Permeability determination and pharmacokinetic study of nobiletin in rat plasma and brain by validated high-performance liquid chromatography method

In the present study, we are reporting permeability and pharmacokinetics of nobiletin in rat plasma and brain, using a validated reverse phase high performance liquid chromatographic method. Protein precipitation method was used for the extraction of nobiletin and coumarin (IS) from rat plasma and brain tissue. The system was run in isocratic mode with mobile phase consisting of potassium dihydrogen ortho-phosphate (pH 4.5; 0.04 mM) and acetonitrile in ratio of 50:50, v/v. The total chromatographic run time was 9.0 min. The method was proved to be accurate and precise at linearity range of 0.05–10 μg/mL with a correlation coefficient (r) of ≥ 0.994 in rat plasma and ≥ 0.995 in rat brain. The intra- and inter-day precision and accuracy values are found to be within the assay variability limits as per the FDA guidelines. Nobiletin was found stable in the battery of stability studies viz., bench-top, auto-sampler, freeze/thaw cycles and long term storage in a freezer at − 70 ± 10 °C. Maximum concentrations of nobiletin in both plasma and brain were observed at 1 h after single oral dosing (50 mg/kg). The maximum concentration in plasma and brain were 1.78 and 4.20 μg/mL, respectively. The AUC_0–t in plasma and brain were 7.49 and 20.66 μg·h/mL, respectively. The mean elimination half life (t_1/2) in plasma and brain were 1.80 and 11.42 h, respectively. The Parallel Artificial Membrane Permeability Assay (PAMPA) permeability of nobiletin was found to be high at both pH 4.0 and 7.0.     

Indole acetic acid differently changes growth and nitrogen metabolism in Pisum sativum L. seedlings under chromium (VI) phytotoxicity: Implication of oxidative stress

The present study investigated impact of exogenous application of indole acetic acid (IAA; 10 and 100 μM) in pea seedlings under hexavalent chromium (Cr VI; 50, 100 and 250 μM). Cr and 100 μM IAA alone as well as in combination decreased seed germination rate compared to control. However, under Cr phytotoxicity, addition of 10 μM IAA recovered seed germination rate to the level of control. Exposure of pea seedlings to Cr and 100 μM IAA during their early stage caused decrease in fresh mass, length, protein and nitrogen contents of roots and shoots compared to control. Treatment of pea seedlings with Cr resulted in a rapid accumulation of this metal in roots and shoots. Moreover, addition of 100 μM IAA together with Cr, further increased accumulation of this metal in roots and shoots compared to Cr treatments alone. Treatment of pea seedlings with Cr and 100 μM IAA, resulted in a marked decrease in nitrate reductase, nitrite reductase, glutamine synthetase and glutamate synthase (GOGAT) activities (except 50 μM Cr alone for GOGAT), and an increase in ammonium content and glutamate dehydrogenase activity. Parameters related with oxidative stress, i.e. superoxide radicals and reactive carbonyl groups (protein oxidation) were increased by Cr and 100 μM IAA compared to control. By contrast, addition of 10 μM IAA together with Cr, alleviated negative effect of Cr on growth, protein, nitrogen and nitrogen metabolism, and led to decrease in oxidative injuries caused by Cr. The data indicate that 10 μM IAA protects pea seedlings during the early growth period against Cr phytotoxicity by regulating Cr accumulation and oxidative damage. However, addition of 100 μM IAA together with Cr showed opposite responses.     

Increase in growth,productivity and nutritional status of rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L. cv. Basmati) and enrichment in soil fertility applied with an organic matrix entrapped urea

Field experiments were conducted to evaluate the effects of eco-friendly organic matrix entrapped urea (OMEU) on growth, productivity, and yield of rice (Oryza sativa L. cv. Basmati) and soil enrichment in the paddy field at Rohtak (Haryana) located near Delhi. The OMEU prepared in granular form contained cow dung, rice bran (grain cover of Oryza sativa), powder of neem leaves (Azadirachta indica), and clay soil (diameter of particles < 0.02 mm) in 1:1:1:1 ratios and saresh (plant gum of Acacia sp.) as binder along with half of the recommended dose of commercially available soluble urea (free urea; FU). Single basal application of OMEU showed an increase in plant growth in terms of fresh and dry weights, root length, root, leaf and tiller numbers, soluble protein, total N and ammonium in leaves, productivity in terms of grain and straw yield, and nutritional and microbial activities of field soil over free form of urea and no fertilizer application. Nutritional status of rice grains was also improved over the free urea and no fertilizer controls. Our data indicate that OMEU, which is low cost and based on bio-degradable, non-toxic, and locally available agro-waste, can be attempted to replace the conventional use of soluble urea in rice.     

Effects of extraction methods on phenolic contents and antioxidant activity in aerial parts of Potentilla atrosanguinea Lodd. and quantification of its phenolic constituents by RP-HPLC

The effects of different solvent systems (methanol, ethanol, acetone, and their 50% aqueous concentrations) and extraction procedures (microwave, ultrasound, Soxhlet and maceration) on the antioxidant activity of aerial parts of Potentilla atrosanguinea were investigated by three different bioassays: 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), 2,2'-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assays and ferric reducing antioxidant potential (FRAP). The 50% aqueous ethanol extracts exhibited strong antioxidant activity measured in terms of Trolox equivalent antioxidant capacity (TEAC) [(54.34 to 122.96, 29.82 to 101.22 and 13.64 to 41.43) mg of Trolox/g] with ABTS (*+), DPPH (*) and FRAP assays, respectively. In general, TEAC of Soxhlet extracts was found to be 1.8 and 3 times higher than ultrasound and maceration but slightly (1.2 times) higher than microwave. A positive correlation (r(2) = 0.931 to 0.982) was observed between total polyphenol (TPC) and total flavonoid (TFC) contents which ranged between 26.7 to 30.7 mg/g gallic acid equivalent and 16.8 to 20.8 mg/g quercetin equivalent respectively, with antioxidant activity. In addition, some of its bioactive phenolic constituents which contribute largely toward antioxidant potential such as chlorogenic acid, catechin, caffeic acid, p-coumaric acid and quercetin were also quantified in different extracts by RP-HPLC.     

Identification of potential sources of mungbean yellow mosaic India virus resistance in black gram (Vigna mungo) and expression of antioxidants and R-genes modulating resistance response in cultivated and its two wild relatives

Black gram is one of the most important short duration grain legume, which contributes significantly towards nutritional security and environmental sustainability. The virus specific primers confirms the presence of mungbean yellow mosaic India virus (MYMIV) in representative samples. A total of 27 cultivated and two wild species were found as highly resistant (HR) to MYMIV and validated through molecular markers. The start codon target (SCoT) markers analysis revealed that the SCoT loci, namely, SCoT-4 (2200 bp), SCot-9 (1150/ 1200 bp), SCoT-15 (1150/1100 bp), SCoT-16 (700 bp), SCoT-24 (2500 bp), SCoT-25 (700 bp), SCoT-33 (900/1000 bp), and SCoT-34 (600 bp), were found unique, able to distinguish HR and highly susceptible (HS) genotypes. Biochemical characterization and gene expression profiling revealed the higher expression of antioxidants and R-genes just after pathogen inoculation indicated the activation of defence mechanism in both cultivated and its wild relatives, which modulates the resistant responses in cultivated and wild accessions. These information will be really helpful in accelerating resistance breeding in black gram.     

Assessment of the Nutritive,Biochemical, Antioxidant and Antibacterial Potential of Eight Tropical Macro algae Along Kachchh Coast,India as Human Food Supplements

Eight tropical marine macro algae were investigated for in-vitro antioxidant, antibacterial, and biochemical properties. The moisture content [% DW (dry weight)] ranged from 7.21 to 14.72%; ash, 24.92 to 47.04%; lipids, 0.73 to 2.67%; protein, 4.56 to 12.59%; and carbohydrate, 30.1 to 48.51%. The % sum of polyunsaturated fatty acids (PUFAs) ranged from 21.71 to 78.22%, while omega-6/omega-3 (ω6/ω3) ratio was from 0 to 2.08, which remained within the prescribed World Health Organization (WHO) standards (<10). The % sum of essential amino acids (EAAs) ranged from 86.18 to 204.66, and Na/K ratio ranged from 0.37 to 2.85. The extracts exhibited significant (P < 0.05) values for total phenol (7.25–16.0 µg PGE mg^–1 DW) and total flavonoid (4.06–15.63 µg QE mg^–1 DW) at 200 µg/ml. Antibacterial activity was tested against the selected food spoilage and pathogenic bacteria. The principal component analysis (PCA) confirmed a positive correlation between total phenolic content, 1, 1-diphenyl-2-picryl-hydrazyl (DPPH) radical-scavenging activities, and ferric reducing antioxidant power (FRAP), which supports the integration of seaweeds as ingredients in functional foods.     

Effect of organic and inorganic sources of nutrients on soil microbial activity and soil organic carbon build-up under rice in west coast of India

The effect of medium-term (5 years) application of organic and inorganic sources of nutrients (as mineral or inorganic fertilizers) on soil organic carbon (SOC), SOC stock, carbon (C) build-up rate, microbial and enzyme activities in flooded rice soils was tested in west coast of India. Compared to the application of vermicompost, glyricidia (Glyricidia maculate) (fresh) and eupatorium (Chromolaena adenophorum) (fresh) and dhaincha (Sesbania rostrata) (fresh), the application of farmyard manure (FYM) and combined application of paddy straw (dry) and water hyacinth (PsWh) (fresh) improved the SOC content significantly (p < 0.05). The lowest (p < 0.05) SOC content (0.81%) was observed in untreated control. The highest (p < 0.05) SOC stock (23.7 Mg C ha^?1) was observed in FYM-treated plots followed by recommended dose of mineral fertilizer (RDF) (23.2 Mg C ha^?1) and it was lowest (16.5 Mg C ha^?1) in untreated control. Soil microbial biomass carbon (C_mb) (246 µg g^?1 soil) and C_mb/SOC (1.92%) were highest (p < 0.05) in FYM-treated plot. The highest (p < 0.05) value of metabolic quotient (qCO₂) was recorded under RDF (19.7 µg CO₂-C g^?1 C_mb h^?1) and untreated control (19.6 µg CO₂-C g^?1 C_mb h^?1). Application of organic and inorganic sources of nutrients impacted soil enzyme activities significantly (p < 0.05) with FYM causing highest dehydrogenase (20.5 µg TPF g^?1 day^?1), phosphatase (659 µg PNP g^?1 h^?1) and urease (0.29 µg urea g^?1 h^?1) activities. Application of organic source of nutrients especially FYM improved the microbial and enzyme activities in flooded and transplanted rice soils. Although the grain yield was higher with the application of RDF, but the use of FYM as an organic agricultural practice is more useful when efforts are intended to conserve more SOC and improved microbial activity.     

Soil Carbon Dynamics Under Changing Climate——A Research Transition from Absolute to Relative Roles of Inorganic Nitrogen Pools and Associated Microbial Processes: A Review

It is globally accepted that soil carbon (C) dynamics are at the core of interlinked environmental problems,deteriorating soil quality and changing climate.Its management remains a complex enigma for the scientific community due to its intricate relationship with soil nitrogen (N) availability and moisture-temperature interactions.This article reviews the management aspects of soil C dynamics in light of recent advances,particularly in relation to the availability of inorganic N pools and associated microbial processes under changing climate.Globally,drastic alterations in soil C dynamics under changing land use and management practices have been primarily attributed to the variation in soil N availability,resulting in a higher decomposition rate and a considerable decline in soil organic C (SOC) levels due to increased soil CO2 emissions,degraded soil quality,and increased atmospheric CO2 concentrations,leading to climate warming.Predicted climate warming is proposed to enhance SOC decomposition,which may further increase soil N availability,leading to higher soil CO2 efflux.However,a literature survey revealed that soil may also act as a potential C sink,if we could manage soil inorganic N pools and link microbial processes properly.Studies also indicated that the relative,rather than the absolute,availability of inorganic N pools might be of key importance under changing climate,as these N pools are variably affected by moisture-temperature interactions,and they have variable impacts on SOC turnover.Therefore,multi-factorial studies are required to understand how the relative availability of inorganic N pools and associated microbial processes may determine SOC dynamics for improved soil C management.     

Investigation of transferrin polymorphism in Garole sheep

The aim of this study was to determine the genetics of polymorph systems of Transferrin in Garole sheep breed. The present study was conducted on 95 adult Garole sheep comprising 52 ewes and 43 rams, maintained at Sheep and Goat Breeding Farm of West Bengal University of Animal and Fishery Sciences, West Bengal, during the period from April–September, 2009. The polymorphism of transferrin was determined through SDS-Polyacrylamide gel electrophoresis technique. It was found that the transferrin type was controlled by five codominant alleles (TfA, TfB, TfC TfD and TfE) in Garole sheep. These five alleles, because of co-dominant nature of inheritance, determined the occurrence of nine transferrin genotypes in the analyzed flock. Four (TfAA, TfBB, TfCC and TfDD) of these were homozygous and the remaining five (TfAD, TfBC, TfBD, TfCD and TfDE) heterozygous. It was found that the TfDD genotype (0.263) was predominant while TfDE genotype (0.042) was least common in the analyzed flock. Frequencies of other genotypes were as: TfCD(0.242), TfBD(0.126), TfCC(0.084), TfBB(0.074), TfAA(0.063), TfAD and TfBC (0.053 for each genotype ) in whole population. From the result it was found that in whole population combined, the heterozygotic genotypic frequency (0.516) was more than that of homozygotic genotypic frequency (0.484). Considerable variations were recognized in the frequencies of transferrin alleles. In the whole population frequencies of transferrin alleles were found to be TfA = 0.089, TfB = 0.163, TfC = 0.232, TfD = 0.495 and TfE = 0.021. Transferrin system has shown an absence of genetic equilibrium among the analyzed herd (χ2 value = 51.31). In conclusion, there were polymorphism in Transferrin types and the presence of differences among the frequencies of the five alleles by categories could be a source of genetic variation in Garole sheep.     

Modulation of manganese toxicity in Pisum sativum L. seedlings by kinetin

In the present study, the effects of kinetin (KN; 10 and 100 μM) application under manganese toxicity (Mn; 50, 100 and 250 μM) were investigated, on growth, photosynthetic pigments, total protein, total nitrogen, ammonium (NH₄⁺) content, NH₄⁺ assimilating enzymes and antioxidant system in pea seedlings. The exposure of pea seedlings to Mn and 100 μM of KN alone and in combination, caused decrease in growth, photosynthetic pigments, total protein and total nitrogen contents, and an increase in NH₄⁺ content. However, application of 10 μM of KN together with Mn reduced the Mn toxicity symptoms, promoted the growth of seedlings and led to the decrease in NH₄⁺ content compared to Mn treatments alone. The root and shoot activities of glutamine synthetase (GS), glutamate oxoglutarate aminotransferase (GOGAT) and catalase (CAT) were decreased while glutathione reductase (GR) and dehydroascorbate reductase (DHAR) activities exhibited differential responses when pea seedlings were exposed to Mn and 100 μM of KN. However, under similar treatments, activities of glutamate dehydrogenase (GDH), superoxide dismutase (SOD) and ascorbate peroxidase (APX) in root and shoot were increased. It was noticed that addition of 10 μM of KN together with Mn, caused significant stimulation in activities of enzymes of NH₄⁺ assimilation and antioxidant defense system even over their respective control values. Non-enzymatic antioxidants (ascorbate and glutathione) in root and shoot of pea seedlings exposed to Mn stress were significantly increased by the addition of 10 μM of KN. Therefore, ameliorative effect of 10 μM of KN against Mn toxicity was observed. This study thus suggests that 10 μM of KN appreciably improves Mn tolerance of pea seedlings under Mn toxicity while reverse effects were exhibited by 100 μM of KN.