3D QSAR pharmacophore-based virtual screening for the identification of potential inhibitors of tyrosinase

Abstract

Tyrosinase plays an important role in melanin biosynthesis and protects skin against ultraviolet radiations. Functional deficiency of tyrosinase results in serious dermatological diseases. Tyrosinase also participates in neuromelanin formation in the human brain, which leads to neurodegeneration resulting in Parkinson’s disease. In fruits and vegetables, tyrosinase plays a critical role in senescence, causing undesired browning that results in faster deterioration and shorter shelf lines. The only commercially available tyrosinase is mushroom tyrosinase and it shows the highest homology to the mammalian tyrosinase. Although kojic acid is currently used as a tyrosinase inhibitor, they have serious side effects such as dermatitis, carcinogenesis and hepatotoxicity. Therefore, in order to develop a more active and safer tyrosinase inhibitor, 3D QSAR pharmacophore models were generated based on experimentally known inhibitors. The pharmacophore model, Hypo1, was developed with a large cost difference, high correlation coefficient and low RMS deviation. Hypo1 showed a good spatial arrangement; consisting of five-point features including two hydrogen bond acceptor, one hydrogen bond donor and two hydrophobic features. Hypo1 was further validated by cost analysis, test set and Fisher’s randomisation method. Hypo1 was used as a 3D query for screening the in-house drug-like databases, and the hits were further selected by applying ADMET, Lipinski’s rule of five and fit value criteria. To identify binding conformations, the obtained hits were subjected to molecular docking. Finally, molecular dynamics simulations revealed the appropriate binding modes of hit compounds. To conclude, we propose the final three hit compounds with new structural scaffolds as a virtual candidate as tyrosinase inhibitors.

Communicated by Ramaswamy H. Sarma     

Sex-specific paternal age effects on offspring quality in Drosophila melanogaster

Advanced paternal age has been repeatedly shown to modulate offspring quality via male- and/or female-driven processes, and there are theoretical reasons to expect that some of these effects can be sex-specific. For example, sex allocation theory predicts that, when mated with low-condition males, mothers should invest more in their daughters compared to their sons. This is because male fitness is generally more condition-dependent and more variable than female fitness, which makes it less risky to invest in female offspring. Here, we explore whether paternal age can affect the quality and quantity of offspring in a sex-specific way using Drosophila melanogaster as a model organism. In order to understand the contribution of male-driven processes on paternal age effects, we also measured the seminal vesicle size of young and older males and explored its relationship with reproductive success and offspring quality. Older males had lower competitive reproductive success, as expected, but there was no difference between the offspring sex ratio of young and older males. However, we found that paternal age caused an increase in offspring quality (i.e., offspring weight), and that this increase was more marked in daughters than sons. We discuss different male- and female-driven processes that may explain such sex-specific paternal age effects.     

Isolation and identification of a plant growth promotive substance from mixture of essential plant oils

The PCS (commercial products by Field Science Co, Japan, used for air fresheners) was analyzed for the presence of bioactive constituents and their role as root growth promoters. Chromatographic separation of the methanolic solution of PCS resulted in the isolation of an promoting active substance, which was identified using GC-mass spectrometry and NMR spectroscopy as 1,2-propanediol (CH₃CH(OH)CH₂OH). Lettuce seedling growth bioassay as test plant revealed that 1,2-propanediol can act as potent root growth promoter; enhancing the growth of lettuce seedling radicle at a concentration 0.01 ppm. The concentration of 1,2-propanediol in PCS mixture was estimated as 4 g/l. These studies suggest that 1,2-propanediol might play an important role in the plant growth promoting activity of PCS.     

Role of the sequence surrounding predicted transmembrane helix M4 in membrane association and function of the Ca(2+) release channel of skeletal muscle sarcoplasmic reticulum (ryanodine receptor isoform 1)

The role of the sequence surrounding M4 in ryanodine receptors (RyR) in membrane association and function was investigated. This sequence contains a basic, 19-amino acid M3/M4 loop, a hydrophobic 44-49 amino acid sequence designated M4 (or M4a/M4b), and a hydrophilic M4/M5 loop. Enhanced green fluorescent protein (EGFP) was inserted into RyR1 and truncated just after the basic sequence, just after M4, within the M4/M5 loop, just before M5 and just after M5. The A52 epitope was inserted into RyR2 and truncated just after M4a. Analysis of these constructs ruled out a M3/M4 transmembrane hairpin and narrowed the region of membrane association to M4a/M4b. EGFP inserted between M4a and M4b in full-length RyR2 was altered conformationally, losing fluorescence and gaining trypsin sensitivity. Although it was accessible to an antibody from the cytosolic side, tryptic fragments were membrane-bound. The expressed protein containing EGFP retained caffeine-induced Ca(2+) release channel function. These results suggest that M4a/M4b either forms a transmembrane hairpin or associates in an unorthodox fashion with the cytosolic leaflet of the membrane, possibly involving the basic M3/M4 loop. The expression of a mutant RyR1, Delta4274-4535, deleted in the sequence surrounding both M3 and M4, restored robust, voltage-gated L-type Ca(2+) currents and Ca(2+) transients in dyspedic myotubes, demonstrating that this sequence is not required for either orthograde (DHPR activation of sarcoplasmic reticulum Ca(2+) release) or retrograde (RyR1 increase in DHPR Ca(2+) channel activity) signals of excitation-contraction coupling. Maximal amplitudes of L-currents and Ca(2+) transients with Delta4274-4535 were larger than with wild-type RyR1, and voltage-gated sarcoplasmic reticulum Ca(2+) release was more sensitive to activation by sarcolemmal voltage sensors. Thus, this region may act as a negative regulatory module that increases the energy barrier for Ca(2+) release channel opening.     

Cytochemical localization of phosphatase in differentiating secondary vascular cells

Summary Phosphatase activity was studied in the cambium and differentiating vascular cells of beech by using a modified Washstein and Meisel method. After fixation in glutaraldehyde or crotonaldehyde and incubation in a medium containing ATP and lead nitrate at pH 7.2, a deposit of electron-opaque granules was found in the nucleoli, nucleoplasm, nuclear envelope, endoplasmic reticulum, plastids, mitochondria, and at the plasmalemma. Although located at these different sites, the distribution varied both inter- and intra-cellularly. This is thought to be a true reflection of the variation in activity between closely adjacent cells in this part of the stem.Some reaction was obtained when ADP replaced ATP in the reaction mixture, but there was no reaction at all when both ATP and ADP were omitted. Fixation in hydroxyadipaldehyde, or incubation at pH 6.4 both produced very little reaction.     

Functional significance of the activities of glutaminase and ornithine-ω-aminotransferase in rat brain

The activities of glutaminase, glutamine synthetase (GS), arginase and ornithine amino transferase (orn-T) were studied in three regions of rat brain in heightened neuronal activity by producing convulsions by leptazol. These enzymes were studied in preconvulsive, convulsive and postconvulsive phases. Glutaminase activity was found to increase in all the three regions in the preconvulsive and convulsive phases. GS activity decreased in the preconvulsive phase but rose gradually to the control level when the postconvulsive phase was reached. The activity of arginase decreased in the cerebellum in preconvulsive and convulsive phases. However, in the cerebral cortex there was a decrease in the activity of this enzyme only in the convulsive phase. The results suggest that glutamine acts more likely as a precursor for the neurotransmitter pool of glutamate, while ornithine serves more as a precursor for the neurotransmitter pool of GABA.     

The “unguarded‐X” and the genetic architecture of lifespan: Inbreeding results in a potentially maladaptive sex‐specific reduction of female lifespan in Drosophila melanogaster

Sex differences in ageing and lifespan are ubiquitous in nature. The "unguarded‐X” hypothesis (UXh) suggests they may be partly due to the expression of recessive mutations in the hemizygous sex chromosomes of the heterogametic sex, which could help explain sex‐specific ageing in a broad array of taxa. A prediction central to the UX hypothesis is that inbreeding will decrease the lifespan of the homogametic sex more than the heterogametic sex, because only in the former does inbreeding increase the expression of recessive deleterious mutations. In this study, we test this prediction by examining the effects of inbreeding on the lifespan and fitness of male and female Drosophila melanogaster across different social environments. We found that, across social environments, inbreeding resulted in a greater reduction of female than male lifespan, and that inbreeding effects on fitness did not seem to counterbalance sex‐specific effects on lifespan, suggesting the former are maladaptative. Inter‐ and intra‐sexual correlation analyses also allowed us to identify evidence of an underlying joint genetic architecture for inbreeding effects on lifespan. We discuss these results in light of the UXh and other alternative explanations, and suggest that more attention should be paid to the possibility that the “unguarded‐X” may play an important role in the evolution of sex‐specific lifespan.     

Risk assessment of heavy metal toxicity through contaminated vegetable from sewage water: Implications for populace health

The present investigation was carried out to evaluate the levels of metals and metalloids in okra (Abelmoschus esculentus) irrigated with city wastewater. Soil and vegetable samples from two different sites irrigated with wastewater were wet-digested and analyzed. Arsenic (As) was found higher at both sites and Cr was many-fold lower at both sampling sites. Among all heavy metals, Mn and Zn were abundant. Highest value of coefficient factor was found for Cr and the lowest for Cd. The high transfer value was recorded for Cu at site-I and for Ni at site-II. Copper and Se showed negative and significant correlations between soil and vegetable, whereas Mn, Zn, As, Cd, Cr, and Ni showed positive but non-significant correlations. Pollution load index in this vegetable was found to be higher for Cd and lower for Cu. Health risk index at site-I was in the order of As > Mn > Mo > Pb > Cd > Ni > Zn > Se > Fe > Co > Cr > Cu, whereas the same order was observed at site-II of the sampling locations. Thus, the health risks of metals through ingestion of vegetables were of great concern in the study area.