Publications by authors named "João Carlos Palazzo de Mello"

50 Publications

Hydrogel containing (1 → 6)-β-D-glucan (lasiodiplodan) effectively promotes dermal wound healing.

Int J Biol Macromol 2021 Jul 27;183:316-330. Epub 2021 Apr 27.

Programa de Pós-Graduação em Biotecnologia, Universidade Tecnológica Federal do Paraná, Câmpus Dois Vizinhos, CEP 85660-000 Dois Vizinhos, Paraná, Brazil; Departamento de Química, Universidade Tecnológica Federal do Paraná, Campus Pato Branco, CEP 85503-390 Pato Branco, Paraná, Brazil. Electronic address:

A hydrogel containing exocellular (1 → 6)-β-D-glucan (lasiodiplodan, LAS) was developed and its wound healing potential was evaluated. β-Glucans have attracted much interest by the cosmetic industry sector because of their bioactive and functional properties and in promoting skin health. In the present work an β-glucan was studied as a healing biomaterial that has not hitherto been reported in the scientific literature. LAS produced by the ascomycete Lasiodiplodia theobromae MMPI was used in the formulation of a healing hydrogel. Physicochemical and microbiological quality parameters, antioxidant potential and stability of the formulation was evaluated. FTIR, thermal analysis and SEM techniques were also employed in the characterization. Wistar rats were used as a biological model to investigate the wound healing potential. Histological analyses of cutaneous tissue from the dorsal region were conducted after 4, 7, 10 and 14 days of treatment, and evaluated re-epithelialization, cell proliferation and collagen production. Physicochemical stability, microbiological quality and antioxidant potential, especially in relation to its ability to scavenge hydroxyl radicals were found. The hydrogel stimulated cell re-epithelialization and proliferation during all days of the treatment, and stimulated an increase of collagen fibers. Lasiodiplodan showed immunomodulatory activity in wound healing and this biomacromolecule could be an alternative compound in wound care.
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http://dx.doi.org/10.1016/j.ijbiomac.2021.04.169DOI Listing
July 2021

Antileishmanial activity of -clerodane diterpenes from .

Nat Prod Res 2020 Nov 28:1-7. Epub 2020 Nov 28.

Department of Chemistry, Universidade Estadual de Maringá, Maringá, Paraná, Brazil.

Nowadays, new leishmanicidal drugs are needed and natural products arise as a promising alternative source. Therefore, bioguided fractionation of a hydroethanolic extract from the stem bark of Baill. were conducted based on its antileishmanial activity. Two novel -clerodane diterpenoids methyl-15,16-epoxy-3,13(16),14--clerodatrien-17,18-dicarboxylate () and dimethyl-3-oxo-15,16-epoxy-13(16),14--clerodadien-17,18-dicarboxylate () were isolated, as well as four known compounds () and lupeol, from the hexane fraction. Their structures were established by NMR analysis. The crude extract, fractions and the compounds ( and ) were evaluated for their antileishmanial activity and cytotoxicity against macrophages J774A.1. The selectivity index (SI) were calculated. The most active compound against promastigote forms of was the clerodane diterpene , with IC values of 8.3 µM and SI value of 80.9. Our results highlighted stem bark of Baill. as a promising source for the development of a new chemotherapeutic agent to combat leishmaniasis.
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http://dx.doi.org/10.1080/14786419.2020.1851221DOI Listing
November 2020

The plant Stryphnodendron adstringens (Mart.) Coville as a neutralizing source against some toxic activities of Bothrops jararacussu snake venom.

Toxicon 2020 Oct 19;186:182-190. Epub 2020 Aug 19.

Department of Molecular and Cellular Biology, Institute of Biology, Federal Fluminense University, Niterói, 24020-141, RJ, Brazil. Electronic address:

Snakebites produce several toxic effects in victims, such as hemorrhage, tissue necrosis, hemostatic, renal, or cardiotoxic alterations, inflammation, and death. To counteract these symptoms, antivenom is the official treatment. Although such therapy prevents death, it does not efficiently neutralize necrosis or other local effects, leading to amputation or morbidities of the affected limb. Therefore, the search for better and more efficient therapies deserves attention; further, plants have been used to ameliorate a number of diseases and medical conditions, including snakebites, for many years. Thus, the aim of this work was to evaluate the antivenom effect of the crude extract, fractions (aqueous and diethyl acetate), and subfractions derived from the aqueous fraction (P1, P2, P3, and P4) of the plant Stryphnodendron adstringens against in vitro (coagulation and proteolytic) and in vivo (edema, hemorrhage, and myotoxic) activities caused by Bothrops jararacussu venom. Overall, all extracts inhibited the toxic effect of B. jararacussu venom, but with different potencies, regardless of whether plant samples were incubated together with venom or injected before or after venom injection into animals; the crude extract and aqueous fraction were found to be the most effective. Indeed, phytochemical and mass spectrometry analysis of S. adstringens samples revealed the presence of flavonols, tannins, and saponins. In conclusion, the plant S. adstringens may represent a promising natural source of molecules to treat the toxic effects associated with envenomation by B. jararacussu snakebites.
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http://dx.doi.org/10.1016/j.toxicon.2020.08.011DOI Listing
October 2020

Stem bark extract of Poincianella pluviosa incorporated in polymer film: Evaluation of wound healing and anti-staphylococcal activities.

Injury 2020 Apr 11;51(4):840-849. Epub 2020 Feb 11.

Department of Pharmacy, Laboratory of Pharmaceutical Biology - Palafito, Universidade Estadual de Maringá, BR-87020-900, Maringá, Brazil.. Electronic address:

Poincianella pluviosa has already been described as capable of healing skin wounds. In an attempt to prolong contact of the drug with the wound, it was proposed in this study to evaluate wound healing using a crude extract (CE) of P. pluviosa incorporated in carboxymethylcellulose polymer films. The chromatographic profile of the semipurified fraction of P. pluviosa was evaluated by ultra-high performance liquid chromatography (UHPLC), confirming the compounds gallic acid, geraniin, and ellagic acid. The films were evaluated for their physical and mechanical properties, water vapor permeability, moisture absorption capacity, and FTIR spectroscopy. For in vivo experiments, wounds were made on the back of rats and treated daily for 4, 7, 10, or 14 days with film containing CE or control film. At the end of each period, skin permeation analysis and histological analysis were made using re-epithelialisation, cell proliferation, and collagen formation. Statistical significance was determined by GraphPad Prism using t test and Mann-Whitney test. Anti-staphylococcal activity was evaluated with standard strains of Staphylococcus aureus, methicillin-resistant, and coagulase negative. It was demonstrated that the presence of CE in the films increased the capacity to absorb water and decreased resistance and permeability. The CE of the film permeated the skin, reaching the dermis and was able to influence re-epithelisation, cell proliferation, and collagen formation. Satisfactory results were observed against S. aureus strains, particularly coagulase negative. Films with CE of P. pluviosa can be an alternative in the wound healing, protecting against opportunistic infections and giving comfort to the patient.
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http://dx.doi.org/10.1016/j.injury.2020.02.027DOI Listing
April 2020

Dimeric glycosylated flavan-3-ol and antimicrobial evaluation of extracts.

Nat Prod Res 2019 Dec 9:1-8. Epub 2019 Dec 9.

Laboratory of Pharmaceutical Biology, Palafito, Department of Pharmacy, Universidade Estadual de Maringá, Maringá, Brazil.

is a tree known as "catuaba", widely distributed in Brazil. Studies carried out with barks suggest that plant has antidepressant, antidiabetic, antimicrobial, antioxidant, antiviral, and preventive against brain damage. The aim of this work was to isolate and characterise compounds from the semipurified fraction of barks, and to conduct microbiological screening against bacteria and fungi. The crude extract (CE) of "catuaba" was produced by turbo extraction with acetone-water, and later, partitioned to yield ethyl-acetate (EAF) and aqueous (AqF) fractions. From AqF the new catechin-3--α-L-rhamnoside-(4α→8)-epicatechin was isolated, identified, and described here for the first time. Regarding antimicrobial activity, the extracts presented impressive results, mainly for Vancomycin Resistant (VRE) with MIC of 156.5 μg/mL. The results suggest that extract of could potentially be used as an adjuvant to treatment and is a promising candidate for the development of new antimicrobial drugs.
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http://dx.doi.org/10.1080/14786419.2019.1698569DOI Listing
December 2019

Ethyl Acetate Fraction from Trichilia catigua Confers Partial Neuroprotection in Components of the Enteric Innervation of the Jejunum in Diabetic Rats.

Cell Physiol Biochem 2019 ;53(1):76-86

Department of Morphological Sciences, Universidade Estadual de Maringá, Paraná, Brasil.

Background/aims: Diabetes causes damage to the enteric nervous system. The enteric nervous system consists of neurons and enteric glial cells (EGCs). The present study evaluated the effects of an ethyl-acetate fraction (EAF) from Trichilia catigua (T. catigua; 200 mg/kg) on the total population of enteric neurons (HuC/D-immunoreactive [IR]) and EGCs (S100-IR and glial fibrillary acidic protein [GFAP]-IR) in the total preparation and jejunal mucosa in diabetic rats.

Methods: The animals were distributed into four groups: normoglycemic rats (N), diabetic rats (D), normoglycemic rats that received the EAF (NC), and diabetic rats that received the EAF (DC). The jejunum was processed for immunohistochemistry to evaluate HuC/D, S100, and GFAP immunoreactivity. The expression of S100 and GFAP proteins was also quantified by Western blot.

Results: The D group exhibited a decrease in the number of neurons and EGCs, an increase in the area of cell bodies, an increase in S100 protein expression, a decrease in GFAP protein expression, and a decrease in S100-IR and GFAP-IR EGCs in the jejunal mucosa. The DC group exhibited a decrease in the number of neurons and EGCs, a decrease in the area of cell bodies, a decrease in S100 and GFAP protein expression, and a decrease in S100-IR and GFAP-IR EGCs in the jejunal mucosa. The NC group exhibited maintenance of the number of neurons and EGCs, an increase in the area of cell bodies, and a decrease in S100 and GFAP protein expression.

Conclusion: The EAF from T. catigua partially conferred protection against diabetic neuropathy in the enteric nervous system.
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http://dx.doi.org/10.33594/000000122DOI Listing
June 2019

In vitro evaluation of the protective effects of plant extracts against amyloid-beta peptide-induced toxicity in human neuroblastoma SH-SY5Y cells.

PLoS One 2019 14;14(2):e0212089. Epub 2019 Feb 14.

Programa de Pós-Graduação em Ciências Farmacêuticas, Department of Pharmacy, Universidade Estadual de Maringá, Maringá, Paraná, Brazil.

Alzheimer's disease (AD) is the most common form of dementia and has no cure. Therapeutic strategies focusing on the reduction of oxidative stress, modulation of amyloid-beta (Aβ) toxicity and inhibition of tau protein hyperphosphorylation are warranted to avoid the development and progression of AD. The aim of this study was to screen the crude extracts (CEs) and ethyl-acetate fractions (EAFs) of Guazuma ulmifolia, Limonium brasiliense, Paullinia cupana, Poincianella pluviosa, Stryphnodendron adstringens and Trichilia catigua using preliminary in vitro bioassays (acetylcholinesterase inhibition, antioxidant activity and total polyphenol content) to select extracts/fractions and assess their protective effects against Aβ25-35 toxicity in SH-SY5Y cells. The effect of the EAF of S. adstringens on mitochondrial membrane potential, lipid peroxidation, superoxide production and mRNA expression of 10 genes related to AD was also evaluated and the electropherogram fingerprints of EAFs were established by capillary electrophoresis. Chemometric tools were used to correlate the in vitro activities of the samples with their potential to be evaluated against AD and to divide extracts/fractions into four clusters. Pretreatment with the EAFs grouped in cluster 1 (S. adstringens, P. pluviosa and L. brasiliense) protected SH-SY5Y cells from Aβ25-35-induced toxicity. The EAF of S. adstringens at 15.62 μg/mL was able completely to inhibit the mitochondrial depolarization (69%), superoxide production (49%) and Aβ25-35-induced lipid peroxidation (35%). With respect to mRNA expression, the EAF of S. adstringens also prevented the MAPT mRNA overexpression (expression ratio of 2.387x) induced by Aβ25-35, which may be related to tau protein hyperphosphorylation. This is the first time that the neuroprotective effects of these fractions have been demonstrated and that the electropherogram fingerprints for the EAFs of G. ulmifolia, L. brasiliense, P. cupana, P. pluviosa and S. adstringens have been established. The study expands knowledge of the in vitro protective effects and quality control of the evaluated fractions.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0212089PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6375598PMC
November 2019

Chemical analysis of the semipurified extract of and evaluation of inhibitory effects against .

Nat Prod Res 2020 Aug 2;34(16):2332-2335. Epub 2019 Jan 2.

Laboratory of Pharmaceutical Biology, Palafito, Department of Pharmacy, State University of Maringá - UEM, Maringá, PR, Brazil.

Kunth, commonly known as guarana, is a native Brazilian plant species from the Amazon area that presents various biological effects, including antimicrobial action. The aim of this study was to chemically analyse the semipurified aqueous extract (AqF) of the plant and to evaluate the activity of crude (CE), ethyl-acetate (EAF), and AqF extracts against . The chemical profile of AqF was determined based on solid analysis C-NMR, direct infusion mass spectrometry (ESI-MS), and MALDI-TOF. The C-NMR spectrum showed characteristics of flavan-3-ol and oligomeric proanthocyanidins. ESI-MS revealed the presence of procyanidin, caffeic acid and its derivatives. MALDI-TOF analysis detected procyanidins of up to 6 units and profisetinidins of up to 5 units. Whereas CE and EAF showed inhibitory activity against , CE, EAF, and AqF presented not high inhibitory activity against urease. The results demonstrate the potential of to control and prevent infection.
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http://dx.doi.org/10.1080/14786419.2018.1533825DOI Listing
August 2020

Maternal diet-induced obesity during suckling period programs offspring obese phenotype and hypothalamic leptin/insulin resistance.

J Nutr Biochem 2018 11 25;61:24-32. Epub 2018 Jul 25.

Department of Morphological Sciences, State University of Maringá, Maringá, PR, Brazil.

During the early post-natal period, offspring are vulnerable to environmental insults, such as nutritional and hormonal changes, which increase risk to develop metabolic diseases later in life. Our aim was to understand whether maternal obesity during lactation programs offspring to metabolic syndrome and obese phenotype, in addition we aimed to assess the peripheral glucose metabolism and hypothalamic leptin/insulin signaling pathways. At delivery, female Wistar rats were randomly divided in two groups: Control group (CO), mothers fed a standard rodent chow (Nuvilab); and Diet-induced obesity group (DIO), mothers who had free access to a diet performed with 33% ground standard rodent chow, 33% sweetened condensed milk (Nestlé), 7% sucrose and 27% water. Maternal treatment was performed throughout suckling period. All offspring received standard rodent chow from weaning until 91-day-old. DIO dams presented increased total body fat and insulin resistance. Consequently, the breast milk from obese dams had altered composition. At 91-day-old, DIO offspring had overweight, hyperphagia and higher adiposity. Furthermore, DIO animals had hyperinsulinemia and insulin resistance, they also showed pancreatic islet hypertrophy and increased pancreatic β-cell proliferation. Finally, DIO offspring showed low ObRb, JAK2, STAT-3, IRβ, PI3K and Akt levels, suggesting leptin and insulin hypothalamic resistance, associated with increased of hypothalamic NPY level and decreased of POMC. Maternal obesity during lactation malprograms rat offspring to develop obesity that is associated with impairment of melanocortin system. Indeed, rat offspring displayed glucose dyshomeostasis and both peripheral and central insulin resistance.
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http://dx.doi.org/10.1016/j.jnutbio.2018.07.006DOI Listing
November 2018

RNAm expression profile of cancer marker genes in HepG2 cells treated with different concentrations of a new indolin-3-one from Pseudomonas aeruginosa.

Sci Rep 2018 08 24;8(1):12781. Epub 2018 Aug 24.

Laboratory of Toxicological Genetics, Department of General Biology, State University of Londrina, Londrina, PR, Brazil.

The present study tested the effects of a newly identified indolin-3-one compound (compound 1), produced by Pseudomonas aeruginosa, on HepG2 cells. The MTT assays demonstrated decreased metabolic activities in HepG2 cells treated with compound 1, with dose- and time-dependent intensifying effect, starting at a concentration of 40 µM. The IC after 24, 48, 72, and 96 h treatments were 41.35, 52.7, 92.79 and 66.65 μM of compound 1, respectively. Below 80 µM, no significative damage on erythrocytes membranes was observed by the hemolytic assays. The RT-qPCR revealed that the compound modulated key genes involved in carcinogenesis process, indicating possible indolin-3-one mechanisms of action. The data showed that gene expression alterations promoted by compound 1, in concentrations up to 60 μM after 48 h, led to a decrease in cellular progression and there was no direct cellular damage. In addition, non-cytotoxic concentrations of compound 1 halved the concentration of the chemotherapeutic doxorubicin, maintaining similar therapeutic effect against HepG2 cells. The novelty of the molecule and the biological activities observed in the present study emphasize the potential of the compound 1 in cancer therapy research.
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http://dx.doi.org/10.1038/s41598-018-30893-wDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6109079PMC
August 2018

Treatment with Trichilia catigua ethyl-acetate fraction improves healing and reduces oxidative stress in TNBS-induced colitis in rats.

Biomed Pharmacother 2018 Nov 5;107:194-202. Epub 2018 Aug 5.

Laboratory of Animal Histology, Department of Morphological Sciences, State University of Maringá, Av. Colombo 5790, 87020-900, Maringá, PR, Brazil. Electronic address:

Beverages containing Trichilia catigua are commonly employed in folk medicine. T. catigua bark extracts possess antioxidant, anti-inflammatory, and bactericidal properties. These properties suggest T. catigua bark extracts as a potential treatment for inflammatory bowel diseases (IBD). Using the 2,4,6-trinitrobenzenesulphonic acid (TNBS)-induced model of colitis in rats we evaluated the effect of an ethyl-acetate fraction (EAF) of T. catigua (200 mg/kg) administered by daily oral gavage or intrarectally at different time points after TNBS challenge. TNBS treatment evoked severe colonic inflammation after 24 h that persisted for 7 days, characterized by weight loss, high levels of myeloperoxidase activity, histological and macroscopic damage, and elevated index of oxidative stress in the blood. T. catigua EAF treatment prevented the oxidative stress within 24 h and enhanced tissue recovery observed at day 7, returning histological and macroscopic damage levels to that of the control group. TNBS treatment led to loss of myenteric neurons after 28 days. T. catigua EAF was unable to prevent the neuronal loss. Oral delivery of T. catigua EAF was more effective than intrarectal administration of the extract. In conclusion, T. catigua EAF treatment normalized oxidative stress parameters in blood and reduced the degree of acute inflammation in TNBS colitis.
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http://dx.doi.org/10.1016/j.biopha.2018.07.160DOI Listing
November 2018

Larvicidal Activity against and Chemical Characterization of the Inflorescences of .

Evid Based Complement Alternat Med 2017 7;2017:9602368. Epub 2017 Dec 7.

Programa de Pós-Graduação em Ciências Farmacêuticas, Department of Pharmacy, Laboratory of Pharmaceutical Biology (Palafito), Universidade Estadual de Maringá, Avenida Colombo 5790, Maringá, PR, Brazil.

The crude acetone extract (CAE) of defatted inflorescences of was partitioned into five semipurified fractions: -hexane (HF), dichloromethane (DF), ethyl acetate (EAF), -butanol (BF), and aqueous (AQF). BF was fractionated by reversed-phase polyamide column chromatography, obtaining 34 subfractions, which were subjected to HSCCC, where patuletin and patulitrin were isolated. CAE and the fractions BF, EAF, DF, and AQF were analyzed by LC-DAD-MS, and patuletin and patulitrin were determined as the major substances in EAF and BF, respectively. BF was also analyzed by HPLC and capillary electrophoresis (CE), and patulitrin was again determined to be the main substance in this fraction. CAE and the semipurified fractions (750, 500, 300, 100, and 50 mg/L) were assayed for larvicidal activity against , with mortality rate expressed as percentage. All fractions except AQF showed insecticidal activity after 24 h exposure of larvae to the highest concentration. However, EAF showed the highest activity with more than 50% reduction in larval population at 50 mg/L. The insecticidal activity observed with EAF might have been due to the higher concentration of patuletin present in this fraction.
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http://dx.doi.org/10.1155/2017/9602368DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5738574PMC
December 2017

Proanthocyanidin polymeric tannins from Stryphnodendron adstringens are effective against Candida spp. isolates and for vaginal candidiasis treatment.

J Ethnopharmacol 2018 Apr 8;216:184-190. Epub 2018 Jan 8.

Departamento de Microbiologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, SP, Brazil. Electronic address:

Ethnopharmacological Relevance: The stem bark of Stryphnodendron adstringens (Mart.) Coville is popularly used as anti-inflammatory, astringent and in the treatment of wounds and vaginal infections. Several pharmacological activities have been scientifically proven by in vitro and in vivo experimental assays for antibacterial, antiviral, antiprotozoan, anti-inflammatory and antioxidant.

Aim Of The Study: We investigated whether proanthocyanidin polymeric tannins from the Stryphnodendron adstringens stem bark with antifungal activity against Candida albicans in vitro are also active against planktonic and biofilm cells of Candida non-albicans (CNA, including fluconazole-resistant isolates) and are capable of controlling Candida vaginitis in vivo.

Materials And Methods: A total of 46 clinical isolates and 5 reference Candida spp. strains were used in this study. The antifungal effects in vitro of tannins (F2 and sub-fraction F2.4) from S. adstringens stem bark were evaluated using a broth microdilution assay (for planktonic yeasts and biofilm dispersion cells) or by XTT assay (for biofilm sessile cells). For in vivo antifungal activity analysis, mice with vaginal infection by C. albicans or C. glabrata were treated with a topical gel containing F2 (alone or combined with oral fluconazole), and the vaginal histopathology and fungal burden (by CFU counts from vaginal homogenates) were analyzed.

Results: F2 and F2.4 inhibited the proliferation of planktonic cells of Candida spp., especially that of fluconazole- and/or amphotericin B-resistant isolates. F2 and F2.4 also inhibited the proliferation of Candida biofilm dispersion cells. Moreover, a gel containing F2 efficiently controlled vaginal infection by C. albicans and C. glabrata in mice, with no noticeable toxicity to vaginal tissue.

Conclusions: Our data show that proanthocyanidin polymeric tannins obtained from S. adstringens have antifungal activity in vitro against C. albicans and CNA (including fluconazole-resistant isolates) and presented efficacy in the control of candidiasis in murine model. Therefore, these tannins have potential use in the treatment of vaginal candidiasis, representing interesting alternatives to current antifungals.
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http://dx.doi.org/10.1016/j.jep.2018.01.008DOI Listing
April 2018

Anti-Diabetic Effects of the Ethyl-Acetate Fraction of Trichilia catigua in Streptozo-tocin-Induced Type 1 Diabetic Rats.

Cell Physiol Biochem 2017 29;42(3):1087-1097. Epub 2017 Jun 29.

Department of Morphological Sciences, State University of Maringá, Maringá, Brazil.

Background/aims: Trichilia catigua A. Juss., known as "catuaba" in Brazil, has been popularly used as a tonic for fatigue, impotence and memory deficits. Previously, our group demonstrated that the ethyl-acetate fraction (EAF) of T. catigua has antioxidant and anti-inflammatory effects. The present study evaluated the anti-diabetic activity of EAF in type 1 diabetic rats.

Methods: Male Wistar rats were divided into four groups (N: non-diabetic group, D: type 1 diabetic group, NC: non-diabetic + EAF group and DC: type 1 diabetic + EAF group). The latter two groups were treated with 200 mg/kg EAF. Type 1 diabetes was induced by intravenous streptozotocin (STZ) injection (35 mg/kg). Starting two days after STZ injection, EAF was administered daily by gavage for 8 weeks.

Results: EAF attenuated body mass loss and reduced food and water intake. EAF improved hyperglycaemia and other biochemical parameters, such as alkaline phosphatase (ALP), alanine aminotransferase (ALT) and aspartate aminotransferase (AST). Furthermore, the number of pancreatic β-cells and the size of the islets had increased by β-cell proliferation in the DC group. EAF promoted reduction in kidney tissue damage in STZ-induced diabetic rats by reduction of renal fibrosis.

Conclusion: The present study showed that EAF improves glucose homeostasis and endocrine pancreas morphology and inhibits the development of diabetic nephropathy in STZ-induced diabetic rats.
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http://dx.doi.org/10.1159/000478761DOI Listing
November 2017

Pheophorbide a, a compound isolated from the leaves of Arrabidaea chica, induces photodynamic inactivation of Trypanosoma cruzi.

Photodiagnosis Photodyn Ther 2017 Sep 3;19:256-265. Epub 2017 Jun 3.

Post-Graduate Program in Pharmaceutical Sciences, Laboratory of Technological Innovation in the Development of Drugs and Cosmetics, State University of Maringá, Maringá, Paraná, Brazil; Department of Basic Sciences of Health, State University of Maringá, Maringá, Paraná, Brazil. Electronic address:

Background: Approximately 6-7 million people are infected with Trypanosoma cruzi, the etiological agent of Chagas' disease. Only two therapeutic compounds have been found to be useful against this disease: nifurtimox and benznidazole. These drugs have been effective in the acute phase of the disease but less effective in the chronic phase; they also have many side effects. Thus, the search for new compounds with trypanocidal action is necessary. Natural products can be the source of many important substances for the development of drugs to treat this infection. The present study evaluated the biological activity of an extract and fractions of Arrabidaea chica against T. cruzi and observed morphological and ultrastructural characteristics of parasites exposed to the isolated compound pheophorbide a.

Methods: The crude hydroethanolic extract of A. chica was prepared. Fractions were obtained by partition and separated by liquid chromatography.

Results: We observed a progressive increase in activity against epimastigote, trypomastigote, and amastigote forms of the parasite over the course of the fractionation process. Interestingly, we isolated a compound known as a photosensitizer that is used in photodynamic therapy. This method of treatment involving a photosensitizer, activation light and molecular oxygen is of great importance due to its selectivity. Pheophorbide a had activity against the protozoan in the presence of light and caused morphological and ultrastructural changes, demonstrating its potential in photodynamic therapy.

Conclusions: Based on the ability of pheophorbide a to eliminate bloodstream forms of T. cruzi, we suggest its use in blood banks for hemoprophylaxis.
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http://dx.doi.org/10.1016/j.pdpdt.2017.05.004DOI Listing
September 2017

Evaluation of Antibacterial, Antineoplastic, and Immunomodulatory Activity of Seeds Crude Extract and Ethyl-Acetate Fraction.

Evid Based Complement Alternat Med 2016 7;2016:1203274. Epub 2016 Dec 7.

Laboratory of Immunomodulation and New Therapeutical Approaches, Research Centre for Therapeutic Innovation Suely Galdino (NUPIT-SG), Federal University of Pernambuco, Recife, PE, Brazil.

(Guarana) is a native plant of Amazon region that has very traditional importance. Its seeds are rich in bioactive compounds, including tannins, which exhibit relevant properties. This study aimed to evaluate antibacterial, antineoplastic, and immunomodulatory activity of seeds crude extract (CE) and ethyl-acetate fraction (EAF). Antibacterial activity was evaluated by determination of minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC). Antineoplastic activity was evaluated by MTT assays in hepatocellular carcinoma (HepG2), breast adenocarcinoma (MCF-7), ductal carcinoma (T47-D), non-Hodgkin's B cell lymphoma (Toledo), T cell leukemia (Jukart), and Acute Leukemia (HL-60) cell lines. BALB/c mice splenocytes were treated to assess IFN-, IL-6, IL-17, and IL-10 levels by sandwich ELISA. CE and EAF were not toxic to peripheral blood cells and splenocytes. CE and EAF fractions showed a bacteriostatic activity (MIC = 250 g/mL) and presented IC values of 70.25 g/mL and 61.18 g/mL in HL-60 leukemia cell line. All cytokines evaluated had their levels reduced after treatment, following dose-response model. Different biological activities were observed for both CE and EAF, suggesting as a source of bioactive substances, especially tannins that may be used for several diseases treatments.
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http://dx.doi.org/10.1155/2016/1203274DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5174172PMC
December 2016

Capillary Electrophoresis as Tool for Diastereomeric Separation in a Trichilia catigua Fraction.

Phytochem Anal 2017 May 2;28(3):144-150. Epub 2016 Dec 2.

Programa de Pós-Graduação em Ciências Farmacêuticas, Department of Pharmacy, Universidade Estadual de Maringá, Maringá, PR. Avenida Colombo, 5790, BR-87020-900, Maringá, PR, Brazil.

Introduction: The tree Trichilia catigua, popularly known as "catuaba", shows several biological activities and has emerged as a potential source of new drugs. Considering that more than 10 species are known under the same popular name, regulatory agencies require more rigorous quality control of this medicinal plant.

Objective: To develop and validate a methodology using capillary electrophoresis (CE) with ultraviolet (UV) detection for analysing polyphenols in the ethyl-acetate fraction (EAF) of Trichilia catigua.

Methodology: Different electrophoretic conditions (such as wavelength of UV detection, voltage, buffer concentration and pH, cyclodextrin type and concentration) were investigated. After optimisation, borate buffer 80 mmol/L at pH 8.80 with 2-hydroxypropyl-β-cyclodextrin 10 mmol/L was selected as background electrolyte. A voltage reduction was used to improve the separation of a diastereomeric pair of cinchonains.

Results: The method proved to be simple, sensitive, accurate, linear, precise and reproducible. For the first time in natural products analysis, a voltage reduction and hydroxypropyl-β-cyclodextrin were used to improve the separation of diastereomeric pairs. Until now, this is the only described methodology able to separate catechin, epicatechin, cinchonains Ia, Ib, IIa, and IIb from Trichilia catigua samples on the same run in less than 12 min. When compared to the high performance liquid chromatography with photo-diode array detection (HPLC-PDA) method previously developed by our research group, the CE method was more efficient, faster, less expensive and less polluting.

Conclusion: Our results demonstrate that this method could be employed in a quality-control laboratory for the quantification of polyphenols in EAF of Trichilia catigua. Copyright © 2016 John Wiley & Sons, Ltd.
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http://dx.doi.org/10.1002/pca.2653DOI Listing
May 2017

Antiviral Activity of Crude Hydroethanolic Extract from Schinus terebinthifolia against Herpes simplex Virus Type 1.

Planta Med 2017 Apr 5;83(6):509-518. Epub 2016 Oct 5.

Programa de Pós-Graduação em Ciências Farmacêuticas, Universidade Estadual de Maringá, Maringá, PR, Brazil.

virus infections persist throughout the lifetime of the host and affect more than 80 % of the humans worldwide. The intensive use of available therapeutic drugs has led to undesirable effects, such as drug-resistant strains, prompting the search for new antiherpetic agents. Although diverse bioactivities have been identified in , its antiviral activity has not attracted much attention. The present study evaluated the antiherpetic effects of a crude hydroethanolic extract from the stem bark of against virus type 1 and as well as its genotoxicity in bone marrow in mammals and established the chemical composition of the crude hydroethanolic extract based on liquid chromatography-diode array detector-mass spectrometry and MS/MS. The crude hydroethanolic extract inhibited all of the tested virus type 1 strains and was effective in the attachment and penetration stages, and showed virucidal activity, which was confirmed by transmission electron microscopy. The micronucleus test showed that the crude hydroethanolic extract had no genotoxic effect at the concentrations tested. The crude hydroethanolic extract afforded protection against lesions that were caused by virus type 1 . Liquid chromatography-diode array detector-mass spectrometry and MS/MS identified 25 substances, which are condensed tannins mainly produced by a B-type linkage and prodelphinidin and procyanidin units.
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http://dx.doi.org/10.1055/s-0042-117774DOI Listing
April 2017

Guaraná (Paullinia cupana) seeds: Selective supercritical extraction of phenolic compounds.

Food Chem 2016 Dec 15;212:703-11. Epub 2016 Jun 15.

Department of Pharmacy, State University of Maringá, Av. Colombo 5790, BR-87020-900 Maringá, Paraná, Brazil. Electronic address:

Approximately 70% of the Brazilian production of guaraná (Paullinia cupana) seeds is absorbed by the beverage industries. Guaraná has several pharmacological properties: energy stimulant, antimicrobial, chemoprophylactic, antigenotoxic, antidepressive, anxiolytic, and anti-amnesic effects. Supercritical carbon dioxide extraction of bioactive compounds from guaraná seeds was carried out and optimized by an orthogonal array design (OA9(3(4))). The factors/levels studied were: modifier(s) (ethanol and/or methanol), extraction time (20, 40, and 60min), temperature (40, 50, and 60°C), and pressure (100, 200, and 300bar). The statistical design was repeated with increasing proportions of modifiers. The percentage of modifier used was proportional to the amount of polar compounds extracted. The best conditions for the supercritical extraction, based on the content of polyphenols, epicatechin/catechin quantification, yield and operating cost, proved to be: 40% ethanol:methanol during 40min, under 40°C, and 100bar. The temperature had a significant effect on the total phenolic content.
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http://dx.doi.org/10.1016/j.foodchem.2016.06.028DOI Listing
December 2016

Pyrostegia venusta (Ker Gawl.) Miers Crude Extract and Fractions: Prevention of Dental Biofilm Formation and Immunomodulatory Capacity.

Pharmacogn Mag 2016 May 11;12(Suppl 2):S218-22. Epub 2016 May 11.

Department of Post-Graduation Program in Pharmaceutical Sciences, Institute of Health Sciences, Federal University of Pará, Augusto Corrêa Avenue, No. 01, University Campus of Guamá, Belém, Pará CEP 67150-110, Brazil; Pharmacy Course, Federal University of Rio de Janeiro, Campus Macaé, Av. Aluízio da Silva Gomes, 50 Granja dos Cavaleiros, CEP 27930-560, Macaé, Rio de Janeiro, CEP 87020-900, Brazil.

Background: Caries and periodontal diseases remain as important diseases in the Brazilian population. One important pathogen associated with this situation is Streptococcus mutans and other important factor is this pathogen's ability to adhere firmly to the tooth surface leading to dental biofilm formation and caries development.

Objectives: Determine the antibacterial and other biological activities of P. venusta related to its potential to be used in the treatment of caries and periodontal disease.

Methods: The growth inhibition by P. venusta of Streptococcus mutans, S. mitis, S. oralis and Candida albicans was determined using the broth microdilution method. In addition, the effect of the samples in adherence and reducing production of acids by S. mutans, and germ-tube formation of C. albicans was analysed. The Nitric Oxide (NO) production and cytotoxicity of P. venusta to peripheral blood mononuclear cells (PBMC) and RAW 264.7 Cell Line Murine Macrophage from Blood were assessed.

Results: The crude extract (CE) and ethyl-acetate (AF) and n-butanol (BF) fractions showed antibacterial activity. The ethyl-acetate (AF) fraction showed the highest inhibition percentage against the adherence of S. mutans and C. albicans cells without budding, beyond NO production inhibition. There was not any cytotoxicity in the murine macrophages RAW 264.7 cells.

Conclusion: Our results suggest that P. venusta presents potential to be used as a preliminary source of compounds that can provide helpful activity when used in prophylaxis or treatment of caries or periodontal disease.

Summary: Biological activities of Pyrostegia venusta and its potential for use in formulations for the prevention of oral diseases. Abbreviations used: NO: Nitric oxide, PBMC: Peripheral blood mononuclear cells, CE: Crude extract, AF: Ethyl-acetate fraction, BF: n-butanol fraction, HF: Hexane fraction, WF: Water fraction, MIC: Minimum inhibitory concentration, MBC: Minimum bactericidal concentration, ATCC: American Type Culture Collection, CFU: Colony-forming units, BHI: Brain heart infusion, RPMI: Roswell Park Memorial Institute, MOPS: 3-(N-morpholino)propanesulfonic acid, DMEM: Dulbecco's modified Eagle's médium, LPS: Lipopolysacharide, MTT: 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide, OD: Optical density, AC: Acteoside.
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http://dx.doi.org/10.4103/0973-1296.182150DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4883082PMC
May 2016

In vitro Cytotoxicity and Anti-herpes Simplex Virus Type 1 Activity of Hydroethanolic Extract, Fractions, and Isolated Compounds from Stem Bark of Schinus terebinthifolius Raddi.

Pharmacogn Mag 2016 Apr-Jun;12(46):160-4

Post-Graduate Program in Pharmaceutical Sciences, State University of Maringá, Maringá, Paraná, Brazil; Department of Health Basic Sciences, State University of Maringá, Av. Colombo 5790, BR-87020-900, Maringá, Paraná, Brazil.

Background: Herpes simplex virus type 1 (HSV-1) is associated with orofacial infections and is transmitted by direct contact with infected secretions. Several efforts have been expended in the search for drugs to the treatment for herpes. Schinus terebinthifolius is used in several illnesses and among them, for the topical treatment of skin wounds, especially wounds of mucous membranes, whether infected or not.

Objective: To evaluate the cytotoxicity and anti-HSV-1 activity of the crude hydroethanolic extract (CHE) from the stem bark of S. terebinthifolius, as well as its fractions and isolated compounds.

Materials And Methods: The CHE was subjected to bioguided fractionation. The anti-HSV-1 activity and the cytotoxicity of the CHE, its fractions, and isolated compounds were evaluated in vitro by SRB method. A preliminar investigation of the action of CHE in the virus-host interaction was conducted by the same assay.

Results: CHE presented flavan-3-ols and showed anti-HSV-1 activity, better than its fractions and isolated compounds. The class of substances found in CHE can bind to proteins to form unstable complexes and enveloped viruses, as HSV-1 may be vulnerable to this action. Our results suggest that the CHE interfered with virion envelope structures, masking viral receptors that are necessary for adsorption or entry into host cells.

Conclusion: The plant investigated exhibited potential for future development treatment against HSV-1, but further tests are necessary, especially to elucidate the mechanism of action of CHE, as well as preclinical and clinical studies to confirm its safety and efficacy.

Summary: Crude hydroethanolic extract (CHE) presents promising activity against herpes simplex virus type 1 (HSV 1), with selectivity index (SI) = 22.50CHE has flavan-3-ols in its composition, such as catechin and gallocatechinThe fractions and isolated compounds obtained from CHE by bioguided fractionation are less active than the CHE against HSV-1CHE interferes with viral entry process in the host cell and acts directly on the viral particle. Abbreviations used: HSV: Herpes simplex virus, CHE: Crude hydroethanolic extract, WF: Water fraction, AF: Ethyl-acetate fraction, MPLC: Medium-performance liquid chromatography, TLC: Thin-layer chromatography, NMR: Nuclear magnetic resonance, ESI-MS: Electrospray ionization mass spectrometry, SRB: Sulforhodamine B, CPE: Cytopathic effect, CC50: 50% cytotoxic concentration, EC50: 50% effective concentration, PBS: Phosphate-buffered saline.
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http://dx.doi.org/10.4103/0973-1296.177903DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4809172PMC
April 2016

Enhanced Cutaneous Wound Healing In Vivo by Standardized Crude Extract of Poincianella pluviosa.

PLoS One 2016 3;11(3):e0149223. Epub 2016 Mar 3.

Departamento de Farmácia, Palafito, Programa de Pós Graduação em Ciências Farmacêuticas, Universidade Estadual de Maringá, Maringá, Paraná, Brazil.

Wound healing is a complex process that involves several biological events, and a delay in this process may cause economic and social problems for the patient. The search continues for new alternative treatments to aid healing, including the use of herbal medicines. Members of the genus Caesalpinia are used in traditional medicine to treat wounds. The related species Poincianella pluviosa (DC.) L.P. Queiroz increases the cell viability of keratinocytes and fibroblasts and stimulates the proliferation of keratinocytes in vitro. The crude extract (CE) from bark of P. pluviosa was evaluated in the wound-healing process in vivo, to validate the traditional use and the in vitro activity. Standardized CE was incorporated into a gel and applied on cutaneous wounds (TCEG) and compared with the formulation without CE (Control) for 4, 7, 10, or 14 days of treatment. The effects of the CE on wound re-epithelialization; cell proliferation; permeation, using photoacoustic spectroscopy (PAS); and proteins, including vascular endothelial growth factor (VEGF), superoxide dismutase 2 (SOD-2) and cyclooxygenase 2 (COX-2) were evaluated. The TCEG stimulated the migration of keratinocytes at day 4 and proliferation on the following days, with a high concentration of cells in metaphase at 7 days. Type I collagen formed more rapidly in the TCEG. PAS showed that the CE had permeated through the skin. TCEG stimulated VEGF at day 4 and SOD-2 and COX-2 at day 7. The results suggest that the CE promoted the regulation of proteins and helped to accelerate the processes involved in healing, promoting early angiogenesis. This led to an increase in the re-epithelialized surface, with significant mitotic activity. Maturation of collagen fibers was also enhanced, which may affect the resistance of the extracellular matrix. PAS indicated a correlation between the rate of diffusion and biological events during the healing process. The CE from P. pluviosa appears promising as an aid in healing.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0149223PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4777426PMC
July 2016

Development and characterization of multiple emulsions for controlled release of Trichilia catigua (Catuaba) extract.

Pharm Dev Technol 2016 Dec 11;21(8):933-942. Epub 2015 Sep 11.

a Post-Graduate Program in Pharmaceutical Sciences, Department of Pharmacy.

Considering the antioxidant activity of the Trichilia catigua extract (TCE), the aim of the current study was to develop and characterize W/O/W multiple emulsions containing different vegetable oils as a platform to deliver a TCE. The extract displayed antioxidant activity (IC) of 4.59 µg/mL and total phenol content (TPC) of 50.84%. Formulations were prepared by the phase-inversion emulsification method and analyzed for morphological appearance, pH, conductivity, droplet size and distribution, content of active, rheological properties, in vitro release, skin permeation, and stability. Formulations prepared with canola oil were selected and displayed regular morphology, mean diameter 2.77 µm (without TCE), 3.07 µm with 0.5% and 3.23 µm with 1.0% TCE. Rheometry (flow) showed pseudoplastic behavior with minimal thixotropy for both systems. TCE could be released from emulsions containing 1.0% and 0.5% TCE in a controlled manner for 16 and 23 h, respectively. The emulsions allowed good retention of TCE in the skin (stratum corneum, epidermis, and dermis). In a 180-d assessment of accelerated chemical stability, TPC was more reduced for the emulsions at 40 °C; other parameters remained stable. Multiple emulsions containing TCE were developed, exhibited good characteristics, and may be considered for future investigations as anti-aging formulations for the skin.
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http://dx.doi.org/10.3109/10837450.2015.1081611DOI Listing
December 2016

Antifungal Properties of Crude Extracts, Fractions, and Purified Compounds from Bark of Curatella americana L. (Dilleniaceae) against Candida Species.

Evid Based Complement Alternat Med 2015 11;2015:673962. Epub 2015 Aug 11.

Laboratory of Microbiology Applied to Natural and Synthetic Products, State University of Maringá, Colombo Avenue 5790, 87020-900 Maringá, PR, Brazil.

The ethnomedicinal plant Curatella americana L. (Dilleniaceae) is a common shrub in the Brazilian cerrado, in which crude extract showed antifungal activity in a preliminary study. In this work, the antifungal and cytotoxic properties of the crude extract, fractions, and isolated compounds from C. americana were evaluated against the standard yeast strains Candida albicans, C. tropicalis, and C. parapsilosis, clinical isolates, and fluconazole-resistant strains. The combinatory effects between subfractions and isolated compounds and effects on cell morphology, virulence factors, and exogenous ergosterol were also evaluated. The MIC obtained against the Candida species including fluconazole-resistant strain ranged from 15.3 to 31.3 µg/mL for crude extract, 3.9 to 15.6 µg/mL for ethyl acetate fraction, and 7.8 to 31.3 µg/mL for subfractions. The isolated compounds identified as 4'-O-methyl-catechin, epicatechin-3-O-gallate, and 4'-O-methyl-catechin-3-O-gallate showed lower antifungal activity than the crude extract and fractions (MIC ranging from 31.3 to 125.0 µg/mL). The addition of exogenous ergosterol to yeast culture did not interfere in the antifungal activity of the extract and its fractions. Synergistic antifungal activity was observed between subfractions and isolated compounds. The effects on virulence factors and the different mechanisms of action compared to fluconazole and nystatin suggest that this ethnomedicinal plant may be an effective alternative treatment for candidiasis.
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http://dx.doi.org/10.1155/2015/673962DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4548135PMC
September 2015

Stryphnodendron adstringens: Clarifying Wound Healing in Streptozotocin-Induced Diabetic Rats.

Planta Med 2015 Aug 28;81(12-13):1090-6. Epub 2015 Jul 28.

Departamento de Farmácia, Laboratório de Biologia Farmacêutica, Palafito, Universidade Estadual de Maringá, Maringá, Brazil.

Diabetes mellitus is a serious public health problem in which a major complication is impaired wound healing. Among the strategies developed to foster tissue repair is the use of medicinal plants. The bark of Stryphnodendron adstringens, which is popularly used as an aid in wound healing, has a documented effect on wound repair in normal rats. This study evaluated the healing action of the crude extract of S. adstringens in diabetic rats, and its chemical content. Compounds present in the crude extract were characterized by mass spectrometry. In diabetic rats (streptozotocin 35 mg/kg), two wounds made in the skin were treated daily for 4, 7, 10, and 14 days with gel containing 1 % crude extract or with base gel. Histological analyses involved the measurement of the length and thickness of the re-epithelialized surface, quantification of the number of cells in mitosis, and types I and III collagen fibers. Also, cutaneous permeation by photoacoustic spectroscopy, and the expression of cyclooxygenase-2 and vascular endothelial growth factor by Western blot were assessed. The crude extract fingerprint showed masses indicating proanthocyanidins. The crude extract mainly stimulated cell migration and proliferation of keratinocytes at the beginning of the treatment in addition to stimulating the replacement of type III collagen fibers by type I collagen fibers at 10 and 14 days. The photoacoustic spectroscopy technique showed that the gel containing 1 % of crude extract permeated through the skin to the dermis, where the crude extract was found. Vascular endothelial growth factor was stimulated after 7 days of treatment with the crude extract and cyclooxygenase-2 at 4, 7, and 10 days. The crude extract of S. adstringens acted in tissue repair in wounds in diabetic rats by stimulating the production of collagen fibers at the wound site. The crude extract favored the formation of a more organized extracellular matrix and filled the entire extent of the wound, and also fostered the upregulation of cyclooxygenase-2 and vascular endothelial growth factor, which are essential to this process. These crude extract actions in diabetic wounds are probably due to the presence of proanthocyanidins.
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http://dx.doi.org/10.1055/s-0035-1546209DOI Listing
August 2015

Proanthocyanidins polymeric tannin from Stryphnodendron adstringens are active against Candida albicans biofilms.

BMC Complement Altern Med 2015 Mar 19;15:68. Epub 2015 Mar 19.

Laboratório de Quimioterapia Antifúngica, Departamento de Microbiologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, Av. Prof. Lineu Prestes 1374, 05508-900, SP, Brazil.

Background: Biofilm formation is important in Candida albicans pathogenesis and constitutes a mechanism of antifungal resistance. Thus, we evaluated the effect of proanthocyanidins polymer-rich fractions from Stryphnodendron adstringens (fraction F2 and subfraction F2.4) against C. albicans biofilms.

Methods: Firstly, the antifungal activity of F2 and F2.4 against planktonic cells of Candida albicans (ATCC 10231) was determined using broth microdilution method. Anti-biofilm effect of F2 and F2.4 was evaluated during biofilm formation or on mature biofilm of C. albicans and compared with standard antifungals amphotericin B and fluconazole. Metabolic activity of sessile and dispersion cells from biofilms after antifungal treatments were measured using a tetrazolium reduction assay and the biofilm total biomass was quantified by crystal violet-based assay. Morphological alterations after treatments were observed using scanning electron microscopy.

Results: The anti-biofilm effect of F2 and F2.4 were comparable to standard antifungals (amphotericin B and fluconazole). F2 and F2.4 treatments reduced biofilm metabolic activity (in sessile and in dispersion cells) during biofilm formation, and in mature biofilms, unlike fluconazole, which only prevents the biofilm formation. Treatments with F2, F2.4 or fluconazole reduced biofilm biomass during biofilm formation, but not in mature biofilm. Amphotericin B presented higher inhibitory effect on biofilm formation and on mature biofilm of C. albicans. F2 and F2.4 treatments led to the appearance of dumbbell-shaped blastoconidia and of blastoconidia clusters in biofilms.

Conclusion: Proanthocyanidins polymer-rich fractions from S. adstringens successfully inhibited C. albicans planktonic growth and biofilm development, and they represent a potential new agent for the treatment of biofilm-associated candidiasis.
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http://dx.doi.org/10.1186/s12906-015-0597-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4369060PMC
March 2015

The exposure to Trichilia catigua (catuaba) crude extract impairs fertility of adult female rats but does not cause reproductive damage to male offspring.

J Ethnopharmacol 2015 May 16;166:86-91. Epub 2015 Mar 16.

Department of Physiological Sciences, State University of Londrina, BR-86051-980 Londrina, Paraná, Brazil. Electronic address:

Ethnopharmacological Relevance: Trichilia catigua is broadly used in folk medicine due to its mental and physical tonic activities and stimulant effects. In animal models, its antidepressant-like effects have been associated with the dopaminergic (DA) system modulation, which has an important role on maternal behavior and male offspring reproductive development.

Aim Of The Study: Since little is known about the adverse effects of the exposure to T. catigua crude extract (CAT) in rats, specially regarding maternal homeostasis and offspring development, the aim of the present study was to evaluate whether CAT exposure may influence maternal toxicity parameters and behavior or disrupt male offspring physical and reproductive development.

Material And Methods: Dams were treated daily (by gavage) with 400mg/kg of CAT or vehicle (control=CTR) throughout pregnancy and lactation. Fertility and maternal behavior tests were conducted in dams. Male offspring reproductive and behavioral parameters were analyzed.

Results: Dams exposed to CAT showed increased pre- and post-implantation losses rates when compared to CTR group. No significant changes regarding maternal behavior or male offspring parameters were observed.

Conclusion: In conclusion, maternal exposure to CAT interfered with implantation during the initial phases of pregnancy but did not induce changes on maternal behavior or male offspring reproductive and behavioral parameters.
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http://dx.doi.org/10.1016/j.jep.2015.03.018DOI Listing
May 2015

In vitro effect of Paullinia cupana (guaraná) on hydrophobicity, biofilm formation, and adhesion of Candida albicans' to polystyrene, composites, and buccal epithelial cells.

Arch Oral Biol 2015 Mar 9;60(3):471-8. Epub 2014 Jun 9.

Department of Dentistry, State University of Maringá, Av. Mandacaru 1550, CEP 87080-000, Maringá, PR, Brazil. Electronic address:

Objective: In vitro evaluation of the effect of guaraná (GUAR) on cell surface hydrophobicity (CSH), on biofilm formation, and on adhesion of C. albicans to polystyrene, to composite resins, and to buccal epithelial cells (BEC).

Materials And Methods: Lyophilised aqueous extract of GUAR was tested on C. albicans ATCC (90028). The effect of GUAR was evaluated by examining the CSH of C. albicans, as determined by microbial adhesion to hydrocarbons test, by assessing biofilm production and through adhesion assays (microplates of polystyrene, BEC and composites). One nanoparticle (Z350(®)) and two microhybrid (LLis(®), Opallis(®)) composites were tested. Scanning electron microscopy (SEM) was used to analyse adhesion of C. albicans composites. Assays were performed in triplicate and the results analysed by Chi-square test, Kruskal-Wallis test and Dunn's Multiple Comparison post hoc test at 5% significance level.

Results: GUAR did not inhibit growth of C. albicans at any concentration, but it reduced adhesion to polystyrene surface (p < 0.001). Exposure to GUAR did not change CSH and biofilm formation, but it increased adhesion of C. albicans to the nanoparticle composite (p = 0.042) and reduced its adhesion to BEC (p < 0.001). SEM confirmed an aggregatory pattern of adhesion of C. albicans to composites.

Conclusion: GUAR increased the adhesion of C. albicans to the surface of the nanoparticle composite. However, it reduced the adhesion of C. albicans to BEC and to polystyrene, which reveals its potential use in prevention of oral diseases.
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http://dx.doi.org/10.1016/j.archoralbio.2014.05.026DOI Listing
March 2015

Hydrolyzable tannins from hydroalcoholic extract from Poincianella pluviosa stem bark and its wound-healing properties: phytochemical investigations and influence on in vitro cell physiology of human keratinocytes and dermal fibroblasts.

Fitoterapia 2014 Dec 16;99:252-60. Epub 2014 Oct 16.

University of Münster, Institute of Pharmaceutical Biology and Phytochemistry, Corrensstr. 48, D-48149 Münster, Germany. Electronic address:

Extracts from Poincianella pluviosa stem bark are used in traditional medicine of South America for its wound healing properties. For validation of this traditional use and for rationalizing a potential pharmaceutical development towards standardized preparations bioassay-guided fractionation of EtOH-water (1:1v/v) extract (crude extract, CE) of P. pluviosa bark was performed. HaCaT keratinocytes cell line and human primary dermal fibroblasts (pNHDF) were used as in vitro systems. Significant stimulation of mitochondrial activity was found for CE on both cell types, which caused a strong increase of cell proliferation of keratinocytes. Fractionation of CE over Sephadex LH20 revealed two inactive fractions (FA and FB) and an active fraction FC, which was further fractionated by MPLC into 4 subfractions. Subfraction FC1 increased mitochondrial activity and proliferation of keratinocytes and dermal fibroblasts in a dose dependent manner (10 to 100 μg/mL) and did not show necrotic cytotoxicity on keratinocytes (LDH release assay). FC1 was investigated by ESI-MS/MS and solid-state (13)C NMR which confirmed the presence of various polyphenols and hydrolyzable tannins. MS studies suggest the presence of pyrogallol (1), gallic acid (2), gallic acid methyl ester (3), ellagic acid (4), corilagin (5), 1,4,6-tri-O-galloyl-glucose (6), tellimagrandin I (7), 1,2,3,6-tetra-O-galloyl-glucose (8), mallotinic acid (9), tellimagrandin II (10), 1,2,3,4,6-penta-O-galloyl-glucose (11), geraniin (12), and mallotusinic acid (13).
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http://dx.doi.org/10.1016/j.fitote.2014.10.007DOI Listing
December 2014

Direct proof by (13)C-nuclear magnetic resonance of semi-purified extract and isolation of ent-Catechin from leaves of Eucalyptus cinerea.

Pharmacogn Mag 2014 Jul;10(39):191-4

Postgraduate Program in Pharmaceutical Sciences, Universidade Federal do Paraná, Av. Lothário Meissner, 632, BR-80210-170, Curitiba, PR, Brazil.

Background: Eucalyptus cinerea F. Muell. ex Benth. is native to Australia and acclimatized to Southern Brazil. Its aromatic leaves are used for ornamental purposes and have great potential for essential oil production, although reports of its use in folk medicine are few.

Objective: This study evaluated the composition of E. cinerea leaves using the solid state (13)C-nuclear magnetic resonance (NMR) and isolation of the compound from the semipurified extract (SE).

Materials And Methods: The SE of E. cinerea leaves was evaluated in the solid state by (13)C-NMR spectrum, and the SE was chromatographed on a Sephadex LH-20 column, followed by high-speed counter-current chromatography to isolate the compound. The SE was analyzed by (13)C-NMR and matrix-assisted laser desorption/ionization-time-of-flight spectra.

Results: Flavan-3-ol units were present, suggesting the presence of proanthocyanidins as well as a gallic acid unit. The uncommon ent-catechin was isolated.

Conclusion: The presence of ent-catechin is reported for the first time in this genus and species.
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http://dx.doi.org/10.4103/0973-1296.137355DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4159908PMC
July 2014
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