Studies on the antioxidant activities of natural vanilla extract and its constituent compounds through in vitro models

Vanilla extract was prepared by extraction of cured vanilla beans with aqueous ethyl alcohol (60%). The extract was profiled by HPLC, wherein major compounds, viz., vanillic acid, 4-hydroxybenzyl alcohol, 4-hydroxy-3-methoxybenzyl alcohol, 4-hydroxybenzaldehyde and vanillin, could be identified and separated. Extract and pure standard compounds were screened for antioxidant activity using beta-carotene-linoleate and DPPH in vitro model systems. At a concentration of 200 ppm, the extract showed 26% and 43% of antioxidant activity by beta-carotene-linoleate and DPPH methods, respectively, in comparison to corresponding values of 93% and 92% for BHA. Interestingly, 4-hydroxy-3-methoxybenzyl alcohol and 4-hydroxybenzyl alcohol exhibited antioxidant activity of 65% and 45% by beta-carotene-linoleate method and 90% and 50% by DPPH methods, respectively. In contrast, pure vanillin exhibited much lower antioxidant activity. The present study points toward the potential use of vanilla extract components as antioxidants for food preservation and in health supplements as nutraceuticals.     

Mitochondrial phylogeography reveals high haplotype diversity and unique genetic lineage in Indian dugongs (Dugong dugon)

1. India plays a significant role in dugong conservation by having the largest population within South Asia. The status of dugongs in India is largely unknown due to a paucity of reliable ecological data. This study generated mitochondrial control region sequences from ~10% of dugong individuals from existing populations within India. Furthermore, data generated in this study were compared with the global data to assess genetic lineages, population structure, and genetic diversity of Indian populations.
2. Multiple analyses suggest that the Indian dugong populations are part of a single genetic cluster, comprising South Asia, North-west Indian Ocean, and South-west Indian Ocean populations. Despite small population size, they retain high genetic diversity with unique mitochondrial DNA haplotypes within South Asia. Within India, novel haplotypes are observed from all dugong habitats sampled, with overall high haplotype diversity (0.85 ± 0.04) but low nucleotide diversity (0.005 ± 0.001). Indian populations exhibit genetic differentiation with higher within-population variance (63.41%) than among populations (36.59%). Two of the haplotypes observed in India are shared with Sri Lanka, implying genetic connectivity between these populations.
3. The genetic data from Indian dugong populations provide critical insights into the identification of dugong corridors and important dugong conservation zones in India. We suggest site-specific interventions, including the creation of new marine protected areas and boundary reorganization and expansion of other existing protected areas, to ensure population connectivity. In addition, simultaneous efforts towards seagrass meadow restoration, reduction of dugong mortalities, and community participation in dugong conservation are recommended for population recovery of this threatened marine herbivore.

     

The neu gene: an erbB-homologous gene distinct from and unlinked to the gene encoding the EGF receptor

The neu oncogene, identified in ethylnitrosourea-induced rat neuroglioblastomas, had strong homology with the erbB gene that encodes the epidermal growth factor receptor. This homology was limited to the region of erbB encoding the tyrosine kinase domain. It was concluded that the neu gene is a distinct novel gene, as it is not coamplified with sequences encoding the EGF receptor in the genome of the A431 tumor line and it maps to human chromosome 17.     

<Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated transformation of safflower and the efficient recovery of transgenic plants via grafting

Background

Safflower (Carthamus tinctorius L.) is a difficult crop to genetically transform being susceptible to hyperhydration and poor in vitro root formation. In addition to traditional uses safflower has recently emerged as a broadacre platform for the production of transgenic products including modified oils and pharmaceutically active proteins. Despite commercial activities based on the genetic modification of safflower, there is no method available in the public domain describing the transformation of safflower that generates transformed T₁ progeny.

Results

An efficient and reproducible protocol has been developed with a transformation efficiency of 4.8% and 3.1% for S-317 (high oleic acid content) and WT (high linoleic acid content) genotypes respectively. An improved safflower transformation T-DNA vector was developed, including a secreted GFP to allow non-destructive assessment of transgenic shoots. Hyperhydration and necrosis of Agrobacterium-infected cotyledons was effectively controlled by using iota-carrageenan, L-cysteine and ascorbic acid. To overcome poor in vitro root formation for the first time a grafting method was developed for safflower in which ~50% of transgenic shoots develop into mature plants bearing viable transgenic T₁ seed. The integration and expression of secreted GFP and hygromycin genes were confirmed by PCR, Southern and Western blot analysis. Southern blot analysis in nine independent lines indicated that 1-7 transgenes were inserted per line and T₁ progeny displayed Mendelian inheritance.

Conclusions

This protocol demonstrates significant improvements in both the efficiency and ease of use over existing safflower transformation protocols. This is the first complete method of genetic transformation of safflower that generates stably-transformed plants and progeny, allowing this crop to benefit from modern molecular applications.

     

Synthesis of new compounds related to the commercial fungicide tricyclazole

BACKGROUND: Tricyclazole is a commercial fungicide used to control rice blast. As part of re‐registration activities, samples of metabolites and process impurities are required. In addition, isotopically labeled tricyclazole samples are also required. RESULTS: Four new compounds related to tricyclazole are reported. An isotopically labeled sample of tricyclazole was prepared that contained two ¹⁵N atoms and one ¹³C atom. Radiolabeled tricyclazole with ¹⁴C at the triazole C3 position was also synthesized. A new process impurity in technical tricyclazole was identified and synthesized. A new metabolite of tricyclazole was identified, independently synthesized and characterized by X‐ray crystallography. CONCLUSION: A previously unreported metabolite of tricyclazole has been identified and structurally characterized. In addition, a new process impurity has been identified by independent synthesis. Identification of these new compounds has facilitated the continued registration of this important fungicide. Copyright © 2011 Society of Chemical Industry     

Isolation and characterization of neutral lipids of desilked eri silkworm pupae grown on castor and tapioca leaves

The neutral lipid of desilked eri silkworm pupae (Samia cynthia ricini) grown on two different host plants, castor (Ricinus communis Linn.) and tapioca (Manihot utilizsima Phol.) leaves, was extracted with hexane. The oil content in pupae was estimated to be in the range of 18-20% (dry basis). The pupal oil was found to be enriched with alpha-linolenic acid (ALA) with palmitic acid as the second major fatty acid. The level of ALA in the oil of silkworm pupae was found to be significantly higher (P < 0.001) when grown on tapioca (58.3%) as compared to those grown on castor (42.9%). Other chemical parameters such as percent free fatty acid, peroxide value, phosphorus content, percent unsaponifiable matter, and composition of sterols were also determined in both of the oils and compared. Reversed-phase high-performance liquid chromatography analysis of triacylglycerol molecular species showed that the pupal oil is rich in molecular species with equivalent carbon numbers (ECN) C36, C40, C42, C44, and C48. There was a significantly higher level (P < 0.001) of trilinolenin (C36) in the oil of tapioca-based silkworm as compared to castor-based silkworm pupae. Regiospecific analysis of the oil showed a higher level of ALA at the sn-2 position of silkworm pupae grown on tapioca (60.2%) as compared to those grown on castor (47.3%) oil. Thus, the presence of a large amount of ALA and their predominance at the sn-2 position make the eri pupal oil highly nutritious, provided that the oxidative stability is ensured.     

Molecular characterization of Hepatozoon spp. infection in endangered Indian wild felids and canids

Hepatozoon species are parasites that infect a wide variety of domestic and wild animals. The objective of this study was to perform the molecular detection and characterization of Hepatozoon spp. in Asiatic lion, Indian tiger, Indian leopard, Indian wild dog, Indian domestic dog and cat based on partial 18S rRNA gene sequences from Hepatozoon spp. in the naturally infected animals. Hepatozoon spp. could be detected in blood samples of 5 out of 9 Asiatic lions, 2 out of 5 Indian tigers, 2 out of 4 Indian leopards and 2 out of 2 Indian wild dogs and, 2 out of 4 domestic cats and 2 out of 3 domestic dog samples by PCR. Sequencing of PCR amplicon and BLAST analysis of partial 18S rRNA gene sequences indicated that the Hepatozoon spp. in Asiatic lion, Bengal tiger, Indian leopard and domestic cat was Hepatozoon felis (98-99% similarity) and in the Indian wild and domestic dog the phylogenetic neighbour was Hepatozoon canis (97-100% similarity). Presence of H. felis and H. canis in both domestic and wild animals suggested that they are not host specific and the same parasite causes infection in domestic and wild felids and canids in India and from different parts of the world. To our knowledge, this is the first report on detection and molecular characterization of H. felis infection in Asiatic lions, Indian tigers, Indian leopards and H. canis in Indian wild dog. Hepatozoon spp. may be a potential pathogen and an opportunistic parasite in immuno-compromised animals and could thus represent a threat to endangered Indian wild felids and canids.     

Effect of tillage management on energy-use efficiency and economics of soybean (Glycine max) based cropping systems under the rainfed conditions in North-West Himalayan Region

The study on energy-use efficiency and economics of soybean based cropping system, viz., soybean–wheat (Glycine max 9 L., Triticum aestivum L. emend. Fiori & Paol.), soybean–lentil (Lens culinaris Medicus) and soybean–field pea (Pisum sativum L., sensu lato), was carried out at the Hawalbagh experimental farm of Vivekananda Institute of Hill Agriculture, Almora, Uttarakhand, India during 2001–2003 (29°36′ N, 79°40′ E). These cropping systems, under rainfed conditions, were evaluated with different tillage management practices, viz., zero tillage (ZT), minimum tillage (MT) and conventional tillage (CT). Each tillage management practice, under each cropping system was evaluated for total energy output, energy input–output ratio, gross income, net income and marginal income, to variable cost of cultivation. Results revealed that the maximum energy was consumed in terms of chemical fertilizers, followed by seed and plant protection chemicals, in all cropping systems. Equivalent energy was used from literature for conversion purpose. The maximum output energy was observed in CT (44,253 MJ/ha), followed by MT for soybean–lentil cropping system (43,450 MJ/ha). The output–input energy ratio was maximum in ZT for soybean–lentil (4.9) followed by MT for soybean–pea cropping system (4.6). The economic analysis also revealed that the maximum benefits could be obtained from these sequences. Conventional tillage for all cropping sequences was found to be a better option as compared to minimum tillage and zero tillage. Benefit–cost ratios were higher in conventional tillage in all the three cropping systems. However, from the point of energy saving or cost reduction, zero tillage and minimum tillage may be considered depending on resources.     

Western blot detection of PrP Sc in archived paraffin-embedded brainstem from scrapie-affected sheep

Scrapie is a naturally occurring fatal neurodegenerative disease of adult sheep and goats, one of a group of mammalian diseases known as transmissible spongiform encephalopathies (TSE) or prion diseases. Immunoassays that identify disease-associated prion protein (PrP Sc) are integral to the diagnosis of scrapie and other prion diseases. Results obtained by either immunohistochemistry (IHC) or Western blot (WB) assay are generally adequate for the definitive diagnosis. Approved or accepted methods for WB diagnosis of TSEs requires the use of fresh or frozen nonfixed tissue samples, whereas formalin-fixed, paraffin-embedded tissue is required for the localization of PrP Sc by IHC. Because disparate processing methods are used for these accepted diagnostic techniques, separate tissue samples are collected from the same animal. Occasions arise in which there is either insufficient quantity of tissue available to complete analysis by both techniques or initial tissue processing is incompatible with one of the assays. Also, results between the assays may differ because of the vagaries of sampling, especially in case material that contains moderate-to-low levels of PrP Sc. The present article describes a method to conduct a WB assay from the same paraffin-embedded brainstem sample used for the IHC diagnosis of experimentally induced sheep scrapie.