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Thakur, A. K.

Publications and source records attributed to Thakur, A. K..

5 recordsLinked to original sources

Discerning amyloid network in plants

Amyloids are proteinaceous fibrillar structures and are known for their pathogenic and functional roles across the kingdoms. Besides proteinaceous deposits, amyloid-like structures are present in small metabolite assemblies and fibrillar hydrogels. Recent cryoelectron microscopy studies have shed light on the heterogeneous nature of the amyloid structures and their association with carbohydrate or lipid molecules, suggesting that amyloids are not exclusively proteinaceous. The association of amyloids with carbohydrates is further supported because the gold-standard dye of amyloid detection, Congo red, also binds to carbohydrates, probably due to similar stacking interactions. We name the association between amyloids, carbohydrates and other biomolecules as amyloid-network and propose that plants might contain such structures. Specifically, we hypothesize that cereal seeds containing glutamine-repeat-rich granules of storage proteins may have amyloid-like structures. This is because, polyQ repeats are associated with protein aggregation and amyloid formation in humans and are linked to multiple neurodegenerative conditions. Also seed storage proteins and seed cell wall proteins possess carbohydrate affinity. Thus, plant seeds might contain an intercalated network of proteins and carbohydrates, lending strength, stability and dynamics to these structures. In this paper, we show that, plant seeds have a mesh-like network that shows apple-green birefringence on staining with Congo red, a characteristic of amyloids. This congophilic network is more prominent in protein-rich seed sections of wheat and lentils, as compared to starch-rich compartments of potato. The findings suggest an amyloid network in the seeds and might be extended to other plant tissues. Further investigation with mass spectrometry and other techniques would detail the exact compositional analysis of these networks.

plant biology↗

Proliferation of lung epithelial cells is regulated by the mechanisms of autophagy upon exposure of soots

Soots are known to cause many diseases in humans but their underlying mechanisms of toxicity are still not known. Here, we report that, soots induce cell proliferation of lung epithelial cells via modulating autophagic pathways. Fullerene soot and diesel exhaust particles (DEP) induced cell proliferation of lung epithelial, A549 cells, via distinct autophagic mechanisms and did not cause cell death. Exposure of fullerene soot protected cell death of A549 cells, caused by hydrogen peroxide and inhibited LPS-induced autophagy. Fullerene soot co-localize with the autophagic proteins and inhibited starvation-induced autophagy (downregulated ATG-5, beclin-1, p62 and LC3 expressions) independent of its antioxidant properties. Similarly, it decreased expression profile of autophagic genes and upregulated proliferation responsive gene, Ki-67, in mice. We observed that, expressions of fullerene soot responsive genes (Beclin-1, ATG-5 and p62) were reverted by Akt Inhibitor X, indicating an important role of Akt pathway. On the other hand, DEP up-regulated expressions of autophagy genes. Akt Inhibitor X and Etoricoxib, did not attenuate DEP-induced cell proliferation and autophagic response. However, autophagic inhibiter 3-MA and chloroquine has significantly inhibited DEP-induced cell roliferation. In conclusion, distinct autophagic mechanisms are operational in cell proliferation of lung epithelial cells in response to soots and may have implication in diseases. The proliferation inducing potential of fullerene soot may be utilized as a regenerative agent in autophagy-associated disorders.

cell biology↗

Antibiofilm activity of Fmoc-phenylalanine against Gram-positive and Gram-negative bacterial biofilms

BackgroundBiofilm associated infections are the major contributor of mortality, morbidity and financial burden in patients with bacterial infection. Molecules with surfactant behaviour are known to show significant antibiofilm effect against these infections. Thus, newly discovered antibacterial Fmoc-phenylalanine (Fmoc-F) and other Fmoc-amino acids (Fmoc-AA) with surfactant properties, could have potential antibiofilm properties. ObjectivesTo evaluate and characterise the antibiofilm activity of Fmoc-F and some Fmoc-AA against various clinically relevant bacteria. MethodsBiofilm inhibition and eradication was evaluated by crystal violet staining procedure along with scanning electron microscopy (SEM). Attenuated Total Reflection - Fourier Transform Infrared Spectroscopy (ATR-FTIR), Biochemical assays and Congo red staining were employed to investigate mechanism of antibiofilm action. ResultsWe showed that Fmoc-F not only inhibits the biofilm formation in S. aureus and P. aeruginosa, but also eradicates the already formed biofilms over the surface. Further, Fmoc-F coated glass surface resists S. aureus and P. aeruginosa biofilm formation and attachment, when biofilm is grown over the surface. The mechanistic investigation suggests that Fmoc-F reduces the ECM components such as proteins carbohydrates and eDNA in the biofilm and affect its stability via direct interactions with ECM components and/ or indirectly through reducing bacterial cell population. Finally, we showed that Fmoc-F treatment in combination with other antibiotics such as vancomycin and ampicillin synergistically inhibit biofilm formation. ConclusionsOverall, the study demonstrates the potential application of Fmoc-F and other Fmoc-AA molecules individually as well as in combination as antibiofilm agents and antibiofilm coating material for treating biofilm associated infections.

microbiology↗

Nucleation-dependent aggregation kinetics of Yeast Sup35 fragment GNNQQNY

An N-terminal hepta-peptide sequence of yeast prion protein Sup35 with the sequence GNNQQNY serves as an ideal model for structural understanding of amyloid assembly and kinetics. In this study, we used a reproducible solubilisation protocol that allows the generation of homogenous monomeric solution of GNNQQNY to understand the molecular details of its self-assembly mechanism. The aggregation kinetics data show that the GNNQQNY sequences follow nucleation-dependent aggregation kinetics with a critical nucleus of size ~7 monomers and that the size and efficiency of nucleation was found to be inversely related to the reaction temperature. The generated nucleus reduces the thermodynamic energy barrier by acting as a template for further self-assembly and results in highly ordered amyloid fibrils. The fibers grown at different temperatures showed similar Thioflavin T positivity, Congo red binding and {beta}-sheet rich structures displaying a characteristic cross-{beta} diffraction pattern. These aggregates also share morphological and structural identity with those reported earlier. The mature GNNQQNY fibers exerted no significant oxidative stress or cytotoxicity upon incubating with differentiated SHSY5Y cells. To our knowledge, this is the first study to experimentally validate previous predictions based on theoretical and molecular dynamics simulations. These findings will provide the basis for understanding the kinetics and thermodynamics of amyloid nucleation and elongation of amyloidogenic systems associated with many systemic and neurodegenerative diseases.

biophysics↗

\"Risk Factors for Visceral Leishmaniasis in Selected High Endemic Areas of Morang District, Nepal\": A case control Study

Visceral leishmaniasis is a major public health concern in Nepal. During the last few years, several KA outbreaks have been reported from Tarai region including Morang district. A case control study was conducted to assess the risk factors associated with VL in 5 endemic VDC of Morang district with 62 cases already treated from BPKIHS and Koshi zonal hospital and 248 controls selected randomly from the same village. Data collected using semi structured questionnaire from September to November 2013. This study revealed that people living in thatched house, sleeping in ground floor, ownership of animal, history of migration to India (Bihar and Jharkhand) and proximity to other KA cases within 50 m distance of household were strong risk factors for VL. Education remains protective (OR 0.39,95 % CI 0.19-0.79). The association with socioeconomic status showed clear dose - response effect. The odds for VL consistently decreased as the level of socioeconomic status increased (OR 4.26, 3.81). Strengthening surveillance system for early diagnosis and treatment, awareness programme and further extensive study is needed on risk factor, vector and control measures. Author SummaryThis study aims to explore the risk factors for visceral leishmaniasis. Based on findings there is a need to educate people in high-transmission areas how to realize, reduce or avoid environmental factors that favor the survival of the vector in the community. Similarly introduction of more exact surveillance tools in order to improve morbidity and mortality surveillance by health sector. People living in mud thatched houses need to be aware about cracks and crevices in the mud walls, their function as breeding places and how they can be controlled, for example by plastering with lime and mud.. However, a primary condition is that people need to understand the purpose of all these efforts in order to be motivated to put them into practice.

epidemiology↗