Data on optimization of microprojectile bombardment parameters in development of salinity tolerant transgenic lines.

This data deals with the optimization of microprojectile bombardment particles for efficient genetic transformation in an indica rice involving AmSOD gene for development of salinity tolerant transgenic lines [1]. In this study, various parameters such as effect of genotypes, helium pressure, osmoticum, explants, flight distance, particle size, particle volume, vacuum, carrier DNA and stopping screen properties have been evaluated to determine their role in transformation of indica rice involving AmSOD gene for development of salinity tolerant Pusa Basmati 1 rice variety. To perform the transformation process, plasmid vector pCAMBIA 1305.2 was used, which harbours GUS Plus™ gene, intron from the castor bean catalase gene, pBR322 ori, kanamycin resistant gene and Xho I site. The transformants have been confirmed using slot blot, polymerase chain reaction and Southern hybridization techniques.

Effect of dietary supplementation of hemp (Cannabis sativa) and dill seed (Anethum graveolens) on performance, serum biochemicals and gut health of broiler chickens.

The present study was carried out to study the effect of different doses of hemp seed alone or in combination with dill seed against antibiotic growth promoter on performance, serum biochemicals and gut health of broiler chickens over a period of 42 days. Total 192 broiler chicks were grouped randomly into six treatments and fed with basal diet (BD) along with different levels of seeds, viz., T1 (BD), T2 (BD + 0.2% HS), T3 (BD + 0.2% HS + 0.3 DS), T4 (BD + 0.3% HS) and T5 (BD + 0.3% HS + 0.3 DS) and T6 (BD + 0.025% Bacitracin Methylene Disalicylate-BMD). The performance traits like feed intake, body weight gain and feed conversion ratio (FCR) and carcass traits like cut-up parts, giblet and abdominal fat yield remained unaffected due to dietary treatments for overall trial period; however, the average feed intake in early phase (0-3 weeks) reduced significantly (p < 0.05) in treatment birds than both controls (T1 & T6). Serum protein concentration remained unchanged, whereas significant (p < 0.05) reduction in serum lipids like triglyceride, LDL and total cholesterol concentration was noticed due to dietary inclusion of seeds. Serum enzymes like AST and ALT concentrations depleted significantly (p < 0.05) treated groups, except at higher seed doses (T5); however, alkaline phosphatase levels were unaffected. Coliform count in caecum and jejunum reduced linearly (p < 0.01) due to seed inclusion, whereas dose-dependent proliferation of lactobacilli was evident (p < 0.01) in caecum and jejunum of treated birds. No effect was observed on the villus height and crypt depth of the jejunal mucosa. To conclude, dietary supplementation of hemp and dill seed could not affect the growth performance and carcass traits; however, it positively altered the serum lipid profile of the birds and improved gut health as well, thereby enhanced overall performance of broiler chickens.

Assessment of genetic purity in rice (Oryza sativa L.) hybrids using microsatellite markers.

The objective of the present study is to detect genetic impurity in the seed lots of CMS lines, restorers and hybrids and to identify signature markers to differentiate parents and hybrids through DNA-based assays. Furthermore, attempts have been made to find out an alternative to Grow-Out-Test, which is very tedious, time consuming and used conventionally for seed genetic purity testing since beginning of quality seed multiplication chain. Fifty-one rice-specific sequence tagged microsatellite (STMS) primer pairs distributed throughout the rice genome were employed for fingerprinting of eight rice hybrids and their parental lines with a view to assess variation within parental lines and to test the genetic purity of the commercial seed lots. Among those, 51 markers, 28 microsatellite markers showed polymorphism (54.90 %). A total of 98 alleles were obtained with an average of 1.92 alleles per primer pair and number of alleles amplified for each primer pair ranged from 1 to 4. A set of markers were identified to differentiate parental lines of the hybrids and which emphasizes the immense scope of those molecular markers for their use in the unambiguous identification of hybrid, which would be of great benefit to farmers that depend on the hybrids.

Molecular Level Insights on Collagen-Polyphenols Interaction Using Spin-Relaxation and Saturation Transfer Difference NMR.

Interaction of small molecules with collagen has far reaching consequences in biological and industrial processes. The interaction between collagen and selected polyphenols, viz., gallic acid (GA), pyrogallol (PG), catechin (CA), and epigallocatechin gallate (EGCG), has been investigated by various solution NMR measurements, viz., (1)H and (13)C chemical shifts (δH and δC), (1)H nonselective spin-lattice relaxation times (T1NS) and selective spin-lattice relaxation times (T1SEL), as well as spin-spin relaxation times (T2). Furthermore, we have employed saturation transfer difference (STD) NMR method to monitor the site of GA, CA, PG, and EGCG which are in close proximity to collagen. It is found that -COOH group of GA provides an important contribution for the interaction of GA with collagen, as evidenced from (13)C analysis, while PG, which is devoid of -COOH group in comparison to GA, does not show any significant interaction with collagen. STD NMR data indicates that the resonances of A-ring (H2', H5' and H6') and C-ring (H6 and H8) protons of CA, and A-ring (H2' and H6'), C-ring (H6 and H8), and D-ring (H2″and H6″) protons of EGCG persist in the spectra, demonstrating that these protons are in spatial proximity to collagen, which is further validated by independent proton spin-relaxation measurement and analysis. The selective (1)H T1 measurements of polyphenols in the presence of protein at various concentrations have enabled us to determine their binding affinities with collagen. EGCG exhibits high binding affinity with collagen followed by CA, GA, and PG. Further, NMR results propose that presence of gallic acid moiety in a small molecule increases its affinity with collagen. Our experimental findings provide molecular insights on the binding of collagen and plant polyphenols.

Evaluation of an ethnomedicinal combination containing Semecarpus kurzii and Hernandia peltata used for the management of inflammation.

CONTEXT: Scientific validation of an ethnomedicinal combination consisting of Semecarpus kurzii Engler (Anacardeaceae) leaves (SKL) and Hernandia peltata Meisn (Hernandeaceae) stem-bark (HPB), traditionally used in ailments related to inflammation, pain and fever. OBJECTIVE: To validate in vivo and in vitro analgesic and antiinflammatory activities of methanol extract of SKL, HPB and their combination. MATERIALS AND METHODS: Analgesic activity was tested by acetic acid induced writhing reflex and tail flick in Swiss albino mice, while the anti-inflammatory activity was studied in acute, subacute and chronic model on Wistar rats. The vascular permeability, membrane stabilization and protein denaturation were examined to know the possible mode of action. RESULTS: Significant (p < 0.01) analgesic (78.04% inhibition of writhing) and antiinflammatory (72.54% inhibition of paw edema) activity was observed in combination of SKL and HPB extracts at 250 mg/kg each. The SKL extract alone inhibits acetic acid-induced vascular permeability (64.4%) at 500 mg/kg, while in combination at 250 mg/kg each, the inhibition was 69.49% (p < 0.01). Furthermore, SKL in combination with HPB (0.25 mg/mL each) prevent RBC hemolysis (61.91%) and inhibition of protein denaturation (76.52%)-like indomethacin. DISCUSSION AND CONCLUSION: The SKL and HPB extract, alone (500 mg/kg) and in combination, (250 mg/kg each) had significant analgesic and antiinflammatory activity, probably by inhibiting the release of certain inflammatory mediators and membrane stabilization, due to the presence of triterpenes, tannins and related phytochemicals in the extracts. Thus, our results demonstrated that this combination provide the scientific rationale of its folk use.

Osmotic shock augments ethanol stress in Saccharomyces cerevisiae MTCC 2918.

Yeast cells sense and respond to hypertonicity. Saccharomyces cerevisiae MTCC 2918 was tested for its metabolic status in 1 M NaCl by cell viability analysis, intracellular glycerol content and total antioxidant capacity. Yeast cell viability was maximum in 1 M NaCl and 24 h addition of 1 M NaCl was effective in induction of hyperosmolarity. Increased glycerol contents in cells treated with salt indicated adaptation to osmotic stress with a maximum of 240.87 ± 0.38 mg/g dry weight (DW) at 72 h. The total antioxidant status with 1 M NaCl was 9.29 ± 0.39 mM/g DW at 96 h reflecting free radical quenching to overcome stress with increasing growth period. Considering that pre-adaptation to one type of stress evoked a protective response to other stress factors, we have attempted the cross adaptation of osmotic shock to high ethanol concentrations. In effect, we observed that osmotic shock lowered the cell survival by augmentation of cell toxicity by ethanol due to stress induction during exponential phase. Glycerol accumulation to an order of 470.27 ± 0.53 mg/g DW at 48 h in 1 M NaCl and 12% ethanol indicated that both stresses culminated in membrane disruption further leading to cell burst and contributed to the stress overload.

Comparative study on secondary structural changes in diabetic and non-diabetic human finger nail specimen by using FTIR spectra.

BACKGROUND: In human anatomy, a nail is a hornlike envelope covering the dorsal aspect of the terminal phalanges of fingers and toes. Nail disorders are most common among the geriatric population. Diabetes mellitus is also supposed to affect the condition of nails. Acceptable differences in infrared (IR) spectra of chronic and acute diabetes mellitus patient fingernail specimens compared to control normal specimens were investigated in this study. METHODS: Using a Nicolet 360 Fourier Transform Infra Red (FTIR) spectrometer, the spectra of the nails of diabetics and normal specimens were recorded. RESULT: In the case of non-diabetic patients, the amide I band was observed <1640 cm(-1) (1626, 1632, and 1638 cm(-1)). The bands around 1637 cm(-1), were attributable to amide I of ß sheet structures. Amide II bands were absent in all the non-diabetic patients. Amide III bands around 1250 cm(-1) were observed both in diabetic and non-diabetic patients. In all the diabetic patients, a peak of <500 cm(-1), particularly around 468 cm(-1), was observed. CONCLUSION: The proteins in the nails of diabetic patients contain α-helical structure, including the presence of amide II bonds. Alkyl thiolated structures are observed. Nails of non-diabetic patients do not have the amide II structures.

Effect of dietary level of methionine on growth performance and immune response in Japanese quails (Coturnix coturnix japonica).

BACKGROUND: Methionine (Met) being the first limiting amino acid in maize/soybean-based quail diets, its supplementation provides scope for improvement of protein quality and reduction of dietary protein concentration. The question remains to what extent it can be incorporated in the diet of genetically improved quails. Therefore the effect of dietary Met level was assessed on growth performance and immune response in growing Japanese quails (n = 400) divided equally into 20 groups. Five dietary treatments (approximately 230 g kg(-1) crude protein and 12.14 MJ kg(-1) metabolisable energy) were formulated with 3.5, 4.5, 5.0, 5.5 and 6.0 g kg(-1) Met respectively, and each was offered to four groups of birds from 0 to 35 days of age. RESULTS: Live weight at day 35 increased (P < 0.0001) up to 5.0 g kg(-1) dietary Met level but did not improve further at higher Met levels (5.5 and 6.0 g kg(-1)). Improved (P < 0.039) feed conversion ratio was achieved at 5.5 g kg(-1) Met level, which was statistically similar to that at 5.0 g kg(-1) Met level during 0-14 days of age. Cellular (phytohaemagglutinin from Phaseolus vulgaris) immune response increased (P < 0.0001) with increasing dietary Met concentration, whereas humoral (sheep red blood cells) immune response did not differ. CONCLUSION: The optimal requirement of Met was 5.0 g kg(-1) for growth and 5.5 g kg(-1) for maximum cellular immune response.

Preparation, characterization and evaluation of a biopolymeric gold nanocomposite with antimicrobial activity.

The present study describes the antimicrobial activity of C-AuNp-Amp (chitosan-capped gold nanoparticles coupled with ampicillin). C-AuNp-Amp was synthesized using the wet precipitation method and characterized using FTIR (Fourier-transform IR) spectroscopy and AFM (atomic force microscopy) techniques. The optimal level of ampicillin concentration that couples with the C-AuNp nanocomposite was determined by using UV-visible spectroscopy. The agar-well diffusion method was used to evaluate the antimicrobial activity, and the broth dilution assay was used to determine the MIC (minimum inhibitory concentration). The size of the ellipsoidal C-AuNp-Amp particles was found to be in the range of 50-100 nm. The FTIR spectrum confirms the bonding between amino groups of chitosan and carboxylic groups of ampicillin. The maximum coupling of ampicillin with C-AuNp was found to be 4.07 mg/10 ml. These results revealed the antimicrobial efficacy of C-AuNp-Amp and a 2-fold increase in activity was achieved when compared with that of free ampicillin. By reducing the antibiotic dosage to 50%, the side effects produced by antibiotics can be minimized.

Sperm motility inhibiting activity of a phytosterol from Alstonia macrophylla Wall ex A. DC. leaf extract: a tribal medicine.

The role of methanolic extract and n-butanol fraction of A. macrophylla leaves was investigated on the forward motility of goat spermatozoa. The methanol extract (600 micro/g/ml) and one n-butanol fraction (Fraction A; 100 microg/ml) showed marked inhibition of sperm forward motility, tested by microscopic and spectrophotometric methods. Approximately, 50-60% of the spermatozoa lost their motility when treated with 600 microg/ml of methanol extract or 100 microg/ml of Fraction A. The Fraction A at 400 microg/ml concentration showed complete inhibition of sperm forward motility at 0 min. The inhibitory activity increased with the increasing concentrations of the fraction. The motility inhibitory activity of the Fraction A was stable to heat treatment at 100 degrees C for 2 min. The compound showed high inhibitory effect in the pH range 6.7-7.6. Fraction A also showed high efficacy for inhibiting human sperm motility, assessed by the microscopic method. The phytochemical analysis of methanolic extract of A. macrophylla leaves revealed the presence of sterols, triterpene, flavonoid, alkaloid, tannin and reducing sugar, while the Fraction A contains beta-sitosterol, a common phytosterol. The results demonstrate that Fraction A (beta-sitosterol) is a potent inhibitor of sperm motility and thus it has the potential to serve as a vaginal contraceptive.

A potent sperm motility-inhibiting activity of bioflavonoids from an ethnomedicine of Onge, Alstonia macrophylla Wall ex A. DC, leaf extract.

The methanol extract (ME) and the n-butanol fractions of methanolic extract of Alstonia macrophylla Wall ex A. DC leaves were investigated on the forward motility (FM) of mammalian (goat and human) spermatozoa. The ME at 600 microg mL-1 as well as fraction B at 100 microg mL-1 concentrations showed marked inhibition of sperm FM in both goat and human species when tested by microscopic and spectrophotometric methods. Approximately 60-80% of the goat spermatozoa lost their FM when treated with 600 microg mL-1 of ME and 100 microg mL-1 of fraction B. At 100 microg mL-1 concentration, fraction B showed 90% loss of FM in human spermatozoa, while fraction B at 400 microg mL-1 concentration showed complete inhibition of sperm FM at 0 min. The inhibitory activity of fraction B increases with increasing concentration in a dose-dependent manner. Phytochemical study of the extract revealed that the leaf contains tannins, flavonoids, sterols, triterpenes, alkaloids and reducing sugars. Further fractionation and purification of the bioactive n-butanol part of ME showed the presence of ursolic acid (fraction B), beta-sitosterol (fraction A), beta-sitosterol glucoside and a mixture of minor compounds (fraction C, detected on thin-layer chromatography). The results reveal that fraction B (ursolic acid), a pentacyclic triterpene, has the potential of sperm motility inhibition and can serve as a topical vaginal contraceptive.

Antipyretic activity of Alstonia macrophylla Wall ex A. DC: an ethnomedicine of Andaman Islands.

PURPOSE: Alstonia macrophylla Wall ex A. DC. Leaf, used in different ailments by the Onge tribes of Little Andaman Island, India, was investigated for its antipyretic potential. METHODS: The methanol extract and its fractions were tested on normal body temperature and yeast-induced pyrexia in Wistar Albino rats. RESULTS: The leaf extract at oral doses of 200 and 300 mg/kg, and the n-butanol fractions of the extract at 50 mg/kg showed significant reduction in normal body temperature and yeast-provoked elevated temperature in a dose-dependent manner comparable to that of standard antipyretic drug paracetamol. The antipyretic effect was started at 1 h and extended for at least 5 h after the drug administration. CONCLUSIONS: The antipyretic effect was more pronounced when the fraction A and B was administered together, indicating that both the fractions may contain antipyretic compounds which produce an additive effect in combination. Phytochemically these fractions contain beta-sitosterol and ursolic acid.

CNS activity of Alstonia macrophylla leaf extracts: an ethnomedicine of Onge of Bay Islands.

Methanol extract at 100-200 mg/kg p.o. and major nonpolar fraction B at 50 mg/kg of Alstonia macrophylla leaves caused a significant reduction in spontaneous activity, remarkable decrease in exploratory behavioural pattern, a reduction in muscle relaxant activity and also significantly potentiated phenobarbitone sodium-induced sleeping time. The phytochemical study of crude leaf extract revealed the presence of tannin, triterpenoid, flavonoid, sterol, alkaloid and reducing sugars. Further fractionation and purification of the n-butanol part of methanol extract yielded fraction A, fraction B and fraction C along with some minor fatty acids as the major compounds.

CNS activity of the methanol extract of Mallotus peltatus (Geist) Muell Arg. leaf: an ethnomedicine of Onge.

The aim of the present study was to investigate several neuropharmacological effects of the methanol extract and different fractions of Mallotus peltatus (Geist) Muell Arg. var acuminatus (Euphorbiaceae) leaves in Wistar albino rats and Swiss albino mice. General behavior, exploratory behavior, muscle relaxant activity and phenobarbitone sodium-induced sleeping time were studied. The results revealed that the crude extract at 200-300 mg kg(-1) p.o. and its fractions A and B at 50 mg kg(-1) caused a significant reduction in spontaneous activity (general behavioral profile), remarkable decrease in exploratory behavioral pattern (Y-maze and head dip tests), a reduction in muscle relaxant activity (rotarod, 30 degrees inclined screen and traction tests), and also significantly potentiated phenobarbitone sodium-induced sleeping time. The phytochemical study of crude leaf extract revealed the presence of tannin, triterpenoid, flavonoid, sterol, alkaloid and reducing sugar. Further fractionation and purification yielded two major fractions A (ursolic acid) and B (beta-sitosterol) with some fatty acids as the major compounds. The psychopharmacological activity of the crude leaf extracts appeared to be either due to fraction A (50 mg kg(-1)) or a combination of fractions A and B (50 mg kg(-1)) along with some fatty acids present in the n-butanol part of methanol extract of M. peltatus leaf (MEMPL).

Antimicrobial and anti-inflammatory activity of folklore: Mallotus peltatus leaf extract.

Since ages Mallotus peltatus (Geist) Muell. Arg. var acuminatus (Euphorbiaceae) leaf and stem bark is used in folk medicine to cure intestinal ailments and skin infections. In several intestinal ailments, localized inflammation is of common occurrence and hence we have evaluated the antimicrobial as well as anti-inflammatory activity of M. peltatus leaf extract. The crude methanol extract of M. peltatus leaves was found to be active against Staphylococcus aureus, Staphylococcus saprophyticus, Streptococcus faecalis, Bacillus subtilis, Escherichia coli and Proteus mirabilis and the dermatophytic fungi Microsporum gypseum. The minimum inhibitory concentration (MIC) ranges from 128 to 2000 microg ml(-1) for bacteria and 128 mg ml(-1) for fungi, while the minimum bactericidal concentration (MBC) was 2-4-fold higher than MIC. The methanol-water fraction of the extract showed similar activity against Staphylococcus, Streptococcus, Bacillus and Proteus isolates. The anti-inflammatory activity of the extract against carrageenan (acute model) and dextran-induced (subacute model) rat paw oedema and cotton pellet-induced granuloma (chronic model) in rats were studied using indomethacin (10 mg kg(-1)), a nonsteroid anti-inflammatory drug, as standard. The methanol extract at 200 and 400 mg kg(-1), and the n-butanol fractions A and B at 25 mg kg(-1), exhibited significant anti-inflammatory activity in Albino rats, compared with indomethacin. Phytochemical study revealed the presence of tannins, saponins, terpenoids, steroids and reducing sugars in the crude extract while the n-butanol fractions showed the presence of ursolic acid, beta-sitosterol and some fatty acids as major compounds. Further study with fractions showed that the antibacterial and anti-inflammatory activity is due to either fraction A (ursolic acid) alone or the combination of fractions A and B (beta-sitosterol and fatty acids) of the extract.

Feeding standards for lactating riverine buffaloes in tropical conditions.

Data from 33 feeding trials, conducted on lactating riverine buffaloes from different institutes across India, were subjected to multiple regression analysis to derive nutritional requirements for dry matter (DM), total digestible nutrients (TDN), crude protein (CP) and digestible crude protein (DCP) for maintenance, milk production and body weight gain. Maintenance requirements for DM, TDN, CP and DCP were 59.9, 35.3, 5.43 and 3.14 g/kgW(0.75), respectively; corresponding requirements for producing 1 kg 6% FCM were 688, 406, 90.3 and 55.2 g and for 1 g gain in body weight were 3.37, 1.97, 0.327 and 0.23 g. Regression equations had high R2 values (061. 0.66, 0.84 and 0.68 for prediction of DM, TDN, CP and DCP, respectively) and the equations (F-value) as well as coefficients were highly significant (P <0.001). Regressed values were used to derive feeding standards. Derived values matched well with the actual intake versus performance of animals under diverse feeding conditions. New standards predicted requirements and intake of nutrients for different production levels better than existing feeding standards. Because they are based on a more thorough analysis of data, the new feeding standards will be appropriate for use widely in India.