Sarita Jaiswal

Barley Grain Constituents, Starch Composition, and Structure Affect Starch in Vitro Enzymatic Hydrolysis

The relationship between starch physical properties and enzymatic hydrolysis was determined using ten different hulless barley genotypes with variable carbohydrate composition. The ten barley genotypes included one normal starch (CDC McGwire), three increased amylose starches (SH99250, SH99073, and SB94893), and six waxy starches (CDC Alamo, CDC Fibar, CDC Candle, Waxy Betzes, CDC Rattan, and SB94912). Total starch concentration positively influenced thousand grain weight (TGW) (r(2) = 0.70, p < 0.05). Increase in grain protein concentration was not only related to total starch concentration (r(2) = -0.80, p < 0.01) but also affected enzymatic hydrolysis of pure starch (r(2) = -0.67, p < 0.01). However, an increase in amylopectin unit chain length between DP 12-18 (F-II) was detrimental to starch concentration (r(2) = 0.46, p < 0.01). Amylose concentration influenced granule size distribution with increased amylose genotypes showing highly reduced volume percentage of very small C-granules (<5 μm diameter) and significantly increased (r(2) = 0.83, p < 0.01) medium sized B granules (5-15 μm diameter). Amylose affected smaller (F-I) and larger (F-III) amylopectin chains in opposite ways. Increased amylose concentration positively influenced the F-III (DP 19-36) fraction of longer DP amylopectin chains (DP 19-36) which was associated with resistant starch (RS) in meal and pure starch samples. The rate of starch hydrolysis was high in pure starch samples as compared to meal samples. Enzymatic hydrolysis rate both in meal and pure starch samples followed the order waxy > normal > increased amylose. Rapidly digestible starch (RDS) increased with a decrease in amylose concentration. Atomic force microscopy (AFM) analysis revealed a higher polydispersity index of amylose in CDC McGwire and increased amylose genotypes which could contribute to their reduced enzymatic hydrolysis, compared to waxy starch genotypes. Increased β-glucan and dietary fiber concentration also reduced the enzymatic hydrolysis of meal samples. An average linkage cluster analysis dendrogram revealed that variation in amylose concentration significantly (p < 0.01) influenced resistant starch concentration in meal and pure starch samples. RS is also associated with B-type granules (5-15 μm) and the amylopectin F-III (19-36 DP) fraction. In conclusion, the results suggest that barley genotype SH99250 with less decrease in grain weight in comparison to that of other increased amylose genotypes (SH99073 and SH94893) could be a promising genotype to develop cultivars with increased amylose grain starch without compromising grain weight and yield.

Evidence based herbal drug standardization approach in coping with challenges of holistic management of diabetes: a dreadful lifestyle disorder of 21st century

Plants by virtue of its composition of containing multiple constituents developed during its growth under various environmental stresses providing a plethora of chemical families with medicinal utility. Researchers are exploring this wealth and trying to decode its utility for enhancing health standards of human beings. Diabetes is dreadful lifestyle disorder of 21st century caused due to lack of insulin production or insulin physiological unresponsiveness. The chronic impact of untreated diabetes significantly affects vital organs. The allopathic medicines have five classes of drugs, or otherwise insulin in Type I diabetes, targeting insulin secretion, decreasing effect of glucagon, sensitization of receptors for enhanced glucose uptake etc. In addition, diet management, increased food fiber intake, Resistant Starch intake and routine exercise aid in managing such dangerous metabolic disorder. One of the key factors that limit commercial utility of herbal drugs is standardization. Standardization poses numerous challenges related to marker identification, active principle(s), lack of defined regulations, non-availability of universally acceptable technical standards for testing and implementation of quality control/safety standard (toxicological testing). The present study proposed an integrated herbal drug development & standardization model which is an amalgamation of Classical Approach of Ayurvedic Therapeutics, Reverse Pharmacological Approach based on Observational Therapeutics, Technical Standards for complete product cycle, Chemi-informatics, Herbal Qualitative Structure Activity Relationship and Pharmacophore modeling and, Post-Launch Market Analysis. Further studies are warranted to ensure that an effective herbal drug standardization methodology will be developed, backed by a regulatory standard guide the future research endeavors in more focused manner.

A reliable and rapid method for soluble sugars and RFO analysis in chickpea using HPAEC-PAD and its comparison with HPLC-RI.

A high performance anion exchange chromatography (HPAEC) coupled with pulsed amperometric detection (PAD) was optimised to separate with precision, accuracy and high reproducibility soluble sugars including oligosaccharides present in pulse meal samples. The optimised method within 20min separated myo-inositol, galactinol, glucose, fructose, sucrose, raffinose, stachyose and verbascose in chickpea seed meal extracts. Gradient method of eluting solvent (sodium hydroxide) resulted in higher sensitivity and rapid detection compared to similar analytical methods. Peaks asymmetry equivalent to one and resolution value ⩾1.5 support columns precision and accuracy for quantitative determinations of soluble sugars in complex mixtures. Intermediate precision determined as relative standard deviation (1.8-3.5%) for different soluble sugars confirms reproducibility of the optimised method. The developed method has superior sensitivity to detect even scarcely present verbascose in chickpea. It also quantifies myo-inositol and galactinol making it suitable both for RFO related genotype screening and biosynthetic studies.

Genome-specific granule-bound starch synthase I (GBSSI) influences starch biochemical and functional characteristics in near-isogenic wheat ( Triticum aestivum L.) lines.

Near-isogenic wheat ( Triticum aestivum L.) lines differing at the Waxy locus were studied for the influence of genome-specific granule-bound starch synthase I (GBSSI/Waxy; Wx-A, Wx-B, Wx-D) on starch composition, structure, and in vitro starch enzymatic hydrolysis. Grain composition, amylose concentration, amylopectin unit-chain length distribution, and starch granule size distribution varied with the loss of functional GBSSI. Amylose concentration was more severely affected in genotypes with GBSSI missing from two genomes (double nulls) than from one genome (single nulls). Unit glucan chains (DP 6-8) of amylopectin were reduced with the complete loss of GBSSI as compared to wheat starch with a full complement of GBSSI. Wx-A and Wx-B had an additive effect toward short-chain phenotype of waxy amylopectin. Loss of Wx-D isoprotein alone significantly (p < 0.05) reduced the C-type starch granules. However, the absence of Wx-D in combination with Wx-A or Wx-B increased the B-type and C-type starch granules but decreased the volume of A-type starch granules. The rate of in vitro starch enzymatic hydrolysis was highest in completely waxy grain meal and purified starch. However, the presence of Wx-D reduced wheat starch hydrolysis as it increased the large A-type starch granule content (volume %) and reduced short chains (DP 6-8) in amylopectin. Factors such as small C-type starch granules, amylose concentration, and long chains of amylopectin (DP 23-45) also influenced wheat starch hydrolysis.

Himalayan Bioresource Rhodiola imbricata as a promising radioprotector for nuclear and radiological emergencies

In recent years, a lot of interest has been generated world over in the area of radioprotection for first responders going to work in the hot zones at the incident site. A large number of molecular drugs have been screened for radioprotective efficacy, but with little success. The requirement of differential radioprotection necessitates a holistic approach, which can be realized using herbs in view of their multifaceted mode of action. Our earlier studies showed the radioprotective potential of Rhodiola imbricata, a Himalayan high-altitude plant. In this study, our focus has been to compare the pro-oxidant/antioxidant activities of three fractionated extracts of R. imbricata. The aqueous fraction exhibited significant (P < 0.05) pro-oxidant activity (up to 100 μg/ml) under metal ion-induced stress ± flux [transition metal (Fe/Cu) ± 0.25 kGy]. A decrease in the dielectric constant of the solvent system utilized for extraction, exhibited a significant (P < 0.05) negative correlation (–0.955) with mean protection potential of lipid against radiation flux. Such an effect was visualized as a significant shift from pro-oxidant to antioxidant activity in methanolic fraction (dielectric constant = 33), as compared to aqueous fraction (dielectric constant = 80). Aqueous fraction is predominantly pro-oxidant at maximal concentrations, indicating its anticancer potential. The presence of transition metals modulates such a biphasic activity differentially in various fractions, i.e., the conversion of Fe(III) or Cu(II) to Fe(II) or Cu(I), respectively, due to the presence of certain bioactive constituents (electron donation at lower concentrations), favors pro-oxidant activity. On the other hand, certain other active constituents involved in metal ion chelation contributed to the overall antioxidant activity. The methanolic fraction exhibited significant antioxidant activity up to 250 μg/ml, which contributed to its radioprotective efficacy. The aquo-methanolic fraction exhibited (disparate properties), i.e., concentration-dependant cytotoxicity (up to 250 μg/ml) and cytoprotection at 1000 μg/ml. R. imbricata, in general, exhibited a significant solvent-dependant variation in radioprotective efficacy. In conclusion, solvent extraction and dose are crucial in bioactivity modulation and R. imbricata could be developed as a potential prophylactic radiation countermeasure for use in nuclear and radiological emergencies.

Galactinol synthase enzyme activity influences raffinose family oligosaccharides (RFO) accumulation in developing chickpea (Cicer arietinum L.) seeds

To understand raffinose family oligosaccharides (RFO) metabolism in chickpea (Cicer arietinum L.) seeds, RFO accumulation and corresponding biosynthetic enzymes activities were determined during seed development of chickpea genotypes with contrasting RFO concentrations. RFO concentration in mature seeds was found as a facilitator rather than a regulating step of seed germination. In mature seeds, raffinose concentrations ranged from 0.38 to 0.68 and 0.75 to 0.99 g/100 g, whereas stachyose concentrations varied from 0.79 to 1.26 and 1.70 to 1.87 g/100 g indicating significant differences between low and high RFO genotypes, respectively. Chickpea genotypes with high RFO concentration accumulated higher concentrations of myo-inositol and sucrose during early seed developmental stages suggesting that initial substrate concentrations may influence RFO concentration in mature seeds. High RFO genotypes showed about two to three-fold higher activity for all RFO biosynthetic enzymes compared to those with low RFO concentrations. RFO biosynthetic enzymes activities correspond with accumulation of individual RFO during seed development.

Wheat genome specific granule-bound starch synthase I differentially influence grain starch synthesis.

Wheat grain development is a complex process and is characterized by changes in physicochemical and structural properties of starch. The present study deals with endosperm starch physicochemical properties and structure during development in different granule-bound starch synthase I (GBSSI) null also known as waxy (Wx) genotypes. The study was conducted with pure starch isolated from wheat grains at 3-30 days post anthesis (DPA), at 3-day intervals. Amylose concentration increased throughout grain development in non-waxy (7.2-30.5%) and partial waxy genotypes (6.0-26.8%). Completely waxy genotype showed 7.0% amylose at 3 and 6 DPA, which declined during development and reached non-detectable quantities by 30 DPA. Amylopectin structure had a higher content of short chains at 3 DPA, which decreased continuously until 12 DPA, after which there were only minor changes in amylopectin chain length distribution. Similarly, the average degree of polymerization (DP) increased from 3 DPA (12.3) to 12 DPA (15.0), and then did not differ significantly up to 30 DPA (15.0). This suggests the formation of basic amylopectin architecture in wheat by 12 DPA. Wx-B and Wx-D affected amylopectin short chains mostly of DP 6-8 at 3 and 6 DPA. Wx-A affected the same fraction of chains at 9 and 12 DPA, and Wx-D affected DP 18-25 chains from 18 to 30 DPA, suggesting differential effect of waxy isoproteins on amylopectin structure formation.

Medical radiation countermeasures for nuclear and radiological emergencies: Current status and future perspectives.

Nuclear and radiological emergencies (NREs) occurred globally and recent incidences in India are indicating toward the need for comprehensive medical preparedness required both at incident site and hospitals. The enhanced threat attributed toward insurgency is another causative factor of worry. The response capabilities and operational readiness of responders (both health and non-health service providers) in contaminated environment need to be supported by advancement in R & D and technological efforts to develop prophylactics and radiation mitigators. It is essential to develop phase 1 alternatives of such drugs for unseen threats as a part of initial preparedness. At the incident site and hospital level, external decontamination procedures need to be standardized and supported by protective clothing and Shudika kits developed by INMAS. The medical management of exposure requires systematic approach to perform triage, resuscitation and curative care. The internal contamination requires decorporation agents to be administered based on procedural diagnostics. Various key issues pertaining to policy decisions, R & D promotion, community awareness, specialized infrastructure for NREs preparedness has been discussed. The present review is an attempt to provide vital information about the current status of various radiation countermeasures and future perspective(s) ahead.

Development of barley (Hordeum vulgare L.) lines with altered starch granule size distribution.

Microscope analysis of starches prepared from 139 barley genotypes identified a Japanese genotype, Kinai Kyoshinkai-2 (KK-2), with altered starch granule size distribution. Compared to normal barley starch, KK-2 produced consistently higher volumes of starch granules with 5-15 μm diameter and reduced volumes of starch granules with >15 μm diameter when grown in different environments. A cross between KK-2 and normal starch cultivar CDC Kendall was made and led to the production of 154 F5 lines with alterations to the normal 7:3:1 distribution for A-:B-:C-type starch granule volumes. Three F5 lines showed unimodal starch granule size distribution due to apparent lack of very small (<5.0 μm diameter) C-type starch granules, but the phenotype was accompanied by reduced grain weight and total starch concentration. Five F5 lines produced a significantly larger population of large (>15 μm diameter) A-type starch granules as compared to normal starch and showed on average a 10:4:1 distribution for A-:B-:C-type starch granule volumes. The unusual starch phenotypes displayed by the F5 lines confirm starch granule size distribution in barley can be genetically altered.

Targeting Antibiotic Resistant Salmonella enterica: Bio-matrix Based Selectionand Bioactivity Prediction of Potential Nutraceuticals

Aim: Potential herbal leads as novel therapeutic alternatives against Multi-Drug Resistant Salmonella enterica serovar typhimurium were investigated and validated by mol inspiration. Methodology: The present study utilizes an in silico ‘Herbal Informatics’ model which deploys dynamic search protocols, priority indexing and systemic categorization for rationale based selection of nutraceuticals targeting the critical virulence factors of Multi-Drug Resistant Salmonella enterica serovar typhimurium. Furthermore, in silico biochemical activity prediction was conducted using ‘Mol inspiration’ chemiinformatics tool, so as to propose the drug likeliness of selected natural plant products. Results: Out of the 05 selected bioactivity parameters of Salmonella enterica, lipo-polysaccharide inhibition exhibited maximum relevance as physiological target i.e., 65%, followed by other parameters like enterochelin inhibition, Type III secretory system inhibition, Superoxide dismutase inhibition and Symptomatic relief provision. The binary matrix analysis of database of 50 plants identified using classical bioprospection filtered 28 herbals which exhibited probable potential to mitigate 03 or more virulence factors. The weightage matrix analysis further scrutinized the selection upto 10 herbals having a score more than median weightage matrix score i.e., 14.98. The optimization of data on a scale of 0-1 using fuzzy score matrix analysis led to the final selection of 10 herbals i.e., Abrus precatorius, Azadirachta indica, Camellia sinensis, Holarrhena antidysenterica, Andrographis paniculata, Adhatoda vasica, Euphorbia hirta, Ocimum sanctum, Terminalia arjuna and Terminalia belerica. In silico Bioactivity prediction analysis of predominant phytoconstituents of selected herbals revealed Holarrhena antidysenterica (Conessine), Andrographis paniculata (Andrographolide), Euphorbia hirta (Amyrin) and Terminalia arjuna (Arjunolic acid) exhibiting drug likeliness with their targeted action as a GPCR Ligand, Nuclear Receptor Ligand or Protease inhibitor.