The genus Odontomachus is widely distributed in neotropical areas throughout Central and South America. It is a stinging ant that subdues its prey (insects) by injecting them a cocktail of toxic ...molecules (venom). Ant venoms are generally composed of formic acid, alkaloids, hydrocarbons, amines, peptides, and proteins. Odontomachus chelifer is an ant that inhabits neotropical regions from Mexico to Argentina. Unlike the venom of other animals such as scorpions, spiders and snakes, this ant venom has seldom been analyzed comprehensively, and their compositions are not yet completely known. In the present study, we performed a partial investigation of enzymatic and functional activities of O. chelifer ant venom, and we provide a global insight on the transcripts expressed in the venom gland to better understand their properties. The crude venom showed phospholipase A2 and antiparasitic activities. RNA sequencing (Illumina platform) of the venom gland of O. chelifer generated 61, 422, 898 reads and de novo assembly Trinity generated 50,220 contigs. BUSCO analysis against Arthropoda_db10 showed that 92.89% of the BUSCO groups have complete gene representation (single-copy or duplicated), while 4.05% are only partially recovered, and 3.06% are missing. The 30 most expressed genes in O. chelifer venom gland transcriptome included important transcripts involved in venom function such as U-poneritoxin (01)-Om1a-like (pilosulin), chitinase 2, venom allergen 3, chymotrypsin 1 and 2 and glutathione S-transferase. Analysis of the molecular function revealed that the largest number of transcripts were related to catalytic activity, including phospholipases. These data emphasize the potential of O. chelifer venom for prospection of molecules with biotechnological application.
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•This is the first toxinological report of O. chelifer ant venom.•O. chelifer venom has phospholipase A2 and antiparasitic activities.•The transcriptome of O. chelifer was performed, analyzed and annotated.•Different transcripts coding for phospholipases were identified in the transcriptome.
Oranges are among the most consumed fruits in Brazil, with over 80 million tons production per year. Currently, one of the main uses of orange residues is to supplement animal feeds. However, orange ...residues can be used in other ways, including for obtaining enzymes. However, for the pyridoxal 5-phosphate (PLP)-dependent enzyme group, which catalyze transamination reactions, no work has been described. The use of orange-peel residues for solid-state fermentation using endophytic microorganisms is a new and interesting approach to produce these enzymes at lower cost compared to the use of purified enzymes associated to PLP co-factor. In the batch process developed in this study, it was possible to achieve a maximum conversion of 49% after 48 h of incubation, an enantiomeric excess of 99% (R)-1,2,3,4-tetra-hydro-1-naphthylamine, and productivity of 0.0102 mg biocatalyst.h−1, using the solid enzyme preparation from Alternaria arborescens as biocatalyst. In a continuous-flow system, 51% conversion and >99% ee of (R)-1,2,3,4-tetra-hydro-1-naphthylamine were achieved, with a six-fold increase productivity (0.0625 mg biocatalyst.h−1). We have screened different fungi samples for transaminase activity and we found a hit for solid-state fermentation. The endophytic fungus genetically identified as Alternaria arborescens showed promising results for the transaminase activity on the kinetic resolution of chiral amine blocks. Therefore, along with other improvement of the biocatalytic toolbox for chiral amines synthesis, we have found that the solid enzymatic preparation of the endophytic fungus Alternaria arborescens could accept bulky substrates with reasonable activity, compared to the wild-type transaminase already published over literature, and, with high enantioselectivity.
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•First report of transaminase activity for whole cells and solid enzyme preparation of Alternaria arborescens and Sordaria sp.•Higher ω-transaminase activity on solid enzymatic preparation of A. arborescens.•High conversions in (R)-1,2,3,4-tetra-hydro-1-naphthylamine with solid enzyme preparation in a continuous-flow system.
This study aimed to evaluate endophytic fungi isolated from Tocoyena bullata and Humiria balsamifera plant species for their antimycobacterial and anti-inflammatory activities, focusing on severe ...pulmonary tuberculosis cases which are often associated with exacerbated inflammation.
Mycobacterium suspensions were incubated with the samples for 5 days. RAW 264.7 macrophages stimulated with LPS were also incubated with them for 24 h to assess the inhibition of inflammatory mediator production and cytotoxicity. C57BL/6 mice were infected with Mtb M299 and treated for 15 days with lasiodiplodin (Lasio).
Endophytic fungus Sordaria tamaensis, obtained from T. bullata, was the most promising. Its ethanolic extract impaired mycobacterial growth with MIC50 (µg/ml): 1.5 ± 0.6 (BCG), 66.8 ± 0.1 (H37Rv) and 80.0 ± 0.1 (M299). (R)-(+)-Lasio showed MIC50 92.2 ± 1.8 µg/ml (M299). In addition, Lasio was able to inhibit NO, IL-1β and TNF-α production and was not cytotoxic for macrophages. M. tuberculosis-infected C57BL/6 animals treated by Lasio reduced the number of acid-fast bacilli, lung pathology, leucocyte influx and proinflammatory cytokine production in the lungs. The class IIa fructose 1,6-bisphosphate aldolase was the predicted hypothetical target of Lasio.
(R)-(+)-Lasio stood out as a promising anti-TB compound, exhibiting anti-inflammatory and antimycobacterial effects, as well as low cytotoxicity.
This research aimed to determine the upper limit level of crystalline amino acid (CAAs) inclusion in tambaqui nutrition. Additionally, we investigated the possibility of total replacement of fish ...meal (FM) by soy protein concentrate (SPC). An FM‐based control diet was formulated to meet the known nutrient requirements of tambaqui, and three additional diets were prepared by replacing FM protein with 250, 500, and 750 g/kg nitrogen of CAA mixture. The fifth diet was formulated to completely replace the FM by SPC. A growth and a digestibility trial were performed in a completely randomized design with five treatments and three and four replicates, respectively. The optimum replacement level of FM protein by CAA was 280 g/kg based on daily weight gain (DWG). The apparent digestibility coefficients of the nutrients were higher in fish fed the SPC diet than the fish fed the FM diet. The whole‐body lipid and adiposity significantly declined with the increase of CAA in tambaqui diets. Protein and amino acid retention of tambaqui linearly declined with the increase of CAA. Blood chemistry metabolites were not affected by CAA inclusion. We concluded that the SPC could replace FM in tambaqui diets without affecting growth and nutrient digestibility. CAAs could replace FM intact protein up to 250 g/kg (or 97 g/kg inclusion level) without showing any adverse effect on growth and metabolism of tambaqui.
The ability of Stemphylium lycopersici immobilized in MCM 48 zeolite structure as biocatalyst for ω-transamination reactions was evaluated. The conversion capacity for rac-1-methylbenzylamine, and ...also rac-1,2,3,4-tetrahydro-1-naphthylamine was verified in order to ascertain the potential of this biocatalyst in relation to the bioconversion of amines with different side chains when using immobilized crude enzyme extract in zeolite. In this first reaction (after 24 h), a conversion of 23% to acetophenone and 27% to tetralone was obtained. Observing the promising values obtained in the kinetic resolution reactions, an experimental design was carried considering the amount (mg) out of biocatalyst present in the support and the amount of ionic liquid used Central Compound Rotational Design (CCRD2 complete). After optimization, values close to 36% conversion were achieved. In the present work, there were performed 25 cycles of 18 h with Zeolite MCM48 with the addition of ionic liquids (LI). The presence of LI's helps with stability and still acts as structure drivers, which may have helped by providing a large number of recycles. In this analysis, 87% of the initial activity was maintained.
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•Stemphylium lycopersici immobilized in MCM 48 zeolite structure showed potential as a biocatalyst for ω-transamination reactions.•Conversion capacities for different amines were evaluated, with promising results obtained.•An experimental design was carried out to optimize the reaction conditions, achieving close to 36% conversion.•The addition of ionic liquids (LI) helped maintain stability and allowed for a high number of recycles.•Overall, 87% of the initial activity of the biocatalyst was maintained throughout the analysis.
The genus Ocotea (Lauraceae) is distributed mainly in tropical and subtropical regions. Some species of this genus as O. puberula and O. quixos have been described in the literature, showing ...antibacterial activity. And Ocotea macrophylla showed anti-inflammatory activity with inhibition of COX-1, COX-2, and LOX-5. The purpose of this study was the phytochemical investigation of the plant species Ocotea notata from Restinga Jurubatiba National Park, Macaé, RJ, Brazil, and the search for antimycobacterial fractions and compounds. The crude extract was evaluated for antimycobacterial activity and presented 95.75±2.53% of growth inhibition at 100 µg/mL. Then, it was subjected to a liquid-liquid partition and subsequently was chemically investigated by HPLC, revealing the major presence of flavonoids. In this process the partition fractions hexane, ethyl acetate, and butanol are shown to be promising in the antimycobacterial assay. In addition, ethyl acetate fraction was chromatographed and afforded two flavonoids identified by MS and NMR as afzelin and isoquercitrin. The isolated flavonoids afzelin and isoquercitrin were evaluated for their antimycobacterial activity and for their ability to inhibit NO production by macrophages stimulated by LPS; both flavonoids isoquercitrin (Acet22) and afzelin (Acet32) were able to inhibit the production of NO by macrophages. The calculated IC50 of Acet22 and Acet32 was 1.03 and 0.85 µg/mL, respectively.
Abstract Tuberculosis (TB) still constitutes a threat to public health in various regions of the world. The existing treatment is long and has many side effects. The need to identify new anti-TB ...compounds and also adjuvants to control exacerbated inflammation in severe TB cases is relevant. Therefore, the objective of this study was to evaluate the anti-mycobacterial activity of extracts and fractions in vitro from plant species collected in the Restinga of Jurubatiba, in Rio de Janeiro state, Brazil. In addition, to verify their immunomodulatory action and cytotoxicity on macrophages. The dichloromethane fraction of Kielmeyera membranacea and Eremanthus crotonoides showed the lowest MIC50 against Mycobacterium bovis BCG (0.95 ± 1.08 and 2.17 ± 1.11 μg/mL, respectively) and M. tuberculosis H37Rv (4.38 ± 1.19 and 15.28 ± 1.21 μg/mL, respectively). They were also able to inhibit the NO and TNF-α production in LPS-stimulated macrophages, without being toxic to cells. Using gas chromatography analysis coupled with mass spectrometer it was possible to suggest the presence of fatty acids and terpenes in the most promising fractions. Those compounds have been described for their anti-mycobacterial activity. These results have enabled identifying Kielmeyera membranacea and Eremanthus crotonoides as the most promising studied species in searching for new anti-TB compounds with dual activity.
Resumo A tuberculose (TB) ainda representa um problema de Saúde Pública em várias regiões do mundo. O tratamento existente é longo e apresenta diversos efeitos adversos. Neste contexto, é relevante a necessidade de identificar novas substancias anti-TB e complementares ao controle do processo inflamatório exacerbado em quadros severos da doença pulmonar. O objetivo deste trabalho foi avaliar, in vitro, extratos e frações de espécies vegetais coletadas na Restinga de Jurubatiba, quanto a sua atividade antimicobacteriana, assim como verificar a ação imunomoduladora e citotóxica em macrófagos. As frações em diclorometano de Kielmeyera membranacea e Eremanthus crotonoides apresentaram os menores CIM50 contra Mycobacterium bovis BCG (0,95 ± 1,08 e 2,17 ± 1,11 μg/mL; respectivamente) e M. tuberculosis H37Rv (4,38 ± 1,19 e 15,28 ± 1,21 μg/mL; respectivamente). Essas também foram capazes de inibir a produção de NO e TNF-α em macrófagos estimulados por LPS, sem serem tóxicas para as células. Através de análise por cromatografia em fase gasosa acoplada ao espectrômetro de massas foi possível sugerir a presença de ácidos graxos e terpenos nas frações mais promissoras, substancias estas descritas por apresentarem atividade antimicobacteriana. Estes resultados permitiram a identificação de Kielmeyera membranacea e Eremanthus crotonoides como as espécies mais promissoras desse estudo, tendo em vista a busca de novos fármacos anti-TB com dupla atividade.
Natural products extracted from plants represent a valuable source of new bioactive substances. Many studies describe the potential of plant products for the treatment of cardiovascular diseases. ...Species of the
genus have been studied for their biological activities, mainly as antioxidant, anti-inflammatory, and vasorelaxant. However, the phytochemical and pharmacological profiles of
have not been investigated yet. The aim of this study was to evaluate the vasodilator effect of the hydroalcoholic extract of the leaves of
, as well as its chemical profile. Chemical analysis and quantification of major compounds were performed by HPLC analysis. Total flavonoid content was quantified based on rutin equivalents, and major compounds were identified based on HPLC-DAD-MS analysis.
leaf extract-induced vasodilation was investigated in rat aortic rings precontracted with phenylephrine. The total flavonoids were quantified as 3.25 ± 0.11 % w/w of the hydroalcoholic leaf extract, and HPLC-DAD-MS allowed for the identification of luteolin and quercetin glycosides. The maximal relaxant effect of the hydroalcoholic leaf extract was 86.07 ± 1.68 % at a concentration of 30 µg/mL (p < 0.05; n = 6). The concentration of hydroalcoholic extract of the leaves of
necessary to reduce phenylephrine-induced contractions of the endothelium-intact aorta by 50 % was 0.82 ± 0.10 µg/mL.
leaf extract-induced vasodilation was abolished in aortas pretreated with NG-nitro-L-arginine methyl ester and 1H-1,2,4oxadiazolo-4,3-
quinoxalin-1-one. In addition, diphenhydramine partially inhibited the effect of the hydroalcoholic extract of the leaves of
. Thus,
-induced relaxation depends on the endothelium and on the activation of the nitric oxide/cyclic GMP pathway, with the involvement of endothelial histamine H
receptors. Luteolin and quercetin glycosides seem to contribute to the extract activity.
The new outbreak of coronavirus disease 2019 (COVID-19) has infected and caused the death of millions of people worldwide. Intensive efforts are underway around the world to establish effective ...treatments. Immunoglobulin from immunized animals or plasma from convalescent patients might constitute a specific treatment to guarantee the neutralization of the virus in the early stages of infection, especially in patients with risk factors and a high probability of progressing to severe disease. Worldwide, a few clinical trials using anti-SARS-CoV-2 immunoglobulins from horses immunized with the entire spike protein or fragments of it in the treatment of patients with COVID-19 are underway. Here, we describe the development of an anti-SARS-CoV-2 equine F(ab')
immunoglobulin using a newly developed SARS-CoV-2 viral antigen that was purified and inactivated by radiation. Cell-based and preclinical assays showed that the F(ab')
immunoglobulin successfully neutralizes the virus, is safe in animal models, and reduces the severity of the disease in a hamster model of SARS-CoV-2 infection and disease.
Breast cancer is a highly malignant carcinoma and remains the second leading cause of mortality among women. The antitumor effects of metalloproteinases and disintegrins from snake venom on various ...types of cancer cells have been investigated. In this study, we evaluated the antitumor and antiangiogenic effects on MDA-MB-231 human breast cancer cells and endothelial cells induced by Bothropoidin, a disintegrin-like metalloproteinase isolated from Bothrops pauloensis snake venom. At 24h after treatment at 100μg/mL, Bothropoidin exerted a moderate cytotoxic effect of 30% on MDA-MB-231 versus 10% cytotoxicity against MCF10A (a non-tumorigenic breast cell line), a significant difference that suggests a possible preference by this protein for targets in cancer cells. Early and late apoptosis of MDA-MB-231 was observed after Bothropoidin treatment (10μg/mL and 40μg/mL). Furthermore, this toxin inhibited not only the adhesion of MDA-MB-231 cells in a dose-dependent manner but also cell migration by approximately 45%. In addition, Bothropoidin decreased endothelial cells viability and adhesion in Matrigel and inhibited in vitro angiogenesis in Matrigel stimulated by bFGF, showing significantly fewer formed vessels. The results demonstrated that Bothropoidin has potent in vitro antitumor and antiangiogenic effect and represents a biotechnological tool for elucidating the antitumor effect of disintegrins-like metalloproteinases in cancer cells.
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