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1.
Int J Food Microbiol ; 388: 110083, 2023 Mar 02.
Artículo en Inglés | MEDLINE | ID: mdl-36708610

RESUMEN

This is proven for a long that the incorporation of probiotics and prebiotics in diet exhibits beneficial effects on intestinal and intrinsic health. Nevertheless, this may encounter loss of vitality all along the absorption in the gastrointestinal tract, leading to meager intestinal delivery of probiotic active ingredients. In recent times, nanotechnology has been passionately used to escalate the bioavailability of active ingredients. Versatile forms of nanoparticles (NPs) are devised to be used with probiotics/prebiotics/synbiotics or their different combinations. The NPs currently in trend are constituted of distinctive organic compounds like carbohydrates, proteins, fats, or inorganics such as oxides of silver and titanium or magnesium etc. This review critically explicates the emerging relationship of nanotechnology with probiotics and prebiotics for different applications in neutraceuticals. Here in this review, formulations of nanoprobiotics and nanoprebiotics are discussed in detail, which behave as an effective drug delivery system. In addition, these formulations exhibit anti-cancerous, anti-microbial, anti-oxidant and photo-protective properties. Limited availability of scientific research on nanotechnology concerning probiotics and prebiotics implies dynamic research studies on the bioavailability of loaded active ingredients and the effective drug delivery system by including the safety issues of food and the environment.


Asunto(s)
Probióticos , Simbióticos , Prebióticos , Probióticos/metabolismo , Nanotecnología , Dieta
2.
J Food Sci Technol ; 58(8): 3019-3029, 2021 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-34294964

RESUMEN

The effect of photoperiod durations (16 h light:8 h dark vs 22 h light:2 h dark) and different doses (0.5x and 1x) of Murashige and Skoog medium on the yield and antioxidant characteristics of wheatgrass from hard, medium-hard and soft wheat varieties were analyzed. The average wheatgrass height and wheatgrass yield increased in MS media both under normal photoperiod as well as in water under prolonged photoperiod. An increase in total phenolic content (TPC) and ferric reducing antioxidant power (FRAP) of wheatgrass in different strengths of MS media under normal photoperiod was observed. Whereas, increase in protein content, chlorophyll (Chl) a, Chl b, total Chl, average TPC, DPPH inhibition and FRAP values were observed for wheatgrass of different varieties grown in water under prolonged photoperiod. The accumulation of polypeptides (PPs) of 92 kDa, 33 kDa, 23 kDa, 14 kDa, 12 kDa, and 10 kDa for wheatgrass shoot powder of different varieties was affected by strength of MS media and duration of photoperiod. On the contrary, wheatgrass juice powder showed major changes in the accumulation of PPs 33 kDa and 23 kDa PPs under varied strength of MS media and prolonged photoperiod.

3.
Food Chem ; 345: 128861, 2021 May 30.
Artículo en Inglés | MEDLINE | ID: mdl-33348134

RESUMEN

Effect of different degrees of milling (DoM: 2%, 4%, 6% and 8%) and defatting on the proximate composition, protein characteristics, functional properties of bran of long grain rice cultivars and texture characteristics of bran (RB) supplemented muffins were evaluated. Protein, ash content, redness and yellowness increased while fat content decreased for RB by extended DoM and defatting. A higher proportion of ß sheets, random coils, α-helix and ß-turns for all fractions of RB of both cultivars after defatting were also observed. Defatting and extended DoM both improved the essential amino acid content in RB. A higher level of prolamines (15-18 kDa) in RB and DF-RB of PUSA1121 than PR111 was observed. Muffins made from 2% DoM bran from PUSA1121 showed improved texture characteristics and achieved the highest score for sensory attributes. Therefore, DoM and defatting improved the proximate, protein profiling, and functionality of the different fraction of RB.


Asunto(s)
Dieta Sin Gluten , Ácidos Grasos/aislamiento & purificación , Manipulación de Alimentos/métodos , Oryza/química , Calidad de los Alimentos
4.
Food Chem ; 344: 128725, 2021 May 15.
Artículo en Inglés | MEDLINE | ID: mdl-33279352

RESUMEN

The physicochemical and functional characteristics of grain, meal and flour of timely sown wheat (TSW) and delayed sown wheat (DSW) were compared to see the effects of heat stress (HS). TSW and DSW of different lines were sown as per the approved timings. DSW experienced higher temperature during flowering and had shorter vegetative and maturation period than TSW. Pasting and dough rheological properties were measured using Rapid Visco-Analyser and Farinograph, respectively, while gliadins and glutenins profiling was done by SDS-PAGE. Delayed sowing decreased grain yield and diameter while increased protein and all categories of gliadins and high molecular weight glutenins. DSW showed higher peak viscosity, breakdown-viscosity and dough stability and lower setback viscosity, damaged starch, arabinoxylans and water absorption than TSW. HS in DSW appeared to lower starch synthesis causing proportionate increase in grain hardness and proteins content leading to changes in milling and rheological characteristics.


Asunto(s)
Fenómenos Químicos , Harina/análisis , Respuesta al Choque Térmico , Triticum/química , Granos Enteros/química , Glútenes/química , Dureza , Reología , Almidón/química , Viscosidad , Agua/química
5.
J Food Sci ; 85(1): 65-76, 2020 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-31869859

RESUMEN

The effect of lipids extraction on protein salvation, pasting, and dough rheological behavior of flours dough from hard wheat (HW), extraordinarily soft wheat (Ex-SW), and medium-hard wheat (MHW) flour was analyzed. The varieties selected had wide variation in grain hardness index (17 to 95). Ex-SW revealed lower tryptophan fluorescent emission and water absorption (WA) than MHW and HW varieties. The change in pasting parameters on defatting was the highest for Ex-SW varieties. Native flour (NF) of HW varieties showed high protein content, pasting and dough strength, and fluorescence intensity in comparison to Ex-SW varieties, while on defatting this was reversed. Protein pattern of defatted flour and NF did not differ significantly. Defatting of flours increased WA and decreased dough stability (DS). The decrease in DS on defatting was more for Ex-SW varieties than others. DS for HW and MHW varieties reduced upon defatting. Defatting significantly increased all mixographic and rheological properties except peak time. Overall, results showed that defatting of flour improved paste and dough strength. PRACTICAL APPLICATION: Higher gluten strength of defatted wheat flour of varied grain hardness has wide applications. Most of the Indian bread wheat varieties possess GluD1 high-molecular-weight glutenin subunit (HMW-GS) allelic composition of (2 + 12), which is not suitable for bread making due to weak gluten strength. Defatting of flours improved the gluten strength of strong and weak flour dough of different wheat varieties irrespective of GluD1 allelic composition for HMW-GS. Defatted flour may be used to improve the baking and cooking performance of dough made from weak wheat. These findings are highly suitable for wheat milling and baked product manufacturing industries.


Asunto(s)
Harina/análisis , Manipulación de Alimentos/métodos , Triticum/química , Pan/análisis , Culinaria , Glútenes/química , Dureza , Reología , Triticum/clasificación
6.
Food Res Int ; 126: 108675, 2019 12.
Artículo en Inglés | MEDLINE | ID: mdl-31732053

RESUMEN

Brown rice of different long-grain Indica cultivars was polished to variable degree of milling (DoM) to see the difference in proteins and starches characteristics in head (HR) and broken rice (BR). Study revealed differential accumulation of starch, fat and proteins in both HR and BR. Extended DoM of brown rice resulted in a progressive decrease in HR yield and increase in BR yield. The extended DoM caused a decrease in protein and fat content in both HR and BR, whereas; an increase in peak viscosity and final viscosity was observed. On the contrary, the setback viscosity of HR and BR of different rice cultivars was influenced by cultivars and extended DoM. Milled rice from different cultivars milled to 6% DoM showed higher levels of 59 kDa, 54 kDa, 51 kDa, 32 kDa, 31 kDa, 30 kDa, 28 kDa, 24 kDa, 23 kDa, 15 kDa, 13 and 12 kDa PPs, while 28 kDa, 24 kDa, 23 kDa, 15 kDa, 13 kDa and 12 kDa PPs was the least or not observed in BR. The major quantitative changes were observed in 28 kDa, 24 kDa, and 23 kDa PPs. MALDI-ToF/MS analysis revealed the identity of 28 kDa PP as 60S ribosomal protein L10a and glutelin type-D 1 proteins. Whereas, the identity of 24 and 23 kDa PP, respectively was established as pathogenesis-related protein 1 (Oryza sativa Japonica Group) and Oryza sativa 1-Cys peroxiredoxin A. HR showed the presence of highly condensed packaged starch granules with smooth edges, which were tightly imbibed in the proteins matrix. However, the inter-cultivar differences in the starch structure and packaging were also observed. On the contrary, BR revealed lesser accumulation of starch particles with abnormal protein filling and several fissures and cracks in the starch granules of BR of different cultivars.


Asunto(s)
Oryza/química , Proteínas de Plantas/análisis , Proteínas de Plantas/química , Almidón/análisis , Almidón/química , Manipulación de Alimentos , India , Microscopía Electrónica de Rastreo , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción
7.
Food Res Int ; 123: 327-339, 2019 09.
Artículo en Inglés | MEDLINE | ID: mdl-31284983

RESUMEN

The objective of this work was to evaluated the effects of debranning (2% and 4%) of Indian and exotic wheat varieties on pasting, protein profile and rheological properties of meal. Exotic varieties grains showed significant difference in cooking properties despite having similar protein content. Debranned grains of exotic varieties showed significantly lower water uptake (WU) than those from Indian varieties. Ash content, protein content and mixographic properties (mixing time, dough strength) of meal decreased with increase in extent of debranning. With increase in debranning level unextractable polymeric proteins (UnEx-PP) decreased while unextractable monomeric proteins (UnEx-MP) increased. Exotic varieties had lower extractable PP as compared to Indian wheat varieties. With increase in debranning level pasting parameters (final, breakdown and peak viscosity) increased. Exotic varieties showed the higher gluten strength indicated by mixographic parameters. Wheat grains debranned to 4% debranning level showed better cooking properties. Exotic wheat varieties showed better pasting and mixing properties as compared to Indian wheat varieties.


Asunto(s)
Grano Comestible/química , Glútenes/análisis , Triticum/química , Triticum/clasificación , Pan/análisis , Fenómenos Químicos , Color , Culinaria , Fibras de la Dieta/análisis , Electroforesis en Gel de Poliacrilamida , Harina/análisis , India , Proteínas de Plantas/química , Reología , Viscosidad , Agua
8.
J Food Sci Technol ; 55(11): 4661-4674, 2018 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-30333663

RESUMEN

In the present study coarse fraction (CF), medium fine fraction (MFF) and fine fraction (FF) were separated from flours milled from medium-hard and extraordinarily soft wheat varieties and were evaluated for various quality characteristics. Grain hardness of medium-hard and extraordinarily soft wheat varieties varied from 77 to 80 and 17 to 18, respectively. Ash and protein content was the highest for FF and the lowest for CF. Varieties with greater hardness produced higher CF and lower of FF. FF showed higher unextractable polymeric protein (UnEx-PP) and dough stability as compared to MFF and CF. FF showed lower damage starch content as related by lower Sodium SRC (NaSRC) as compared to MFF and FF. CF showed higher paste viscosities than FF and difference were greater amongst fractions from varieties with lower grain hardness. FF with greater proportion of small size particles showed greater accumulation of 98 kDa and 85 kDa PPs than CF. This study demonstrated that fractionation of flours can be employed to produce fractions with varied gluten strength required for production of various products.

9.
J Food Sci Technol ; 55(4): 1256-1269, 2018 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-29606740

RESUMEN

The particle size distribution, protein profile, pasting and dough rheological properties of meal from forty-two Indian durum wheat accessions were evaluated. Meal from accessions with higher grain hardness index (GHI) showed a high proportion of large size particles with higher protein content and lower paste viscosities. Elastic and viscous modulii (G' and G″) of dough were negatively correlated with paste viscosities, which was associated with the presence/absence of LMW-GS and HMW-GS. Wheat accessions with allelic combinations of (13 + 16) with 97 + 91 kDa polypeptides (PPs) had higher G' and G″. The accession with 35 kDa PP showed higher while those with 35 and 62 kDa PPs showed lower paste viscosity. Among all accessions, 25 accessions possess 7 + 8 (97 and 88 kDa) type HMW-GS allelic combination. Durum accessions with diverse GHI, particle size distribution, protein profile, paste and dough rheology indicates their variation in milling and processing behaviour.

10.
Food Res Int ; 100(Pt 2): 306-317, 2017 10.
Artículo en Inglés | MEDLINE | ID: mdl-28888455

RESUMEN

Hard wheat (HW), medium-hard wheat (MHW) and extraordinarily soft wheat (Ex-SW) varieties with grain hardness index (GHI) of 83 to 95, 72 to 80, 17 to 29 were evaluated for pasting, protein molecular weight (MW) distribution, dough rheology and baking properties. Flours from varieties with higher GHI had more protein content, ash content and paste viscosities. Ex-SW had more glutenins proportion as compared to HW and MHW. Flours from Ex-SW varieties showed lower NaSRC, WA and mixographic parameters as compared to HW and MHW. Dough from flours milled from Ex-SW had higher Intermolecular-ß-sheets (IM-ß-sheets) than those from MHW and HW. Muffins volume increased with decrease in GHI, Ex-SW varieties had more muffin volume and less air space. The accumulation of polypeptides (PPs) varied significantly in different varieties. Ex-SW variety (QBP12-10) showed accumulation of 98, 90, 81 and 79kDa PPs, which was unique and was different from other varieties.


Asunto(s)
Pan/análisis , Manipulación de Alimentos , Glútenes/análisis , Proteínas de Plantas/análisis , Triticum/química , Grano Comestible/química , Harina/análisis , Dureza , India , Peso Molecular , Tamaño de la Partícula , Reología , Viscosidad
11.
Biochimie ; 137: 115-123, 2017 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-28322928

RESUMEN

Imposition of different biotic and abiotic stress conditions results in an increase in intracellular levels of Ca2+ which is sensed by various sensor proteins. Calmodulin (CaM) is one of the best studied transducers of Ca2+ signals. CaM undergoes conformational changes upon binding to Ca2+ and interacts with different types of proteins, thereby, regulating their activities. The present study reports the cloning and characterization of a sorghum cDNA encoding a protein (SbGRBP) that shows homology to glycine-rich RNA-binding proteins. The expression of SbGRBP in the sorghum seedlings is modulated by heat stress. The SbGRBP protein is localized in the nucleus as well as in cytosol, and shows interaction with CaM that requires the presence of Ca2+. SbGRBP depicts binding to single- and also double-stranded DNA. Fluorescence spectroscopic analyses suggest that interaction of SbGRBP with nucleic acids may be modulated after binding with CaM. To our knowledge, this is the first study to provide evidence for interaction of a stress regulated glycine-rich RNA-binding protein with CaM.


Asunto(s)
Proteínas de Unión a Calmodulina/metabolismo , Calmodulina/metabolismo , Regulación de la Expresión Génica de las Plantas , Glicina/química , Proteínas de Plantas/metabolismo , Sorghum/metabolismo , Calcio , Proteínas de Unión a Calmodulina/genética , ADN Complementario/genética , ADN de Plantas , Proteínas de Plantas/genética , Unión Proteica , Sorghum/genética , Sorghum/crecimiento & desarrollo , Temperatura
12.
J Food Sci Technol ; 53(8): 3186-3196, 2016 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-27784913

RESUMEN

The effects of transplantation date on milled rice (physicochemical, amino acids composition and protein profiling) of different cultivars and their starch characteristics (granules size distribution, pasting and thermal) were investigated. Head rice yield increased (2.0-4.1 %) and chalky grains (5-10 %) decreased with delaying the paddy transplantation of different cultivars by 20 days. Delayed transplantation of paddy significantly increased asparagine, glutamine, threonine, cysteine, methionine, tryptophan, lysine and proline content in milled rice. Early transplantation of paddy showed higher accumulation of glutelin and prolamines than that in milled rice from delayed transplantation. The change in amino acid composition of milled rice with delay in transplantation was related to variation in accumulation of glutelin and prolamines. Starch from delayed transplanted paddy showed higher peak viscosity and lower breakdown viscosity than those from early transplanted paddy. These differences were due to higher accumulation of amylose in starch from delayed transplanted paddy than that from early transplanted paddy due to exposure of former to lower night air temperature during starch synthesis.

13.
J Food Sci Technol ; 53(5): 2452-62, 2016 May.
Artículo en Inglés | MEDLINE | ID: mdl-27407212

RESUMEN

Effect of different level (60, 120 and 180 kg N/ha) of nitrogen (N) application on protein profiling, pasting and cooking quality characteristics of milled rice from different paddy cultivars was evaluated. N dose showed positive correlation with protein content and negative correlation with L*, whiteness and amylose content. N application significantly affected the protein profile, textural and pasting properties of different cultivars. All the cultivars expect PR120 and PAU201, showed an increase in the amount of accumulation of 60 kDa polypeptide with increase in N application. Accumulation of prolamines (16 and 14 kDa) and polypeptides of 38 and 35 kDa increased in all the cultivars. Size exclusion chromatography revealed decrease in low molecular weight subunits and increase in medium molecular weight subunits in all the cultivars upon N application. However, high molecular weight subunits increased in IET21214 and decreased in PR120 and PAU20 upon N application. N application resulted in increase in glutelins and decrease in peak and breakdown viscosity. PAU201 and PR120 showed lower AAC due to low accumulation of 60 kDa granule-bound starch synthase (GBSS), in response to N application. Gumminess and hardness of cooked rice increased with the increase in N dose and the increase was significant at 60 kg N/ha.

14.
Food Res Int ; 89(Pt 1): 526-532, 2016 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-28460947

RESUMEN

The aim of the present study was to evaluate the effect of canning on color, protein and phenolic profile of grains of kidney bean, field pea and chickpea varieties/accession. Color of grains of different pulses was enhanced after canning. Grains L* (lightness) decreased while a* (redness to yellowness) and b* (greenness to blueness) increased after canning in all the pulses. Protein profiling of grains of different pulses after canning revealed that kidney bean and chickpea, respectively, had the least and the most thermally susceptible polypeptides. Kidney bean and chickpea showed higher Percentage washed drained weight (PWDW) than field pea. Pulse with more grain hardness and PWDW showed higher degree of grain splitting during canning. Grain splitting was also higher in dark colored accessions/varieties as compared to the light colored. Ferulic acid was the most predominant compound present in raw grains of different pulses. Raw kidney bean grains showed higher accumulation of catechin, chlorogenic, protocatechuic acid, p-coumaric acid and ferulic acid than those of chickpea and field pea. Canning caused reduction in all the phenolic compounds except gallic acid and most prominent effect of canning on protocatechuic acid, chlorogenic and ferulic acid was observed.

15.
Food Chem ; 194: 337-44, 2016 Mar 01.
Artículo en Inglés | MEDLINE | ID: mdl-26471563

RESUMEN

The relationships of polymeric as well as monomeric proteins (unextractable and extractable) with various flour properties amongst Indian wheat varieties were evaluated. Unextractable polymeric proteins and unextractable monomeric proteins in flours ranged from 23.83% to 51.97% and 48.03% to 76.17%, respectively. Varieties with higher grain hardness index resulted into flours with higher a(∗), ash content and protein content. Unextractable polymeric and monomeric proteins were related to grain hardness index. Unextractable polymeric proteins showed a positive correlation with gluten index and LASRC. Majority of varieties with HMW-GS combinations of 91kDa+80kDa+78kDa+74kDa PPs showed very high grain hardness index (97-100).


Asunto(s)
Grano Comestible/química , Harina/análisis , Glútenes/química , Triticum/química , India
16.
Food Chem ; 188: 517-26, 2015 Dec 01.
Artículo en Inglés | MEDLINE | ID: mdl-26041226

RESUMEN

The grain, flour, dough and noodle making properties of Indian durum wheat varieties were evaluated. Varieties having higher grain weight had lower hardness and higher yellow pigment content. Gluten performance index showed positive correlation with α-helix and negative with intermolecular+antiparallel-ß-sheets in gluten. The proportion of extracted polymeric proteins was related to dough strength. Elastic (G') and loss (G″) modulus of dough were positively correlated to intermolecular+antiparallel-ß-sheets and negatively with ß-turn+ ß-sheets proportion of dough and gluten. PDW291 with exceptionally higher G' and G″ and best noodle making properties showed the presence 90 kDa and 88 kDa polypeptides corresponding to 14+15 and type 2 banding pattern.


Asunto(s)
Cromatografía Líquida de Alta Presión/métodos , Harina/análisis , Triticum/química
17.
J Food Sci ; 80(6): C1144-55, 2015 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-25943010

RESUMEN

Dry milling of different corn types resulted in varied proportions of germ, pericarp, grit and flour. Grit and flour produced during different reduction stages varied in particle size and chemical constituents, hence applications in food industry. In this study, recovery of different fractions and variation in physicochemical and pasting properties of grit and flour fractions obtained during 3 successive reduction dry millings of 2 normal (African tall, HQPM1) and 1 waxy corn (IC 550353) were evaluated. Waxy corn grains had the highest L*, a*, b*, ash, fat, and protein content and the lowest weight. Waxy and African tall gave the highest recovery of germ and pericarp, respectively. Waxy corn showed lower grit and flour recovery as compared to normal corn. Flour fractions showed higher L* and lower a* and b* values than grit fractions. Particle size of grit and flour fractions ranged from 840 to 982 µm and 330 to 409 µm, respectively. Fractions with larger particle size showed lower L* value. The b* value showed positive correlation with yellow pigment content. Grit and flour from the 1st reduction stage showed higher ash and fat content. Protein content was correlated positively with ash content and negatively with L* value. Grit and flour fractions with higher protein content had lower pasting viscosities. Pasting viscosities were higher for flours than their corresponding grits. Protein profiling of grit and flour fractions from different stages showed quantitative and qualitative differences in medium (22, 28, and 35 kDa) and low molecular weight (16, 17, and 19 kDa) polypeptides and were related to grit and flour yield.


Asunto(s)
Grano Comestible/química , Harina/análisis , Industria de Procesamiento de Alimentos , Tamaño de la Partícula , Zea mays/química , Color , Dieta , Grasas de la Dieta/análisis , Proteínas en la Dieta/análisis , Humanos , Pigmentación , Especificidad de la Especie , Viscosidad , Ceras , Zea mays/clasificación
18.
Food Chem ; 172: 528-36, 2015 Apr 01.
Artículo en Inglés | MEDLINE | ID: mdl-25442588

RESUMEN

The grain and flour characteristics of different field pea (FP) accessions were evaluated. Accessions with higher grain weight had less compact structure with a greater proportion of large-sized starch granules. Accessions with higher protein content had lower starch content, blue value and λ(max) whereas accessions with higher amylose showed higher resistant starch (RS) and final viscosity and lower rapidly digestible starch (RDS). Ca, Zn, K and Fe content vary significantly amongst different accessions and creamish green and white seeds accessions showed higher Fe and Zn content. Yellow coloured accessions (1.36-3.71%) showed lower antioxidant activity as compared to brownish and green coloured accessions (4.06-9.30%). Out of 21 major polypeptides observed (9-100 kDa), 11 showed differential trypsin inhibitory activity (TIA) under non-reducing conditions. Polypeptides of 68, 46, 33 and 22 kDa showed prominent TIA.


Asunto(s)
Pisum sativum/metabolismo , Proteínas de Plantas/metabolismo , Amilosa/análisis , Antioxidantes/química , Electroforesis en Gel de Poliacrilamida , Harina/análisis , Metales/análisis , Pisum sativum/química , Fenoles/química , Semillas/química , Semillas/metabolismo , Almidón/química , Almidón/aislamiento & purificación , Inhibidores de Tripsina/análisis , Viscosidad
19.
Gene ; 501(1): 85-8, 2012 Jun 10.
Artículo en Inglés | MEDLINE | ID: mdl-22521868

RESUMEN

PCR is usually performed on purified DNA. However, the extraction of DNA from whole blood is time consuming and involves the risk of contamination at every step. Hence, it is desirable to amplify DNA directly from whole blood. Earlier, investigators tried to achieve this target by either pretreatment of whole blood samples with different agents or by altering the conventional thermal cyclic conditions. This would make the technique cumbersome and time consuming. Here, we describe a simple protocol to amplify DNA directly from whole blood without the need of pretreatment. PCR buffer system was optimized in the laboratory and Apolipoprotein B gene was used as a model for this experiment. 480 bp was the target site for amplification. Fresh whole blood samples were used both from healthy and diseased individuals (coronary artery disease patients). Successful amplification was achieved with 1 µl volume of whole blood and it was comparable to that of genomic DNA. No pretreatment of whole blood samples was required with the optimized buffer system. 3mM concentration of MgCl(2) was observed to be optimal and hence used in the reaction mixture. Amplification was relatively better with this buffer system as compared to that of commercially available PCR buffer. With the present technique, amplicon detection did not require the centrifugation/dilution of the PCR products which further saves time. Successful amplification was achieved in both the healthy and diseased blood samples, indicating the robustness of the technique as changed blood composition and presence of increased inhibitory molecules in the diseased state did not seem to affect the efficacy of the present technique. In conclusion, as compared to the existing protocols for whole blood PCR, the present technique is relatively novel, simple, requires minimal steps and eliminates the need for additional standardizations.


Asunto(s)
Sangre , ADN/aislamiento & purificación , Reacción en Cadena de la Polimerasa/métodos , Apolipoproteínas B/genética , Estudios de Casos y Controles , Enfermedad de la Arteria Coronaria/genética , Humanos
20.
Plant Signal Behav ; 6(3): 393-9, 2011 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-21336025

RESUMEN

Pharmacological studies, using Ca(2+) channel blockers (LaCl 3 and verapamil) and calmodulin (CaM) antagonists (CPZ and W7), were carried out to understand the role of Ca(2+)/CaM in the regulation of heat shock-induced expression of Hsp90 (Hsp87 and Hsp85) and Hsp70 (Hsp75 and Hsp73) members in sorghum. It was observed that the expression of both Hsp87 and Hsp85 proteins was decreased in presence of Ca ( 2+) channel blockers and CaM antagonists, under both control and heat stress conditions, as contrary to the steady state levels of Hsp75 and Hsp73, which were not affected significantly under similar conditions. Further, the exposure of sorghum seedlings to geldanamycin, a specific inhibitor of Hsp90, resulted in induction of Hsp87 and Hsp85 in the absence of heat shock also. This study provides the first evidence suggesting that in plants, the in vivo expression of Hsp90 (Hsp87 and Hsp85) is likely to be modulated by Ca(2+)/CaM under normal and thermal stress conditions. The likely implications of these findings are discussed.


Asunto(s)
Calmodulina/metabolismo , Proteínas HSP90 de Choque Térmico/metabolismo , Proteínas de Plantas/metabolismo , Sorghum/metabolismo , Calmodulina/genética , Proteínas HSP90 de Choque Térmico/genética , Immunoblotting , Modelos Biológicos , Proteínas de Plantas/genética , Sorghum/genética
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