Neospora caninum, the causative agent of neosporosis, is an obligate intracellular parasite consi... more Neospora caninum, the causative agent of neosporosis, is an obligate intracellular parasite considered to 29 be a major cause of abortion in cattle throughout the world. Most studies concerning N. caninum have 30 focused on life cycle, seroepidemiology, pathology and vaccination, while data on host-parasite interaction, 31 such as host cell migration, mechanisms of evasion and dissemination of this parasite during the early phase 32 of infection are still poorly understood. Here we show the ability of excreted/secreted antigens from 33 N. caninum (NcESAs) to attract monocytic cells to the site of primary infection in both in vitro and in vivo 34 assays. Molecules from the family of cyclophilins present on the NcESAs were shown to work as chemo-35 kine-like proteins and NcESA-induced chemoattraction involved G i protein signaling and participation of 36 CC-chemokine receptor 5 (CCR5). Additionally, we demonstrate the ability of NcESAs to enhance the expres-37 sion of CCR5 on monocytic cells and this increase occurred in parallel with the chemotactic activity of 38 NcESAs by increasing cell migration. These results suggest that during the first days of infection, N. caninum 39 produces molecules capable of inducing monocytic cell migration to the sites of infection, which will 40 consequently enhance initial parasite invasion and proliferation. Altogether, these results help to clarify 41 some key features involved in the process of cell migration and may reveal virulence factors and therapeutic 42 targets to control neosporosis. 43
Suramin (Sur) acts as an -NTPDase inhibitor in and a P2-purinoceptor antagonist in mammalian cell... more Suramin (Sur) acts as an -NTPDase inhibitor in and a P2-purinoceptor antagonist in mammalian cells. Although the potent antitrypanosomal effect of Sur has been shown , limited evidence suggests that this drug can be dangerous to -infected hosts. Therefore, we investigated the dose-dependent effect of Sur-based chemotherapy in a murine model of Chagas disease. Seventy uninfected and -infected male C57BL/6 mice were randomized into five groups: SAL = uninfected; INF = infected; SR5, SR10, and SR20 = infected treated with 5, 10, or 20 mg/kg Sur. In addition to its effect on blood and heart parasitism, the impact of Sur-based chemotherapy on leucocytes myocardial infiltration, cytokine levels, antioxidant defenses, reactive tissue damage, and mortality was analyzed. Our results indicated that animals treated with 10 and 20 mg/kg Sur were disproportionally susceptible to , exhibiting increased parasitemia and cardiac parasitism (amastigote nests and parasite load ( DNA)), intense protein...
Dengue virus (DENV) is an arbovirus that belongs to the Flaviviridae family. Studies reveal that ... more Dengue virus (DENV) is an arbovirus that belongs to the Flaviviridae family. Studies reveal that peptides secreted by amphibians have many functions, such as antiviral and antimicrobial activities. As there is no antiviral drug effective against the DENV, the antiviral activity of a synthetic peptide called HS-1, derived from the secretion of the anuran Hypsiboas semilineatus, has been evaluated. The assays of neutralization in the Vero cells show a complete inhibition of infection of the serotypes 2 and 3. Furthermore, the direct action of peptides on the viral particle can be observed through atomic force microscopy. In vivo tests display 80% protection against the dengue-2 virus due to the presence of HS-1, which reveals its potential as an antiviral against the DENV.
We aimed to evaluate the postprandial secretion of inflammatory markers induced by SFA or MUFA hi... more We aimed to evaluate the postprandial secretion of inflammatory markers induced by SFA or MUFA high-fat meal consumption and whether orange juice intake could modulate this induction. This study included 55 healthy women (aged 20 to 40 years): 33 participants received an SFA high-fat meal (≈1000 kcal, 37.6% of energy intake (E) from SFA) and 22 participants received an MUFA high-fat meal (≈1000 kcal, 56.3% E from MUFA). Both interventions were accompanied by 500 ml of orange juice (test) or water (control). The plasma concentrations of inflammatory cytokines (IL-2, IL-4, IL-6, IL-10, IL-17A, IFN-γ, and TNF-α) and CRP were determined before (fasting) and 2, 3 and 5 hours after the test meal intake. The SFA high-fat meal induced a significant increase in AUC values (for TNF-α, IL-12, IL-10, IL-6 and IL-2 adjusted for baseline concentrations) in comparison with MUFA high-fat meal intervention. The results were independent of the drink which accompanied the meal (water or orange juice)....
International Journal of Food and Nutritional Science, 2016
The coconut oil is popularly recognized for its anti-inflammatory properties, but research has sh... more The coconut oil is popularly recognized for its anti-inflammatory properties, but research has shown conflicting results. Thus, our aim was to assess the postprandial inflammatory response of moderate coconut oil intake. We evaluated the acute consumption of a breakfast, offered in a single day, containing 25 mL of coconut oil (test) or 25 mL of olive oil (control)on postprandial serum inflammatory cytokines (IL-2, IL-4, IL-6, IL-10, IL-17A, IFN-γ and TNF) in healthy overweight women. No change was observed in relation to the effect of fatty meals, the time and the interaction of both factors (meal x time). Thus, despite being a saturated fat source, coconut oil consumption did not promote an inflammatory response. Further studies are needed considering a larger intervention period to check long-term effect.
International Journal of Biological Macromolecules, 2017
Lectins are involved in a wide range of biological mechanisms, like immunomodulatory agent able t... more Lectins are involved in a wide range of biological mechanisms, like immunomodulatory agent able to activate the innate immunity. In this study, we purified and characterized a new lectin from cauliflower (Brassica oleracea ssp. botrytis-BOL) by three sequential chromatographic steps and confirmed the purity by SDS-PAGE. Additionally, we evaluated the role of the lectin in innate immunity by a phagocytosis assay, production of H 2 O 2 and NO. BOL was characterized like a non-glycosylated protein that showed a molecular mass of ∼34 kDa in SDS-PAGE. Its N-terminal sequence (ETRAFREERPSSKIVTIAG) did not reveal any similarity to the other lectins; nevertheless, it showed 100% homology to a putative TRAFlike protein from Brassica rapa and Brassica napus. This is a first report of the TRAF-protein with lectinic activity. The BOL retained its complete hemagglutination activity from 4 • C up to 60 • C, with stability being more apparent between pH 7.0 and 8.0. Moreover, the lectin was able to stimulate phagocytosis and induce the production of H 2 O 2 and NO. Therefore, BOL can be explored as an immunomodulatory agent by being able to activate the innate immunity and favor antigen removal.
The mosquito Aedes aegypti transmit virus that cause diverse human diseases and the vector contro... more The mosquito Aedes aegypti transmit virus that cause diverse human diseases and the vector control is an important strategy to avoid their propagation. Plants in Annonaceae are recognized as source to molecules with use in medical and agriculture fields. Molecules of secondary metabolites of Annonaceae plants exhibit insecticidal potential against insect pest and vectors, with highlight to acetogenins that show high toxicity with low doses, which encouraged research to producing new insecticide molecules. Herein, we identify an acetogenin from Annona mucosa seeds (chemical analysis) and provide toxicity test against larvae of A. aegypti (target insect), its predators Culex bigoti and Toxorhynchites theobaldi (non-target insects) and cytotoxicity to human leukocytes. We identify squamocin (C37 H66 O7 ) a fatty acid with presence of bis-tetrahydrofuran ring. In A. aegypti, this compound exhibited behavioral disturb before larval death, high mortality and require low concentrations to ...
Kefir supplementation in rats with induced metabolic syndrome was able to lower fasting glucose, ... more Kefir supplementation in rats with induced metabolic syndrome was able to lower fasting glucose, fasting insulin levels, and reduce insulin resistance.
Podisus nigrispinus Dallas (Hemiptera: Pentatomidae) is a predator insect with potential applicat... more Podisus nigrispinus Dallas (Hemiptera: Pentatomidae) is a predator insect with potential applications in biological control because both nymphs and adults have been shown to prey on other insect pests by injection of toxic salivary gland contents. This study identified non-proteinaceous compounds with insecticidal activity from the saliva of P. nigrispinus in Anticarsia gemmatalis. In particular, the ether extract from P. nigrispinus saliva led to mortality in A. gemmatalis larvae, with a LC50 = 2.04 μL and LC90 = 3.27 μL. N,N-dimethylaniline and 1,2,5-trithiepane fractions were identified as non-proteinaceous extract components. N,N-dimethylaniline had a LC50 = 136.1 nL and LC90 = 413.8 nL, suggesting that it could be responsible for toxicity in P. nigrispinus saliva.
Comparative Biochemistry and Physiology Part A: Molecular & Integrative Physiology, 2008
A thrombin-like enzyme named BjussuSP-I, isolated from B. jararacussu snake venom, is an acidic s... more A thrombin-like enzyme named BjussuSP-I, isolated from B. jararacussu snake venom, is an acidic single chain glycoprotein with approximately 6% sugar, Mr = 61,000 under reducing conditions and pI ∼ 3.8, representing 1.09% of the chromatographic A 280 recovery. BjussuSP-I is a glycosylated serine protease containing both N-linked carbohydrates and sialic acid in its structure. BjussuSP-I showed a high clotting activity upon human plasma, which was inhibited by PMSF, leupeptin, heparin and 1,10-phenantroline. This enzyme showed high stability regarding coagulant activity when analyzed at different temperatures (−70 to 37°C), pHs (4.5 to 8.0), and presence of two divalent metal ions (Ca 2+ and Mg 2+ ). It also displayed TAME esterase and proteolytic activities toward natural (fibrinogen and fibrin) and synthetic (BAPNA) substrates, respectively, being also inhibited by PMSF and leupeptin. BjussuSP-I can induce production of polyclonal antibodies able to inhibit its clotting activity, but unable to inhibit its proteolytic activity on fibrinogen. The enzyme also showed crossed immunoreactivity against 11 venom samples of Bothrops, 1 of Crotalus, and 1 of Calloselasma snakes, in addition of LAAO isolated from B. moojeni venom. It displayed neither hemorrhagic, myotoxic, edema-inducing profiles nor proteolytic activity on casein. BjussuSP-I showed an N-terminal sequence (VLGGDECDINEHPFLA FLYS) similar to other thrombin-like enzymes from snake venoms. Based on its biochemical, enzymatic and pharmacological characteristics, BjussuSP-I was identified as a new thrombin-like enzyme isoform from Bothrops jararacussu snake venom.
We report the identification and characterization of a new AMP named Hs-1.Hs-1 is the first AMP d... more We report the identification and characterization of a new AMP named Hs-1.Hs-1 is the first AMP described for the Brazilian endemic frog Hypsiboas semilineatus.Hs-1 is selective against Gram-positive bacteria, especially against MRSA.Hs-1 has a direct antibacterial killing by membrane disruption.Hs-1 is predominantly structured as an amphipathic alpha helix.The multidrug-resistant bacteria have become a serious problem to public health. In this scenery the antimicrobial peptides (AMPs) derived from animals and plants emerge as a novel therapeutic modality, substituting or in addition to the conventional antimicrobial. The anurans are one of the richest natural sources of AMPs. In this work several cycles of cDNA cloning of the skin of the Brazilian treefrog Hypsiboas semilineatus led to isolation of a precursor sequence that encodes a new AMP. The sequence comprises a 27 residue signal peptide, followed by an acidic intervening sequence that ends in the mature peptide at the carboxy terminal. The AMP, named Hs-1, has 20 amino acids residues, mostly arranged in an alpha helix and with a molecular weight of 2144.6 Da. The chemically synthesized Hs-1 showed an antimicrobial activity against all Gram-positive bacteria tested, with a range of 11–46 μM, but it did not show any effect against Gram-negative bacteria, which suggest that Hs-1 may have a selective action for Gram-positive bacteria. The effects of Hs-1 on bacterial cells were also demonstrated by transmission electron microscopy. Hs-1 is the first AMP to be described from H. semilineatus.
Antimicrobial agents and chemotherapy, Jan 13, 2015
Although Suramin is suggested as a potential drug candidate in the management of Chagas disease, ... more Although Suramin is suggested as a potential drug candidate in the management of Chagas disease, this issue has not been objectively tested. In this study, we examined the applicability of the concomitant treatment with Benznidazole (Bz) and Suramin (Sur) in mice infected with a virulent strain of Trypanosoma cruzi. Eighty 12-week-old male C57BL/6 mice were equally randomized in seven groups: 1) non-infected (negative control) and infected with T. cruzi Y strain receiving 2) no treatment (positive control); 3) Bz 100 mg/kg/day; 4) Sur 20 mg/kg/day; 5 to 8) Sur 20 mg/kg/day combined with Bz 100; 50; 25 or 5 mg/kg/day. Bz was administered by gavage and Sur intraperitoneally. Sur dramatically increased the parasitemia, cardiac content of parasite DNA, inflammation, oxidative tissue damage and mortality. In response to high parasitic load in cardiac tissue, Sur stimulated the immune system in a manner typical of the acute phase of Chagas disease, increasing tissue levels of IFN-γ and TN...
The Journal of pharmacy and pharmacology, Jan 23, 2015
This study investigated the combined effects of benznidazole (BZ) and ibuprofen (IB) on the oxida... more This study investigated the combined effects of benznidazole (BZ) and ibuprofen (IB) on the oxidative and inflammatory status of the cardiac tissue in vivo. Swiss mice were randomized in groups receiving BZ (100 mg/kg) and IB (400 mg/kg) alone or combined (BZ + IB 200 or 400 mg/kg). Control animals were concurrently treated with 1% carboxymethyl cellulose. All treatments were administered orally for 7 days. BZ treatment increased cardiac production of nitrogen/oxygen-reactive species, malondialdeyde, carbonyl proteins, prostaglandins as well as the activities of catalase, superoxide dismutase and glutathione peroxidase. These parameters were attenuated by IB, with the best results at higher dose. Individually, BZ and IB significantly reduced the tissue levels of chemokine ligand 2, tumour necrosis factor-α and IL-10, but no reduction was observed when the treatments were combined. BZ triggers an oxidative and nitrosative route, which is associated with increased prostaglandin synthe...
The search of new drugs and the use of animal experimentation had been contributed excessively to... more The search of new drugs and the use of animal experimentation had been contributed excessively to the development of the health, of the science and technology, promoting along the years the discovery of prophylactic measured and treatments of illnesses that affect the human beings. Animals of several species have been used from the origins of the civilization for the scientific development, but among all of the experimental models to the which peptic ulcers can be induced, the rats Wistar, doubtless, is the used. The objective of this work was to accomplish an anatomical and histological description. Anatomically, the stomach of the Wistar rat (Rattus norvegicus) is located in the abdominal cavity in the ventral midline in the left portion, caudal to the liver. This body can be divided into two regions glandular and non glandular. These regions are separated by the folded edge that makes the division between the keratinized stratified squamous epithelium that lines the portion of ex...
This study used a murine model of Chagas disease to investigate the isolated and combined impact ... more This study used a murine model of Chagas disease to investigate the isolated and combined impact of Trypanosoma cruzi infection and benznidazole (BZ) therapy on liver structure and function. Male C57BL/6 mice were challenged with T. cruzi and BZ for 15 days. Serum levels of cytokines and hepatic enzymes, liver oxidative stress, morphology, collagen, and glycogen content were monitored. Separately, T. cruzi infection and BZ treatment resulted in a pro-oxidant status and hepatic reactive damage. Concurrently, both T. cruzi infection and BZ treatment induced upregulation of antioxidant enzymes and pathological reorganization of the liver parenchyma and stroma. T. cruzi infection increased serum levels of Th1 cytokines, which were reduced by BZ in both infected and non-infected animals. BZ also induced functional organ damage, increasing serum levels of liver enzymes. When combined, T. cruzi infection and BZ therapy elicited intense hepatic reactive damage that was not compensated by antioxidant enzymatic reaction, subsequently culminating in more severe morphofunctional hepatic injury. Taken together, these findings indicate that during specific treatment of Chagas disease, hepatic pathology may be a result of an interaction between BZ metabolism and specific mechanisms activated during the natural course of T. cruzi infection, rather than an isolated toxic effect of BZ on liver structure and function.
In this work, we report the evaluation of two DNA vaccines against dengue-3 virus (DENV-3). The f... more In this work, we report the evaluation of two DNA vaccines against dengue-3 virus (DENV-3). The first construction, called pVAC3DEN3, was engineered inserting the pre-membrane (prM) and envelope (E) gene of DENV-3 truncated with a restriction site between them, as previously described. The second construction was developed cloning the full gene sequence of prM and E from DENV-3 virus in pCI plasmid for mammalian expression and was denominated pVAC1WDEN3. The results showed that both constructions were capable of expressing the prM and E proteins, as demonstrated by ELISA and immunoblotting detection in cell culture transfected with the plasmids. After positive "in vitro" results, the vaccine candidates were used to immunize BALB/c mice and the elicited response was investigated. After immunization by intramuscular inoculation with three doses of each vaccinal clone the animals were sacrificed, the cytokine levels and T cell response were analyzed in the spleens, after three days of culture with stimulus, our analysis showed that the two constructions elicited T cell responses measured by BrdU incorporation assay and high levels of IFN-γ, detected in the supernatant of the cultures. Moreover, both constructions induced detectable titers of neutralizing antibodies in mice. And finally the survival rate of the immunized animals after intracerebral challenge was analyzed, showing a better result in the pVAC3DEN3 group with an 80% survival compared with a 50% survival of the pVAC1 WDEN3.
In calves, neonatal mortality and disease susceptibility are greatly influenced by failure in pas... more In calves, neonatal mortality and disease susceptibility are greatly influenced by failure in passive immunization, normally provided by colostrum ingestion just after birth. Formulations projected to replace natural colostrum have not been successful, and one of the possible reasons for such failure is that orally administered Ig are probably digested in the gastrointestinal tract, so they are not absorbed as intact functional molecules. With the aim of finding an adequate colostrum substitute, we used columns of immobilized jacalin, a lectin known by its ability to bind O-linked oligosaccharides, to obtain a colostral Ig population putatively protected against enzymatic cleavage by the presence of sugar chains. Immunoglobulin G1 is a major constituent of colostrum Ig bound to jacalin (JB-Ig). This preparation contains 10% of the total colostral Ig and is typically 3 to 6 times more resistant to pepsin digestion than the Ig contained in the fraction that is not bound to jacalin, wh...
Bovine mastitis is a major threat to animal health and the dairy industry. Staphylococcus aureus ... more Bovine mastitis is a major threat to animal health and the dairy industry. Staphylococcus aureus is a contagious pathogen that is usually associated with persistent intramammary infections, and biofilm formation is a relevant aspect of the outcome of these infections. Several biological activities have been described for snake venoms, which led us to screen secretions of Bothrops jararacussu for antibiofilm activity against S. aureus NRS155. Crude venom was fractionated by size-exclusion chromatography, and the fractions were tested against S. aureus. Biofilm growth, but not bacterial growth, was affected by several fractions. Two fractions (15 and 16) showed the best activities and were also assayed against S. epidermidis NRS101. Fraction 15 was identified by TripleTOF mass spectrometry as a galactose-binding C-type lectin with a molecular weight of 15 kDa. The lectin was purified from the crude venom by D-galactose affinity chromatography, and only one peak was observed. This pure lectin was able to inhibit 75% and 80% of S. aureus and S. epidermidis biofilms, respectively, without affecting bacterial cell viability. The lectin also exhibited a dose-dependent inhibitory effect on both bacterial biofilms. The antibiofilm activity was confirmed using scanning electron microscopy. A pre-formed S. epidermidis biofilm was significantly disrupted by the C-type lectin in a time-dependent manner. Additionally, the lectin demonstrated the ability to inhibit biofilm formation by several mastitis pathogens, including different field strains of S. aureus, S. hyicus, S. chromogenes, Streptococcus agalactiae, and Escherichia coli. These findings reveal a new activity for C-type lectins. Studies are underway to evaluate the biological activity of these lectins in a mouse mastitis model.
The cabbage (Brassica oleraceae var. capitata) is an herbaceous and leafy plant which belongs to ... more The cabbage (Brassica oleraceae var. capitata) is an herbaceous and leafy plant which belongs to the Brassicaceae family, native to coastal southern and Western Europe. Used in cooking for its nutritional value also has known anti-inflammatory activity. OBJECTIVE We studied the antiulcerogenic activity of aqueous extract of Brassica oleracea var. capitata (AEB) in order to validate ethnobotanical claims regarding the plant use in the gastric disorders. Acute gastric ulcers were induced in rats by the oral administration of acetylsalicylic acid. The gastroprotective potential of the AEB (0.250, 0.500 and 1.000 mg.kg-1/body weight) was compared with omeprazole (20 mg.kg-1/body weight). The stomach analysis indicated that treatment with AEB inhibited the gastric damage. The gastroprotective activity as evidenced by its significant inhibition in the formation of ulcers induced by chemical agent with a maximum of 99.44% curation (250 mg.kg-1 body weight) in acetylsalicylic acid-induced u...
Neospora caninum, the causative agent of neosporosis, is an obligate intracellular parasite consi... more Neospora caninum, the causative agent of neosporosis, is an obligate intracellular parasite considered to 29 be a major cause of abortion in cattle throughout the world. Most studies concerning N. caninum have 30 focused on life cycle, seroepidemiology, pathology and vaccination, while data on host-parasite interaction, 31 such as host cell migration, mechanisms of evasion and dissemination of this parasite during the early phase 32 of infection are still poorly understood. Here we show the ability of excreted/secreted antigens from 33 N. caninum (NcESAs) to attract monocytic cells to the site of primary infection in both in vitro and in vivo 34 assays. Molecules from the family of cyclophilins present on the NcESAs were shown to work as chemo-35 kine-like proteins and NcESA-induced chemoattraction involved G i protein signaling and participation of 36 CC-chemokine receptor 5 (CCR5). Additionally, we demonstrate the ability of NcESAs to enhance the expres-37 sion of CCR5 on monocytic cells and this increase occurred in parallel with the chemotactic activity of 38 NcESAs by increasing cell migration. These results suggest that during the first days of infection, N. caninum 39 produces molecules capable of inducing monocytic cell migration to the sites of infection, which will 40 consequently enhance initial parasite invasion and proliferation. Altogether, these results help to clarify 41 some key features involved in the process of cell migration and may reveal virulence factors and therapeutic 42 targets to control neosporosis. 43
Suramin (Sur) acts as an -NTPDase inhibitor in and a P2-purinoceptor antagonist in mammalian cell... more Suramin (Sur) acts as an -NTPDase inhibitor in and a P2-purinoceptor antagonist in mammalian cells. Although the potent antitrypanosomal effect of Sur has been shown , limited evidence suggests that this drug can be dangerous to -infected hosts. Therefore, we investigated the dose-dependent effect of Sur-based chemotherapy in a murine model of Chagas disease. Seventy uninfected and -infected male C57BL/6 mice were randomized into five groups: SAL = uninfected; INF = infected; SR5, SR10, and SR20 = infected treated with 5, 10, or 20 mg/kg Sur. In addition to its effect on blood and heart parasitism, the impact of Sur-based chemotherapy on leucocytes myocardial infiltration, cytokine levels, antioxidant defenses, reactive tissue damage, and mortality was analyzed. Our results indicated that animals treated with 10 and 20 mg/kg Sur were disproportionally susceptible to , exhibiting increased parasitemia and cardiac parasitism (amastigote nests and parasite load ( DNA)), intense protein...
Dengue virus (DENV) is an arbovirus that belongs to the Flaviviridae family. Studies reveal that ... more Dengue virus (DENV) is an arbovirus that belongs to the Flaviviridae family. Studies reveal that peptides secreted by amphibians have many functions, such as antiviral and antimicrobial activities. As there is no antiviral drug effective against the DENV, the antiviral activity of a synthetic peptide called HS-1, derived from the secretion of the anuran Hypsiboas semilineatus, has been evaluated. The assays of neutralization in the Vero cells show a complete inhibition of infection of the serotypes 2 and 3. Furthermore, the direct action of peptides on the viral particle can be observed through atomic force microscopy. In vivo tests display 80% protection against the dengue-2 virus due to the presence of HS-1, which reveals its potential as an antiviral against the DENV.
We aimed to evaluate the postprandial secretion of inflammatory markers induced by SFA or MUFA hi... more We aimed to evaluate the postprandial secretion of inflammatory markers induced by SFA or MUFA high-fat meal consumption and whether orange juice intake could modulate this induction. This study included 55 healthy women (aged 20 to 40 years): 33 participants received an SFA high-fat meal (≈1000 kcal, 37.6% of energy intake (E) from SFA) and 22 participants received an MUFA high-fat meal (≈1000 kcal, 56.3% E from MUFA). Both interventions were accompanied by 500 ml of orange juice (test) or water (control). The plasma concentrations of inflammatory cytokines (IL-2, IL-4, IL-6, IL-10, IL-17A, IFN-γ, and TNF-α) and CRP were determined before (fasting) and 2, 3 and 5 hours after the test meal intake. The SFA high-fat meal induced a significant increase in AUC values (for TNF-α, IL-12, IL-10, IL-6 and IL-2 adjusted for baseline concentrations) in comparison with MUFA high-fat meal intervention. The results were independent of the drink which accompanied the meal (water or orange juice)....
International Journal of Food and Nutritional Science, 2016
The coconut oil is popularly recognized for its anti-inflammatory properties, but research has sh... more The coconut oil is popularly recognized for its anti-inflammatory properties, but research has shown conflicting results. Thus, our aim was to assess the postprandial inflammatory response of moderate coconut oil intake. We evaluated the acute consumption of a breakfast, offered in a single day, containing 25 mL of coconut oil (test) or 25 mL of olive oil (control)on postprandial serum inflammatory cytokines (IL-2, IL-4, IL-6, IL-10, IL-17A, IFN-γ and TNF) in healthy overweight women. No change was observed in relation to the effect of fatty meals, the time and the interaction of both factors (meal x time). Thus, despite being a saturated fat source, coconut oil consumption did not promote an inflammatory response. Further studies are needed considering a larger intervention period to check long-term effect.
International Journal of Biological Macromolecules, 2017
Lectins are involved in a wide range of biological mechanisms, like immunomodulatory agent able t... more Lectins are involved in a wide range of biological mechanisms, like immunomodulatory agent able to activate the innate immunity. In this study, we purified and characterized a new lectin from cauliflower (Brassica oleracea ssp. botrytis-BOL) by three sequential chromatographic steps and confirmed the purity by SDS-PAGE. Additionally, we evaluated the role of the lectin in innate immunity by a phagocytosis assay, production of H 2 O 2 and NO. BOL was characterized like a non-glycosylated protein that showed a molecular mass of ∼34 kDa in SDS-PAGE. Its N-terminal sequence (ETRAFREERPSSKIVTIAG) did not reveal any similarity to the other lectins; nevertheless, it showed 100% homology to a putative TRAFlike protein from Brassica rapa and Brassica napus. This is a first report of the TRAF-protein with lectinic activity. The BOL retained its complete hemagglutination activity from 4 • C up to 60 • C, with stability being more apparent between pH 7.0 and 8.0. Moreover, the lectin was able to stimulate phagocytosis and induce the production of H 2 O 2 and NO. Therefore, BOL can be explored as an immunomodulatory agent by being able to activate the innate immunity and favor antigen removal.
The mosquito Aedes aegypti transmit virus that cause diverse human diseases and the vector contro... more The mosquito Aedes aegypti transmit virus that cause diverse human diseases and the vector control is an important strategy to avoid their propagation. Plants in Annonaceae are recognized as source to molecules with use in medical and agriculture fields. Molecules of secondary metabolites of Annonaceae plants exhibit insecticidal potential against insect pest and vectors, with highlight to acetogenins that show high toxicity with low doses, which encouraged research to producing new insecticide molecules. Herein, we identify an acetogenin from Annona mucosa seeds (chemical analysis) and provide toxicity test against larvae of A. aegypti (target insect), its predators Culex bigoti and Toxorhynchites theobaldi (non-target insects) and cytotoxicity to human leukocytes. We identify squamocin (C37 H66 O7 ) a fatty acid with presence of bis-tetrahydrofuran ring. In A. aegypti, this compound exhibited behavioral disturb before larval death, high mortality and require low concentrations to ...
Kefir supplementation in rats with induced metabolic syndrome was able to lower fasting glucose, ... more Kefir supplementation in rats with induced metabolic syndrome was able to lower fasting glucose, fasting insulin levels, and reduce insulin resistance.
Podisus nigrispinus Dallas (Hemiptera: Pentatomidae) is a predator insect with potential applicat... more Podisus nigrispinus Dallas (Hemiptera: Pentatomidae) is a predator insect with potential applications in biological control because both nymphs and adults have been shown to prey on other insect pests by injection of toxic salivary gland contents. This study identified non-proteinaceous compounds with insecticidal activity from the saliva of P. nigrispinus in Anticarsia gemmatalis. In particular, the ether extract from P. nigrispinus saliva led to mortality in A. gemmatalis larvae, with a LC50 = 2.04 μL and LC90 = 3.27 μL. N,N-dimethylaniline and 1,2,5-trithiepane fractions were identified as non-proteinaceous extract components. N,N-dimethylaniline had a LC50 = 136.1 nL and LC90 = 413.8 nL, suggesting that it could be responsible for toxicity in P. nigrispinus saliva.
Comparative Biochemistry and Physiology Part A: Molecular & Integrative Physiology, 2008
A thrombin-like enzyme named BjussuSP-I, isolated from B. jararacussu snake venom, is an acidic s... more A thrombin-like enzyme named BjussuSP-I, isolated from B. jararacussu snake venom, is an acidic single chain glycoprotein with approximately 6% sugar, Mr = 61,000 under reducing conditions and pI ∼ 3.8, representing 1.09% of the chromatographic A 280 recovery. BjussuSP-I is a glycosylated serine protease containing both N-linked carbohydrates and sialic acid in its structure. BjussuSP-I showed a high clotting activity upon human plasma, which was inhibited by PMSF, leupeptin, heparin and 1,10-phenantroline. This enzyme showed high stability regarding coagulant activity when analyzed at different temperatures (−70 to 37°C), pHs (4.5 to 8.0), and presence of two divalent metal ions (Ca 2+ and Mg 2+ ). It also displayed TAME esterase and proteolytic activities toward natural (fibrinogen and fibrin) and synthetic (BAPNA) substrates, respectively, being also inhibited by PMSF and leupeptin. BjussuSP-I can induce production of polyclonal antibodies able to inhibit its clotting activity, but unable to inhibit its proteolytic activity on fibrinogen. The enzyme also showed crossed immunoreactivity against 11 venom samples of Bothrops, 1 of Crotalus, and 1 of Calloselasma snakes, in addition of LAAO isolated from B. moojeni venom. It displayed neither hemorrhagic, myotoxic, edema-inducing profiles nor proteolytic activity on casein. BjussuSP-I showed an N-terminal sequence (VLGGDECDINEHPFLA FLYS) similar to other thrombin-like enzymes from snake venoms. Based on its biochemical, enzymatic and pharmacological characteristics, BjussuSP-I was identified as a new thrombin-like enzyme isoform from Bothrops jararacussu snake venom.
We report the identification and characterization of a new AMP named Hs-1.Hs-1 is the first AMP d... more We report the identification and characterization of a new AMP named Hs-1.Hs-1 is the first AMP described for the Brazilian endemic frog Hypsiboas semilineatus.Hs-1 is selective against Gram-positive bacteria, especially against MRSA.Hs-1 has a direct antibacterial killing by membrane disruption.Hs-1 is predominantly structured as an amphipathic alpha helix.The multidrug-resistant bacteria have become a serious problem to public health. In this scenery the antimicrobial peptides (AMPs) derived from animals and plants emerge as a novel therapeutic modality, substituting or in addition to the conventional antimicrobial. The anurans are one of the richest natural sources of AMPs. In this work several cycles of cDNA cloning of the skin of the Brazilian treefrog Hypsiboas semilineatus led to isolation of a precursor sequence that encodes a new AMP. The sequence comprises a 27 residue signal peptide, followed by an acidic intervening sequence that ends in the mature peptide at the carboxy terminal. The AMP, named Hs-1, has 20 amino acids residues, mostly arranged in an alpha helix and with a molecular weight of 2144.6 Da. The chemically synthesized Hs-1 showed an antimicrobial activity against all Gram-positive bacteria tested, with a range of 11–46 μM, but it did not show any effect against Gram-negative bacteria, which suggest that Hs-1 may have a selective action for Gram-positive bacteria. The effects of Hs-1 on bacterial cells were also demonstrated by transmission electron microscopy. Hs-1 is the first AMP to be described from H. semilineatus.
Antimicrobial agents and chemotherapy, Jan 13, 2015
Although Suramin is suggested as a potential drug candidate in the management of Chagas disease, ... more Although Suramin is suggested as a potential drug candidate in the management of Chagas disease, this issue has not been objectively tested. In this study, we examined the applicability of the concomitant treatment with Benznidazole (Bz) and Suramin (Sur) in mice infected with a virulent strain of Trypanosoma cruzi. Eighty 12-week-old male C57BL/6 mice were equally randomized in seven groups: 1) non-infected (negative control) and infected with T. cruzi Y strain receiving 2) no treatment (positive control); 3) Bz 100 mg/kg/day; 4) Sur 20 mg/kg/day; 5 to 8) Sur 20 mg/kg/day combined with Bz 100; 50; 25 or 5 mg/kg/day. Bz was administered by gavage and Sur intraperitoneally. Sur dramatically increased the parasitemia, cardiac content of parasite DNA, inflammation, oxidative tissue damage and mortality. In response to high parasitic load in cardiac tissue, Sur stimulated the immune system in a manner typical of the acute phase of Chagas disease, increasing tissue levels of IFN-γ and TN...
The Journal of pharmacy and pharmacology, Jan 23, 2015
This study investigated the combined effects of benznidazole (BZ) and ibuprofen (IB) on the oxida... more This study investigated the combined effects of benznidazole (BZ) and ibuprofen (IB) on the oxidative and inflammatory status of the cardiac tissue in vivo. Swiss mice were randomized in groups receiving BZ (100 mg/kg) and IB (400 mg/kg) alone or combined (BZ + IB 200 or 400 mg/kg). Control animals were concurrently treated with 1% carboxymethyl cellulose. All treatments were administered orally for 7 days. BZ treatment increased cardiac production of nitrogen/oxygen-reactive species, malondialdeyde, carbonyl proteins, prostaglandins as well as the activities of catalase, superoxide dismutase and glutathione peroxidase. These parameters were attenuated by IB, with the best results at higher dose. Individually, BZ and IB significantly reduced the tissue levels of chemokine ligand 2, tumour necrosis factor-α and IL-10, but no reduction was observed when the treatments were combined. BZ triggers an oxidative and nitrosative route, which is associated with increased prostaglandin synthe...
The search of new drugs and the use of animal experimentation had been contributed excessively to... more The search of new drugs and the use of animal experimentation had been contributed excessively to the development of the health, of the science and technology, promoting along the years the discovery of prophylactic measured and treatments of illnesses that affect the human beings. Animals of several species have been used from the origins of the civilization for the scientific development, but among all of the experimental models to the which peptic ulcers can be induced, the rats Wistar, doubtless, is the used. The objective of this work was to accomplish an anatomical and histological description. Anatomically, the stomach of the Wistar rat (Rattus norvegicus) is located in the abdominal cavity in the ventral midline in the left portion, caudal to the liver. This body can be divided into two regions glandular and non glandular. These regions are separated by the folded edge that makes the division between the keratinized stratified squamous epithelium that lines the portion of ex...
This study used a murine model of Chagas disease to investigate the isolated and combined impact ... more This study used a murine model of Chagas disease to investigate the isolated and combined impact of Trypanosoma cruzi infection and benznidazole (BZ) therapy on liver structure and function. Male C57BL/6 mice were challenged with T. cruzi and BZ for 15 days. Serum levels of cytokines and hepatic enzymes, liver oxidative stress, morphology, collagen, and glycogen content were monitored. Separately, T. cruzi infection and BZ treatment resulted in a pro-oxidant status and hepatic reactive damage. Concurrently, both T. cruzi infection and BZ treatment induced upregulation of antioxidant enzymes and pathological reorganization of the liver parenchyma and stroma. T. cruzi infection increased serum levels of Th1 cytokines, which were reduced by BZ in both infected and non-infected animals. BZ also induced functional organ damage, increasing serum levels of liver enzymes. When combined, T. cruzi infection and BZ therapy elicited intense hepatic reactive damage that was not compensated by antioxidant enzymatic reaction, subsequently culminating in more severe morphofunctional hepatic injury. Taken together, these findings indicate that during specific treatment of Chagas disease, hepatic pathology may be a result of an interaction between BZ metabolism and specific mechanisms activated during the natural course of T. cruzi infection, rather than an isolated toxic effect of BZ on liver structure and function.
In this work, we report the evaluation of two DNA vaccines against dengue-3 virus (DENV-3). The f... more In this work, we report the evaluation of two DNA vaccines against dengue-3 virus (DENV-3). The first construction, called pVAC3DEN3, was engineered inserting the pre-membrane (prM) and envelope (E) gene of DENV-3 truncated with a restriction site between them, as previously described. The second construction was developed cloning the full gene sequence of prM and E from DENV-3 virus in pCI plasmid for mammalian expression and was denominated pVAC1WDEN3. The results showed that both constructions were capable of expressing the prM and E proteins, as demonstrated by ELISA and immunoblotting detection in cell culture transfected with the plasmids. After positive "in vitro" results, the vaccine candidates were used to immunize BALB/c mice and the elicited response was investigated. After immunization by intramuscular inoculation with three doses of each vaccinal clone the animals were sacrificed, the cytokine levels and T cell response were analyzed in the spleens, after three days of culture with stimulus, our analysis showed that the two constructions elicited T cell responses measured by BrdU incorporation assay and high levels of IFN-γ, detected in the supernatant of the cultures. Moreover, both constructions induced detectable titers of neutralizing antibodies in mice. And finally the survival rate of the immunized animals after intracerebral challenge was analyzed, showing a better result in the pVAC3DEN3 group with an 80% survival compared with a 50% survival of the pVAC1 WDEN3.
In calves, neonatal mortality and disease susceptibility are greatly influenced by failure in pas... more In calves, neonatal mortality and disease susceptibility are greatly influenced by failure in passive immunization, normally provided by colostrum ingestion just after birth. Formulations projected to replace natural colostrum have not been successful, and one of the possible reasons for such failure is that orally administered Ig are probably digested in the gastrointestinal tract, so they are not absorbed as intact functional molecules. With the aim of finding an adequate colostrum substitute, we used columns of immobilized jacalin, a lectin known by its ability to bind O-linked oligosaccharides, to obtain a colostral Ig population putatively protected against enzymatic cleavage by the presence of sugar chains. Immunoglobulin G1 is a major constituent of colostrum Ig bound to jacalin (JB-Ig). This preparation contains 10% of the total colostral Ig and is typically 3 to 6 times more resistant to pepsin digestion than the Ig contained in the fraction that is not bound to jacalin, wh...
Bovine mastitis is a major threat to animal health and the dairy industry. Staphylococcus aureus ... more Bovine mastitis is a major threat to animal health and the dairy industry. Staphylococcus aureus is a contagious pathogen that is usually associated with persistent intramammary infections, and biofilm formation is a relevant aspect of the outcome of these infections. Several biological activities have been described for snake venoms, which led us to screen secretions of Bothrops jararacussu for antibiofilm activity against S. aureus NRS155. Crude venom was fractionated by size-exclusion chromatography, and the fractions were tested against S. aureus. Biofilm growth, but not bacterial growth, was affected by several fractions. Two fractions (15 and 16) showed the best activities and were also assayed against S. epidermidis NRS101. Fraction 15 was identified by TripleTOF mass spectrometry as a galactose-binding C-type lectin with a molecular weight of 15 kDa. The lectin was purified from the crude venom by D-galactose affinity chromatography, and only one peak was observed. This pure lectin was able to inhibit 75% and 80% of S. aureus and S. epidermidis biofilms, respectively, without affecting bacterial cell viability. The lectin also exhibited a dose-dependent inhibitory effect on both bacterial biofilms. The antibiofilm activity was confirmed using scanning electron microscopy. A pre-formed S. epidermidis biofilm was significantly disrupted by the C-type lectin in a time-dependent manner. Additionally, the lectin demonstrated the ability to inhibit biofilm formation by several mastitis pathogens, including different field strains of S. aureus, S. hyicus, S. chromogenes, Streptococcus agalactiae, and Escherichia coli. These findings reveal a new activity for C-type lectins. Studies are underway to evaluate the biological activity of these lectins in a mouse mastitis model.
The cabbage (Brassica oleraceae var. capitata) is an herbaceous and leafy plant which belongs to ... more The cabbage (Brassica oleraceae var. capitata) is an herbaceous and leafy plant which belongs to the Brassicaceae family, native to coastal southern and Western Europe. Used in cooking for its nutritional value also has known anti-inflammatory activity. OBJECTIVE We studied the antiulcerogenic activity of aqueous extract of Brassica oleracea var. capitata (AEB) in order to validate ethnobotanical claims regarding the plant use in the gastric disorders. Acute gastric ulcers were induced in rats by the oral administration of acetylsalicylic acid. The gastroprotective potential of the AEB (0.250, 0.500 and 1.000 mg.kg-1/body weight) was compared with omeprazole (20 mg.kg-1/body weight). The stomach analysis indicated that treatment with AEB inhibited the gastric damage. The gastroprotective activity as evidenced by its significant inhibition in the formation of ulcers induced by chemical agent with a maximum of 99.44% curation (250 mg.kg-1 body weight) in acetylsalicylic acid-induced u...
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Papers by Leandro L Oliveira