Intracultural Cognizance of Medicinal Plants of Warangal North Forest Division, Northern Telangana, India

Differences in the traditional botanical knowledge of Koya communities inhabiting Eturnagaram Wildlife Sanctuary (Warangal North Forest Division) are investigated. Eighteen villages (16 within the wildlife sanctuary and two outside it) were selected to test the null hypothesis that there exist no cognitive differences among the ethnic inhabitants in their ability to recognize the plants and recall the vernacular names and medicinal uses since they are recipients of the same dry deciduous forest ecosystem services. The Koyas were found to use as medicine 237 species in 66 angiosperm families. Analyses of data gathered from villagers showed that there is significant intracultural diversity in terms of taxonomic groups and growth forms in regard to utilizing the proximate plant resource for their primary healthcare and disease treatment of pets

High quality factor 1-D Er 3+ -activated dielectric microcavity fabricated by RF-sputtering

Rare earth-activated 1-D photonic crystals were fabricated by RF-sputtering technique. The cavity is constituted by an Er3+-doped SiO2 active layer inserted between two Bragg reflectors consisting of ten pairs of SiO2/TiO2 layers. Scanning electron microscopy is employed to put in evidence the quality of the sample, the homogeneities of the layers thickness and the good adhesion among them. Near infrared transmittance and variable angle reflectance spectra confirm the presence of a stop band from 1500 nm to 2000 nm with a cavity resonance centered at 1749 nm at 0° and a quality factor of 890. The influence of the cavity on the 4I13/2 -> 4I15/2 emission band of Er3+ ion is also demonstrated

Tryptophan fluorescence quenching as a binding assay to monitor protein conformation changes in the membrane of intact mitochondria

Intrinsic protein fluorescence is due to aromatic amino acids, mainly tryptophan, which can be selectively measured by exciting at 295 nm. Changes in emission spectra of tryptophan are due to the protein conformational transitions, subunit association, ligand binding or denaturation, which affect the local environment surrounding the indole ring. In this study, tryptophan fluorescence was monitored in intact mitochondria at 333 nm following excitation at 295 nm in presence of insecticides using spectrofluorometer. Methyl-parathion, carbofuran, and endosulfan induced Trp fluorescence quenching and release of cytochrome c when incubated with the mitochondria, except fenvalarate. Mechanism of insecticide-induced mitochondrial toxicity for the tested insecticides has been discussed. Reduction in the intensity of tryptophan emission spectra of mitochondrial membrane proteins in presence of an increasing concentration of a ligand can be used to study the interaction of insecticides/drugs with the intact mitochondria. Furthermore, this assay can be readily adapted for studying protein–ligand interactions in intact mitochondria and in other cell organelles extending its implications for pesticide and pharma industry and in drug discovery

Monitoring biosafety of pharmaceutical drugs, insecticides, and other bioactive molecules to mitochondria

Mitochondria are large cell organelles in plant and animal cells. They convert chemical energy from food in the cell to usable energy using oxygen. This process is called oxidative phosphorylation, which takes place inside the mitochondria. Reactions of Krebs cycle produce a chemical, NADH (Nicotinamide Adenine Dinucleotide, reduced form), which is then used by enzymes present in the inner mitochondrial membrane to generate energy rich adenosine triphosphate (ATP) molecules. These molecules are utilized by the cells whenever energy is required. Therefore, mitochondria are also referred as power houses of the cell. Mitochondria are abundantly present in muscle and brain cells, and sperms to supply ATPs during demand for energy. Monitoring biosafety of pharmaceutical drugs, insecticides, and other bioactive molecules to mitochondria is necessary because some of them interact with the mitochondrial membrane or membrane proteins, thereby affecting the energy levels of cells or even induces mitochondrial-dependent apoptosis (programmed cell death)

Elimination of Gut Microbes with Antibiotics Confers Resistance to Bacillus thuringiensis Toxin Proteins in Helicoverpa armigera (Hubner)

Helicoverpa armigera is one of the most important pests worldwide. Transgenic crops with toxin genes from Bacillus thuringiensis (Bt) have been deployed on a large scale to control this pest. The insecticidal activity of Bt is probably influenced by the insect midgut microbes, which vary across crop hosts and locations. Therefore, we examined the role of gut microbes in pathogenicity of Bt toxins in the H. armigera. Antibiotic cocktail was used for the complete elimination of the H. armigera gut microbes. Activated Cry1Ac, Bt formulation, and transgenic cotton resulted in larval weight loss and increase in mortality, but pretreatment of larvae with antibiotic cocktail significantly decreased larval mortality and increased the larval weight gain. Activated Cry1Ac and Bt formulation inhibited the activity of proteases in midgut of H. armigera larvae but showed no such effect in the larvae pretreated with antibiotic cocktail. Five protease bands in activated Cry1Ac and two in Bt formulation-treated larvae were inhibited but no such effect in the larvae pretreated with antibiotic cocktail. Cry1Ac protein was detected in Bt/Cry1Ac protoxin-fed larval gut extract in the absence of antibiotic cocktail, but fewer in larvae pretreated with antibiotic cocktail. The activity of antioxidant enzymes and aminopeptidases increased in larvae fed on Bt toxin, but there was no significant increase in antioxidant enzymes in larvae reared on toxin protein in combination with antibiotic cocktail. The results suggest that gut microbes exercise a significant influence on the toxicity of Cry1Ac and Bt formulation in H. armigera larvae. The implications of these results have been discussed in relation to development of insect resistance to Bt transgenic crops deployed for pest management

Influence of bacillus thuringiensis toxins on the development and midgut proteases in different larval instars of helicoverpa armigera

Helicoverpa armigera (Hubner) (Lepidoptera: Noctuidae) is one of the most important pest worldwide. Bacillus thuringiensis (Bt) toxins have used as a biopesticide or deployed in transgenic plants for controlling this pest. We examined the biological activity of Cry1Ac, Cry1Ab and Bt formulation in different larval instars of H. armigera with respect to larval development and proteolytic activity to pinpoint the most susceptible instar, and the insect response to ingestion of Bt toxins. In the presence of Bt toxins, the larval mortality and weight loss increased in a dose-dependent manner, and the maximum effect was observed in neonates. Active Cry1Ac toxin resulted in greater mortality and weight loss in all the larval instars. Total protease, trypsin and chymotrypsin activities declined in the presence of Bt toxins as compared to the untreated control in all the larval instars. Ten protease isozymes were observed in the untreated control larvae in second, third and fourth instars. Maximum protease isozymes inhibition was observed in Bt toxin fed neonates. Inhibition of protease activity increased with the concentration of Bt toxins. In presence of Bt toxins, aminopeptidase activity increased from II to IV instar and alkaline phosphatase activity decreased from II to III and then increased in IV instar. The pathogenicity of Bt was greater in early larval instars of H. armigera than the later instars, suggesting that application of Bt formulation or deployment of Bt toxins in transgenic plants should be directed against the early larval instars

Methylparathion- and carbofuran-induced mitochondrial dysfunction and oxidative stress in Helicoverpa armigera (Noctuidae: Lepidoptera)

The cotton bollworm, Helicoverpa armigera is a polyphagous pest of several crops in Asia, Africa, and the Mediterranean Europe. Organophosphate and carbamate insecticides are used on a large-scale to control Helicoverpa. Therefore, we studied the effect of methylparathion and carbofuran, an organophosphate and carbamate insecticide, respectively, on oxidative phosphorylation and oxidative stress in H. armigera larvae to gain an understanding of the different target sites of these insecticides. It was observed that state III and state IV respiration, respiratory control index (RCI), and P/O ratios were inhibited in a dose-dependent manner by methylparathion and carbofuran under invitro and in vivo conditions. Methylparathion and carbofuran inhibited complex II by ∼ 45% and 30%, respectively. Lipid peroxidation, H2O2 content, and lactate dehydrogenase (LDH) activity increased and glutathione reductase (GR) activity decreased in a time- and dose-dependent manner in insecticide-fed larvae. However, catalase activity was not affected in insecticide-fed larvae. Larval growth decreased by ∼ 64 and 67% in larvae fed on diets with sub-lethal doses of methylparathion and carbofuran. The results suggested that both the insecticides impede the mitochondrial respiratory functions and induced lipid peroxidation, H2O2, and LDH leak, leading to oxidative stress in cells, which contribute to deleterious effects of these insecticides on the growth of H. armigera larvae, along with their neurotoxic effects

Characterization of dihydrolipoamide dehydrogenase from the mitochondria of Helicoverpa armigera, a pest resistant to insecticides

Dihydrolipoamide dehydrogenase (DHLDH) was isolated from the mitochondria of Helicoverpa armigera, a destructive pest which has developed resistance to commonly used insecticides. The flavoenzyme was purified 17.98-fold to homogeneity with an overall yield of 10.53% by employing ammonium sulfate precipitation, hydroxylapatite chromatography and CM-Sephadex chromatography. The purified enzyme exhibited the specific activity of 18.7 U/mg and was characterized as a dimer with a subunit mass of 66 kDa. The enzyme showed specificity for nicotinamide adenine dinucleotide – hydrogen (NADH) and lipoamide, as substrates, with Michaelis-Menten constants (Km) of 0.083 mmol/L and 0.4 mmol/L, respectively. The reduction reaction of lipoamide by the enzyme could be explained by ping-pong mechanism. The spectra of DHLDH showed the maximum absorbance at 420 nm, 455 nm and 475 nm. The enzyme activity was strongly inhibited by mercurial and arsenical compounds. The N-terminal sequence of Ha-DHLDH showed homology with those of mammalian and arthropod DHLDH. Since H. armigera has developed high levels of resistance to commonly used insecticides, biochemical properties of the metabolic enzymes such as DHLDH, could be helpful to develop insecticidal molecules for the control of H. armigera, with a different mode of action

Mitochondrial P-Glycoprotein ATPase Contributes to Insecticide Resistance in the Cotton Bollworm, Helicoverpa armigera (Noctuidae: Lepidoptera)

Cotton bollworm, Helicoverpa armigera, is one of the most damaging polyphagous pests worldwide, which has developed high levels of resistance to commonly applied insecticides. Mitochondrial P-glycoprotein (Pgp) was detected in the insecticide-resistant strain of H. armigera using C219 antibodies, and its possible role was demonstrated in the efflux of xenobiotic compounds using spectrofluorometer. The TMR accumulated in mitochondria in the absence of ATP, and effluxed out in presence of ATP; the process of efflux was inhibited in the presence of ortho-vandate, an inhibitor of Pgp, in insecticide-resistant larvae of H. armigera. The mitochondria isolated from insecticide-resistant larvae were resistant to insecticide-induced inhibition of oxygen consumption and cytochrome c release. Membrane potential decreased in a dose-dependent manner in the presence of higher concentration of insecticides (>50 µM) in mitochondria of insecticide-resistant larvae. In conclusion, mitochondrial Pgp ATPase detected in the insecticide-resistant larvae influenced the efflux of xenobiotic compounds. Pgp might be involved in protecting the mitochondrial DNA and the components of the electron transport chain from damage due to insecticides, and contributing to the resistance to the deleterious effects of insecticides on the growth of insecticide-resistant H. armigera larvae

Interaction of plant cell signaling molecules, salicylic acid and jasmonic acid, with the mitochondria of Helicoverpa armigera

The cotton bollworm, Helicoverpa armigera is a polyphagous pest in Asia, Africa, and the Mediterranean Europe. Salicylic acid (SA) and jasmonic acid (JA) are the cell signaling molecules produced in response to insect attack in plants. The effect of these signaling molecules was investigated on the oxidative phosphorylation and oxidative stress of H. armigera. SA significantly inhibited the state III and state IV respiration, respiratory control index (RCI), respiratory complexes I and II, induced mitochondrial swelling, and cytochrome c release in vitro. Under in vivo conditions, SA induced state IV respiration as well as oxidative stress in time- and dose-dependent manner, and also inhibited the larval growth. In contrast, JA did not affect the mitochondrial respiration and oxidative stress. SA affected the growth and development of H. armigera, in addition to its function as signaling molecules involved in both local defense reactions at feeding sites and the induction of systemic acquired resistance in plants