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1.
Exp Parasitol ; 236-237: 108258, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-35421387

RESUMEN

Glycyrrhetinic acid (GA) is one of the important Pentacyclic Triterpenoids (PT) found in the roots of licorice. This study aimed to evaluate the in vitro growth inhibitory effect of 18ß-GA (18ß-Glycyrrhetinic acid) and C-30 esters against Theileria annulata, the causative agent of Tropical Bovine Theileriosis. C-30 esters of 18ß-GA were synthesized and their structures were elucidated using spectroscopy. The pharmacodynamic properties of 18ß-GA and its C-30 esters were predicted using DataWarrior and Swiss ADME tools. Cattle isolates of T. annulata schizont-infected bovine lymphoblastoid cells were cultured using standard conditions and the growth inhibitory effect of GA and its esters were evaluated using MTT assay. The isopropyl ester of 18ß-GA (GI50- 1.638 µM; R2- 0.818) showed improved anti-theileriosis efficacy than other 18ß-GA derivatives. The propyl (GI50 - 5.549 µM), ethyl (GI50 - 5.638 µM), and benzyl (GI50 - 7.431 µM) esters also showed considerable inhibitory effect. The GI50 value for 18ß-GA was recorded as 6.829 µM. This study throws light on the usefulness of 18ß-GA and its esters for the treatment of Tropical Bovine Theileriosis.


Asunto(s)
Ácido Glicirretínico , Theileriosis , Animales , Bovinos , Ésteres/farmacología , Ácido Glicirretínico/análogos & derivados , Ácido Glicirretínico/química , Ácido Glicirretínico/farmacología , Extractos Vegetales , Theileriosis/tratamiento farmacológico
2.
Planta ; 254(5): 90, 2021 Oct 05.
Artículo en Inglés | MEDLINE | ID: mdl-34609619

RESUMEN

MAIN CONCLUSION: Identification of molecular markers and characterization of nutrient transporters could help to improve the tolerance under abiotic and low nutrient stresses in sorghum ensuring higher yield to conserve food security Sorghum is an important cereal crop delivering food and energy security in the semi-arid tropics of the world. Adverse climatic conditions induced by global warming and low input agriculture system in developing countries demand for the improvement of sorghum to tolerate various abiotic stresses. In this review, we discuss the application of marker-assisted breeding and nutrient transporter characterization studies targeted towards improving the tolerance of sorghum under drought, salinity, cold, low phosphate and nitrogen stresses. Family members of some nutrient transporters such as nitrate transporter (NRT), phosphate transporter (PHT) and sulphate transporter (SULTR) were identified and characterized for improving the low nutrient stress tolerance in sorghum. Several quantitative trait loci (QTL) were identified for drought, salinity and cold stresses with an intention to enhance the tolerance of sorghum under these stresses. A very few QTL and nutrient transporters have been identified and validated under low nitrogen and phosphorus stresses compared to those under drought, salinity and cold stresses. Marker-assisted breeding and nutrient transporter characterization have not yet been attempted in sorghum under other macro- and micro-nutrient stresses. We hope this review will raise awareness among plant breeders, scientists and biotechnologists about the importance of sorghum and need to conduct the studies on marker-assisted breeding and nutrient transporter under low nutrient stresses to improve the sorghum production.


Asunto(s)
Sorghum , Grano Comestible , Nutrientes , Fitomejoramiento , Sorghum/genética , Estrés Fisiológico
3.
Biotechnol Lett ; 43(5): 1005-1018, 2021 May.
Artículo en Inglés | MEDLINE | ID: mdl-33515159

RESUMEN

Streptomyces is one of the most prolific producers of economically important bioactive compounds used against several illnesses; it has also been found to produce industrially useful enzymes. In this study, Streptomyces sp. (ERINLG-201) was isolated from the soil sample of Kodanad forest (Southern Western Ghats), The Nilgiris, Tamil Nadu, India. ERINLG-201 isolate showed promising antibacterial activity against tested Gram-positive and Gram-negative bacteria which was confirmed by perpendicular 'T' streak method. Secondary metabolites of ERINLG-201 isolate exhibited promising antibacterial activity against tested Gram-positive and Gram-negative bacteria which was confirmed by disc diffusion method using the ethylacetate extract. Further, the ethylacetate extract of ERINLG-201 (15 g) was packed in column chromatography over silica gel and eluted; it resulted in isolation of a new naphthoquinone derivative named bluemomycin from the active fraction. Bluemomycin showed promising antibacterial activity against Gram-negative bacteria and clinical isolates at least concentration (6.25 µg/mL). Cytotoxic studies of bluemomycin showed promising activity against A549, Skvo-3 and HepG2 cell lines with IC50 values of 5.9, 24.2 and 11 µM, respectively.


Asunto(s)
Antiinfecciosos/farmacología , Antineoplásicos/farmacología , Naftoquinonas/farmacología , Streptomyces/metabolismo , Antiinfecciosos/química , Antiinfecciosos/metabolismo , Antineoplásicos/química , Antineoplásicos/metabolismo , Bacterias/efectos de los fármacos , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , ADN Bacteriano/genética , Bosques , Hongos/efectos de los fármacos , Humanos , India , Pruebas de Sensibilidad Microbiana , Naftoquinonas/química , Naftoquinonas/metabolismo , Filogenia , ARN Ribosómico 16S/genética , Microbiología del Suelo , Streptomyces/clasificación , Streptomyces/genética , Streptomyces/aislamiento & purificación
4.
Planta ; 252(6): 98, 2020 Nov 07.
Artículo en Inglés | MEDLINE | ID: mdl-33159589

RESUMEN

MAIN CONCLUSION: This is a first comprehensive study to analyze the 12 PHT1 family phosphate transporter genes in 20 foxtail millet genotypes for the improvement of millets and other crops for phosphate use efficiency. Phosphorus (P), absorbed from soil solutions as inorganic phosphate (Pi), is a limiting nutrient for plant growth and yield. Twenty genotypes of foxtail millet (Setaria italica) with contrasting degree of growth and Pi uptake responses under low Pi (LP) and high Pi (HP) supply were chosen based on a previous study. To gain molecular insights, expression dynamics of 12 PHosphate Transporter 1 (PHT1) family (SiPHT1;1 to 1;12) genes were analyzed in these 20 genotypes and compared with their Pi and total P (TP) contents. SiPHT1;1, 1;2, 1;3 and 1;8 genes were expressed in shoot tissues of three (ISe 1209, ISe 1305 and Co-6) of the LP best performing genotypes (LPBG); however, they were expressed in only one of the LP worst performing genotype (LPWG) (ISe 748). More importantly, this is correlating with higher shoot Pi and TP contents of the LPBG compared to LPWG. Apart from this condition, expression of SiPHT1 genes and their Pi and TP contents do not correlate directly for many genotypes in other conditions; genotypes with low Pi and TP contents induced more SiPHT1 genes and vice versa. Promoter analysis revealed that genotype ISe 1888 with a high level of SiPHT1;8 expression possesses two additional root box motifs compared to other genotypes. The PHT1 family genes seem to play a key role for LP stress tolerance in foxtail millet and further studies will help to improve the P-use efficiency in foxtail millet and other cereals.


Asunto(s)
Regulación de la Expresión Génica de las Plantas , Proteínas de Transporte de Fosfato , Setaria (Planta) , Estrés Fisiológico , Genotipo , Proteínas de Transporte de Fosfato/genética , Fosfatos/toxicidad , Setaria (Planta)/efectos de los fármacos , Setaria (Planta)/genética , Estrés Fisiológico/efectos de los fármacos , Estrés Fisiológico/genética
5.
Biochim Biophys Acta ; 1863(9): 2333-44, 2016 09.
Artículo en Inglés | MEDLINE | ID: mdl-27350235

RESUMEN

The clustered, regularly interspaced, short palindromic repeat (CRISPR) and CRISPR associated protein 9 (Cas9) system discovered as an adaptive immunity mechanism in prokaryotes has emerged as the most popular tool for the precise alterations of the genomes of diverse species. CRISPR/Cas9 system has taken the world of genome editing by storm in recent years. Its popularity as a tool for altering genomes is due to the ability of Cas9 protein to cause double-stranded breaks in DNA after binding with short guide RNA molecules, which can be produced with dramatically less effort and expense than required for production of transcription-activator like effector nucleases (TALEN) and zinc-finger nucleases (ZFN). This system has been exploited in many species from prokaryotes to higher animals including human cells as evidenced by the literature showing increasing sophistication and ease of CRISPR/Cas9 as well as increasing species variety where it is applicable. This technology is poised to solve several complex molecular biology problems faced in life science research including cancer research. In this review, we highlight the recent advancements in CRISPR/Cas9 system in editing genomes of prokaryotes, fungi, plants and animals and provide details on software tools available for convenient design of CRISPR/Cas9 targeting plasmids. We also discuss the future prospects of this advanced molecular technology.


Asunto(s)
Sistemas CRISPR-Cas/genética , Edición Génica , Genoma , Genómica/métodos , Animales , ARN Guía de Kinetoplastida/metabolismo , Programas Informáticos
6.
Biochem Soc Trans ; 44(5): 1541-1548, 2016 10 15.
Artículo en Inglés | MEDLINE | ID: mdl-27911737

RESUMEN

The plant PHosphate Transporter 1 (PHT1) family of membrane proteins belongs to the major facilitator super family and plays a major role in the acquisition of inorganic phosphate (Pi) from the soil and its transport within the plant. These transporters have been well characterized for expression patterns, localization, and in some cases affinity. Furthermore, the crystal structure of a high-affinity eukaryotic phosphate transporter from the fungus Piriformospora indica (PiPT) has revealed important information on the residues involved in Pi transport. Using multiple-sequence alignments and homology modelling, the phosphate-binding site residues were shown to be well conserved between all the plant PHT1 proteins, Saccharomyces cerevisiae PHO84 and PiPT. For example, Asp 324 in PiPT is conserved in the equivalent position in all plant PHT1 and yeast transporters analyzed, and this residue in ScPHO84 was shown by mutagenesis to be important for both the binding and transport of Pi. Moreover, Asp 45 and Asp 149, which are predicted to be involved in proton import, and Lys 459, which is putatively involved in Pi-binding, are all fully conserved in PHT1 and ScPHO84 transporters. The conserved nature of the residues that play a key role in Pi-binding and transport across the PHT1 family suggests that the differing Pi affinities of these transporters do not reside in differences in the Pi-binding site. Recent studies suggest that phosphate transporters could possess dual affinity and that post-translational modifications may be important in regulating affinity for phosphate.


Asunto(s)
Proteínas de Arabidopsis/metabolismo , Proteínas de Transporte de Fosfato/metabolismo , Fosfatos/metabolismo , Simportadores de Protón-Fosfato/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Proteínas de Arabidopsis/genética , Secuencia de Bases , Sitios de Unión/genética , Unión Competitiva , Evolución Molecular , Proteínas de Transporte de Fosfato/genética , Unión Proteica , Simportadores de Protón-Fosfato/genética , Proteínas de Saccharomyces cerevisiae/genética , Homología de Secuencia de Aminoácido
7.
Biotechnol Lett ; 38(4): 689-700, 2016 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-26712371

RESUMEN

OBJECTIVES: Involvement of the outer membrane protein C (OmpC) of Escherichia coli in neurodegeneration was investigated using a mouse model. RESULTS: OmpC formed protease-resistant fibres that exhibited the diagnostic features of an amyloid. The spectral shift in the Congo Red and the thioflavin T assays produced features similar to neurotoxic peptides. Intramuscular administration of OmpC in mice resulted in spongiform neurodegeneration of the brain through calcium-dependent apoptosis and also showed upregulation of apoptosis related genes. Immunolocalization of OmpC in brain demonstrated the direct involvement of the porin in neurodegeneration and formation of spongiform encephalopathy. CONCLUSION: We have demonstrated the ability of OmpC of E. coli to induce neurodegeneration in mice.


Asunto(s)
Encéfalo/patología , Enfermedades Neurodegenerativas/etiología , Porinas/toxicidad , Animales , Apoptosis , Encéfalo/metabolismo , Calcio/metabolismo , Modelos Animales de Enfermedad , Regulación de la Expresión Génica , Inyecciones Intramusculares , Ratones , Enfermedades Neurodegenerativas/metabolismo , Enfermedades Neurodegenerativas/patología , Porinas/administración & dosificación
8.
Pharm Biol ; 53(2): 174-84, 2015 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-25289526

RESUMEN

CONTEXT: Delonix elata (L.) Gamble (Fabaceae) has been used in the Indian traditional medicine system to treat rheumatism and inflammation. AIM: To assess the anti-inflammatory effect of Delonix elata flowers and to isolate the active principle. MATERIALS AND METHODS: The prompt anti-inflammatory constituent was isolated from Delonix elata flower extracts using bioassay guided fractionation in liposaccharide (LPS) stimulated RAW 264.7 macrophage cell line. The anti-inflammatory activity of extracts/fractions/sub-fractions/compounds (10, 25, and 50 µg/ml) was evaluated by estimating the levels of nitric oxide (NO), TNF-α, and IL-1ß after 24 h of LPS induction (1 µg/ml). The isolated active compound was subjected to NMR, IR, and UV analyses for structure determination. RESULTS: In an attempt to search for anti-inflammatory constituents, the active pure principle was isolated and crystallized as a white compound from Delonix elata flowers methanol extract. This active compound (50 µg/ml) decreased the release of inflammatory mediators levels such as NO (0.263 ± 0.03 µM), TNFα (160.20 ± 17.57 pg/ml), and IL-1ß (285.79 ± 15.16 pg/ml) significantly (p < 0.05); when compared to the levels of NO (0.774 ± 0.08 µM), TNFα (501.71 ± 25.14 pg/ml), and IL-1ß (712.68 ± 52.25 pg/ml) from LPS-stimulated macrophage cells. The active compound was confirmed as hesperidin with NMR, IR, and UV spectroscopy data. This is the first report of this compound from Delonix elata flowers. CONCLUSION: The findings of the study support the traditional use of Delonix elata flowers to treat inflammation.


Asunto(s)
Antiinflamatorios no Esteroideos/aislamiento & purificación , Fabaceae/química , Hesperidina/aislamiento & purificación , Macrófagos/efectos de los fármacos , Extractos Vegetales/química , Animales , Antiinflamatorios no Esteroideos/farmacología , Antiinflamatorios no Esteroideos/toxicidad , Bioensayo , Línea Celular , Fraccionamiento Químico , Relación Dosis-Respuesta a Droga , Flores/química , Hesperidina/farmacología , Hesperidina/toxicidad , Interleucina-1beta/metabolismo , Macrófagos/inmunología , Macrófagos/metabolismo , Ratones , Óxido Nítrico/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo
9.
Inflamm Res ; 63(6): 451-62, 2014 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-24492951

RESUMEN

OBJECTIVE AND DESIGN: Rheumatoid arthritis is a chronic inflammatory and autoimmune disease that leads to aggressive joint cartilage and bone destruction. Swertiamarin is a secoiridoid glycoside found in Enicostema axillare (Lam) A. Raynal, a medicinal plant used in the Indian system of traditional medicine. In the present study, the potential of swertiamarin was evaluated in IL-1ß induced fibroblast-like synoviocytes (FLS). METHODS: The FLS were isolated from Freund's Complete Adjuvant induced arthritic (AA) rats and cultured with IL-1ß. The normal FLS and AA-FLS were cultured and used for subsequent experiment in fibroblastic morphology form. The efficacy of swertiamarin (10-50 µg/ml) was evaluated on mRNA and protein expression levels of inflammatory and osteoclastogenesis mediators. The efficacy was also evaluated on p38 MAPKα levels with time course studies (2, 4, 6, 8 and 12 h). RESULTS: IL-1ß induced cell proliferation (149.46 ± 13.73 %) and NO production (162.03 ± 11.03%) in AA-FLS; treatment with swertiamarin controlled proliferation (82.77 ± 4.22%) and NO production (82.06 ± 3.91% at 50 µg/ml) in a dose-dependent manner. It also significantly (P < 0.05) modulated the expression of apoptotic marker (caspase 3), proinflammation mediators (TNFα, IL-6, PGE2, COX-2, iNOS, MMPs) and osteoclastogenic mediator (RANKL) at both the mRNA and protein levels. Treatment with swertiamarin inhibited the levels of p38 MAPKα in a dose-dependent manner and also significantly (P < 0.05) attenuated the release of the same in time dependent mode. CONCLUSION: These findings suggest that treatment with swertiamarin attenuated IL-1ß induced FLS, and it revealed anti-inflammatory potential by attenuating aggressive FLS.


Asunto(s)
Antiinflamatorios/farmacología , Artritis Experimental/inmunología , Artritis Reumatoide/inmunología , Glucósidos Iridoides/farmacología , Pironas/farmacología , Animales , Apoptosis/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Citocinas/genética , Citocinas/inmunología , Fibroblastos/citología , Fibroblastos/efectos de los fármacos , Fibroblastos/inmunología , Ratones Endogámicos C57BL , Óxido Nítrico/inmunología , Osteoclastos/citología , Ligando RANK/inmunología , Ratas Sprague-Dawley , Membrana Sinovial/citología , Proteínas Quinasas p38 Activadas por Mitógenos/inmunología
10.
Inflamm Res ; 63(2): 127-38, 2014 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-24202617

RESUMEN

INTRODUCTION: The leaves of Clerodendrum phlomidis L.f. have been used in the Indian traditional system of medicine to treat several inflammatory diseases and arthritis. The aim of the present study was to assess the anti-inflammatory and anti-arthritic activities of the leaves of C. phlomidis and to isolate the active principle by bioactivity guided fractionation. MATERIALS AND METHODS: To find the anti-inflammatory constituents from this plant, fractionations were performed with concurrent bioassays. Carrageenan-induced inflammation and Freund complete adjuvant (FCA)-induced arthritic rat models were used. The anti-inflammatory and anti-arthritic activities of the isolated compound were studied by assessing the histology of the joints, levels of lysosomal enzymes, protein-bound carbohydrates, acute phase protein, etc., in plasma, as well as by estimating the levels and expression of pro-inflammatory cytokines in the joints. RESULTS: Repeated fractionations and bioassays yielded a novel bioactive compound: 3-hydroxy, 2-methoxy-sodium butanoate. Treatment with this compound reduced the paw edema induced by carrageenan and FCA dose dependently. The levels of lysosomal enzymes and protein-bound carbohydrates decreased significantly upon treatment with the compound. The level of plasma acute phase protein was also decreased compared with control animals. Protein levels and mRNA expression of pro-inflammatory cytokines TNF, IL-1 and IL-6 in the joints were decreased significantly in a dose-dependent manner and the histopathological data also added evidence of the anti-arthritic property of the compound. CONCLUSION: The 3-hydroxy,2-methoxy sodium butanoate isolated from plant leaves displays considerable potency in anti-inflammatory action and has a prominent anti-arthritic effect. This is the first report of this natural compound with bioactivity.


Asunto(s)
Artritis Experimental/tratamiento farmacológico , Butiratos/uso terapéutico , Clerodendrum , Edema/tratamiento farmacológico , Fitoterapia , Animales , Antiinflamatorios/farmacología , Antiinflamatorios/uso terapéutico , Artritis Experimental/inmunología , Artritis Experimental/patología , Butiratos/farmacología , Carragenina , Citocinas/genética , Citocinas/inmunología , Edema/inducido químicamente , Edema/inmunología , Edema/patología , Femenino , Articulaciones del Pie/inmunología , Articulaciones del Pie/patología , India , Medicina Tradicional de Asia Oriental , Neutrófilos/inmunología , Peroxidasa/inmunología , Extractos Vegetales/química , Hojas de la Planta/química , ARN Mensajero/metabolismo , Ratas , Ratas Wistar , Especies Reactivas de Oxígeno/inmunología
11.
Dig Dis Sci ; 59(6): 1121-33, 2014 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-24395383

RESUMEN

BACKGROUND AND AIM: Probiotics are live microorganisms that confer a health benefit on the host when administered in adequate amounts. In the present study, the putative probiotic strain was identified from the gut of Drosophila melanogaster and assessed for its protective effect in inflammatory bowel disease. METHODS: Active probiotics were screened from the Drosophila melanogaster gut by the selection criteria of gastric juice tolerance, hydrophobic property, antimicrobial potential, adhesion, and invasion properties. The active probiotics were identified by 16s rDNA sequencing and the effect of these active probiotics was evaluated in a Dextran sulphate sodium (DSS)-induced mice model by estimating inflammatory markers and histopathological changes. RESULTS: Nine Gram-positive and bile salt tolerant bacterial isolates were obtained from the gut samples. The isolates PTH 2, PTH 4, and PTH 7 clearly showed significant activity in antimicrobial potential, hydrophobic (>74%) property, and intestinal juice tolerance. Among these, PTH 7 was selected for further studies due to its significant low-invasion ability and it proved capable of reducing the secretion of proinflammatory cytokines. The 16s rDNA studies revealed that PTH 7 was Myroides pelagicus. Administration of M. pelagicus to the DSS-induced colitic animals significantly suppressed myeloperoxidase, ALP, and malondialdehyde levels, and also lowered levels of proinflammatory cytokine expression. Further, the recovery from the disease by the probiotic treatment was supported by histopathological and macroscopic observation. The treated animals did not show any adverse signs in their physiology or behavior. CONCLUSION: Myroides pelagicus successfully prohibited inflammatory markers and acted as a potent probiotic. Future studies with this stain might prove its efficacy as a drug for the management of colitis.


Asunto(s)
Colitis/terapia , Sulfato de Dextran/toxicidad , Drosophila melanogaster/microbiología , Flavobacteriaceae/fisiología , Tracto Gastrointestinal/microbiología , Probióticos/uso terapéutico , Animales , Peso Corporal , Células CACO-2 , Colitis/inducido químicamente , Colitis/patología , Colon/patología , Flavobacteriaceae/clasificación , Flavobacteriaceae/aislamiento & purificación , Humanos , Masculino , Ratones , Ratones Endogámicos C57BL , Probióticos/aislamiento & purificación
12.
World J Microbiol Biotechnol ; 30(1): 9-18, 2014 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-23821127

RESUMEN

Actinomycetes play an essential role in producing several bioactive compounds. In the present study, microbicidal and anti-inflammatory effects of metabolites from actinomycetes were investigated. Actinomycetes were isolated from north eastern Himalayan soil samples, India. The actinomycetes were investigated for their microbicidal property by conventional method and the active actinomycetes were identified by 16s rDNA sequence analyses. Further the metabolites were extracted and fractionated to evaluate the antimicrobial potency; they were subjected to GC-MS analysis. The active fraction was evaluated for selective toxicity and anti-inflammatory potential. Among isolated actinomycetes, EHA-2 showed potent antimicrobial activity and was identified as Actinomadura spadix. Fraction-8 from ethyl acetate extract of EHA-2 showed 100% inhibition against Candida sp. (MIC-80 µg/mL) and Enterococcus faecalis (MIC-80 µg/mL). The expression of GAPDH in primary cells and 16s rRNA levels on E. faecalis treated with fraction-8 revealed no toxicity to the primary cells. Fraction-8 also suppressed the paw thickness on carrageenan induced animals and also controlled the release of NO, TNFα and IL-1ß levels on LPS induced RAW 264.7 cell lines. GC-MS profile of fraction-8 showed the presence of an antimicrobial agent 3,6 di-isobutyl 2,5 piperazinedione, which is the first report in A. spadix. The actinomycetes isolate EHA-2 can be proceed further to produce antibiotics.


Asunto(s)
Actinomycetales/clasificación , Actinomycetales/metabolismo , Antiinfecciosos/metabolismo , Antiinflamatorios/metabolismo , Microbiología del Suelo , Actinomycetales/aislamiento & purificación , Animales , Antiinfecciosos/aislamiento & purificación , Antiinflamatorios/aislamiento & purificación , Bacterias/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , ADN Bacteriano/química , ADN Bacteriano/genética , Femenino , Hongos/efectos de los fármacos , Cromatografía de Gases y Espectrometría de Masas , Células Precursoras de Granulocitos , India , Macrófagos/efectos de los fármacos , Macrófagos/inmunología , Ratones , Pruebas de Sensibilidad Microbiana , Datos de Secuencia Molecular , ARN Ribosómico 16S/genética , Ratas Sprague-Dawley , Análisis de Secuencia de ADN
13.
Bioorg Med Chem Lett ; 23(6): 1839-45, 2013 Mar 15.
Artículo en Inglés | MEDLINE | ID: mdl-23395665

RESUMEN

Novel dispirooxindole-pyrrolidine derivatives have been synthesized through 1,3-dipolar cycloaddition of an azomethine ylide generated from isatin and sarcosine with the dipolarophile 3-(1H-indol-3-yl)-3-oxo-2-(2-oxoindolin-3-ylidene)propanenitrile, and also spiro compound of acenaphthenequinone obtained by the same optimized reaction condition. Synthesized compounds were evaluated for their antimicrobial activity and all the compounds shown significant activity. Anticancer activity was evaluated against A549 human lung adenocarcinoma cancer cell lines. Compounds 7b, 7g, 7i and 7r exhibit very good anticancer activity 62.96%, 62.03%, 67.67% and 60.22%, respectively, at the dose of 200µg/mL and compound 7i shows IC50 value in 50µg/mL.


Asunto(s)
Antiinfecciosos/síntesis química , Antineoplásicos/síntesis química , Indoles/química , Pirrolidinas/química , Adenocarcinoma/tratamiento farmacológico , Adenocarcinoma del Pulmón , Antiinfecciosos/química , Antiinfecciosos/farmacología , Antineoplásicos/uso terapéutico , Antineoplásicos/toxicidad , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Cristalografía por Rayos X , Bacterias Gramnegativas/efectos de los fármacos , Bacterias Grampositivas/efectos de los fármacos , Humanos , Indoles/farmacología , Indoles/toxicidad , Neoplasias Pulmonares/tratamiento farmacológico , Pruebas de Sensibilidad Microbiana , Conformación Molecular , Oxindoles , Pirrolidinas/farmacología , Pirrolidinas/toxicidad , Compuestos de Espiro/química , Relación Estructura-Actividad
14.
Bioorg Med Chem Lett ; 23(9): 2708-13, 2013 May 01.
Artículo en Inglés | MEDLINE | ID: mdl-23522833

RESUMEN

Cu(OTf)2 catalyzed efficient synthesis of spiropyrano[3,2-b]pyran-4(8H)-ones is accomplished via one-pot three component reaction between isatin, kojic acid and active methylenes. This synthetic protocol is operationally simple and affords product with good to excellent yields at a short reaction time. The synthesized compounds were evaluated for their tumor cell growth inhibitory activity against the human lung cancer cell line (A549) and found that 13 compounds exhibited moderate to good anticancer potency. Molecular docking studies were performed for all the synthesized compounds and the results showed that compound 4e showed greater affinity for anaplastic lymphoma kinase (ALK) receptor.


Asunto(s)
Antineoplásicos/síntesis química , Mesilatos/química , Piranos/química , Compuestos de Espiro/química , Quinasa de Linfoma Anaplásico , Antineoplásicos/química , Antineoplásicos/toxicidad , Sitios de Unión , Catálisis , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Cristalografía por Rayos X , Humanos , Isatina/química , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patología , Conformación Molecular , Simulación del Acoplamiento Molecular , Estructura Terciaria de Proteína , Piranos/síntesis química , Piranos/toxicidad , Pironas/química , Proteínas Tirosina Quinasas Receptoras/química , Proteínas Tirosina Quinasas Receptoras/metabolismo
15.
Plant Physiol Biochem ; 196: 724-730, 2023 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-36812799

RESUMEN

Genome editing tools based on CRISPR/Cas system have been posed to solve many issues in agriculture and improve food production. Genetic engineering by Agrobacterium-mediated transformation has helped to impart specific traits straightaway in many crops. Many GM crops have also reached the field for commercial cultivation. Genetic engineering requires mostly a transformation protocol often mediated by Agrobacterium to insert a specific gene at a random locus. Genome editing with CRISPR/Cas system is a more precise technique for the targeted modification of genes/bases in the host plant genome. Unlike the conventional transformation system, wherein elimination of marker/foreign gene was possible only post-transformation, CRISPR/Cas system could generate transgene-free plants by delivering CRISPR/Cas reagents such as the Cas protein and guide RNAs gRNA(s) preassembled to form ribonucleoproteins (RNPs) into plant cells. CRISPR reagent delivery might be helpful to overcome issues with plants that are recalcitrant to Agrobacterium transformation and the legal hurdles due to the presence of the foreign gene. More recently, the grafting of wild-type shoots to transgenic donor rootstocks developed by the CRISPR/Cas system has reported transgene-free genome editing. CRISPR/Cas system also requires only a small piece of gRNA besides Cas9 or other effectors to target a specific region in the genome. So this system has been projected to be a key contributor to future crop breeding. In this article, we recap the main events of plant transformation, compare the difference between genetic transformation and CRISPR/Cas-mediated genome editing, and draw insights into the future application of the CRISPR/Cas system.


Asunto(s)
Sistemas CRISPR-Cas , Edición Génica , Edición Génica/métodos , Sistemas CRISPR-Cas/genética , Plantas Modificadas Genéticamente/genética , Agrobacterium/genética , Fitomejoramiento , Genoma de Planta/genética , Productos Agrícolas/genética
16.
Biotechnol Lett ; 34(6): 995-1002, 2012 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-22350290

RESUMEN

Fungal diseases damage crop plants and affect agricultural production. Transgenic plants have been produced by inserting antifungal genes to confer resistance against fungal pathogens. Genes of fungal cell wall-degrading enzymes, such as chitinase and glucanase, are frequently used to produce fungal-resistant transgenic crop plants. In this review, we summarize the details of various transformation studies to develop fungal resistance in crop plants.


Asunto(s)
Resistencia a la Enfermedad , Hongos/inmunología , Hongos/patogenicidad , Enfermedades de las Plantas/genética , Plantas Modificadas Genéticamente/enzimología , Glicósido Hidrolasas/genética , Glicósido Hidrolasas/metabolismo , Ingeniería Metabólica/métodos , Enfermedades de las Plantas/inmunología , Enfermedades de las Plantas/microbiología
17.
Pharm Biol ; 50(6): 698-706, 2012 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-22129271

RESUMEN

CONTEXT: Enicostema axillare A. Raynal (Gentianaceae) has been used in the traditional Indian system of medicine as a depurative and for the treatment of skin diseases, tumors, intermittent fever, and helminthiasis. OBJECTIVE: E. axillare was investigated for antimutagenic and antioxidant effects. MATERIALS AND METHODS: The antioxidant and antimutagenic activities of E. axillare fractions were determined by 2,2-diphenyl-1-picryl-hydrazyl (DPPH) radical scavenging assay and Ames test using Salmonella typhimurium tester strains TA98 and TA100 against direct-acting mutagens, such as sodium azide (NaN3), 4-nitro-O-phenylene diamine (NPD), and mutagen needing activation, such as 2-aminofluorene (2AF). Toxicity study of these fractions was also performed. RESULTS AND DISCUSSION: The ethyl acetate fraction showed maximum antimutagenic effect by 88.25 and 84.46% (preincubation) and 85.13 and 84.47% (coincubation) of inhibition against NaN3 and NPD-induced mutagenicity, respectively. Inhibition of S9-dependent mutagens such as 2AF was higher than direct-acting mutagens by the ethyl acetate fraction of E. axillare. It showed 90.25 and 92.00% of inhibition in the preincubation and coincubation experiments. The ethyl acetate fraction showed higher total antioxidant capacity (24.79 ± 0.29 µg) and low IC50 value for DPPH radical scavenging assay (192.27 ± 3.67 µg). The overall effect of E. axillare fractions was found to be in the order: ethyl acetate > methanol > hexane in these assays. In subacute toxicity study, with oral administration of these fractions, no marked biochemical and histopathologic changes were observed. CONCLUSION: The significant antimutagenic and antioxidant activities of E. axillare might provide a scientific validation for the traditional use of this plant.


Asunto(s)
Antimutagênicos/farmacología , Depuradores de Radicales Libres/farmacología , Gentianaceae/química , Extractos Vegetales/farmacología , Hojas de la Planta/química , Salmonella typhimurium/efectos de los fármacos , Salmonella typhimurium/genética , Animales , Antimutagênicos/efectos adversos , Antimutagênicos/química , Antimutagênicos/aislamiento & purificación , Flavonoides/análisis , Depuradores de Radicales Libres/efectos adversos , Depuradores de Radicales Libres/química , Depuradores de Radicales Libres/aislamiento & purificación , India , Masculino , Medicina Tradicional , Ratones , Microsomas Hepáticos/efectos de los fármacos , Microsomas Hepáticos/metabolismo , Pruebas de Mutagenicidad , Mutágenos/efectos adversos , Mutágenos/aislamiento & purificación , Mutágenos/metabolismo , Mutágenos/farmacología , Fenoles/análisis , Extractos Vegetales/efectos adversos , Extractos Vegetales/química , Extractos Vegetales/aislamiento & purificación , Ratas , Ratas Wistar , Solventes/química , Pruebas de Toxicidad Subaguda
18.
Int J Med Mushrooms ; 24(11): 35-47, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-36374947

RESUMEN

Non-alcoholic fatty liver disease (NAFLD) is one of the most frequent, chronic liver diseases worldwide and currently has no specific therapy. Our previous study indicated the anti-NAFLD effect of Macrocybe gigantea (Massee) Pegler & Lodge in high-fat diet-fed animals. This study aimed to isolate and identify the active hepatoprotective constituents from M. gigantea using fatty acid induced steatotic HepG2 cells as in vitro model. The effect of the test materials on the viability of HepG2 cells was analyzed using MTT assay. The HepG2 cells were treated with a mixture of palmitate-oleate to induce steatosis; after 24 h of treatment with the test materials, the intracellular lipid content was estimated using Oil Red O staining. The levels of transaminases were also estimated in the spent media. Bioassay-guided isolation of hepatoprotective constituents from M. gigantea yielded two compounds viz., ergosterol and linoleic acid; their structures were confirmed using spectroscopic data. Among these two compounds, ergosterol significantly lowered the levels of intracellular triglyceride content of fatty acid induced HepG2 cells; it also lowered the leakage of transaminases. The reductions caused by linoleic acid were not statistically significant at the tested concentrations. Detailed investigations on efficacy and safety of these compounds and M. gigantea might yield some useful leads for the management of NAFLD.


Asunto(s)
Agaricales , Enfermedad del Hígado Graso no Alcohólico , Animales , Humanos , Dieta Alta en Grasa , Ergosterol/farmacología , Ácidos Grasos/farmacología , Células Hep G2 , Ácido Linoleico/farmacología , Hígado , Transaminasas/farmacología , Agaricales/química
19.
Plant Cell Rep ; 30(9): 1759-70, 2011 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-21584677

RESUMEN

A new Agrobacterium-mediated transformation system was developed for finger millet using shoot apex explants. The Agrobacterium strain LBA4404 harboring binary vector pCAMBIA1301, which contained hygromycin phosphotransferase (hptII) as selectable marker gene and ß-glucuronidase (GUS) as reporter gene, was used for optimization of transformation conditions. Two finger millet genotypes, GPU 45 and CO 14, were used in this study. The optimal conditions for the Agrobacterium-mediated transformation of finger millet were found to be the co-cultivation of explants obtained on the 16th day after callus induction (DACI), exposure of explants for 30 min to agrobacterial inoculum and 3 days of co-cultivation on filter paper placed on medium supplemented with 100 µM acetosyringone (AS). Addition of 100 µM L: -cysteine in the selection medium enhanced the frequency of transformation and transgenic plant recovery. Both finger millet genotypes were transformed by Agrobacterium. A frequency of 19% transient expression with 3.8% stable transformation was achieved in genotype GPU 45 using optimal conditions. Five stably transformed plants were fully characterized by Southern blot analysis. A segregation analysis was also performed in four R(1) progenies, which showed normal Mendelian pattern of transgene segregation. The inheritance of transgenes in R(1) progenies was also confirmed by Southern blot analysis. This is the first report on Agrobacterium-mediated transformation of finger millet. This study underpins the introduction of numerous agronomically important genes into the genome of finger millet in the future.


Asunto(s)
Eleusine/genética , Brotes de la Planta/citología , Transformación Genética , Acetofenonas/metabolismo , Agrobacterium/genética , Agrobacterium/metabolismo , Biomarcadores , Southern Blotting , Medios de Cultivo/química , Cisteína/metabolismo , Eleusine/crecimiento & desarrollo , Eleusine/metabolismo , Genes Reporteros , Vectores Genéticos , Patrón de Herencia , Brotes de la Planta/genética , Brotes de la Planta/crecimiento & desarrollo , Técnicas de Embriogénesis Somática de Plantas/métodos , Plantas Modificadas Genéticamente/genética , Plantas Modificadas Genéticamente/crecimiento & desarrollo , Plantas Modificadas Genéticamente/metabolismo , Regeneración , Transgenes
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