Relatedness of Indian Flax Genotypes (<Emphasis Type="Italic">Linum usitatissimum</Emphasis> L.): An Inter-Simple Sequence Repeat (ISSR) Primer Assay

The objective of this study was to analyze the genetic relationships, using PCR-based ISSR markers, among 70 Indian flax (Linum usitatissimum L.) genotypes actively utilized in flax breeding programs. Twelve ISSR primers were used for the analysis yielding 136 loci, of which 87 were polymorphic. The average number of amplified loci and the average number of polymorphic loci per primer were 11.3 and 7.25, respectively, while the percent loci polymorphism ranged from 11.1 to 81.8 with an average of 63.9 across all the genotypes. The range of polymorphism information content scores was 0.03–0.49, with an average of 0.18. A dendrogram was generated based on the similarity matrix by the Unweighted Pair Group Method with Arithmetic Mean (UPGMA), wherein the flax genotypes were grouped in five clusters. The Jaccard’s similarity coefficient among the genotypes ranged from 0.60 to 0.97. When the omega-3 alpha linolenic acid (ALA) contents of the individual genotypes were correlated with the clusters in the dendrogram, the high ALA containing genotypes were grouped in two clusters. This study identified SLS 50, Ayogi, and Sheetal to be the most diverse genotypes and suggested their use in breeding programs and for developing mapping populations.     

Cloning of a Novel Omega-6 Desaturase from Flax (<Emphasis Type="Italic">Linum usitatissimum</Emphasis> L.) and Its Functional Analysis in <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis>

The Δ12 desaturase represents a diverse gene family in plants and is responsible for conversion of oleic acid (18:1) to linoleic acid (18:2). Several members of this family are known from plants like Arabidopsis and Soybean. Using primers from conserved C- and N-terminal regions, we have cloned a novel Δ12 desaturase gene amplified from flax genomic DNA, denoted as LuFAD2-2. This intron-less gene is 1,149-base pair long encoding 382 amino acids—putative membrane-bound Δ12 desaturase protein. Sequence comparisons show that the novel sequence has 85% similarity with previously reported flax Δ12 desaturase at amino acid level and shows typical features of membrane-bound desaturase such as three conserved histidine boxes along with four membrane-spanning regions that are universally present among plant desaturases. The signature amino acid sequence ‘YNNKL’ was also found to be present at the N terminus of the protein, which is necessary and sufficient for ER localization of enzyme. Neighbor-Joining tree generated from the sequence alignment grouped LuFAD2-2 among the other FAD2 sequences from Ricinus, Hevea, Jatropha, and Vernicia. When LuFAD2-2 and LuFAD2 were expressed in Saccharomyces cerevisiae, they could convert the oleic acid to linoleic acid, with an average conversion rate of 5.25 and 8.85%, respectively. However, exogenously supplied linoleic acid was feebly converted to linolenic acid suggesting that LuFAD2-2 encodes a functional FAD2 enzyme and has substrate specificity similar to LuFAD2.     

An ecofriendly synthesis and DNA binding interaction study of some pyrazolo [1,5-a]pyrimidines derivatives

The DNA molecule is a target for plethora of anticancer and antiviral drugs that forms covalent and non-covalent adducts with major or minor groove of DNA. In present study we synthesized series of novel Pyrazolo [1,5-a]pyrimidine derivatives. The newly synthesized compounds were characterized by elemental analysis, IR, ¹H NMR, and mass spectral data. The selected compounds were studied for interaction with Calf thymus DNA (CT-DNA) using electronic spectra, viscosity measurement and thermal denaturation studies. Further, molecular interactions were revealed for compound IIIa and IVa by computational methodologies. The preferred mode of ligand binding with double helical DNA as well as preferable DNA groove were explored by molecular docking in different DNA models.     

Oxidized phospholipid content destabilizes the structure of reconstituted high density lipoprotein particles and changes their function

High density lipoprotein (HDL) particles are made up of lipid and protein constituents and apolipoprotein A-I (apoA-I) is a principal protein component that facilitates various biological activities of HDL particles. Increase in Ox-PL content of HDL particles makes them 'dysfunctional' and such modified HDL particles not only lose their athero-protective properties but also acquire pro-atherogenic and pro-inflammatory functions. The details of Ox-PL-induced alteration in the molecular properties of HDL particles are not clear. Paraoxonase 1 (PON1) is an HDL-associated enzyme that possesses anti-inflammatory and anti-atherogenic properties; and many of the athero-protective functions of HDL are attributed to the associated PON1. In this study we have characterized the physicochemical properties of reconstituted HDL (rHDL) particles containing varying amounts of Ox-PL and have compared their PON1 stimulation capacity. Our results show that increased Ox-PL content (a) modifies the physicochemical properties of the lipid domain of the rHDL particles, (b) decreases the stability and alters the conformation as well as orientation of apoA-I molecules on the rHDL particles, and (c) decreases the PON1 stimulation capacity of the rHDL particles. Our data indicate that the presence of Ox-PLs destabilizes the structure of the HDL particles and modifies their function.     

Evaluation of metal contamination and risk assessment to human health in a coal mine region of India: A case study of the North Karanpura coalfield

The present study aimed to evaluate metal contamination level in coal mine water of the North Karanpura coalfields and assess the possible health risk due to the intake of untreated mine water. Fourteen coal mine water samples were collected from the study area and analyzed for Al, As, Ba, Cr, Cu, Fe, Mn, Ni, Se, and Zn using inductively coupled plasma-mass spectroscopy (ICP-MS). The heavy metal pollution index (HPI) was used to calculate metal pollution level in the coal mine water. The hazard quotient (HQ) and hazard index (HI) were estimated for health risk to child and adult by using the United States Environmental Protection Agency (USEPA) methods. The concentrations of Al, Fe, Mn, and Ni in coal mine water were exceeding the acceptable as well as the maximum permissible limits specified for drinking and domestic uses. The HPI values were below the critical pollution index level of 100 except at one location. The estimated HQ and HI values for adult and children were greater than the standard limits in nearly half of the water samples. High HQ and HI values suggest that suitable treatment of coal mine water will require before its utilization in domestic and drinking purposes.     

Increased frequency of micronucleated exfoliated cells among humans exposed in vivo to mobile telephone radiations

The health concerns have been raised following the enormous increase in the use of wireless mobile telephones throughout the world. This investigation had been taken, with the motive to find out whether mobile phone radiations cause any in vivo effects on the frequency of micronucleated exfoliated cells in the exposed subjects. A total of 109 subjects including 85 regular mobile phone users (exposed) and 24 non-users (controls) had participated in this study. Exfoliated cells were obtained by swabbing the buccal-mucosa from exposed as well as sex–age-matched controls. One thousand exfoliated cells were screened from each individual for nuclear anomalies including micronuclei (MN), karyolysis (KL), karyorrhexis (KH), broken egg (BE) and binucleated (BN) cells. The average daily duration of exposure to mobile phone radiations is 61.26 min with an overall average duration of exposure in term of years is 2.35 years in exposed subjects along with the 9.84 ± 0.745 micronucleated cells (MNCs) and 10.72 ± 0.889 total micronuclei (TMN) as compared to zero duration of exposure along with average 3.75 ± 0.774 MNC and 4.00 ± 0.808 TMN in controls. The means are significantly different in case of MNC and TMN at 0.01% level of significance. The mean of KL in controls is 13.17 ± 2.750 and in exposed subjects is 13.06 ± 1.793. The value of means of KH in exposed subjects (1.84 ± 0.432) is slightly higher than in controls (1.42 ± 0.737). Mean frequency of broken egg is found to be more in exposed subjects (0.65 ± 0.276) as compared to controls (0.50 ± 0.217). Frequency of presence of more than one nucleus in a cell (binucleated) is also higher in exposed (2.72 ± 0.374) in comparison to controls (0.67 ± 0.231). Although there is a slight increase in mean frequency of KH, BE and BN in exposed subjects but the difference is not found statistically significant. Correlation between 0–1, 1–2, 2–3 and 3–4 years of exposure and the frequency of MNC and TMN has been calculated and found to be positively correlated.     

Bioefficacy of plant essential oils against pulse beetles Callosobruchus spp. (Coleoptera: Bruchidae) in pigeon pea seeds with particular reference to Clausena pentaphylla (Roxb.) DC.

Thirty essential oils from higher plants of Gorakhpur Division (India) were evaluated at 0.36?μl/ml against two pulse beetles, Callosobruchus chinensis L. and C. maculatus F., causing infestation of pigeon pea seeds during storage. Clausena pentaphylla oil was more effective and exhibited absolute repellency against both the insects followed by Ocimum canum, Salvia plebeia and Zingiber zerumbet oils. Among these four oils, C. pentaphylla oil was most toxic and showed 100% mortality of both the insects at 10-μl dosage and 24-h exposure (LD₅₀?=?2.7?μl for C. chinensis & 2.4?μl for C. maculatus). Physical factors, viz. temperature, storage and autoclaving, did not cause any adverse effect on the toxicity of Clausena oil. During in vivo investigation, the oil protected 1?kg of pigeon pea seeds completely without reducing weight loss and seed damage up to 6?months when stored in gunny bags and glass containers. The oil was standardised by determining its various physicochemical properties. Thus, C. pentaphylla oil can be judiciously exploited as herbal insecticide against pulse beetles of pigeon pea seeds during storage.     

Cutting edge: macrophage migration inhibitory factor is necessary for progression of experimental autoimmune encephalomyelitis

Macrophage migration inhibitory factor (MIF) has been implicated in the pathogenesis of inflammatory and autoimmune diseases. The role of MIF in the progression of experimental autoimmune encephalomyelitis (EAE) was explored using MIF-/- mice. Wild-type mice showed a progressive disease course, whereas MIF-/- mice exhibited acute signs but no further progression of clinical disease. MIF-/- mice displayed markedly elevated corticosterone levels and significant decreases in the inflammatory cytokines TNF-alpha, IFN-gamma, IL-2, and IL-6 before, during, and after EAE onset. Taken together, these findings support that MIF is an important mediator of EAE progression through glucocorticoid antagonism and up-regulation of the inflammatory response.