Ayurvedic <Emphasis Type="Italic">Amalaki Rasayana</Emphasis> promotes improved stress tolerance and thus has anti-aging effects in <Emphasis Type="BoldItalic">Drosophila melanogaster</Emphasis>

Amalaki Rasayana (AR) is a common Ayurvedic herbal formulation of Phyllanthus emblica fruits and some other ingredients, and is used for general good health and healthy aging. We reported it to improve life history traits and to suppress neurodegeneration as well as induced apoptosis in Drosophila. The present study examines responses of Drosophila reared on AR-supplemented food to crowding, thermal or oxidative stresses. Wild-type larvae/flies reared on AR-supplemented food survived the various cell stresses much better than those reared on control food. AR-fed mutant park ¹³ or DJ-1β ^Delta93 (Parkinson’s disease model) larvae/flies, however, showed only partial or no protection, respectively, against paraquat-induced oxidative stress, indicating essentiality of DJ-1β for AR-mediated oxidative stress tolerance. AR feeding reduced the accumulation of reactive oxygen species (ROS) and lipid peroxidation even in aged (35-day-old) wild-type flies while enhancing superoxide dismutase (SOD) activity. We show that while Hsp70 or Hsp83 expression under normal or stress conditions was not affected by AR feeding, Hsp27 levels were elevated in AR-fed wild-type control as well as heat-shocked larvae. Therefore, besides the known anti-oxidant activity of Phyllanthus emblica fruits, dietary AR also enhances cellular levels of Hsp27. Our in vivo study on a model organism shows that AR feeding significantly improves tolerance to a variety of cell stresses through reduced ROS and lipid peroxidation on the one hand, and enhanced SOD activity and Hsp27 on the other. The resulting better homeostasis improves life span and quality of organism’s life.     

3D QSAR,pharmacophore and molecular docking studies of known inhibitors and designing of novel inhibitors for M18 aspartyl aminopeptidase of <Emphasis Type="Italic">Plasmodium falciparum</Emphasis>

Background

The Plasmodium falciparum M18 Aspartyl Aminopeptidase (PfM18AAP) is only aspartyl aminopeptidase which is found in the genome of P. falciparum and is essential for its survival. The PfM18AAP enzyme performs various functions in the parasite and the erythrocytic host such as hemoglobin digestion, erythrocyte invasion, parasite growth and parasite escape from the host cell. It is a valid target to develop antimalarial drugs. In the present work, we employed 3D QSAR modeling, pharmacophore modeling, and molecular docking to identify novel potent inhibitors that bind with M18AAP of P. falciparum.

Results

The PLSR QSAR model showed highest value for correlation coefficient r² (88 %) and predictive correlation coefficient (pred_r2) =0.6101 for external test set among all QSAR models. The pharmacophore modeling identified DHRR (one hydrogen donor, one hydrophobic group, and two aromatic rings) as an essential feature of PfM18AAP inhibitors. The combined approach of 3D QSAR, pharmacophore, and structure-based molecular docking yielded 10 novel PfM18AAP inhibitors from ChEMBL antimalarial library, 2 novel inhibitors from each derivative of quinine, chloroquine, 8-aminoquinoline and 10 novel inhibitors from WHO antimalarial drugs. Additionally, high throughput virtual screening identified top 10 compounds as antimalarial leads showing G-scores -12.50 to -10.45 (in kcal/mol), compared with control compounds(G-scores -7.80 to -4.70) which are known antimalarial M18AAP inhibitors (AID743024). This result indicates these novel compounds have the best binding affinity for PfM18AAP.

Conclusion

The 3D QSAR models of PfM18AAP inhibitors provided useful information about the structural characteristics of inhibitors which are contributors of the inhibitory potency. Interestingly, In this studies, we extrapolate that the derivatives of quinine, chloroquine, and 8-aminoquinoline, for which there is no specific target has been identified till date, might show the antimalarial effect by interacting with PfM18AAP.

     

Galectin 3 acts as an enhancer of survival responses in <Emphasis Type="Italic">H. pylori</Emphasis>-infected gastric cancer cells

Galectin 3 (Gal-3) is upregulated in gastric epithelial cells as a host response to Helicobacter pylori infection. However, the significance of Gal-3 expression in H. pylori-infected cells is not well established. We analyzed Gal-3 intracellular expression, localization, and its effects in H. pylori-infected gastric epithelial cells. The predominantly nuclear confined Gal-3 was shown to be upregulated and exported out to the cytoplasm in H. pylori-infected AGS cells. The nuclear export was channeled through CRM-1 (exportin-1) protein. Interestingly, knock down of Gal-3 expression led to reduced NF-κB promoter activity and interleukin-8 (IL-8) secretion, suggesting its pro-inflammatory roles. Furthermore, Gal-3 was found to be pro-proliferative and anti-apoptotic in nature, as its knock down caused a reduction in cell proliferation and an increase in apoptosis, respectively. Taken together, our data suggest the expression and upregulation of Gal-3 as a critical endogenous event in H. pylori infection that interferes with various intracellular events, causing prolonged cell survival, which is characteristic in carcinogenesis.     

Polyamine Biosynthetic Enzymes and the Effect of their Inhibition on the Growth of Some Phytopathogenic Fungi

Studies were conducted on the distribution of two polyaminebiosynthetic enzymes, or-nithine decarboxylase (ODC) and argininedecarboxylase (ADC), and the effect of their inhibitors on growthand polyamine biosynthesis in four phytopathogenic fungi, namely,Helminthosporium maydis, H. carbonum, Fusarium oxysporum f.sp. lycopersici and Ceratocystis ulmi. Three species had highlevel of ODC as compared to ADC activity; in C. ulmi on theother hand, ADC was predominant with very little or no ODC activity.DL--difluoromethylornithine (DFMO) significantly inhibited ODCactivity in all species in vitro with little effect on ADC activity.ADC in all cases was inhibited by DL--difluoromethylarginine(DFMA) but not by DFMO. Mycelial growth of all fungi was inhibitedby 1 to 5 mM concentrations of either DFMO or DFMA within twodays except in H. maydis which remained unaffected even by thehighest concentration (5 mM) of DFMA. In general, the inhibitionwas more pronounced with DFMO as compared to DFMA. Putrescinecompletely reversed the inhibitory effects of DFMO and DFMAin all species. Among the polyamines, spermidine was predominantin all fungi. The cellular concentrations of putrescine andspermidine were considerably lower in the presence of eitherof the inhibitors while spermine levels were higher than thecontrol. ¹Scientific contribution number 1529 from the New HampshireAgricultural Experiment Station. (Received November 25, 1988; Accepted April 11, 1989)     

Phytochelatin accumulation and stress tolerance in tomato cells exposed to cadmium

Cell suspension cultures of tomato (Lycopersicon esculentum) adapted to growing continuously in the presence of 0.1 mM CdCl₂ and accumulated phytochelatins (PCs, poly(-Glu-Cys)n-Gly). The highest level of PCs was measured 4 days after inoculation and coincided with the peak of cellular cadmium concentration. At this time there was an 8-fold molar excess of PC (-Glu-Cys) over Cd. PCs could not be detected after 12 days when the cellular concentration of cadmium was 0.2 mM. These results indicate that PCs are produced in excess of that required to bind the cellular cadmium in the early stage of the culture period followed by degradation of PCs during the stationary phase. Adaptation to 0.1 mM CdCl₂ did not increase tolerance to higher concentrations of cadmium when compared with control cells, but did significantly enhance tolerance to both anaerobiosis and heat shock. Exposure of tomato cells to 0.1 mM CdCl₂ resulted in several changes in proteins synthesized.     

The role of eucaryotic cell surface glycoconjugates in DNA replication

Multiple forms of ricin have been isolated from castor bean seeds. Two forms, ricin-1 and ricin-2, differ in their isoelectric pI values and toxicity towards IMR-32 cells. Inhibition of IMR-32 DNA polymerase α₂ is more pronounced with ricin-1 (65%) than with ricin-2 (10%). Ricin B chain (pI = 5.2) isolated from ricin-1 binds to IMR-32 cell surfaces as well as inhibits DNA polymerase α₂ activity when studiedin vitro. The presence of galβ-linked glycoconjugates near the active site of IMR-32 DNA polymerase α₂ has been proposed. Replication modulators which bind to the glycose portion of the enzymes involved in the replication system may need a mandatory binding to cell surface glycoconjugates for their activity.     

Induction of virulence response in Agrobacterium tumefaciens by tissue explants of various plant species

Forty-four plant species belonging to different taxa were tested for their ability to induce the expression of the virulence E gene in Agrobacterium tumefaciens containing virE:lacZ fusion constructs. With the exception of 6 algae, one fern and 2 monocots, tissue explants of all other plants (2 Algae, 3 Bryophytes, 2 Pteridophytes, 15 Gymnosperms, 8 Monocots and 5 Dicots) induced the expression of the virE gene as detected by the presence of -galactosidase activity in the bacteria.Abbreviations AS acetosyringone - vir virulence genes Scientific Contribution Number 1734 from the New Hampshire Agricultural Experiment Station     

Effect of 2,4-dichlorophenoxyacetic acid on polysomal profiles and polyadenylated RNA in excised tuber tissue of Jerusalem artichoke (Helianthus tuberosus)

Dormant tuber tissue of Jerusalem artichoke ( Helianthus tuberosus L.) can be stimulated by wounding to initiate RNA and protein synthesis. No DNA synthesis or cell divisions occur unless an auxin is provided. Changes in polysomal profiles and levels of Poly(A)⁺-RNA in response to wounding and auxin treatment were studied. Polysomes were isolated at various times after excision and incubation of tissue in the presence or absence of 10⁻⁵ M 2,4-dichlorophenoxyacetic acid. Polysomal profiles were studied by sucrose density gradient centrifugation. Dormant tissue contained ribosomes mainly in monosome form. Within 4 h of excision, a significant increase in the polysomal fraction was observed both in control and auxin-treated tissue. Increases in polysomes continued during the next 20 h. Poly(A)⁺-RNA was isolated from total polysomal RNA by oligo(dT)-cellulose column chromatography. There was a large increase in the amount of poly(A)⁺-RNA within 4 h of excision. During the first 43 h of incubation, levels of total polysomal RNA as well as poly(A)⁺-RNA in tissue treated with 2,4-dichlorophenoxyacetic acid were significantly higher than those in controls.