RESUMO
To investigate the anti-Saprolegnia activities of chalconic compounds, nine dialkoxychalcones 2â»10, along with their key building block 2',4'-dihydroxychalcone 1, were evaluated for their potential oomycide activities against Saprolegnia australis strains. The synthesis afforded a series of O-alkylated derivatives with typical chalcone skeletons. Compounds 4â»10 were reported for the first time. Interestingly, analogue 8 with the new scaffold demonstrated remarkable in vitro growth-inhibitory activities against Saprolegnia strains, displaying greater anti-oomycete potency than the standard drugs used in the assay, namely fluconazole and bronopol. In contrast, a dramatic loss of activity was observed for O-alkylated derivatives 2, 3, 6, and 7. These findings have highlighted the therapeutic potential of the natural compound 1 scaffold to be exploitable as a drug lead with specific activity against various Saprolegnia strains.
Assuntos
Antifúngicos/farmacologia , Chalconas/farmacologia , Peixes/microbiologia , Saprolegnia/efeitos dos fármacos , Animais , Antifúngicos/química , Chalconas/química , Fluconazol/farmacologia , Ligação de Hidrogênio , Testes de Sensibilidade Microbiana , Propilenoglicóis/farmacologia , Relação Quantitativa Estrutura-Atividade , Reprodutibilidade dos Testes , Análise Espectral/métodos , Relação Estrutura-AtividadeRESUMO
Jeffamines® are a family of polymers containing primary amine groups attached to the extremities of polyether backbone which can be used as biomaterials. They have been used in combination with polyethylenimine (PEI) to improve biocompatibility in drug and gene delivery systems. Despite these facts, very few studies have been done on cytotoxicity and genotoxicity of pure Jeffamines® or compared with PEI. The present study aimed to evaluate and compare the cytotoxic and genotoxic effects of Jeffamines® and PEI in CHO-K1 cells. Specifically, polypropylene oxide 2000 (PPO 2000, Jeffamine® D series), polyethylene oxide 1900 (PEO 1900, Jeffamine® ED series), branched 25 kDa PEI, and linear 20 kDa PEI were evaluated at different concentrations. Cell viability and proliferation were assessed by 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) and 5-bromo-2'-deoxyuridine (BrdU) assays, respectively. Genotoxicity was evaluated using single cell gel electrophoresis assay and the cytokinesis-blocked micronucleus assay. PPO 2000 was the most cytotoxic Jeffamine® , whereas PEO 1900 did not caused significant cell death at any tested concentration. Branched PEI was more cytotoxic than linear PEI (LPEI) and both were more cytotoxic than Jeffamines® . Only PPO 2000 induced DNA damage when evaluated in comet assay probably due to its cytotoxicity. PPO 2000, PEO 1900, and PEI did not increase the frequency of micronuclei when tested at sub-cytotoxic concentrations. This work provides new insights about biocompatibility of Jeffamines® and PEI and suggests the genotoxicological safety for further investigations of PEO 1900 in drug and gene delivery systems. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 742-750, 2018.
Assuntos
Proliferação de Células/efeitos dos fármacos , Dano ao DNA , Animais , Células CHO , Morte Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Cricetulus , Avaliação Pré-Clínica de Medicamentos , Polietilenoimina/efeitos adversos , Polietilenoimina/farmacologia , Polímeros/efeitos adversos , Polímeros/farmacologia , Propilenoglicóis/efeitos adversos , Propilenoglicóis/farmacologiaRESUMO
A series of novel oxyalkylchalcones substituted with alkyl groups were designed and synthesized, and the antioomycete activity of the series was evaluated in vitro against Saprolegnia strains. All tested O-alkylchalcones were synthesized by means of nucleophilic substitution from the natural compound 2',4'-dihydroxychalcone (1) and the respective alkyl bromide. The natural chalcone (1) and 10 synthetic oxyalkylchalcones (2-11) were tested against Saprolegnia parasitica and Saprolegnia australis. Among synthetic analogs, 2-hydroxy,4-farnesyloxychalcone (11) showed the most potent activity against Saprolegnia sp., with MIC and MOC values of 125 µg/mL (similar to bronopol at 150 µg/mL) and 175 µg/mL, respectively; however, 2',4'-dihydroxychalcone (1) was the strongest and most active molecule, with MIC and MOC values of 6.25 µg/mL and 12.5 µg/mL.
Assuntos
Chalconas/farmacologia , Saprolegnia/efeitos dos fármacos , Animais , Antifúngicos/farmacologia , Testes de Sensibilidade Microbiana , Propilenoglicóis/farmacologiaRESUMO
The objective of the present study was to evaluate the antimicrobial activity of sodium hypochlorite (NaOCl) associated with a surfactant. Seventy single-rooted extracted human teeth were inoculated with Enterococcus faecalis, and incubated for 21 days (37 °C). The groups were distributed according to the irrigation solution used during root canal preparation: 5%, 2.5% and 1% NaOCl; 5%, 2.5% and 1% Hypoclean®, a solution containing a surfactant (cetrimide) associated with NaOCl. Three microbiological samples were collected from each tooth: S1 - before instrumentation; S2 - immediately after instrumentation; and S3 - after a seven-day period. Data were submitted to ANOVA and Tukey test with 5% significance level. The results showed that immediately after root canal preparation (S2), E. faecalis was eliminated in all the experimental groups. However, after 7 days (S3), only the groups in which Hypoclean was used, remained contamination-free, including Hypoclean associated with 1% NaOCl, while the root canals irrigated with 1% NaOCl only, presented the highest percentage of bacterial growth. In conclusion, the addition of surfactant increased the antimicrobial activity of 1% NaOCl to levels similar to 5% NaOCl.
Assuntos
Antibacterianos/farmacologia , Irrigantes do Canal Radicular/farmacologia , Hipoclorito de Sódio/farmacologia , Tensoativos/farmacologia , Antibacterianos/administração & dosagem , Cetrimônio , Compostos de Cetrimônio/farmacologia , Cavidade Pulpar/microbiologia , Enterococcus faecalis/efeitos dos fármacos , Enterococcus faecalis/crescimento & desenvolvimento , Humanos , Teste de Materiais , Polímeros/farmacologia , Propilenoglicóis/farmacologia , Distribuição Aleatória , Irrigantes do Canal Radicular/administração & dosagem , Preparo de Canal Radicular/métodos , Hipoclorito de Sódio/administração & dosagem , Tensoativos/administração & dosagem , Fatores de TempoRESUMO
Although much is described about the molecules involved in neutrophil migration from circulation into tissues, less is known about the molecular mechanisms that regulate neutrophil entry into lymph nodes (LNs) draining a local inflammatory site. In this study, we investigated neutrophil migration toward LNs in a context of inflammation induced by immunization of BALB/c mice with OVA emulsified in CFA. We demonstrated that neutrophils can enter LNs of OVA/CFA-immunized mice not only via lymphatic vessels but also from blood, across high endothelial venules. By adoptive transfer experiments, we showed that this influx was dependent on an inflammatory-state condition and previous neutrophil stimulation with OVA/anti-OVA immune complexes. Importantly, we have demonstrated that, in the migratory pattern to LNs, neutrophils used L-selectin and P-selectin glycoprotein ligand-1, macrophage-1 Ag and LFA-1 integrins, and CXCR4 to get access across high endothelial venules, whereas macrophage-1 Ag, LFA-1, and CXCR4 were involved in their trafficking through afferent lymphatics. Strikingly, we found that stimulation with immune complexes significantly upregulated the expression of sphingosine-1-phosphate receptor 4 on neutrophils, and that treatment with the sphingosine-1-phosphate agonist FTY720 altered neutrophil LN-homing ability. These findings summarized in this article disclose the molecular pattern that controls neutrophil recruitment to LNs.
Assuntos
Complexo Antígeno-Anticorpo/imunologia , Doenças do Sistema Imunitário/imunologia , Transtornos Leucocíticos/imunologia , Linfonodos/imunologia , Neutrófilos/imunologia , Transferência Adotiva , Animais , Movimento Celular/imunologia , Feminino , Cloridrato de Fingolimode , Imunossupressores/farmacologia , Inflamação/imunologia , Selectina L/imunologia , Linfonodos/citologia , Vasos Linfáticos/imunologia , Antígeno-1 Associado à Função Linfocitária/imunologia , Lisofosfolipídeos/agonistas , Antígeno de Macrófago 1/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Neutrófilos/transplante , Selectina-P/imunologia , Propilenoglicóis/farmacologia , Receptores CXCR4/imunologia , Receptores de Lisoesfingolipídeo/metabolismo , Esfingosina/agonistas , Esfingosina/análogos & derivados , Esfingosina/farmacologiaRESUMO
BACKGROUND: Specific protocols for milt cryopreservation have been established for some freshwater fish species. However, cryopreservation reduces sperm quality, giving unsatisfactory results in reproduction. OBJECTIVE: The objective of this work was to evaluate the effect of different cryoprotectants on the quality of Prochilodus lineatus, Brycon orbignyanus and Piaractus mesopotamicus milt after cryopreservation. METHODS: The milt was diluted in different cryoprotectant solutions containing 10% methanol, dimethyl sulfoxide, glycerol, propylene glycol or ethylene glycol combined with the Beltsville Thawing Solution extender (5%), then placed in the vapour of a liquid nitrogen (LN) storage tank for 24 h, after which they were immersed in LN. After rewarming, the rate (%) and duration (s) of milt motility and abnormal morphology were evaluated. RESULTS: All of cryoprotectant solutions tested used maintained the viability of P. lineatus and P. mesopotamicus milt. However, in P. lineatus, glycerol ensured a lower percentage of abnormal morphology. In case of P. mesopotamicus, all of the cryoprotectant solutions tested may be used in the cryopreservation process, with the exception of those containing glycerol. CONCLUSION: For B. orbignyanus, cryoprotectant solutions containing methanol and ethylene glycol are recommended for use in the cryopreservation process, although they reduced the quality of sperm post-rewarming.
Assuntos
Criopreservação , Crioprotetores/farmacologia , Peixes/fisiologia , Mitocôndrias/efeitos dos fármacos , Preservação do Sêmen/métodos , Espermatozoides/efeitos dos fármacos , Animais , Brasil , Sobrevivência Celular/efeitos dos fármacos , Conservação dos Recursos Naturais , Dimetil Sulfóxido/farmacologia , Etilenoglicol/farmacologia , Glicerol/farmacologia , Masculino , Metanol/farmacologia , Mitocôndrias/metabolismo , Polímeros/farmacologia , Propilenoglicóis/farmacologia , Contagem de Espermatozoides , Motilidade dos Espermatozoides/efeitos dos fármacos , Espermatozoides/citologia , Espermatozoides/metabolismoRESUMO
INTRODUCTION: Ischemia-reperfusion (IR) kidney damage is an important factor for allograft survival in kidney transplantation. Recently it has been shown that immune factors from donor-derived cells are important in IR injury. The aim of this article was to evaluate the impact of short-term immunosuppressive treatment of the donor over a time frame relevant to cadaveric transplantation on IR damage to the rat kidney. METHODS: Male Sprague-Dawley rats served as donors and recipients. Three experimental groups were evaluated according to the donor treatment (n = 6); control (no treatment); sirolimus (1 mg/kg orally) or FTY720 (1 mg/kg intravenously) at 6 or 1 hours prior to left nephrectomy. Kidneys were flushed with cold Euro-Collins solution and after 2 hours transplanted using microsurgical techniques concomittant with a left nephrectomy. After 48 hours (day 0), we removed the right kidney. Serum creatinine (SCr) was determined daily thereafter as well as differential leukocyte counts prior to donor nephrectomy and sirolimus plasma levels thereafter. RESULTS: No difference was observed in SCr on day 1: control (3.97 ± 0.73 mg/dL), sirolimus (4.02 ± 1.44 mg/dL) and FTY 720 (3.27 ± 1.79 mg/dL; P = NS), or thereafter. Mortality was 50% in each group. Animals receiving FTY 720 showed a significant reduction in lymphocyte count (8.0 ± 3.1 to 1.1 ± 0.3 (P < .01). Sirolimus levels were 9.3 ± 1.5 ng/mL. CONCLUSION: We concluded that immunosuppressive treament of the donor within a time frame relevant to cadaveric kidney transplantation did not offer a benefit in terms of preventing IR injury.
Assuntos
Imunossupressores/uso terapêutico , Traumatismo por Reperfusão/prevenção & controle , Animais , Cadáver , Creatinina/sangue , Cloridrato de Fingolimode , Sobrevivência de Enxerto , Soluções Hipertônicas/química , Transplante de Rim , Masculino , Propilenoglicóis/farmacologia , Ratos , Ratos Sprague-Dawley , Sirolimo/farmacologia , Esfingosina/análogos & derivados , Esfingosina/farmacologia , Doadores de TecidosRESUMO
Ischemia and reperfusion injury (IR) is an antigen independent inflammatory process that causes tissue damage. After IR, kidneys up-regulate leukocyte adhesion molecules and toll-like receptors (TLRs). Moreover, injured kidneys can also secrete factors (i.e. heat shock protein) which bind to TLRs and trigger intracellular events culminating with the increase in the gene expression of inflammatory cytokines. FTY720 is an immunomodulatory compound and protects at least in part kidneys submitted to IR. The mechanisms associated with FTY720's beneficial effects on kidneys after IR remain elusive. We investigated whether FTY720 administration in mice submitted to kidney IR is associated with modulation of TLR2 and TLR4 expression. C57BL/6 mice submitted to 30min of renal pedicles clamp were evaluated for serum parameters (creatinine, urea and nitric oxide), kidney histology, spleen and kidney infiltrating cells expression of TLR2 and TLR4, resident kidney cells expression of TLR2 and TLR4 and IL-6 protein expression in kidney. FTY720-treated mice presented decrease in serum creatinine, urea and nitric oxide, diminished expression of TLR2 and TLR4 both in spleen and kidney infiltrating cells, and reduced kidney IL-6 protein expression in comparison with IR non-treated mice. However, acute tubular necrosis was present both in IR non-treated and IR+FTY720-treated groups. Also, FTY720 did not prevent TLR2 and TLR4 expression in kidney resident cells. In conclusion, FTY720 can promote kidney function recovery after IR by reducing the inflammatory process. Further studies are needed in order to establish whether TLR2 and TLR4 down regulation should be therapeutically addressed as protective targets of renal function and structure after IR.
Assuntos
Propilenoglicóis/farmacologia , Traumatismo por Reperfusão/genética , Traumatismo por Reperfusão/metabolismo , Esfingosina/análogos & derivados , Receptor 2 Toll-Like/biossíntese , Receptor 2 Toll-Like/genética , Receptor 4 Toll-Like/biossíntese , Receptor 4 Toll-Like/genética , Animais , Creatinina/sangue , Cloridrato de Fingolimode , Interleucina-6/antagonistas & inibidores , Interleucina-6/genética , Interleucina-6/metabolismo , Rim/efeitos dos fármacos , Rim/metabolismo , Rim/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Óxido Nítrico/sangue , Traumatismo por Reperfusão/patologia , Traumatismo por Reperfusão/prevenção & controle , Esfingosina/farmacologia , Baço/efeitos dos fármacos , Baço/metabolismo , Receptor 2 Toll-Like/antagonistas & inibidores , Receptor 2 Toll-Like/metabolismo , Receptor 4 Toll-Like/antagonistas & inibidores , Receptor 4 Toll-Like/metabolismo , Ureia/sangueRESUMO
Uveitis is an intraocular inflammatory disease causing a significant visual impairment. The disease can be idiopathic, associated with infectious and systemic disorders or arisen from an unknown cause. Over the last 20years the model of EAU in mice has contributed significantly for the establishment of parameters for diagnostic evaluations and therapies for posterior uveitis in human. Many studies using recently discovered molecules which present proinflammatory and anti-inflammatory effects have been described. Moreover, new approaches of research provided by the increasing body of knowledge on components of immune responses such as cytokines, T-cell subpopulations and their associated functions have contributed for the further understanding of uveitis and possible treatment.
Assuntos
Doenças Autoimunes/imunologia , Imunomodulação , Mediadores da Inflamação , Uveíte/imunologia , Animais , Modelos Animais de Doenças , Cloridrato de Fingolimode , Humanos , Fatores Imunológicos/farmacologia , Imunomodulação/efeitos dos fármacos , Imunomodulação/imunologia , Mediadores da Inflamação/imunologia , Propilenoglicóis/farmacologia , Esfingosina/análogos & derivados , Esfingosina/farmacologiaRESUMO
This study evaluated the sealing ability of gray MTA-Angelus mixed with propyleneglycol in furcal perforations using a bacterial leakage test. Furcal perforations were created in 30 human mandibular molars using a size 3 round bur The samples were divided randomly into 2 experimental groups (n=10) according to the mixing agent. In G1, the MTA powder was mixed with propyleneglycol, while distilled water was used in G2. A 3:1 powder-liquid ratio was used for both groups. The MTA was placed in the perforation with an MTA carrier and condensed with hand pluggers. Non-repaired (n=5) and totally sealed (n=5) perforations served as positive and negative controls, respectively. Bacterial leakage was assessed daily for 30 days in a double-chamber apparatus with Enterococcus faecalis. Data were analyzed using Fisher exact test (p < 0.05) for three leakage periods: 1st to 10th day (P1); 11th to 20th day (P2); and 21st to 30th day (P3). The positive control presented leakage in all specimens within the first 24 hours, while no leakage was observed in the negative control during the experimental period. Leakage was observed in five (50%) of the 10 samples of the propyleneglycol group (G1) and seven (70%) of the distilled water group (G2) by the 20th day, without significant difference between the groups in periods P1 and P2 (p = 0.137). The leakage was significantly lower for G1 than G2 in period P3 (50% versus 100%, respectively, p = 0.016). In this single aerobic bacterial leakage method, the use of propyleneglycol as a vehicle for gray MTA-Angelus increased its sealing ability in furcal perforations at the end of the 30-day experimental period.