absence of other serological components, for both the insulin-secreting INS1 cell line and primary islets. The addition of adiponectin to cells treated with plasma from obese donors completely restored β-cell functional integrity, indicating the lack of this hormone was causative of the dysfunction. Overall, our results demonstrate that low circulating adiponectin is a key damaging element for β-cells, and suggest strong therapeutic potential for the modulation of the adiponectin signaling pathway in the prevention of age-related β-cell dysfunction.
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    Antarctic yeasts as a source of L-asparaginase: Characterization of a glutaminase-activity free L-asparaginase from psychrotolerant yeast Leucosporidium scottii L115
    (2023-06-01) Sánchez-Moguel, Ignacio; Costa-Silva, Tales A.; Pillaca-Pullo, Omar S.; Flores-Santos, Juan Carlos; Freire, Rominne Karla Barros; Carretero, Gustavo; da Luz Bueno, Júlia; Camacho-Córdova, David I.; Santos, João H.P.M.; Sette, Lara Durães [UNESP]; Pessoa-Jr, Adalberto; Universidade de São Paulo (USP); Universidad Privada Norbert Wiener; Almirante Paulo Moreira Institute of Studies of the Sea; Universidade Estadual Paulista (UNESP); Federal University of ABC
    Microorganisms from extreme environments, such as the Antarctic ecosystems, have a great potential to produce enzymes with novel characteristics. Within this context, L-asparaginase (ASNase) obtained from yeast species has been poorly studied. In this study, yeasts isolated from samples collected at Admiralty Bay (King George Island, Antarctica) were tested to produce ASNase. From an initial screening of 40 strains, belonging to 13 different species, Leucosporidium scottii L115 produced an ASNase activity (LsASNase activity: 6.24 U g-1 of dry cell weight) with the lowest glutaminase activity. The LsASNase was purified 227-fold, with a specific activity of 137.01 U mg-1 at 37 °C, without glutaminase activity. Moreover, the maximum enzyme activity was observed at pH 7.5 and at a temperature of 55 °C. The enzyme is a multimer of 462 kDa, presenting a single band of 53 kDa molecular mass in reduced conditions; after PGNase F treatment, a single band of 45 kDa was observed. The enzymatic kinetic evaluation revealed an allosteric regulation of the enzyme and the kinetic parameters were determined at 37 °C, pH 7.0 as substrate affinity constant, K0.5 = 233 μM, kcat = 54.7 s−1 and Hill coefficient, nH = 1.52, demonstrating positive cooperativity by the enzyme and the substrate. This is the first study to report L. scottii as a source of glutaminase-activity free L-asparaginase, an acute lymphoblastic leukemia drug feature suitable for the treatment of asparagine synthetase negative cancer cells.
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    Computational Strategies Targeting Inhibition of Helicobacter pylori and Cryptococcus neoformans Ureases
    (2023-01-01) Fabris, Marciéli; Nascimento-Júnior, Nailton M. [UNESP]; Bispo, Marcelle L. F.; Camargo, Priscila G.; Universidade Estadual de Londrina (UEL); Universidade Estadual Paulista (UNESP)
    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 ure-ase inhibitors that are critical for inhibitory activity against H. pylori or Cryptococcus spp. Since the three-dimensional 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 bioac-tive compounds.
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    Effects of Supplementation of Zinc, Manganese, or Copper and Different Phytase Levels in Serum and Bone Acid and Alkaline Phosphatases of Broiler Chicks
    (2023-01-01) Santos, L. F.J. [UNESP]; Gonçalves, Adriano Marques [UNESP]; Neira, L. M. [UNESP]; Nakagi, V. S.; Macari, M. [UNESP]; Laurentiz, A. C. [UNESP]; Pizauro, J. M. [UNESP]; Universidade Estadual Paulista (UNESP); Faculdade de Tecnologia de Ribeirão Preto (FATEC); University of Araraquara (Uniara); Universidade de São Paulo (USP)
    The knowledge of biological trace minerals and phytase requirements for modern broiler genotypes is not established and the pressure to reduce their usage in animal feeding due to environmental issues is increasing. Here, the alkaline phosphatase (ALP) and tartarate-resistant acid phosphatase (TRAP) of the tibia and serum of broilers fed with diets containing various levels of phytase and reduced levels of zinc, manganese, and copper was evaluated. The experiment was performed using 1,200 male Cobb broilers raised according to standard commercial husbandry techniques. Data were analyzed as a 4×3 factorial arrangement with four concentrations of zinc (0.34, 0.49, 0.64, and 0.79 ppm), manganese (0.18, 0.43, 0.68, and 0.93 ppm), or copper (0.09, 0.12, 0.15, and 0.18 ppm) and three concentrations of phytase (0, 500, and 1,000 FTU/kg) for age periods of 1-21 and 36-42 days. While the dietary supplementation with copper did not induce a significant effect in bone tissue biochemical markers, serum TRAP activity of 42-day old broilers increased with higher copper levels. Increasing dietary zinc levels linearly increased ALP activity in tibia growth, suggesting that zinc is essential for longitudinal bone growth. Phytase significantly promoted the increase of TRAP and ALP activities, suggesting that manganese increased growth plate activity, accelerated calcification, and remodeled the newly formed tissue into trabecular bone. Although not every enzymatic activity was affected by the treatments, the phytase use, along with trace minerals, improved the animal response to the rapid growth required nowadays and provided the nutrients for adequate bone metabolism.
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    Antimicrobial metabolites produced by endophytic fungi associated with the leaves of Vochysia divergens
    (2023-01-01) Souza, Jéssica A. de M.; Gubiani, Juliana R.; de Siqueira, Kátia A.; de Camargo, Maria J.; Garcez, Walmir S.; de Sousa, Paulo T.; Soares, Marcos A.; Araújo, Ângela R. [UNESP]; Nunes, Emanuel V. dos S. [UNESP]; Vieira, Lucas C. C.; Sampaio, Olívia M.; Goulart, Letícia S.; Biasetto, Carolina R. [UNESP]; de Menezes, Orivaldo T.; de Oliveira, Camila M.; Nogueira, Cláudio R.; Pinto, Luciano da S.; Teles, Helder L.; Federal University of Mato Grosso; Universidade de São Paulo (USP); Military College of Campo Grande; Federal University of Mato Grosso do Sul; Universidade Estadual Paulista (UNESP); Federal University of Rondonópolis; Federal University of Amazonas; Federal University of Grande Dourados; Universidade Federal de São Carlos (UFSCar)
    Investigation of the endophytic fungi Nigrospora sphaerica, Nigrospora oryzae, and Pseudofusicoccum stromaticum MeOH fractions isolated from the leaves of Vochysia divergens, a medicinal species from the Brazilian Pantanal, led to the identification of five compounds, namely a new compound (1E,8Z)-10,11-dihydroxy-5,5,8-trimethyl-4-oxocycloundeca-1,8-diene-1-carbaldehyde (1) and four known compounds: 5-methylmellein (2), sclerone (3), daldinone A (4), and lasiodiplodin (5). All compounds were identified using spectroscopic methods, and 1 was corroborated with mass spectrometry, while the known compounds were compared with data in the literature. The relative configuration of compound 1 was determined based on theoretical conformational studies as well as the J experimental values between the hydroxymethyne hydrogens. The antimicrobial activity of the compounds was evaluated. Promising results were obtained for compounds 2, 4, and 5 since they inhibited the bacterium Pseudomonas aeruginosa, an opportunistic pathogen, suggesting the potential of these microorganisms as a source of new antibacterial agents.
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    Photobiomodulation Reduces the Cytokine Storm Syndrome Associated with COVID-19 in the Zebrafish Model
    (2023-04-01) Rosa, Ivana F. [UNESP]; Peçanha, Ana P. B.; Carvalho, Tábata R. B.; Alexandre, Leonardo S.; Ferreira, Vinícius G.; Doretto, Lucas B. [UNESP]; Souza, Beatriz M. [UNESP]; Nakajima, Rafael T. [UNESP]; da Silva, Patrick; Barbosa, Ana P.; Gomes-de-Pontes, Leticia; Bomfim, Camila G.; Machado-Santelli, Glaucia M.; Condino-Neto, Antonio; Guzzo, Cristiane R.; Peron, Jean P. S.; Andrade-Silva, Magaiver; Câmara, Niels O. S.; Garnique, Anali M. B.; Medeiros, Renata J.; Ferraris, Fausto K.; Barcellos, Leonardo J. G.; Correia-Junior, Jose D.; Galindo-Villegas, Jorge; Machado, Mônica F. R.; Castoldi, Angela; Oliveira, Susana L. [UNESP]; Costa, Camila C. [UNESP]; Belo, Marco A. A. [UNESP]; Galdino, Giovane; Sgro, Germán G.; Bueno, Natalia F.; Eto, Silas F.; Veras, Flávio P.; Fernandes, Bianca H. V.; Sanches, Paulo R. S. [UNESP]; Cilli, Eduardo M. [UNESP]; Malafaia, Guilherme; Nóbrega, Rafael H. [UNESP]; Garcez, Aguinaldo S.; Carrilho, Emanuel; Charlie-Silva, Ives [UNESP]; Universidade Estadual Paulista (UNESP); São Leopoldo Mandic College; Universidade de São Paulo (USP); INCTBio; Oswaldo Cruz Foundation (Fiocruz); Universidade de Passo Fundo; Federal University Minas Gerais; Nord University; Federal University of Jatai; Universidade Federal de Pernambuco (UFPE); Federal University of Alfenas; FIOCRUZ Paraná; Laboratory of Development and Innovation Butantan Institute; Goiano Federal Institute
    Although the exact mechanism of the pathogenesis of coronavirus SARS-CoV-2 (COVID-19) is not fully understood, oxidative stress and the release of pro-inflammatory cytokines have been highlighted as playing a vital role in the pathogenesis of the disease. In this sense, alternative treatments are needed to reduce the level of inflammation caused by COVID-19. Therefore, this study aimed to investigate the potential effect of red photobiomodulation (PBM) as an attractive therapy to downregulate the cytokine storm caused by COVID-19 in a zebrafish model. RT-qPCR analyses and protein–protein interaction prediction among SARS-CoV-2 and Danio rerio proteins showed that recombinant Spike protein (rSpike) was responsible for generating systemic inflammatory processes with significantly increased levels of pro-inflammatory (il1b, il6, tnfa, and nfkbiab), oxidative stress (romo1) and energy metabolism (slc2a1a and coa1) mRNA markers, with a pattern similar to those observed in COVID-19 cases in humans. On the other hand, PBM treatment was able to decrease the mRNA levels of these pro-inflammatory and oxidative stress markers compared with rSpike in various tissues, promoting an anti-inflammatory response. Conversely, PBM promotes cellular and tissue repair of injured tissues and significantly increases the survival rate of rSpike-inoculated individuals. Additionally, metabolomics analysis showed that the most-impacted metabolic pathways between PBM and the rSpike treated groups were related to steroid metabolism, immune system, and lipid metabolism. Together, our findings suggest that the inflammatory process is an incisive feature of COVID-19 and red PBM can be used as a novel therapeutic agent for COVID-19 by regulating the inflammatory response. Nevertheless, the need for more clinical trials remains, and there is a significant gap to overcome before clinical trials can commence.
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    Maternal dietary zinc status alters offspring female mammary gland development and response to acute 7,12-dimethylbenzanthracene insult
    (2023-01-01) da Silva, Flávia R. M.; Zapaterini, Joyce R. [UNESP]; Grassi, Tony F. [UNESP]; Bidinotto, Lucas T. [UNESP]; Fernandes, Ana Angélica H. [UNESP]; Barbisan, Luis F. [UNESP]; Universidade Estadual de Campinas (UNICAMP); Universidade Estadual Paulista (UNESP); Barretos Cancer Hospital; Barretos School of Health Sciences
    We evaluated the effects of prenatal and postnatal dietary zinc (Zn) deficiency or supplementation on mammary gland morphology and on acute response to 7,12-dimethylbenzanthracene (DMBA) in pubertal female rats. On gestational day 10 (GD 10), rat dams were allocated randomly into three experimental groups of 10: a Zn-adequate diet group (ZnA) fed 35 mg Zn/kg chow, a Zn-deficient diet group (ZnD) fed 3 mg ZN/kg chow and a Zn-supplemented diet group (ZnS) fed 180 mg Zn/kg chow. After weaning, female offspring were fed the same diet as their dams until postnatal day 53 (PND 53). All animals received a single 50 mg/kg dose of DMBA on PND 51 and were euthanized on PND 53. Female ZnD offspring exhibited significantly less weight gain compared to the ZnA group and reduced mammary gland development compared to the ZnD and ZnA groups. By PND 53, the Ki-67 labeling index in mammary gland epithelial cells was significantly greater for the ZnS group than for the ZnA and ZnD groups. Apoptosis and ER-α indices did not differ among groups. The ZnD group exhibited significantly increased lipid hydroperoxide (LOOH) levels and decreased catalase and glutathione peroxidase (GSH-Px) activity compared to the ZnA and ZnS groups. The ZnS group exhibited significantly reduced superoxide dismutase (SOD) activity compared to the ZnA and ZnS groups. We observed atypical ductal hyperplasia in the mammary gland of female ZnS group offspring compared to the ZnA and ZnD groups and decreased expression of the Api5 and Ercc1 genes related to apoptosis inhibition and DNA damage repair, respectively. Both the Zn-deficient and Zn-supplemented diet exerted adverse effects on offspring mammary gland morphology and acute response to DMBA.
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    Bioprocess development for levulinic acid production using sugarcane biomass
    (2023-06-01) dos Santos, Miquéias Gomes [UNESP]; Fogarin, Henrique Maziero [UNESP]; Virginio da Silva, Débora Danielle [UNESP]; Dussán, Kelly Johana [UNESP]; Universidade Estadual Paulista (UNESP)
    Developing technologies for converting lignocellulosic biomass to products such as energy, fuels, and value-added chemicals is an important step in making biorefineries a feasible alternative to the current oil-based productive system. In this study, the conversion of lignocellulosic biomass composed of a mixture of bagasse and sugarcane straw into a cellulose pulp and a sulfonated carbonaceous solid catalyst synthesized from lignin fraction were investigated, as well as the use of this solid acid catalyst to convert cellulose to levulinic acid, an important chemical platform. To overcome this challenge, firstly, the fractionation of lignocellulosic biomass was optimized. Then, tests were carried out to find the best condition for isolation of the soluble lignin from black liquor produced in the optimal condition of delignification. The lignin precipitate was subjected to carbonization tests followed by functionalization tests with concentrated sulfuric acid to synthesize the catalyst. This catalyst was then used to convert cellulose into levulinic acid (LA). The experimental design for cellulose recovery and lignin removal showed that under relatively mild reaction conditions of 110 °C, 117 min, and a solid/NaOH (4.4% w/v) ratio of 1:20, more than 91% of the lignin was dissolved into the black liquor, recovering a solid with a cellulose content higher than 78%. Lignin was fully recovered in the form of precipitate by simply reducing the pH of the black liquor to 3 (under this condition 11 g of precipitate were recovered per liter of liquor). Having the recovered lignin, it was then possible to synthesize a solid carbonaceous catalyst with a total acid site density of 1.48 mmol/g which was able to convert up to 17.11% of the cellulose and obtain yields of up to 38.55% of levulinic acid when a catalyst/cellulose ratio of only 8:10 (w/w) was used. Overall, the future of LA production using carbonaceous acid catalysts looks promising. As research in this area continues, there is the potential to further improve the production process and make it more cost-effective and sustainable. This could lead to wider adoption of this technology and increased use of levulinic acid in various industrial applications, including as a platform chemical to produce a range of chemicals and materials.
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    A new complex of silver(I) with probenecid: Synthesis, characterization, and studies of antibacterial and extended spectrum β-lactamases (ESBL) inhibition activities
    (2023-06-01) Lustri, Wilton R.; Lazarini, Silmara C.; Simei Aquaroni, Nayara Ap.; Resende, Flávia A.; Aleixo, Nadia A.; Pereira, Douglas H.; Lustri, Bruna Cardinali [UNESP]; Moreira, Cristiano Gallina [UNESP]; Ribeiro, Camila M. [UNESP]; Pavan, Fernando R. [UNESP]; Nakahata, Douglas H.; Gonçalves, Adriano M. [UNESP]; Nascimento-Júnior, Nailton M. [UNESP]; Corbi, Pedro P.; Laboratory of Bacterial Cellulose and Medicinal Chemistry (CBQUIM); Federal University of Tocantins; Universidade Estadual de Campinas (UNICAMP); Donostia International Physics Center; Universidade Estadual Paulista (UNESP)
    This article describes the in vitro antibacterial and β-lactamase inhibition of a novel silver(I) complex with the sulfonamide probenecid (Ag-PROB). The formula Ag2C26H36N2O8S2·2H2O for the Ag-PROB complex was proposed based on elemental analysis. High-resolution mass spectrometric studies revealed the existence of the complex in its dimeric form. Infrared, nuclear magnetic resonance spectroscopies and Density Functional Theory calculations indicated a bidentate coordination of probenecid to the silver ions by the oxygen atoms of the carboxylate. In vitro antibacterial activities of Ag-PROB showed significant growth inhibitory activity over Mycobacterium tuberculosis, S. aureus, and P. aeruginosa PA01biofilm-producers, B. cereus, and E. coli. The Ag-PROB complex was active over multi-drug resistant of uropathogenic E. coli extended spectrum β-lactamases (ESBL) producing (EC958 and BR43), enterohemorrhagic E. coli (O157:H7) and enteroaggregative E. coli (O104:H4). Ag-PROB was able to inhibit CTX-M-15 and TEM-1B ESBL classes, at concentrations below the minimum inhibitory concentration for Ag-PROB, in the presence of ampicillin (AMP) concentration in which EC958 and BR43 bacteria were resistant in the absence of Ag-PROB. These results indicate that, in addition to ESBL inhibition, there is a synergistic antibacterial effect between AMP and the Ag-PROB. Molecular docking results revealed potential key residues involved in interactions between Ag-PROB, CTX-M-15 and TEM[sbnd]1B, suggesting the molecular mechanism of the ESBL inhibition. The obtained results added to the absence of mutagenic activity and low cytotoxic activity over non-tumor cell of the Ag-PROB complex open a new perspective for future in vivo tests demonstrating its potential of use as an antibacterial agent.
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    Anti-Inflammatory Effect and Toxicological Profile of Pulp Residue from the Caryocar Brasiliense, a Sustainable Raw Material
    (2023-03-01) Fracasso, Julia Amanda Rodrigues [UNESP]; Ibe, Mariana Bittencourt [UNESP]; da Costa, Luísa Taynara Silvério [UNESP]; Guarnier, Lucas Pires; Viel, Amanda Martins; Brito, Gustavo Reis de [UNESP]; Parron, Mariana Conti [UNESP]; Pereira, Anderson Espírito do Santo [UNESP]; Pegorin Brasil, Giovana Sant’Ana [UNESP]; Farias Ximenes, Valdecir [UNESP]; Fraceto, Leonardo Fernandes [UNESP]; Malacrida Mayer, Cassia Roberta [UNESP]; Ribeiro-Paes, João Tadeu [UNESP]; Ferreira, Fernando Yutaka de [UNESP]; Zoppe, Natália Alves [UNESP]; Santos, Lucinéia dos [UNESP]; Universidade Estadual Paulista (UNESP); Universidade de São Paulo (USP); Nossa Senhora do Patrocínio Faculty Center—CEUNSP
    Caryocar brasiliense Cambess is a plant species typical of the Cerrado, a Brazilian biome. The fruit of this species is popularly known as pequi, and its oil is used in traditional medicine. However, an important factor hindering the use of pequi oil is its low yield when extracted from the pulp of this fruit. Therefore, in this study, with aim of developing a new herbal medicine, we an-alyzed the toxicity and anti-inflammatory activity of an extract of pequi pulp residue (EPPR), fol-lowing the mechanical extraction of the oil from its pulp. For this purpose, EPPR was prepared and encapsulated in chitosan. The nanoparticles were analyzed, and the cytotoxicity of the encapsu-lated EPPR was evaluated in vitro. After confirming the cytotoxicity of the encapsulated EPPR, the following evaluations were performed with non-encapsulated EPPR: in vitro anti-inflammatory activity, quantification of cytokines, and acute toxicity in vivo. Once the anti-inflammatory activity and absence of toxicity of EPPR were verified, a gel formulation of EPPR was developed for topical use and analyzed for its in vivo anti-inflammatory potential, ocular toxicity, and previous stability assessment. EPPR and the gel containing EPPR showed effective anti-inflammatory activity and lack of toxicity. The formulation was stable. Thus, a new herbal medicine with anti-inflammatory activity can be developed from discarded pequi residue.
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    Profile of Bioactive Compounds in Orange Juice Related to the Combination of Different Scion/Rootstocks, Packaging and Storage
    (2023-03-01) Martins, Rafaelly Calsavara [UNESP]; Leonel, Sarita [UNESP]; Souza, Jackson Mirellys Azevedo; Lima, Giuseppina Pace Pereira [UNESP]; Leonel, Magali [UNESP]; Putti, Fernando Ferrari [UNESP]; Monteiro, Gean Charles [UNESP]; Züge, Patrícia Graosque Ülguim [UNESP]; Napoleão, Gabriel Maluf [UNESP]; Figueira, Ricardo [UNESP]; Filho, Jaime Duarte; Universidade Estadual Paulista (UNESP); Universidade Federal de Viçosa (UFV); Regional Development Coordination
    Citrus scion/rootstock combinations alter the concentration of bioactive compounds in orange juice. The shelf life of freshly squeezed juice can be maximized through packaging and storage. The profiles of ascorbic (AA), dehydroascorbic acid (DHAA), and phenolic compounds were analyzed in juices of four sweet orange scions, Sanguínea de Mombuca (SM), Rubi (R), Lue Gin Gong (LGG), and Valência Delta Seedless (VDS), grafted onto ‘Rangpur’ lime (RL) and ‘Swingle’ citrumelo (SC) rootstocks. The juices obtained from the combination of the ‘Rubi’ orange in both rootstocks stood out by their higher concentration of ascorbic acid (AA) and dehydroascorbic acid (DHAA). Overall, all SC-grafted scions showed higher AA and DHAA and some phenolic compound concentrations. In all combinations, phenolic compounds showed the highest concentrations in the juices at the time of fruit extraction and decreased during storage. Dark packaging provided higher bioactive compounds in juices stored for longer periods. These findings can contribute to the diversification of scion/rootstock cultivars in order to increase the variety of orchards by choosing the best combinations for pasteurized orange juice with higher nutritional value.
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    Effects of Resistance Exercise on Slow-Twitch Soleus Muscle of Infarcted Rats
    (2023-02-01) Souza, Lidiane Moreira [UNESP]; Gomes, Mariana Janini; Brandao, Bruna Brasil; Pagan, Luana Urbano [UNESP]; Gatto, Mariana [UNESP]; Damatto, Felipe Cesar [UNESP]; Rodrigues, Eder Anderson [UNESP]; Pontes, Thierres Hernani Dias [UNESP]; Borim, Patricia Aparecida [UNESP]; Fernandes, Ana Angelica Henrique [UNESP]; Murata, Gilson Masahiro; Zornoff, Leonardo Antonio Mamede [UNESP]; Azevedo, Paula Schmidt [UNESP]; Okoshi, Katashi [UNESP]; Okoshi, Marina Politi [UNESP]; Universidade Estadual Paulista (UNESP); Texas AM University; Harvard Medical School; Universidade de São Paulo (USP)
    Although current guidelines recommend resistance exercise in combination with aerobic training to increase muscle strength and prevent skeletal muscle loss during cardiac remodeling, its effects are not clear. In this study, we evaluated the effects of resistance training on cardiac remodeling and the soleus muscle in long-term myocardial infarction (MI) rats. Methods: Three months after MI induction, male Wistar rats were assigned to Sham (n = 14), MI (n = 9), and resistance exercised MI (R-MI, n = 13) groups. The rats trained three times a week for 12 weeks on a climbing ladder. An echocardiogram was performed before and after training. Protein expression of the insulin-like growth factor (IGF)-1/protein kinase B (Akt)/rapamycin target complex (mTOR) pathway was analyzed by Western blot. Results: Mortality rate was higher in MI than Sham; in the R-MI group, mortality rate was between that in MI and Sham and did not differ significantly from either group. Exercise increased maximal load capacity without changing cardiac structure and left ventricular function in infarcted rats. Infarction size did not differ between infarcted groups. Catalase activity was lower in MI than Sham and glutathione peroxidase lower in MI than Sham and R-MI. Protein expression of p70S6K was lower in MI than Sham and p-FoxO3 was lower in MI than Sham and R-MI. Energy metabolism did not differ between groups, except for higher phosphofrutokinase activity in R-MI than MI. Conclusion: Resistance exercise is safe and increases muscle strength regardless structural and functional cardiac changes in myocardial-infarcted rats. This exercise modality attenuates soleus glycolytic metabolism changes and improves the expression of proteins required for protein turnover and antioxidant response.
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    Old Meets New: Mass Spectrometry-Based Untargeted Metabolomics Reveals Unusual Larvicidal Nitropropanoyl Glycosides from the Leaves of Heteropterys umbellata
    (2023-03-24) Mannochio-Russo, Helena [UNESP]; Nunes, Wilhan D. G.; Almeida, Rafael F.; Albernaz, Lorena C.; Espindola, Laila S.; Bolzani, Vanderlan S. [UNESP]; Universidade Estadual Paulista (UNESP); Science and Technology of São Paulo (IFSP); Universidade Estadual de Goiás; Universidade de Brasília (UnB)
    The Aedes aegypti (Diptera: Culicidae) mosquito is the vector of several arboviruses in tropical and subtropical areas of the globe, and synthetic pesticides remain the most widely used combat strategy. This study describes the investigation of secondary metabolites with larvicidal activity from the Malpighiaceae taxon using a metabolomic and bioactivity-based approach. The workflow initially consisted of a larvicidal screening of 394 extracts from the leaves of 197 Malpighiaceae samples, which were extracted using solvents of different polarity, leading to the selection of Heteropterys umbellata for the identification of active compounds. By employing untargeted mass spectrometry-based metabolomics and multivariate analyses (PCA and PLS-DA), it was possible to determine that the metabolic profiles of different plant organs and collection sites differed significantly. A bioguided approach led to the isolation of isochlorogenic acid A (1) and the nitropropanoyl glucosides karakin (2) and 1,2,3,6-tetrakis-O-[3-nitropropanoyl]-beta-glucopyranose (3). These nitro compounds exhibited larvicidal activity, possibly potentialized by synergistic effects of their isomers in chromatographic fractions. Additionally, targeted quantification of the isolated compounds in different extracts corroborated the untargeted results from the statistical analyses. These results support a metabolomic-guided approach in combination with classical phytochemical techniques to search for natural larvicidal compounds for arboviral vector control.
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    Effects of Dimerization, Dendrimerization, and Chirality in p-BthTX-I Peptide Analogs on the Antibacterial Activity and Enzymatic Inhibition of the SARS-CoV-2 PLpro Protein
    (2023-02-01) Bitencourt, Natália Vitória [UNESP]; Righetto, Gabriela Marinho; Camargo, Ilana Lopes Baratella Cunha; de Godoy, Mariana Ortiz; Guido, Rafael Victorio Carvalho; Oliva, Glaucius; Santos-Filho, Norival Alves [UNESP]; Cilli, Eduardo Maffud [UNESP]; Universidade Estadual Paulista (UNESP); Universidade de São Paulo (USP)
    Recent studies have shown that the peptide [des-Cys11,Lys12,Lys13-(p-BthTX-I)2K] (p-Bth) is a p-BthTX-I analog that shows enhanced antimicrobial activity, stability and hemolytic activity, and is easy to obtain compared to the wild-type sequence. This molecule also inhibits SARS-CoV-2 viral infection in Vero cells, acting on SARS-CoV-2 PLpro enzymatic activity. Thus, the present study aimed to assess the effects of structural modifications to p-Bth, such as dimerization, dendrimerization and chirality, on the antibacterial activity and inhibitory properties of PLpro. The results showed that the dimerization or dendrimerization of p-Bth was essential for antibacterial activity, as the monomeric structure led to a total loss of, or significant reduction in, bacterial activities. The dimers and tetramers obtained using branched lysine proved to be prominent compounds with antibacterial activity against Gram-positive and Gram-negative bacteria. In addition, hemolysis rates were below 10% at the corresponding concentrations. Conversely, the inhibitory activity of the PLpro of SARS-CoV-2 was similar in the monomeric, dimeric and tetrameric forms of p-Bth. Our findings indicate the importance of the dimerization and dendrimerization of this important class of antimicrobial peptides, which shows great potential for antimicrobial and antiviral drug-discovery campaigns.
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    Enzymatic hydrolysis of Moringa oleifera Lam flour using bromelain and fig by-products as sources of protease
    (2022-10-01) Bustamante, Lady Johana Arismendi [UNESP]; Maisa, Davanso [UNESP]; de Cerqueira e Silva, Mariana Barros; de Paula, Ariela Veloso [UNESP]; Teixeira, Estelamar Maria Borges; Bassan, Juliana Cristina [UNESP]; Universidade Estadual Paulista (UNESP); Science and Technology of Triangulo Mineiro (IFTM)
    Moringa oleifera Lamarck flour (MOF) has been recognized as a source of vegetable proteins; however, around 70.1±0.98% of protein can be called “insoluble protein”, for its resistance to gastrointestinal digestion. For protein to be absorbed in the human intestinal tract, it must be consumed in the form of a soluble protein. Proteases have shown to increase soluble proteins. The objective of this study was, hence, to investigate the increase in soluble proteins of MOF soluble and insoluble water fraction through the hydrolysis with proteases obtained from fig and pineapple by-products, ficin and bromelain, respectively. In conclusion, the protein of MOF had changes in their solubility by enzymatic effect. In particular, the hydrolysis of water insoluble Moringa oleifera Lam flour fraction (IMOF) with ficin contributed to a significant increase in soluble proteins, a result detectable through the different methods applied (colorimetric Bradford method, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and Reversed phase high-performance liquid chromatography (RP-HPLC)).
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    Plant Defense Elicitation by the Hydrophobin Cerato-Ulmin and Correlation with Its Structural Features
    (2023-02-01) Gallo, Mariana; Luti, Simone; Baroni, Fabio; Baccelli, Ivan; Cilli, Eduardo Maffud [UNESP]; Cicchi, Costanza; Leri, Manuela; Spisni, Alberto; Pertinhez, Thelma A.; Pazzagli, Luigia; University of Parma; University of Florence; National Research Council of Italy; Universidade Estadual Paulista (UNESP)
    Cerato-ulmin (CU) is a 75-amino-acid-long protein that belongs to the hydrophobin family. It self-assembles at hydrophobic–hydrophilic interfaces, forming films that reverse the wettability properties of the bound surface: a capability that may confer selective advantages to the fungus in colonizing and infecting elm trees. Here, we show for the first time that CU can elicit a defense reaction (induction of phytoalexin synthesis and ROS production) in non-host plants (Arabidopsis) and exerts its eliciting capacity more efficiently when in its soluble monomeric form. We identified two hydrophobic clusters on the protein’s loops endowed with dynamical and physical properties compatible with the possibility of reversibly interconverting between a disordered conformation and a β-strand-rich conformation when interacting with hydrophilic or hydrophobic surfaces. We propose that the plasticity of those loops may be part of the molecular mechanism that governs the protein defense elicitation capability.