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1.
Ann Pediatr Cardiol ; 14(1): 99-104, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33679072

RESUMO

A 10-years-old boy presented with a history of effort intolerance and palpitations for 4 months. His electrocardiogram showed wide complex tachycardia suggestive of fascicular ventricular tachycardia (VT). The echocardiogram showed moderate-to-severe left ventricular systolic dysfunction without any structural lesion. The tachycardia was unresponsive to adenosine and direct current cardioversion. It responded to oral verapamil. The electrophysiology study confirmed the tachycardia as left posterior fascicular VT. The tachycardia was successfully ablated guided by Purkinje potential on three-dimensional mappings. He showed improvement in ventricular functions before discharge. He is doing well on short-term follow-up.

2.
Carbohydr Polym ; 250: 116950, 2020 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-33049856

RESUMO

Purified gum odina (PGO) from Odina wodier Roxb. was characterized by rheology, AFM, Raman and CD spectroscopy, in vitro antioxidant activity against hydroxyl radical and superoxide radical, and total antioxidant capacity. The PGO dispersions exhibited pseudoplastic behaviour. The viscosity of dispersion increased with increasing PGO concentration, but decreased with increasing temperature and added salt concentration. The loss modulus was higher than storage modulus indicating prevalently viscous characteristics. AFM analysis showed irregular spherical lumps due to inter- and intramolecular interactions. The Raman spectrum of PGO was similar to that of gum arabic. Circular dichroism revealed partial adoption of polyproline II type conformation, suggesting a less compact structure. The PGO was found to scavenge hydroxyl radical (IC50 517.68 ± 3.60 µg/mL) and superoxide radical (IC50 586.21 ± 3.41 µg/mL), and possess total antioxidant capacity (9.64 ± 0.23 mg gallic acid equivalent/g). Overall, this work would exploit PGO as a new hydrocolloid source in the food and pharmaceutical industries.


Assuntos
Anacardiaceae/química , Antioxidantes/metabolismo , Exsudatos e Transudatos/química , Galactanos/metabolismo , Gomas Vegetais/química , Reologia , Dicroísmo Circular/métodos , Galactanos/química , Galactanos/isolamento & purificação , Microscopia de Força Atômica/métodos , Análise Espectral Raman/métodos , Viscosidade
3.
Nat Prod Res ; 34(4): 599-603, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30417669

RESUMO

The methanolic extract of Cycas revoluta cone (MECR) was analyzed by GC-MS and UHPLC for metabolite profiling and was evaluated for anti-colon cancer property by using in vitro assays like Cell Viability Assay, Colony Formation Assay, ROS Determination, Flowcytometry, DAPI staining assay, Tunel assay. GC-MS and HPLC analysis confirmed the presence of different phytochemicals in the extract of Cycas revoluta cone. In-vitro studies showed MECR extract showed significant anti-colon cancer activity by reducing proliferation and inducing apoptosis in colon cancer cell (HCT-8) line, but no such activity was seen in normal colon cell (CCD-18Co) line. The investigation confirms that MECR may be a promising candidate in colon cancer protection.


Assuntos
Neoplasias do Colo/prevenção & controle , Cycas/metabolismo , Extratos Vegetais/química , Apoptose/efeitos dos fármacos , Linhagem Celular , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Cromatografia Líquida de Alta Pressão , Neoplasias do Colo/tratamento farmacológico , Cromatografia Gasosa-Espectrometria de Massas , Humanos , Extratos Vegetais/farmacologia
4.
J Am Coll Nutr ; 38(5): 415-423, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30696389

RESUMO

Objective: Tea (Camellia sinensis Linn.; family: Theaceae) is popular as a stimulant beverage across the globe and is also utilized as a functional antioxidant in alternative medicine. This study has evaluated the impact of seasonal variation on phyto-constituents of tea. Method: The antiproliferative potential of methanolic extracts of tea leaves collected in the rainy season (MECR) was compared with the extract of tea leaves collected in the autumn season (MECA) of the same mother plant. Evaluation of in vivo antitumor activity was carried out in adult female Swiss albino mice groups inoculated with Ehrlich ascites carcinoma (EAC) cells. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to compare efficacy of MECR with that of MECA in the EAC cell line. Both qualitative and quantitative tests for phytochemical constituents present in MECA and MECR were performed. Antitumor efficacy of both the extracts was determined by evaluating different tumor markers showing dose-dependent cytotoxicity. Results: Statistically significant reduction in EAC-induced tumor was observed in MECR treated mice compared to MECA treated ones. Cell decimation was significantly higher with MECR treatment, where restoration of different parameters including tissue structures returned to normal. Moreover, gas chromatography-mass spectrometry (GC-MS) study revealed the presence of cyclobarbital and benzazulene derivative in MECR, which is thought to be a novel source of these chemicals. Conclusions: To our knowledge, there is no report that has attempted to reveal nutritional changes in terms of efficacy and variation in anticancer constituents in tea leaves, plucked in two seasons. This study revealed a novel source of barbital and benzazulene derivative. The unique presence of cyclobarbital and benzazulene, as revealed from GC-MS data, in methanolic extract of tea leaves collected during the rainy season (MECR) may have contributed to its enhanced in vitro (adopting MTT assay) and in vivo (on EAC-infected Swiss albino mice) cytotoxicity vis-à-vis antiproliferative properties compared to methanolic extract of tea leaves collected during the autumn season (MECA). The nature of plucking leaves in the two selected seasons is different.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Metanol/farmacologia , Extratos Vegetais/farmacologia , Folhas de Planta/química , Chá/química , Animais , Antineoplásicos Fitogênicos/química , Barbital/farmacologia , Camellia sinensis , Carcinoma de Ehrlich , Linhagem Celular Tumoral , Feminino , Metanol/química , Camundongos , Extratos Vegetais/química , Estações do Ano
5.
Biomed Pharmacother ; 100: 282-295, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29448205

RESUMO

In this study, we investigated the in vivo antiproliferative activity of 6,7-dimethoxy-1,2,3,4-tetrahydro-isoquinoline-3-carboxylic acid (M1) in dimethylhydrazine (DMH) induced colorectal carcinoma (CRC) using albino Wistar rats. M1 was administered to DMH induced CRC rats at 10 and 25 mg/kg doses for 15 days. Various physiological, oxidative parameters, histopathology, ELISA, gene and protein expression studies were conducted to evaluate the anti-CRC potential of M1. The histopathology and biochemical tests indicated the protective action of M1 in DMH-induced colon cancer. ELISA confirms that M1 reduced the increased concentration of IL-6 more prominently than those of IL-2 and COX-2. Gene expression analysis revealed that M1 attenuated the increased mRNA over-expression of IL-6, JAK2 and STAT3. The result obtained from quantitative western blot analysis demonstrated that the CRC condition was produced by the IL-6 induced activation/phosphorylation of JAK2 and STAT3 and further down-regulated with M1 treatment. This evidence was supported well with the application of data-based mathematical modeling. Applying the fitted model, we predicted the quantitative behavior of STAT3 populations not accessible to experimental measurement. Later, 1H NMR based serum metabolic profiling was carried out using rat sera to investigate the impact of M1 on CRC-induced metabolic alterations. M1 showed its ability to restore the perturbed metabolites in CRC condition. Altogether, our study provided the first time evidence that M1 exhibits anti-CRC potential through the blockade of IL-6/JAK2/STAT3 oncogenic signaling.


Assuntos
Antineoplásicos Fitogênicos/uso terapêutico , Neoplasias Colorretais/tratamento farmacológico , Interleucina-6/antagonistas & inibidores , Tetra-Hidroisoquinolinas/uso terapêutico , Animais , Antineoplásicos Fitogênicos/administração & dosagem , Antineoplásicos Fitogênicos/isolamento & purificação , Proliferação de Células/efeitos dos fármacos , Colo/efeitos dos fármacos , Colo/imunologia , Colo/patologia , Neoplasias Colorretais/imunologia , Neoplasias Colorretais/patologia , Dimetilidrazinas/farmacologia , Relação Dose-Resposta a Droga , Expressão Gênica/efeitos dos fármacos , Interleucina-6/genética , Masculino , Modelos Teóricos , Mucuna/química , Estresse Oxidativo/efeitos dos fármacos , Ratos Wistar , Transdução de Sinais , Tetra-Hidroisoquinolinas/administração & dosagem , Tetra-Hidroisoquinolinas/isolamento & purificação
6.
Life Sci ; 201: 161-172, 2018 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-29486150

RESUMO

AIMS: To potentiate the well-documented tumor protecting ability of paullones, literatures demand for rational modifications in paullone ring structure and exploration of a precise mechanism underlying their antitumor effects. Thus, recently we synthesized novel paullone-like scaffold, 5H-benzo [2, 3][1,4]oxazepino[5,6-b]indoles, where compounds 13a and 14a attenuated the growth of liver cancer specific Hep-G2 cells in vitro and formed stable binding complex with IL-6. Henceforth, we hypothesized that this action is probably due to the blockade of IL-6 mediated JAK2/STAT3 signaling cascade. MAIN METHODS: A preclinical study was conducted using NDEA-induced HCC rat model by oral administration of FOIs at 10 mg/kg dose for 15 days. The molecular insights were confirmed through ELISA, qRT-PCR, western blot analyses. The study was further confirmed by data-based mathematical modeling using the quantitative data obtained from western blot analysis. 1H NMR based metabolomics study was also performed to unveil metabolite discriminations among various studied groups. KEY FINDINGS: We identified that the HCC condition was produced due to the IL-6 induced activation of JAK2 and STAT3 which, in turn, was due to enhanced phosphorylation of JAK2 and STAT3. The treatment with FOIs led to the significant blockade of the IL-6 mediated JAK2/STAT3 signaling pathway. Besides, FOIs showed their potential ability in restoring perturbed metabolites linked to HCC. In particular, the anticancer efficacy of compound 13a was comparable or somewhat better than marketed chemotherapeutics, 5-flurouracil. SIGNIFICANCE: These findings altogether opened up possibilities of developing fused oxazepino-indoles (FOIs) as new candidate molecule for plausible alternative of paullones to treat liver cancer.


Assuntos
Anticarcinógenos/farmacologia , Carcinogênese/efeitos dos fármacos , Indóis/farmacologia , Interleucina-6/antagonistas & inibidores , Janus Quinase 2/antagonistas & inibidores , Neoplasias Hepáticas/prevenção & controle , Oxazepinas/farmacologia , Fator de Transcrição STAT3/antagonistas & inibidores , Transdução de Sinais/efeitos dos fármacos , Animais , Anticarcinógenos/toxicidade , Citocinas/análise , Indóis/toxicidade , Neoplasias Hepáticas/patologia , Masculino , Metabolômica , Modelos Teóricos , Oxazepinas/toxicidade , Ratos , Ratos Wistar
7.
Pharmacol Res ; 132: 188-203, 2018 06.
Artigo em Inglês | MEDLINE | ID: mdl-29229353

RESUMO

1,4-benzothiazines have ameliorative effects through inhibition of COX-2 mediated STAT-3 pathways at G-protein couple receptor site. As per this scenario, we recently prepared and tested novel 1,4-benzothiazine derivatives against HT-29 human colon cancer cell line. Two compounds namely AR13 and AR15 showed higher inhibitions among all the synthesized compounds. In the present context, we conducted the in vivo antiproliferative action and identified the molecular mechanism associated to cytotoxic action of AR13 and AR15 in dimethylhydrazine (DMH) induced colorectal carcinoma (CRC) model. Various physiological, oxidative stress, histopathology, ELISA, qRT-PCR, western blot and NMR-based metabolomics were accomplished to evaluate the anticancer effect of titled compounds. Both compounds were subjected to histological and biochemical tests to observe the protective action of the compounds. ELISA showed potential role of these compounds to normalize increased levels of IL-2, IL-6 and COX-2 mediators. This action was more pronounced for COX-2 rather than IL-2 and IL-6. Gene expression analyses further revealed that both of them attenuated the over-expressed COX-2 gene. Furthermore, it was confirmed that these compounds exerted antitumor potential via preventing COX-2 induced JAK-2 and STAT-3 phosphorylation. This action was substansiated by immunohistochemistry using JAK2, p-JAK2, STAT3 and p-STAT3 targets in colon tissue. Finally, score plots of PLS-DA models exhibited significant metabolic discriminations between the treated and CRC groups, and both compounds showed ability to restore the imbalance of multiple metabolites during CRC. In conclusion, our study provided the evidence towards better antiproliferative effect of AR13 and AR15 in DMH-induced CRC through the blockade of COX-2/JAK-2/STAT-3 signal transduction pathway and could be demonstrated as useful anti-CRC candidate molecules for future anticancer therapy.


Assuntos
Antineoplásicos/farmacologia , Neoplasias Colorretais/metabolismo , Ciclo-Oxigenase 2/metabolismo , Janus Quinase 2/metabolismo , Fator de Transcrição STAT3/metabolismo , Tiazinas/farmacologia , Animais , Antineoplásicos/uso terapêutico , Colo/efeitos dos fármacos , Colo/metabolismo , Colo/patologia , Neoplasias Colorretais/induzido quimicamente , Neoplasias Colorretais/tratamento farmacológico , Neoplasias Colorretais/patologia , Ciclo-Oxigenase 2/genética , Dimetilidrazinas , Janus Quinase 2/genética , Masculino , Ratos Wistar , Fator de Transcrição STAT3/genética , Transdução de Sinais/efeitos dos fármacos , Estresse Fisiológico , Tiazinas/uso terapêutico
8.
Cancer Biol Ther ; 18(5): 304-313, 2017 05 04.
Artigo em Inglês | MEDLINE | ID: mdl-28358223

RESUMO

Pyrazinoic acid (PA) is structurally similar to nicotinic acid which acts on G-protein-coupled receptor (GPR109A). GPR109A expresses in colonic and intestinal epithelial sites, and involves in DNA methylation and cellular apoptosis. Therefore, it may be assumed that PA has similar action like nicotinic acid and may be effective against colorectal carcinoma (CRC). CRC was produced via subcutaneous injection of dimethylhydrazine (DMH) at 40 mg/kg body weight once in a week for 4 weeks. After that, PA was administered orally at 2 doses of 10 and 25 mg/kg daily for 15 d to observe the antiproliferative effect. Various physiologic, oxidative stress, molecular parameters, histopathology, RT-PCR and NMR based metabolomics were performed to evaluate the antiproliferative potential of PA. Our results collectively suggested that PA reduced body weight, tumor volume and incidence no. to normal. It restored various oxidative stress parameters and normalized IL-2, IL-6, and COX-2 as compared with carcinogen control. In molecular level, overexpressed IL-6 and COX-2 genes became normal after PA administration. Again, normal tissue architecture was prominent after PA administration. Score plots of PLS-DA models exhibited that PA treated groups were significantly different from CRC group. We found that CRC rat sera have increased levels of acetate, glutamine, o-acetyl-glycoprotein, succinate, citrulline, choline, o-acetyl choline, tryptophan, glycerol, creatinine, lactate, citrate and decreased levels of 3-hydroxy butyrate, dimethyl amine, glucose, maltose, myoinositol. Further the PA therapy has ameliorated the CRC-induced metabolic alterations, signifying its antiproliferative properties. In conclusion, our study provided the evidence that PA demonstrated good antiproliferative effect on DMH induced CRC and thus demonstrated the potential of PA as a useful drug for future anticancer therapy.


Assuntos
Carcinoma/tratamento farmacológico , Neoplasias do Colo/tratamento farmacológico , Pirazinamida/análogos & derivados , Animais , Apoptose/efeitos dos fármacos , Carcinoma/induzido quimicamente , Carcinoma/patologia , Proliferação de Células/efeitos dos fármacos , Neoplasias do Colo/induzido quimicamente , Neoplasias do Colo/patologia , Dimetilidrazinas/toxicidade , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Proteínas de Neoplasias/genética , Estresse Oxidativo/efeitos dos fármacos , Pirazinamida/administração & dosagem , Ratos , Estresse Fisiológico/efeitos dos fármacos
9.
Food Funct ; 7(7): 3064-72, 2016 Jul 13.
Artigo em Inglês | MEDLINE | ID: mdl-27251027

RESUMO

The use of prebiotics to escalate certain gut flora is a current aspect of research for effective gut ecology. In the present study we appraise the efficacy of gum odina obtained from the bark of Odina wodier (Anacardiaceae), which is not fully degraded (16%) in the upper GI tract and becomes available to the lower region, as a prebiotic. An in vitro prebiotic activity assay established a quantitative score to describe the extent to which gum odina supports the selective growth of probiotics with a maximum of 5.60 ± 0.11 for Lactobacillus plantarum MTCC 6160. The polysaccharide, upon fermentation, also liberates lactic acid (0.46 ± 0.003 mg ml(-1)) and acetic acid (1.03 ± 0.003 mg ml(-1)). In vivo studies revealed that natural gum selectively stimulates Lactobacillus sp., and eliminates enteric pathogens with a C.F.U. of 384.48 ± 0.11 and 40.56 ± 0.17 respectively on the 8(th) day. The changes in the level of ß-galactosidase signify maturation of macrophages in the gut environment. It also boosts the immune system by increasing sIgA upon infection from the 5(th) day in the gut, when incorporated into the feed of mice. Moreover an increase in levels of IFNγ on the 5(th) day also manifest additional protection against various pathogen-induced primary and secondary infections. Thus, gum odina is a potential prebiotic which not only provides nutrition but also improves gut ecology.


Assuntos
Microbioma Gastrointestinal , Trato Gastrointestinal/microbiologia , Gomas Vegetais/química , Prebióticos/administração & dosagem , Ácido Acético/metabolismo , Anacardiaceae/química , Animais , Bacillus coagulans , Fermentação , Galactosidases/metabolismo , Imunoglobulina A Secretora/metabolismo , Interferon gama/metabolismo , Ácido Láctico/metabolismo , Lactobacillus acidophilus , Lactobacillus helveticus , Lactobacillus plantarum , Lacticaseibacillus rhamnosus , Macrófagos/metabolismo , Masculino , Camundongos , Polissacarídeos/química , Probióticos
10.
Nat Prod Res ; 30(4): 460-3, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-25774560

RESUMO

The present study was undertaken to investigate the antiproliferative action of isolated M1 (6,7-dimethoxy-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid) from Mucuna pruriens seeds using human hepatic carcinoma cell line (Huh-7 cells). Initially, docking studies was performed to find out the binding affinities of M1 to caspase-3 and 8 enzymes. Later, cytotoxic action of M1 was measured by cell growth inhibition (MTT), followed by caspase-3 and 8 enzymes assay colorimetrically. Our results collectively suggested that M1 had strong binding affinity to caspase-8 in molecular modelling. M1 possessed antiproliferative activity on Huh-7 cells (EC50 = 13.97 µM) and also inhibited the action of caspase-8 enzyme, signified process of apoptosis. M1 was active against Huh-7 cells that may be useful for future hepatic cancer treatment.


Assuntos
Alcaloides/farmacologia , Carcinoma Hepatocelular/patologia , Isoquinolinas/farmacologia , Neoplasias Hepáticas/patologia , Mucuna/química , Tetra-Hidroisoquinolinas/farmacologia , Alcaloides/isolamento & purificação , Apoptose/efeitos dos fármacos , Caspase 3/metabolismo , Caspase 8/metabolismo , Linhagem Celular Tumoral/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Humanos , Isoquinolinas/isolamento & purificação , Simulação de Acoplamento Molecular , Estrutura Molecular , Sementes/química , Tetra-Hidroisoquinolinas/isolamento & purificação
11.
J Biomed Mater Res A ; 103(5): 1771-83, 2015 May.
Artigo em Inglês | MEDLINE | ID: mdl-25227875

RESUMO

Urease is a nickel-dependent metalloenzyme that catalyzes the hydrolysis of urea to form ammonia and carbon dioxide. Although the enzyme serves a significant role in several detoxification and analytical processes, its usability is restricted due to high cost, availability in small amounts, instability, and a limited possibility of economic recovery from a reaction mixture. Hence, there is a need to develop an efficient, simple, and reliable immobilization strategy for the enzyme. In this study, the carboxyl terminated surface of glutathione-capped gold nanoparticles have been utilized as a solid support for the covalent attachment of urease. The immobilization has been carried out at different pH conditions so as to elucidate its effect on the immobilization efficiency and enzyme bioactivity. The binding of the enzyme has been quantitatively and qualitatively analyzed through techniques like ultraviolet-visible spectroscopy, intrinsic steady state fluorescence, and circular dichorism. The bioactivity of the immobilized enzyme was investigated with respect to the native enzyme under different thermal conditions. Recyclability and shelf life studies of the immobilized enzyme have also been carried out. Results reveal that the immobilization is most effective at pH of 7.4 followed by that in an acidic medium and is least in alkaline environment. The immobilized enzyme also exhibits enhance activity in comparison to the native form at physiological temperature. The immobilized urease (on gold glutathione nanoconjugates surface) can be effectively employed for biosensor fabrication, immunoassays and as an in vivo diagnostic tool in the future.


Assuntos
Enzimas Imobilizadas/metabolismo , Glutationa/farmacologia , Ouro/farmacologia , Nanopartículas Metálicas/química , Urease/metabolismo , Materiais Biocompatíveis/farmacologia , Morte Celular/efeitos dos fármacos , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Dicroísmo Circular , Difusão Dinâmica da Luz , Humanos , Concentração de Íons de Hidrogênio , Nanopartículas Metálicas/ultraestrutura , Tamanho da Partícula , Estrutura Secundária de Proteína , Padrões de Referência , Espectrometria de Fluorescência , Espectrofotometria Ultravioleta , Espectroscopia de Infravermelho com Transformada de Fourier , Temperatura , Urease/química
12.
EMBO Rep ; 15(7): 775-83, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-24878851

RESUMO

A20 has been suggested to limit NF-κB activation by removing regulatory ubiquitin chains from ubiquitinated substrates. A20 is a ubiquitin-editing enzyme that removes K63-linked ubiquitin chains from adaptor proteins, such as RIP1, and then conjugates them to K48-linked polyubiquitin chains to trigger proteasomal degradation. To determine the role of the deubiquitinase function of A20 in downregulating NF-κB signaling, we have generated a knock-in mouse that lacks the deubiquitinase function of A20 (A20-OTU mice). These mice are normal and have no signs of inflammation, have normal proportions of B, T, dendritic, and myeloid cells, respond normally to LPS and TNF, and undergo normal NF-κB activation. Our results thus indicate that the deubiquitinase activity of A20 is dispensable for normal NF-κB signaling.


Assuntos
Cisteína Endopeptidases/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , NF-kappa B/metabolismo , Proteases Específicas de Ubiquitina/metabolismo , Animais , Cisteína Endopeptidases/genética , Análise Mutacional de DNA , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/imunologia , Células Dendríticas/metabolismo , Modelos Animais de Doenças , Ativação Enzimática , Genótipo , Sistema Imunitário/citologia , Sistema Imunitário/imunologia , Sistema Imunitário/metabolismo , Imunofenotipagem , Peptídeos e Proteínas de Sinalização Intracelular/genética , Lipopolissacarídeos/imunologia , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Macrófagos/metabolismo , Camundongos , Camundongos Transgênicos , Mutação , Fenótipo , Choque/induzido quimicamente , Choque/genética , Choque/imunologia , Choque/metabolismo , Choque/mortalidade , Proteína 3 Induzida por Fator de Necrose Tumoral alfa , Fator de Necrose Tumoral alfa/farmacologia
13.
Methods Mol Biol ; 1141: 211-32, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24567142

RESUMO

A protocol for the synthesis of a smart drug delivery system based on gold nanoparticles has been described in this chapter. The synthesized drug delivery system has been shown to release the bioactive material in response to an intracellular stimulus (glutathione concentration gradient) and hence shown to behave in an intelligent manner. Gold nanoparticles have been employed as the core material with the surface functionalities of thiolated PEG. PEG owing to its non-immunogenicity and non-antigenicity would impart considerable stability and longer in vivo circulation time to the gold nanoparticles. The end groups of PEG chains have been derivatized with functional groups like aldehyde (-CHO) and amine (-NH2) which could behave as flexible arms for the attachment of "target specific ligands" and other bioactive substances. Lactose, a liver targeting ligand, has been employed as the target specific moiety. A Coumarin derivative has been synthesized and used as the model fluorescent tag as well as a linker to examine the glutathione-mediated release through fluorescence spectroscopy and for the conjugation of bioactive molecules, respectively. A check for the cytocompatibility of the synthesized nanovehicle on the cultured mammalian cell lines has also been carried out. Finally, in the latter parts of the chapter (mimicking the in vivo conditions), time-dependent in vitro release of the model fluorescent moiety has also been analyzed at different glutathione concentrations.


Assuntos
Sistemas de Liberação de Medicamentos/métodos , Glutationa/química , Ouro/química , Nanopartículas Metálicas/química , Animais , Linhagem Celular , Cumarínicos/química , Liberação Controlada de Fármacos , Células Epiteliais/citologia , Células Epiteliais/metabolismo , Corantes Fluorescentes/química , Humanos , Lactonas/química , Lactose/química , Fígado , Modelos Biológicos , Polietilenoglicóis/química , Compostos de Sulfidrila/química
14.
AAPS PharmSciTech ; 14(3): 1219-26, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23934434

RESUMO

Targeting drug formulations to specific tissues and releasing the bioactive content in response to a certain stimuli remains a significant challenge in the field of biomedical science. We have developed a nanovehicle that can be used to deliver "drugs" to "specific" tissues. For this, we have simultaneously modified the surface of the nanovehicle with "drugs" and "tissue-specific ligands". The "tissue-specific ligands" will target the nanovehicle to the correct tissue and release the "drug" of interest in response to specific stimuli. We have synthesised a "lactose surface-modified gold nanovehicle" to target liver cells and release the model fluorescent drug (coumarin derivative) in response to the differential glutathione concentration (between blood plasma and liver cells). Lactose is used as the liver-specific targeting ligand given the abundance of L-galactose receptors in hepatic cells. The coumarin derivative is used as a fluorescent tag as well as a linker for the attachment of various biologically relevant molecules. The model delivery system is compatible with a host of different ligands and hence could be used to target other tissues as well in future. The synthesised nanovehicle was found to be non-toxic to cultured human cell lines even at elevated non-physiological concentrations as high as 100 µg/mL. We discover that the synthesised gold-based nanovehicle shows considerable stability at low extracellular glutathione concentrations; however coumarin is selectively released at high hepatic glutathione concentration.


Assuntos
Portadores de Fármacos , Ouro/química , Fígado/metabolismo , Nanopartículas Metálicas/química , Corantes Fluorescentes/química , Glutationa/química , Espectroscopia de Infravermelho com Transformada de Fourier
15.
PLoS One ; 7(8): e43078, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22912792

RESUMO

We recently reported a novel synthesis of copper nanoparticles from copper sulphate utilizing the charge-compensatory effect of ionic liquid [bmim]BF(4) and ethylene glycol. The nanoparticles were characterized and found to be stable for one year. Here we hypothesize that the stabilized nanoparticles should be able to catalyze one-pot multicomponent organic reactions. We show that the nanoparticles catalyzed Biginelli reaction at room temperature to give the product 3,4-dihydopyrimidinone (>90% yield in ~15 minutes) from aldehydes, ß-diketoester (ethylacetoacetate) and urea (or thiourea). ). Remarkably, such high yields and rapid kinetics was found to be independent of the electronic density on the reactant aryl-aldehyde. This was probably because even the surface-active particles reacted faster in the presence of ionic liquid as compared to conventional methods. The heterocyclic dihydropyrimidinones (DHPMs) and their derivatives are widely used in natural and synthetic organic chemistry due to their wide spectrum of biological and therapeutic properties (resulting from their antibacterial, antiviral, antitumor and anti-inflammatory activities. Our method has an easy work-up procedure and the nanoparticles could be recycled with minimal loss of efficiency.


Assuntos
Cobre/química , Nanopartículas Metálicas/química , Pirimidinonas/síntese química , Acetoacetatos/química , Aldeídos/química , Catálise , Cinética , Nanopartículas Metálicas/ultraestrutura , Microscopia Eletrônica de Transmissão , Pirimidinonas/química , Ureia/química , Difração de Raios X
16.
Zhong Xi Yi Jie He Xue Bao ; 10(5): 546-54, 2012 May.
Artigo em Inglês | MEDLINE | ID: mdl-22587977

RESUMO

OBJECTIVE: To examine if a homeopathic mother tincture (Phytolacca Decandra) is capable of precipitating silver nanoparticles from silver nitrate (AgNO(3)) and to characterize the biosynthesized nanoparticles for evaluating their biological activities. METHODS: A total of 100 mg of AgNO(3) was added to 20mL of Milli-Q water and stirred vigorously. Then 5mL of the homeopathic mother tincture of Phytolacca Decandra (ethanolic root extract of Phytolacca decandra) was added and stirred continuously. Reduction took place rapidly at 300K and completed in 10 min as shown by stable light greenish-yellow color of the solution which gave colloid of silver nanoparticles. The colloid solution was then centrifuged at 5000×g to separate the nanoparticles for further use. The nanoparticles were characterized by spectroscopic analysis, particle size analysis and zeta potential measurements, and morphology was analyzed by atomic force microscopy. The drug-DNA interaction was determined by circular dichroism spectrophotometry and melting temperature profiles by using calf thymus DNA as the target. The biological activities were determined using a cancer cell line A549 in vitro and using bacteria Escherichia coli and fungus Saccharomyces cerevisiae as test models. RESULTS: Phytolacca Decandra precipitated silver nanoparticles in ambient conditions. The nanoparticles had 91 nm particle size, with polydispersity index of 0.119 and zeta potential of -15.6 mV. The silver nanoparticles showed anticancer and antibacterial properties, but no clear antifungal properties. CONCLUSION: This could be a novel environment-friendly method to biosynthesize silver nanoparticles using a cost-effective, nontoxic manner. The homeopathic mother tincture may utilize this property of nano-precipitation in curing diseases or disease symptoms.


Assuntos
Química Verde , Homeopatia , Nanopartículas Metálicas , Phytolacca/química , Nitrato de Prata/química , Materia Medica/química , Tamanho da Partícula , Extratos Vegetais/química , Prata/química
17.
Zhong Xi Yi Jie He Xue Bao ; 10(3): 337-46, 2012 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-22409925

RESUMO

OBJECTIVE: To examine to what degree an ultra-highly diluted homeopathic remedy, Arnica Montana 30C (AM-30C), used in the treatment of shock and injury, can modulate the expression of nucleotide excision repair genes in Escherichia coli exposed to ultraviolet (UV) irradiation. METHODS: E. coli were cultured to their log phase in a standard Luria-Bertani medium and then exposed to sublethal doses of UV irradiation at 25 and 50 J/m(2) for 22.5 and 45 s, respectively. The UV-exposed bacteria were then supplemented with either AM-30C (drug) or placebo (P-30C). The drug-treated and placebo-treated bacteria were subjected to assay for DNA damage and oxidative stress 90 min after UV exposure. Several protocols like comet assay, gel electrophoresis for DNA ladder and intracellular reactive oxygen species (ROS) generation, and biomarker measurement like superoxide dismutase (SOD), catalase (CAT) and reduced glutathione (GSH) were conducted. The mRNA expressions of the excision repair genes like ultraviolet repair uvrA, B and C genes (or also known as excision repair genes) were estimated by reverse transcription-polymerase chain reaction method. RESULTS: The UV-exposed bacteria showed DNA damage and oxidative stress, as revealed by an increase in ROS generation, and a decrease in SOD, CAT and GSH activities. As compared to placebo, the AM-30C-treated bacteria showed less DNA damage and oxidative stress as manifested by a decrease in ROS generation, and an increase in SOD, CAT and GSH activities. AM-30C also up-regulated the expression of repair genes as compared to the control. CONCLUSION: AM-30C helped repair the DNA damage through up-regulation of repair genes and also ameliorated the oxidative stress through the reduction of ROS generation and suitable modulation of anti-oxidative stress enzymes.


Assuntos
Arnica , Dano ao DNA/efeitos dos fármacos , Reparo do DNA/genética , Escherichia coli/efeitos dos fármacos , Homeopatia , Catalase/metabolismo , Ensaio Cometa , Escherichia coli/efeitos da radiação , Glutationa/metabolismo , Estresse Oxidativo , Espécies Reativas de Oxigênio/metabolismo , Superóxido Dismutase/metabolismo , Raios Ultravioleta
18.
Zhong Xi Yi Jie He Xue Bao ; 10(2): 210-27, 2012 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-22313889

RESUMO

OBJECTIVE: To examine if potentiated homeopathic drug Arsenicum Album 30C (Ars Alb 30C) can reduce sodium arsenite-induced toxicity in Escherichia coli. METHODS: E. coli were exposed to low arsenite insult after they grew up to log phase in standard Luria-Bertani medium. E. coli were treated with 1 or 2 mmol/L sodium arsenite alone (control), or Ars Alb 30C was added to the medium of a subset of sodium arsenite-treated bacteria (drug-treated), or homeopathically agitated alcohol was added to the medium containing a subset of sodium arsenite-treated bacteria (placebo-treated). A sub-set of untreated E. coli served as the negative control. Glucose uptake, specific activities of hexokinase, lipid peroxidase (LPO), superoxide dismutase (SOD) and catalase, intra- and extra-cellular sodium arsenite content, cell growth, cell membrane potential, DNA damage, intracellular reactive oxygen species (ROS), adenosine triphosphate (ATP) and free glutathione content and expressions of arsB and ptsG gene in normal control, sodium arsenite-treated, drug-treated and placebo-treated E. coli were analyzed. Treatments were blinded and randomized. RESULTS: In sodium arsenite-treated E. coli, glucose uptake, intracellular ROS, LPO and DNA damage increased along with decrease in the specific activities of hexokinase, SOD and catalase, intracellular ATP and free glutathione contents and cell membrane potential and growth, and there were increases in expression levels of arsB gene and ptsG gene. Ars Alb 30C administration reduced arsenic toxicity in E. coli by inhibiting generation of ROS and increasing tolerance to arsenite toxicity and cell growth. CONCLUSION: Ars Alb 30C ameliorated arsenic toxicity and DNA damage, validating efficacy of ultra-highly diluted remedies used in homeopathy.


Assuntos
Arsenicais/antagonistas & inibidores , Arsenitos/toxicidade , Escherichia coli/efeitos dos fármacos , Escherichia coli/metabolismo , Homeopatia , Espécies Reativas de Oxigênio/metabolismo , Dano ao DNA , Regulação Bacteriana da Expressão Gênica/efeitos dos fármacos , Peroxidação de Lipídeos , Regulação para Cima
19.
PLoS One ; 7(1): e29131, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22238589

RESUMO

In this work, we present a novel method for the synthesis of copper nanoparticles. We utilize the charge compensatory effect of ionic liquid [bmim]BF(4) in conjunction with ethylene glycol for providing electro-steric stabilization to copper nanoparticles prepared from copper sulphate using hydrazine hydrate as a reducing agent. The formed copper nanoparticles showed extended stability over a period of one year. Copper nanoparticles thus prepared were characterized by powder X-ray diffraction measurements (pXRD), transmission electron microscopy (TEM) and quasi elastic light scattering (QELS) techniques. Powder X-ray diffraction (pXRD) analysis revealed relevant Bragg's reflection for crystal structure of copper. Powder X-ray diffraction plots also revealed no oxidized material of copper nanoparticles. TEM showed nearly uniform distribution of the particles in methanol and confirmed by QELS. Typical applications of copper nanoparticles include uses in conductive films, lubrication and nanofluids. Currently efforts are under way in our laboratory for using these nanoparticles as catalysts for a variety of organic reactions.


Assuntos
Cobre/química , Etilenoglicol/farmacologia , Imidazóis/farmacologia , Líquidos Iônicos/farmacologia , Nanopartículas Metálicas/química , Pós/síntese química , Soluções Tampão , Catálise/efeitos dos fármacos , Etilenoglicol/química , Humanos , Interações Hidrofóbicas e Hidrofílicas , Imidazóis/química , Líquidos Iônicos/química , Nanopartículas Metálicas/ultraestrutura , Microscopia Eletrônica de Transmissão , Modelos Biológicos , Tamanho da Partícula , Pós/química , Fatores de Tempo , Difração de Raios X
20.
Zhong Xi Yi Jie He Xue Bao ; 9(8): 901-12, 2011 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-21849152

RESUMO

OBJECTIVE: Whether ultra-highly diluted homeopathic remedies can affect living systems is questionable. Therefore, this study sees value in the analysis of whether homeopathically diluted glucose 30C has any effect on Escherichia coli exposed to arsenite stress. METHODS: E. coli were cultured to their log phase in standard Luria-Bertani medium and then treated with either 1 mmol/L or 2 mmol/L sodium arsenite, with or without supplementation of either 1% or 3% glucose, an ultra-highly diluted and agitated ethanolic solution (70%) of glucose (diluted 10(60) times), glucose 30C or 70% ethanol (placebo) in the medium. Glucose uptake, specific activities of hexokinase and glucokinase, membrane potential, intracellular adenosine triphosphate (ATP) and expression of glucose permease in E. coli were analyzed at two different time intervals. Arsenic content in E. coli (intracellular) and in the spent medium (extracellular) was also determined. RESULTS: In arsenite-exposed E. coli, the glucose uptake increased along with decreases in the specific activities of hexokinase and glucokinase, intracellular ATP and membrane potential and an increase in the gene expression level of glucose permease. Glucose uptake increased further by addition of 1%, 3% or ultra-highly diluted glucose in the medium, but not by the placebo. CONCLUSION: The results demonstrated the efficacy of the ultra-highly diluted and agitated glucose in mimicking the action of actual glucose supplementation and its ability to modulate expressions of hexokinase and glucokinase enzymes and glucose permease genes, thereby validating the efficacy of ultra-high dilutions used in homeopathy.


Assuntos
Arsenitos/química , Transporte Biológico/efeitos dos fármacos , Escherichia coli/efeitos dos fármacos , Escherichia coli/metabolismo , Glucose/metabolismo , Glucose/farmacologia , Contagem de Colônia Microbiana , Glucose/administração & dosagem , Materia Medica/química
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