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1.
Artigo em Inglês | MEDLINE | ID: mdl-33138766

RESUMO

Amongst the various diseases on global scale, the second leading cause of mortality and morbidity is ischemic stroke due to the unavailability of an effective therapy. With the growing occurrence and its related health risks along with the absence of effective therapeutics, the ischemic stroke demands the continued and intensive research to explore the effective and safe therapeutics. These therapies may positively affect the numerous pathways associated with neuroprotection thus, extending the advantages to a larger population of stroke patients. Several preclinical studies employing neuroprotectants have shown promising outcomes, but failed in clinical trials either because of the lack of safety or efficacy. The blood brain barrier (BBB) restricts delivery of various potent neuroprotectants to the specific areas of the brain. The application of nanovehicles for delivery of drugs in the brain however, could revolutionize the treatment of ischemic stroke. These nanovehicles loaded with the drug could readily traverse the BBB via carrier, receptor and adsorptive-mediated endocytosis into the brain without compromising the integrity of BBB. Recent advances in neuronanotherapeutics have resulted in the improved neuronal regeneration and recovery after the ischemic stroke. In this review, we have attempted to discuss unexploited neuronanotherapeutics potentials to treat and manage ischemic stroke.

3.
BMC Plant Biol ; 19(1): 274, 2019 Jun 24.
Artigo em Inglês | MEDLINE | ID: mdl-31234787

RESUMO

BACKGROUND: miRNAs are major regulators of gene expression and have proven their role in understanding the genetic regulation of biosynthetic pathways. Stevioside and rebaudioside-A, the two most abundant and sweetest compounds found in leaf extract of Stevia rebaudiana, have been used for many years in treatment of diabetes. It has been found that the crude extract is more potent than the purified extract. Stevioside, being accumulated in higher concentration, imparts licorice like aftertaste. Thus, in order to make the sweetener more potent and palatable, there is a need to increase the intrinsic concentration of steviol glycosides and to alter the ratio of rebaudioside-A to stevioside. Doing so would significantly increase the quality of the sweeteners, and the potential to be used on a wider scale. To do so, in previous report, miRNAs associated with genes of steviol glycosides biosynthetic pathway were identified in S. rebaudiana. In continuation to that in this study, the two miRNAs (miR319g and miRStv_11) targeting key genes of steviol glycosides biosynthetic pathway were modulated and their impact was evaluated on steviol glycosides contents. RESULTS: The over-expression results showed that miRStv_11 induced, while miR319g had repressive action on its target genes. The knock-down constructs for miR319g and miRStv_11 were then prepared and it was demonstrated that the expression of anti-miR319g produced inhibitory effect on its target miRNA, resulting in enhanced expression of its target genes. On the other hand, anti-miRStv_11 resulted in down-regulation of miRStv_11 and its target gene. Further miRStv_11 and anti-miR319gwere co-expressed which resulted in significant increase in stevioside (24.5%) and rebaudioside-A (51%) contents. CONCLUSION: In conclusion, the role of miR319g and miRStv_11 was successfully validated in steviol gycosides biosynthetic pathway gene regulation and their effect on steviol gycosides contents. In this study, we found the positively correlated miRNA-mRNA interaction network in plants, where miRStv_11 enhanced the expression of KAH gene. miRNAs knock-down was also successfully achieved using antisense precursors. Overall, this study thus reveals more complex nature and fundamental importance of miRNAs in biosynthetic pathway related gene networks and hence, these miRNAs can be successfully employed to enhance the ratio of rebaudioside-A to stevioside, thus enhancing the sweetening indices of this plant and making it more palatable.


Assuntos
Diterpenos de Caurano/biossíntese , Glucosídeos/biossíntese , MicroRNAs/metabolismo , RNA de Plantas/metabolismo , Stevia/metabolismo , Diterpenos de Caurano/química , Diterpenos de Caurano/genética , Regulação da Expressão Gênica de Plantas , Técnicas de Inativação de Genes , Inativação Gênica , Glucosídeos/química , Glucosídeos/genética , MicroRNAs/genética , Folhas de Planta/química , Regiões Promotoras Genéticas , RNA de Plantas/genética , Stevia/genética , Edulcorantes/química
4.
Nat Prod Res ; 31(14): 1713-1716, 2017 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-28278673

RESUMO

Determination of endogenous levels of jasmonic acid (JA) is essential, as it plays a pivotal role in the physiological processes during a plant's life cycle. A high performance thin layer chromatography (HPTLC) method was developed for the detection and quantification of JA in leaf extracts of medicinal plant, Stevia rebaudiana (Bertoni) Bertoni. The separation was achieved using the solvents ethyl acetate-benzene (1:1, v/v) as the mobile phase, followed by scanning and quantification at 295 nm. Densitometric analysis of leaf extract resulted in compact spots for JA at Rf = 0.45 ± 0.02. The linear regression analysis showed good relationship with r value. The recovery rate of JA indicated good reproducibility and repeatability of the assay. The statistical analysis proved the reproducibility of the method; therefore, it can be employed for routine quantification of JA in different tissue samples of S. rebaudiana and may also be extrapolated to other biological samples.


Assuntos
Ciclopentanos/análise , Oxilipinas/análise , Stevia/química , Acetatos , Benzeno , Cromatografia em Camada Delgada/métodos , Ciclopentanos/normas , Oxilipinas/normas , Extratos Vegetais/análise , Folhas de Planta/química , Reprodutibilidade dos Testes
5.
J Virol ; 91(6)2017 03 15.
Artigo em Inglês | MEDLINE | ID: mdl-28053106

RESUMO

Japanese encephalitis virus (JEV), a mosquito-borne flavivirus, is the leading cause of viral encephalitis in Southeast Asia with potential to become a global pathogen. Here, we identify glucose-regulated protein 78 (GRP78) as an important host protein for virus entry and replication. Using the plasma membrane fractions from mouse neuronal (Neuro2a) cells, mass spectroscopy analysis identified GRP78 as a protein interacting with recombinant JEV envelope protein domain III. GRP78 was found to be expressed on the plasma membranes of Neuro2a cells, mouse primary neurons, and human epithelial Huh-7 cells. Antibodies against GRP78 significantly inhibited JEV entry in all three cell types, suggesting an important role of the protein in virus entry. Depletion of GRP78 by small interfering RNA (siRNA) significantly blocked JEV entry into Neuro2a cells, further supporting its role in virus uptake. Immunofluorescence studies showed extensive colocalization of GRP78 with JEV envelope protein in virus-infected cells. This interaction was also confirmed by immunoprecipitation studies. Additionally, GRP78 was shown to have an important role in JEV replication, as treatment of cells post-virus entry with subtilase cytotoxin that specifically cleaved GRP78 led to a substantial reduction in viral RNA replication and protein synthesis, resulting in significantly reduced extracellular virus titers. Our results indicate that GRP78, an endoplasmic reticulum chaperon of the HSP70 family, is a novel host factor involved at multiple steps of the JEV life cycle and could be a potential therapeutic target.IMPORTANCE Recent years have seen a rapid spread of mosquito-borne diseases caused by flaviviruses. The flavivirus family includes West Nile, dengue, Japanese encephalitis, and Zika viruses, which are major threats to public health with potential to become global pathogens. JEV is the major cause of viral encephalitis in several parts of Southeast Asia, affecting a predominantly pediatric population with a high mortality rate. This study is focused on identification of crucial host factors that could be targeted to cripple virus infection and ultimately lead to development of effective antivirals. We have identified a cellular protein, GRP78, that plays a dual role in virus entry and virus replication, two crucial steps of the virus life cycle, and thus is a novel host factor that could be a potential therapeutic target.


Assuntos
Vírus da Encefalite Japonesa (Espécie)/fisiologia , Proteínas de Choque Térmico/metabolismo , Interações Hospedeiro-Patógeno , Internalização do Vírus , Replicação Viral , Animais , Linhagem Celular , Humanos , Espectrometria de Massas , Camundongos , Microscopia Confocal , Microscopia de Fluorescência , Neurônios/virologia , Ligação Proteica , Proteínas do Envelope Viral/metabolismo
6.
Front Plant Sci ; 8: 2049, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29312363

RESUMO

The root-knot nematode (RKN), Meloidogyne incognita, is an obligate, sedentary endoparasite that infects a large number of crops and severely affects productivity. The commonly used nematode control strategies have their own limitations. Of late, RNA interference (RNAi) has become a popular approach for the development of nematode resistance in plants. Transgenic crops capable of expressing dsRNAs, specifically in roots for disrupting the parasitic process, offer an effective and efficient means of producing resistant crops. We identified nematode-responsive and root-specific (NRRS) promoters by using microarray data from the public domain and known conserved cis-elements. A set of 51 NRRS genes was identified which was narrowed down further on the basis of presence of cis-elements combined with minimal expression in the absence of nematode infection. The comparative analysis of promoters from the enriched NRRS set, along with earlier reported nematode-responsive genes, led to the identification of specific cis-elements. The promoters of two candidate genes were used to generate transgenic plants harboring promoter GUS constructs and tested in planta against nematodes. Both promoters showed preferential expression upon nematode infection, exclusively in the root in one and galls in the other. One of these NRRS promoters was used to drive the expression of splicing factor, a nematode-specific gene, for generating host-delivered RNAi-mediated nematode-resistant plants. Transgenic lines expressing dsRNA of splicing factor under the NRRS promoter exhibited upto a 32% reduction in number of galls compared to control plants.

7.
Syst Synth Biol ; 9(Suppl 1): 27-37, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26702306

RESUMO

Carotenoids represent a diverse group of pigments derived from the common isoprenoid precursors and fulfill a variety of critical functions in plants and animals. Phytoene synthase (PSY), a transferase enzyme that catalyzes the first specific step in carotenoid biosynthesis plays a central role in the regulation of a number of essential functions mediated via carotenoids. PSYs have been deeply investigated in plants, bacteria and algae however in apicomplexans it is poorly studied. In an effort to characterize PSY in apicomplexans especially the malaria parasite Plasmodium falciparum (P. falciparum), a detailed bioinformatics analysis is undertaken. We have analysed the Phylogenetic relationship of PSY also referred to as octaprenyl pyrophosphate synthase (OPPS) in P. falciparum with other taxonomic groups. Further, we in silico characterized the secondary and tertiary structures of P. falciparum PSY/OPPS and compared the tertiary structures with crystal structure of Thermotoga maritima (T. maritima) OPPS. Our results evidenced the resemblance of P. falciparum PSY with the active site of T. maritima OPPS. Interestingly, the comparative structural analysis revealed an unconserved unique loop in P. falciparum OPPS/PSY. Such structural insights might contribute novel accessory functions to the protein thus, offering potential drug targets.

8.
J Pharm Bioallied Sci ; 7(4): 250-3, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26681876

RESUMO

INTRODUCTION: Herbals drugs became a boon for mankind since ancient times and still are used worldwide for the treatment of various human ailments. The safety of alternative medicinal preparations has been questioned due to reports of unwanted side effects. To maintain the quality and efficacy of these drugs, it is essential to standardize them in order to avoid the use of substandard or adulterated medicines in the market. Unani system of medicine mainly focuses on the treatment by natural drugs. Habb-e-Banafsha Qawi is useful in a cough, catarrah, and coryza. MATERIALS AND METHODS: Physiochemical constants of Habb-e-Banafsha Qawi were determined through organoleptic characters, macro- and micro-scopic characters, ash value, solubility, pH values. Chromatographic fingerprints were developed using thin layer chromatography method. Aflatoxin (AF) contamination, heavy metal, and pesticide residues were also evaluated by standard methods. OBJECTIVES: In the present study, an attempt has been made to develop standard operating procedure and physiochemical parameters to monitor the quality of a Unani poly-herbal formulation, Habb-e-Banafsha Qawi. RESULTS: The tablets tasted sweetish bitter with a pleasant odor, water soluble and acidic in nature. R f values were almost same in all the extracts. No AF, heavy metal, and pesticide residues were observed. CONCLUSIONS: It may be concluded that the values and chromatographic fingerprints obtained can be used for quality evaluation and to set new pharmacopoeial standards for the preparation of Habb-e-Banafsha Qawi.

9.
PLoS One ; 10(10): e0139666, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26436554

RESUMO

Seed weight and seed size both are quantitative traits and have been considered as important components of grain yield, thus identification of quantitative trait loci (QTL) for seed traits in lentil (Lens culinaris) would be beneficial for the improvement of grain yield. Hence the main objective of this study was to identify QTLs for seed traits using an intraspecific mapping population derived from a cross between L. culinaris cv. Precoz (seed weight-5.1g, seed size-5.7mm) and L. culinaris cv. L830 (seed weight-2.2g, seed size-4mm) comprising 126 F8-RILs. For this, two microsatellite genomic libraries enriched for (GA/CT) and (GAA/CTT) motif were constructed which resulted in the development of 501 new genomic SSR markers. Six hundred forty seven SSR markers (including 146 previously published) were screened for parental polymorphism and 219 (33.8%) were found to be polymorphic among the parents. Of these 216 were mapped on seven linkage groups at LOD4.0 spanning 1183.7cM with an average marker density of 5.48cM. Phenotypic data from the RILs was used to identify QTLs for the seed weight and seed size traits by single marker analysis (SMA) followed by composite interval mapping (CIM) which resulted in one QTL each for the 2 traits (qSW and qSS) that were co-localized on LG4 and explained 48.4% and 27.5% of phenotypic variance respectively. The current study would serve as a strong foundation for further validation and fine mapping for utilization in lentil breeding programs.


Assuntos
Mapeamento Cromossômico , Lens (Planta)/genética , Locos de Características Quantitativas , Cromossomos de Plantas , DNA de Plantas/genética , Genes de Plantas , Ligação Genética , Biblioteca Genômica , Repetições de Microssatélites , Tamanho do Órgão , Fenótipo , Polimorfismo Genético , Sementes
10.
Artigo em Inglês | MEDLINE | ID: mdl-26421051

RESUMO

A single dose (30 mg/kg body weight) of standardized sea buckthorn leaf extract (SBL-1), administered 30 min before whole body (60)Co-gamma-irradiation (lethal dose, 10 Gy), protected >90% of mice population. The purpose of this study was to investigate the mechanism of action of SBL-1 on jejunum and bone marrow, quantify key bioactive compounds, and analyze chemical composition of SBL-1. Study with 9-week-old inbred male Swiss albino Strain 'A' mice demonstrated that SBL-1 treatment before (60)Co-gamma-irradiation (10 Gy) significantly (p < 0.05) countered radiation induced decreases in jejunum crypts (1.27-fold), villi number (1.41-fold), villus height (1.25-fold), villus cellularity (2.27-fold), cryptal Paneth cells (1.89-fold), and Bcl2 level (1.54-fold). It countered radiation induced increases in cryptal apoptotic cells (1.64-fold) and Bax levels (1.88-fold). It also countered radiation (2 Gy and 3 Gy) induced bone marrow apoptosis (1.59-fold and 1.85-fold) and micronuclei frequency (1.72-fold and 2.6-fold). SBL-1 rendered radiation protection by promoting cryptal stem cells proliferation, by regulating apoptosis, and by countering radiation induced chromosomal damage. Quercetin, Ellagic acid, Gallic acid, high contents polyphenols, tannins, and thiols detected in SBL-1 may have contributed to radiation protection by neutralization of radiation induced oxidative species, supporting stem cell proliferation and tissue regeneration.

11.
Plant Physiol Biochem ; 94: 57-64, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-26042546

RESUMO

miRNAs are emerging as potential regulators of the gene expression. Their proven promising role in regulating biosynthetic pathways related gene networks may hold the key to understand the genetic regulation of these pathways which may assist in selection and manipulation to get high performing plant genotypes with better secondary metabolites yields and increased biomass. miRNAs associated with genes of steviol glycosides biosynthetic pathway, however, have not been identified so far. In this study miRNAs targeting genes of steviol glycosides biosynthetic pathway were identified for the first time whose precursors were potentially generated from ESTs and nucleotide sequences of Stevia rebaudiana. Thereafter, stem-loop coupled real time PCR based expressions of these miRNAs in different tissues of Stevia rebaudiana were investigated and their relationship pattern was analysed with the expression levels of their target mRNAs as well as steviol glycoside contents. All the miRNAs investigated showed differential expressions in all the three tissues studied, viz. leaves, flowers and stems. Out of the eleven miRNAs validated, the expression levels of nine miRNAs (miR319a, miR319b, miR319c, miR319d, miR319e, miR319f, miR319h, miRstv_7, miRstv_9) were found to be inversely related, while expression levels of the two, i.e. miR319g and miRstv_11 on the contrary, showed direct relation with the expression levels of their target mRNAs and steviol glycoside contents in the leaves, flowers and stems. This study provides a platform for better understanding of the steviol glycosides biosynthetic pathway and these miRNAs can further be employed to manipulate the biosynthesis of these metabolites to enhance their contents and yield in S. rebaudiana.


Assuntos
Diterpenos de Caurano/biossíntese , Regulação da Expressão Gênica de Plantas/fisiologia , Glicosídeos/biossíntese , MicroRNAs/biossíntese , RNA de Plantas/biossíntese , Stevia/metabolismo , Perfilação da Expressão Gênica , Glicosídeos/genética , MicroRNAs/genética , RNA de Plantas/genética , Stevia/genética
12.
Environ Toxicol ; 30(11): 1285-96, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-24771359

RESUMO

With the advancement of human race, different anthropogenic activities have heaped the environment with chemicals that can cause alteration in the immune system of exposed organism. As a first line of barrier, the evolutionary conserved innate immunity is crucial for the health of an organism. However, there is paucity of information regarding in vivo assessment of the effect of environmental chemicals on innate immunity. Therefore, we examined the effect of a widely used environmental chemical, Cr(VI), on humoral innate immune response using Drosophila melanogaster. The adverse effect of Cr(VI) on host humoral response was characterized by decreased gene expression of antimicrobial peptides (AMPs) in the exposed organism. Concurrently, a significantly decreased transcription of humoral pathway receptors (Toll and PGRP) and triglyceride level along with inhibition of antioxidant enzyme activities were observed in exposed organism. This in turn weakened the immune response of exposed organism that was manifested by their reduced resistance against bacterial infection. In addition, overexpression of the components of humoral immunity particularly Diptericin benefits Drosophila from Cr(VI)-induced humoral immune-suppressive effect. To our knowledge, this is the first report regarding negative impact of an environmental chemical on humoral innate immune response of Drosophila along with subsequent protection by AMPs, which may provide novel insight into host-chemical interactions. Also, our data validate the utility and sensitivity of Drosophila as a model that could be used for screening the possible risk of environmental chemicals on innate immunity with minimum ethical concern that can be further extrapolated to higher organisms.


Assuntos
Cromo/toxicidade , Drosophila melanogaster/efeitos dos fármacos , Poluentes Ambientais/toxicidade , Imunidade Humoral/efeitos dos fármacos , Imunidade Inata/efeitos dos fármacos , Animais , Antioxidantes/metabolismo , Cromo/farmacocinética , Relação Dose-Resposta a Droga , Drosophila melanogaster/genética , Drosophila melanogaster/imunologia , Poluentes Ambientais/farmacocinética , Expressão Gênica/efeitos dos fármacos , Humanos , Imunidade Humoral/genética , Imunidade Inata/genética , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/genética , Estresse Oxidativo/imunologia , Reação em Cadeia da Polimerase em Tempo Real , Testes de Toxicidade
13.
Appl Biochem Biotechnol ; 175(4): 2206-23, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25471016

RESUMO

Improved breeding for developing fruit quality in bottle gourd (Lagenaria siceraria (Mol.) Standl.) necessitates knowledge regarding its genetic diversity. To achieve this, a set of 108 locus-specific SSR markers has been developed in bottle gourd by cross-species transferability from 995 mapped Cucumis sativus SSR markers. During screening, 280 primer pairs amplified in the bottle gourd germplasm, which were further evaluated in a diverse set of 42 lines, resulting in 19 polymorphic, 89 monomorphic, 15 with multiple bands, and the rest 157 showed no or very non-specific amplification. The 19 polymorphic primer pairs produced a total of 54 alleles. Gene diversity, Shannon's information index, and Nei's coefficient of differentiation were calculated suggesting a moderate genetic variation at the species level. A model-based population structure analysis divided these germplasm into two subpopulations. This marker set will be applicable for evaluating the genetic structure for association mapping, DNA fingerprinting, and mounting linkage maps and will be a practical tool set for further genetics. This study provides one of the first quantitative views of population genetic variation in bottle gourd.


Assuntos
Cucurbitaceae/genética , DNA de Plantas , Genoma de Planta , Repetições de Microssatélites , Filogenia , Polimorfismo Genético , Alelos , Cruzamento , Mapeamento Cromossômico , Cucurbitaceae/classificação , Marcadores Genéticos , Genótipo , Índia , Filogeografia , Análise de Sequência de DNA , Especificidade da Espécie
14.
Mol Biochem Parasitol ; 197(1-2): 15-20, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25261593

RESUMO

Resistance to almost all class of antimalarial drugs has emerged as one of the greatest challenges to malaria control. Strategies to limit the advent and spread of drug-resistant malaria include development of new drugs and its combination with existing drugs. In this work we provide a strong evidence for phytonutrient lycopene, a non-provitamin A carotenoid, to be effective against Plasmodium falciparum growth in vitro. Consistent with the previous findings in mammalian cells, lycopene's prooxidant activity promoted the production of reactive oxygen species (ROS) in P. falciparum. Also a significant loss of mitochondrial functionality and thus, the loss of the membrane potential was observed in lycopene treated schizonts. Taken together, our results indicated that the generation of ROS and loss of mitochondrial membrane potential accounted for lycopene's cytotoxicity against P. falciparum growth in vitro. These insights will help in the design of new treatment strategies to combat malaria.


Assuntos
Antimaláricos/farmacologia , Carotenoides/farmacologia , Eritrócitos/parasitologia , Plasmodium falciparum/efeitos dos fármacos , Plasmodium falciparum/crescimento & desenvolvimento , Eritrócitos/metabolismo , Eritrócitos/patologia , Humanos , Estágios do Ciclo de Vida , Licopeno , Malária Falciparum/tratamento farmacológico , Malária Falciparum/parasitologia , Fatores de Tempo
15.
Pharmacogn Mag ; 10(Suppl 1): S198-205, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-24914304

RESUMO

BACKGROUND: Adhatoda vasica a perennial herb has been used in Ayurvedic and Unani system of medicines since last 2000 years and has been employed for the treatment of respiratory tract ailments. OBJECTIVE: To develop and validate new, rapid, and highly sensitive high throughput ultra-performance liquid chromatography/quadrupole-time-of-flight mass-spectrometry (UPLC/Q-TOF-MS) method for the quantitative estimation of vasicine in the leaves and to establish in vitro cultures of Adhatoda vasica for production of vasicine. MATERIALS AND METHODS: The chromatographic separation was achieved on a Waters ACQUITY UPLC™ BEH C8 (100.0 × 2.1 mm; 1.7 µm) column packing using isocratic mobile phase consisting of acetonitrile: 20 mM ammonium acetate (90:10; v/v) in a multiple reactions monitoring mode using the transitions m/z 189.09 → 171.08 for vasicine. RESULTS: The vasicine was eluted at 2.58 ± 0.05 min and established a dynamic range of linearity over the concentration range of 1-1000 ng/ml (r (2) = 0.999 ± 0.0005). The lower limit of detection and quantification was 0.68 and 1.0 ng/ml, respectively. There was no significant difference observed in the content of vasicine (0.92-1.04%w/w) among the eleven samples collected from different locations of India. The in vitro cultures developed showed that addition of extra 28 mM KNO3 and 100 mM NaCl in MS medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) + benzyladenine (BA) + indole acetic acid (IAA) (1 ppm each) produces faster biomass and higher amount of quinazoline alkaloids. CONCLUSION: Rapid, efficient, and sensitive UPLC/Q-TOF-MS method was developed for the estimation of vasicine and an efficient protocol for development of in vitro cultures was proposed, which can be used at large scale for industrial production of vasicine using bioreactors.

16.
Mol Biol Rep ; 41(9): 5607-25, 2014 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-24893599

RESUMO

Lentil (Lens culinaris Medik.) is an economically important grain legume, yet the genetic and genomic resources remain largely uncharacterized and unexploited in this crop. Microsatellites have become markers of choice for crop improvement applications. Hence, simple sequence repeat (SSR) markers were developed for lentil through the construction of genomic library enriched for GA/CT motifs. As a result 122 functional SSR primer pairs were developed from 151 microsatellite loci and validated in L. culinaris cv. Precoz. Thirty three SSR markers were utilized for the analysis of genetic relationships between cultivated and wild species of Lens and related legumes. A total of 123 alleles were amplified at 33 loci ranging from 2-5 alleles with an average of 3.73 alleles per locus. Polymorphic information content (PIC) for all the loci ranged from 0.13 to 0.99 with an average of 0.66 per locus. Varied levels of cross genera transferability were obtained ranging from 69.70 % across Pisum sativum to 12.12 % across Vigna radiata. The UPGMA based dendrogram was able to establish the uniqueness of each genotype and grouped them into two major clusters clearly resolving the genetic relationships within lentil and related species. The new set of SSR markers reported here were efficient and highly polymorphic and would add to the existing repertoire of lentil SSR markers to be utilized in molecular breeding. Moreover, the improved knowledge about intra- and inter-specific genetic relationships would facilitate germplasm utilization for lentil improvement.


Assuntos
Variação Genética , Genoma de Planta , Lens (Planta)/classificação , Lens (Planta)/genética , Repetições de Microssatélites , Alelos , Primers do DNA , DNA de Plantas/genética , Loci Gênicos , Biblioteca Genômica , Genômica , Família Multigênica , Polimorfismo Genético , Análise de Sequência de DNA , Especificidade da Espécie
17.
Neurobiol Aging ; 35(10): 2419.e1-2419.e16, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-24819147

RESUMO

Parkinson's disease (PD) is a prevalent and devastating neurodegenerative disorder having limited cure options and strong association with the loss of dopaminergic neurons in the substantia nigra region of the mid brain. Etiology of PD includes both genetic and environmental factors. Paraquat (PQ), a widely used herbicide, is known to be associated with pathogenesis of PD. We report that a mutation in Drosophila methuselah (mth(1)), which is associated with aging, has a role in preventing dopaminergic neuronal cell death in PQ-exposed organism. Exposed mth(1) flies exhibit significant resistance against PQ-induced Parkinson's phenotypes and behavior in terms of oxidative stress, dopaminergic neuronal degeneration, locomotor performance, dopamine content, phosphorylated JNK, pFOXO, Hid, and cleaved caspase-3 levels. Conversely, over-expression of mth in dopaminergic neurons makes the exposed organism more vulnerable to oxidative stress, neuronal cell death, and behavioral deficit. The study suggests that lesser activation of JNK-mediated apoptosis in dopaminergic neurons of exposed mth(1) flies protects the organism from PQ-induced damage, which may be causally linked to a common mechanism for PQ-induced neurodegeneration.


Assuntos
Drosophila/genética , Resistência a Herbicidas/genética , Herbicidas/efeitos adversos , Mutação , Paraquat/efeitos adversos , Doença de Parkinson Secundária/induzido quimicamente , Doença de Parkinson Secundária/genética , Doença de Parkinson Secundária/prevenção & controle , Fenótipo , Envelhecimento , Animais , Apoptose , Caspase 3/metabolismo , Dopamina/metabolismo , Neurônios Dopaminérgicos/patologia , Proteínas de Drosophila/metabolismo , Fatores de Transcrição Forkhead/metabolismo , MAP Quinase Quinase 4/metabolismo , Masculino , Atividade Motora , Degeneração Neural/prevenção & controle , Neuropeptídeos/metabolismo , Estresse Oxidativo , Receptores Acoplados a Proteínas-G/metabolismo
18.
Chem Biol Interact ; 214: 33-40, 2014 May 05.
Artigo em Inglês | MEDLINE | ID: mdl-24565947

RESUMO

Glycine propionyl l-carnitine (GPLC) is a propionyl ester of carnitine that includes an additional glycine component. The present study evaluated hepatoprotective effect of GPLC in d-Galactosamine (d-GalN) induced fulminant hepatic failure. Rats were intraperitonially administered d-GalN (700mg/kgBW). GPLC was given as a pre-treatment (35mg/kgBW/day) for 1month followed by a single dose of d-GalN on the 31st day. d-GalN administration resulted in increased mortality and serum ALT and AST activities. These increases were significantly attenuated by GPLC. d-GalN treatment increased hepatic lipid peroxidation and a decrease in reduced glutathione content was observed. GPLC pre-treatment significantly decreased lipid peroxidation and augmented the level of GSH. d-GalN increased the circulating level of TNF-α and ATM-Kinase and MAP-Kinase expression. GPLC supplementation inhibited the increase in serum TNF-α and ATM-Kinase and MAP-Kinase expression. d-GalN treatment increased the level of Bax and Caspase-3 m-RNA while as a decline was observed in Bcl2 m-RNA. GPLC prevented the increase in Caspase-3 and Bax m-RNA and at the same time augmented the expression of Bcl2 m-RNA. Our findings suggest that GPLC alleviates d-GalN induced liver injury by strengthening antioxidative defense system and reducing apoptotic signalling pathways.


Assuntos
Carnitina/análogos & derivados , Galactosamina/toxicidade , Glicina/análogos & derivados , Falência Hepática Aguda/prevenção & controle , Alanina Transaminase/sangue , Animais , Aspartato Aminotransferases/sangue , Proteínas Mutadas de Ataxia Telangiectasia/metabolismo , Sequência de Bases , Carnitina/farmacologia , Ensaio Cometa , Primers do DNA , Glutationa/sangue , Glicina/farmacologia , Peroxidação de Lipídeos , Falência Hepática Aguda/induzido quimicamente , Masculino , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Ratos , Ratos Wistar , Reação em Cadeia da Polimerase em Tempo Real , Fator de Necrose Tumoral alfa/sangue
19.
Biochem Biophys Res Commun ; 445(1): 138-44, 2014 Feb 28.
Artigo em Inglês | MEDLINE | ID: mdl-24491547

RESUMO

Morphological transition (yeast-hyphal and white-opaque) is an important biological process in the life cycle of pathogenic yeast, Candida albicans and is a major determinant of virulence. Earlier reports show that the amino sugar, N-acetylglucosamine (GlcNAc) induces white to opaque switching in this pathogen. We report here a new contributor to this switching phenomenon, namely N-acetylglucosamine kinase or HXK1, the first enzyme of the GlcNAc catabolic cascade. Microarray profile analysis of wild type vs. hxk1 mutant cells grown under switching inducing condition showed upregulation of opaque specific and cell wall specific genes along genes involved in the oxidative metabolism. Further, our qRT-PCR and immunoblot analysis revealed that the expression levels of Wor1, a master regulator of the white-opaque switching phenomenon remained unaltered during this HXK1 mediated transition. Thus the derepression of opaque specific gene expression observed in hxk1 mutant could be uncoupled to the expression of WOR1. Moreover, this regulation via HXK1 is independent of Ras1, a major regulator of morphogenetic transition and probably independent of MTL locus too. These results extend our understanding of multifarious roles of metabolic enzymes like Hxk1 and suggest an adaptive mechanism during host-pathogen interactions.


Assuntos
Candida albicans/genética , Proteínas Fúngicas/genética , Fosfotransferases (Aceptor do Grupo Álcool)/genética , Transcriptoma , Candida albicans/enzimologia , Candida albicans/fisiologia , Parede Celular/genética , Parede Celular/metabolismo , Proteínas Fúngicas/metabolismo , Regulação Fúngica da Expressão Gênica , Immunoblotting , Mutação , Fosfotransferases (Aceptor do Grupo Álcool)/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Regulação para Cima
20.
Mycotoxin Res ; 30(1): 25-32, 2014 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-24326591

RESUMO

The study was designed to investigate the protective effect of esculin against pro-oxidant aflatoxin B1 (AFB1)-induced nephrotoxicity in mice. In this study toxicity was developed by oral administration of AFB1 at a dose of 66.60 µg/kg bw/day for 90 days in male Swiss albino mice. Esculin (150 mg/kg bw/0.2 ml/day) and standard compound ascorbic acid (300 mg/kg bw/0.2 ml/day) was given after 30 min of AFB1 administration for 90 days. Protective efficacy was assessed by measuring the levels of lipid peroxidation (LPO) and non-enzymatic antioxidants such as reduced glutathione (GSH) and also by measuring activities of enzymatic antioxidants such as glutathione peroxidase (GPX), glutathione-S-transferase (GST), glutathione reductase (GR), superoxide dismutase (SOD) and catalase (CAT) in kidney. Results were analysed at the 30(th), 60(th) and 90(th) day of the daily treatments, which showed a decrease in the level of LPO and an increase in the levels of enzymatic and non-enzymatic antioxidants. The protective effect of esculin was further proved by histopathological findings as it exhibited regenerative activities in mice renal tubules against AFB1-induced nephrotoxicity. The results obtained clearly demonstrate that the protective efficacy of esculin against pro-oxidant AFB1-induced nephrotoxicity in mice might be due to its antioxidants and free radical scavenging properties.


Assuntos
Aflatoxina B1/antagonistas & inibidores , Aflatoxina B1/toxicidade , Antioxidantes/administração & dosagem , Esculina/administração & dosagem , Envenenamento/prevenção & controle , Administração Oral , Animais , Antioxidantes/análise , Ácido Ascórbico/administração & dosagem , Histocitoquímica , Rim/patologia , Peroxidação de Lipídeos , Masculino , Camundongos
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