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.     

Intercropping field crops between rows of Leucaena leucocephala under rainfed conditions in northern India

A field study was conducted for six years (1981–1986) on sandy loam soil on intercropping hedgerows of Leucaena leucocephala (Lam) de Wit. with three field crops viz. maize (Zea mays L), black gram (Vigna mungo L) and cluster bean (Cyamopsis tetragonoloba L Taub.). In treatments 1 and 2 Leucaena hedges were planted as pure crops at close (25 cm × 75 cm) and wide (25 cm × 375 cm) spacings. In treatments 3, 4 and 5 the three field crops were intercropped between the hedgerows of Leucaena at the wide spacing, and in treatments 6, 7 and 8 the field crops were raised as pure crops. Leucaena was topped to 75 cm each time it attained a height of 175 cm. The pure crop of Leucaena at close spacing produced an average, over the six years, of 34 t ha⁻¹a⁻¹ of green fodder and 9.4 t ha⁻¹a⁻¹ of air dry fuelwood. The Leucaena at wide spacing produced 18.9 t ha⁻¹a⁻¹ of green fodder and 6.3 t ha⁻¹a⁻¹ of fuelwood. Intercropping with field crops decreased the yield of green fodder and fuelwood. The yield of all the field crops was less when raised as intercrops than as pure crops. Mean maximum net returns were obtained from intercrops of Leucaena and cluster bean (Rs 3540 ha⁻¹a⁻¹) which were significantly higher than the returns from pure crop of Leucaena at wide spacing but similar to the returns from pure crops of cluster bean. Leucaena with maize (Rs 3273 ha⁻¹a⁻¹) and black gram (Rs 3125 ha⁻¹a⁻¹) gave significantly higher net returns over pure crops of Leucaena at wide spacing, maize and black gram. ICRISAT = International Crops Research Institute for the Semi-Arid Tropics- Hyderabad, India. CIAT = Centro International de Agricultura Tropical - Cali - Columbia     

Resynthesis of Brassica juncea through interspecific crosses between B. rapa and B. nigra

The objective of this study was to broaden the genetic base in oleiferous Brassica juncea by resynthesis, using 10 diverse parental lines of oleiferous B. rapa and two lines of B. nigra of both Indian and exotic origin. Out of 14 crosses attempted using B. rapa as the female parent, eight were successful. Embryo rescue was necessary to obtain interspecific plants. A total of 29 fertile interspecific plants were obtained after colchicine treatment. In the S₂ generation, the expression of component characters in the majority of the resynthesized plants showed a negative trend. The resynthesized B. juncea lines are being maintained through repeated selfing and selection at each generation for desirable plant types. This process will continue till the progeny lines of the desirable plants achieve uniformity.     

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.     

Impact of Aeration on the Growth Performance of Silver Barb,Puntius gonionotus,during Fingerling Rearing

Impact of aeration on growth of silver barb, Puntius gonionotus during fingerling rearing was studied through a 100‐d rearing experiment conducted in 18 concrete tanks of 50 m² (10 × 5 × 1.2 m) size. Fry (0.74 ± 0.27 g, 35 ± 6 mm) were stocked in the experimental tanks at three stocking densities (25, 50, and 75 fry/m²) and were evaluated with and without provision of 6 h (2400–0600 h) of night time aeration. Aeration resulted in higher pH and dissolved oxygen regime and increased fingerling length and weight. The results suggest a rearing density of 75/m² to be ideal for rearing fry to fingerling of this species when aeration is provided, whereas, under non‐aerated condition, rearing the fry to fingerling stage at 50/m² was found advantageous over those at 25 and 75/m².     

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.     

Several micronutrients in the rotifer Brachionus sp. may not fulfil the nutritional requirements of marine fish larvae

The current best practice intensive culture of larval Atlantic cod includes feeding rotifers from onset of exogenous feeding until 25–30 days after hatching. These larvae grow considerably slower and develop higher frequencies of deformities than larvae reared in semi‐extensive systems, using copepods as feed. The present study compares the micronutrient concentrations in rotifers with those of copepods, with the aim of identifying nutrients that may be limiting for normal growth and development of cod larvae. An additional criterion used is the nutrient requirements given for fish in general, by NRC (1993) , as nutrient requirements of cod remains to be determined. Rotifers were fed on four different diets, consisting of baker's yeast with cod liver oil (3.3 : 1 dry weight (DW)/v), baker's yeast with Algamac 2000^TM (3.5 : 1 DW), baker's yeast with live algae Chlorella (4.1 : 1 DW), and Culture Selco 3000^TM (CS). CS was a complete commercial diet for rotifers while the other diets are considered as based on raw ingredients. Compared with copepod nutrient levels, rotifers grown on yeast‐based diets supplemented with either cod liver oil, Algamac 2000 or Chlorella were apparently sufficient for covering the requirements in cod larvae for all the B‐vitamins, except thiamine. Rotifers cultured on the CS diet also had sufficient amounts of thiamine. Of the minerals, only calcium and magnesium were sufficient, using this criterion while iron was on the borderline. However, with reference to the requirements given for larger fish ( NRC 1993 ), only thiamine, vitamin A, manganese, selenium and perhaps copper, appear too low in the rotifers cultured without extra micronutrient supplementation. The other nutrients were present at levels intermediate between copepod and fish requirement levels. This study suggests that it is necessary to develop enrichment techniques to produce rotifers with sufficient amounts of all micronutrients. Such techniques will also be important tools for determining which nutrients are present at levels below the actual requirements in cod larvae.     

Comparative changes in water quality and role of pond soil after application of different levels of organic and inorganic inputs

Changes in water parameters were studied in a yard experiment for 7 weeks after application of cow dung at 20, 50 and 100 t ha^?1, poultry manure at 4, 10 and 20 t ha^?1, feed mixture (groundnut oil cake and rice bran at 1:1) at 10, 20 and 30 t ha^?1 and inorganic fertilizers at 100 kg urea+50 kg single super phosphate (SSP) ha^?1, twice this dose (2x ha^?1) and thrice this dose (3x ha^?1). To study the role of soil in the mineralization process, each treatment was divided into two groups – one with and the other without soil substrate. Higher degree of changes in water parameters was observed at higher input levels. Both organic amendment and inorganic fertilization caused significant reduction (P<0.05) in dissolved oxygen and increase in free CO₂, dissolved organic matter, total ammonia, nitrite, nitrate and phosphorus contents of water. Organic inputs significantly decreased (P<0.05) water pH and increased total alkalinity and hardness. In contrast, inorganic fertilization caused a significant increase in pH; alkalinity and hardness increased significantly in the presence of soil, but reduced in its absence. In organic input, presence of soil substrate caused significantly lower value of pH, dissolved oxygen, dissolved organic matter and phosphate‐phosphorus and significantly higher free CO₂, alkalinity, hardness, ammonia, nitrite and nitrate contents, compared with those in the absence of soil, revealing enhanced microbial mineralization in the presence of soil.     

Effect of sub-lethal nitrite on selected haematological parameters in fingerling Catla catla (Hamilton)

A sub‐lethal nitrite toxicity trial was conducted using static conditions for a period of 96 h with fingerlings of Catla catla (21.4±3.6 g). Fingerlings were exposed to five concentrations of nitrite, that is, 1, 2, 4, 8 and 10.4 mg L^?1 and a nitrite‐free control to study changes in haematological parameters. Nitrite caused an increase in immature erythrocyte population (7–24%) in lower concentrations (0–4 mg L^?1) at 6 h while they were absent in higher concentrations. The total erythrocyte count was reduced at 6 h followed by an increase at 12 h with further reduction up to 96 h in all concentrations of nitrite. The 96‐h exposure resulted in 21.2–31.8% reduction in erythrocyte population in 1–10.4 mg L^?1 nitrite. The haemoglobin content decreased progressively with increasing nitrite concentrations as well as exposure periods. Total leukocyte count decreased initially at 6 h in all treatments followed by an increase after 12 h, signifying development of a protective response of the body to nitrite stress. Blood glucose decreased initially up to 24 h followed by an increase through 96 h. Serum protein level decreased continuously with increasing exposure period. The study revealed that exposure to nitrite caused changes in almost all the haematological parameters in the fingerlings depending on the concentration as well as exposure period. Nitrite being one of the important inorganic nutrients often recorded at higher levels in intensively cultured ponds, the present study highlights its adverse impact on fish and stressed the need for the management of this nutrient in culture ponds.     

Nursery rearing of kalbasu, Labeo calbasu (Hamilton), at different stocking densities in outdoor concrete tanks

The growth and survival of kalbasu, Labeo calbasu, was evaluated at stocking densities of 5, 10 and 15 million spawn ha⁻¹ in nursery rearing in concrete tanks of 50 m², each for a period of 25 days. Survival of fry was density dependent and significantly higher (54.5%) at 5 million ha⁻¹ than those at 10 million ha⁻¹ (50.1%) and 15 million ha⁻¹ (46.9%). Similarly, growth and specific growth rate were inversely related to the stocking density and varied significantly among the three densities.