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1.
Ecotoxicol Environ Saf ; 277: 116337, 2024 Jun 01.
Article En | MEDLINE | ID: mdl-38640798

The intricate architecture of the intestinal epithelium, crucial for nutrient absorption, is constantly threatened by environmental factors. The epithelium undergoes rapid turnover, which is essential for maintaining homeostasis, under the control of intestinal stem cells (ISCs). The central regulator, Wnt/ß-catenin signaling plays a key role in intestinal integrity and turnover. Despite its significance, the impact of environmental factors on this pathway has been largely overlooked. This study, for the first time, investigates the influence of Cd on the intestinal Wnt signaling pathway using a mouse model. In this study, male BALB/c mice were administered an environmentally relevant Cd dose (0.98 mg/kg) through oral gavage to investigate the intestinal disruption and Wnt signaling pathway. Various studies, including histopathology, immunohistochemistry, RT-PCR, western blotting, ELISA, intestinal permeability assay, and flow cytometry, were conducted to study Cd-induced changes in the intestine. The canonical Wnt signaling pathway experienced significant downregulation as a result of sub-chronic Cd exposure, which caused extensive damage throughout the small intestine. Increased intestinal permeability and a skewed immune response were also observed. To confirm that Wnt signaling downregulation is the key driver of Cd-induced gastrointestinal toxicity, mice were co-exposed to LiCl (a recognized Wnt activator) and Cd. The results clearly showed that the harmful effects of Cd could be reversed, which is strong evidence that Cd mostly damages the intestine through the Wnt/ß-catenin signalling axis. In conclusion, this research advances the current understanding of the role of Wnt/ß catenin signaling in gastrointestinal toxicity caused by diverse environmental pollutants.


Cadmium , Intestinal Mucosa , Wnt Signaling Pathway , Animals , Male , Mice , beta Catenin/metabolism , Cadmium/toxicity , Inflammation/chemically induced , Inflammation/pathology , Intestinal Mucosa/drug effects , Intestinal Mucosa/metabolism , Intestines/drug effects , Intestines/pathology , Mice, Inbred BALB C , Wnt Signaling Pathway/drug effects
2.
Int J Biol Macromol ; 254(Pt 2): 127717, 2024 Jan.
Article En | MEDLINE | ID: mdl-37923031

Ubiquitination of histone H2B on chromatin is key to gene regulation. E3 ligase Bre1 and E2 Rad6 in Saccharomyces cerevisiae associate together to catalyze mono-ubiquitination at histone H2BK123. Prior studies identified the role of a highly dynamic C-terminal acidic tail of Rad6 indispensable for H2BK123 mono-ubiquitination. However, the mechanistic basis for the Rad6-acidic tail role remained elusive. Using different structural and biophysical approaches, this study for the first time uncovers the direct role of Rad6-acidic tail in interaction with the Bre1 Rad6-Binding Domain (RBD) and recognition of histones surface to facilitate histone H2B mono-ubiquitination. A combination of NMR, SAXS, ITC, site-directed mutagenesis and molecular dynamics studies reveal that RBD domain of Bre1 interacts with Rad6 to stabilize the dynamics of acidic tail. This Bre1-RBD mediated stability in acidic tail of Rad6 could be one of the key factors for facilitating correct recognition of histone surface and ubiquitin-transfer at H2BK123. We provide biophysical evidence that Rad6-acidic tail and a positivity charged surface on histone H2B are involved in recognition of E2:Histones. Taken together, this study uncovers the mechanistic basis for the role of Rad6-acidic in Bre1-RBD mediated recognition of histone surface that ensure the histone H2B mono-ubiquitination.


Histones , Saccharomyces cerevisiae Proteins , Histones/genetics , Ubiquitin-Conjugating Enzymes/genetics , Ubiquitin-Conjugating Enzymes/metabolism , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae/metabolism , Ubiquitin-Protein Ligases/genetics , Scattering, Small Angle , Saccharomyces cerevisiae Proteins/chemistry , X-Ray Diffraction
3.
Microorganisms ; 11(8)2023 Jul 29.
Article En | MEDLINE | ID: mdl-37630494

Biofilms are complex communities of microorganisms that grow on surfaces and are embedded in a matrix of extracellular polymeric substances. These are prevalent in various natural and man-made environments, ranging from industrial settings to medical devices, where they can have both positive and negative impacts. This review explores the diverse applications of microbial biofilms, their clinical consequences, and alternative therapies targeting these resilient structures. We have discussed beneficial applications of microbial biofilms, including their role in wastewater treatment, bioremediation, food industries, agriculture, and biotechnology. Additionally, we have highlighted the mechanisms of biofilm formation and clinical consequences of biofilms in the context of human health. We have also focused on the association of biofilms with antibiotic resistance, chronic infections, and medical device-related infections. To overcome these challenges, alternative therapeutic strategies are explored. The review examines the potential of various antimicrobial agents, such as antimicrobial peptides, quorum-sensing inhibitors, phytoextracts, and nanoparticles, in targeting biofilms. Furthermore, we highlight the future directions for research in this area and the potential of phytotherapy for the prevention and treatment of biofilm-related infections in clinical settings.

4.
Toxicol Res (Camb) ; 11(1): 206-214, 2022 Feb.
Article En | MEDLINE | ID: mdl-35237425

Type 2 diabetes mellitus (T2DM) is a metabolic disease characterized by reduced insulin sensitivity and dysfunction of ß-cells. Although the increasing prevalence of diabetes worldwide is largely attributed to genetic predisposition or lifestyle factors (insufficient physical activity), and caloric intake. Environmental factors, exposure to xenobiotics and heavy metals have also been reported to be causative factors of T2DM. At this juncture, we, through our work unveil a plausible link between Pb2+ exposure and diabetes mellitus, and delineated a comprehensive understanding of the potential mechanisms of Pb2+-induced ß-cells dysfunction. In our in vivo observations, we found that Pb2+ exposure strongly reduced glucose-stimulated insulin secretion and diminished functional pancreatic ß-cell mass. Mechanistically, we found that Pb2+ downregulates intracellular cAMP level via hyper-activating Ca2+/calmodulin-dependent 3',5'-cyclic nucleotide phosphodiesterase 1C and thereby reduces glucose-stimulated insulin secretion. Further, we report that Pb2+ inhibited mitochondrial adenosine triphosphate production and also identified Pb2+ as a negative regulator of ß-cell proliferation via Ca2+/calmodulin-dependent protein kinase kinases-pAMPK-pRaptor axis. Together, our findings strongly reinforce Pb2+ to hijack the physiological role of calcium ions, by mimicking Ca2+ within pancreatic ß-cell and thereby stands as a diabetogenic xenobiotic.

5.
J Biol Chem ; 296: 100714, 2021.
Article En | MEDLINE | ID: mdl-33930463

Overconsumption of sucrose and other sugars has been associated with nonalcoholic fatty liver disease (NAFLD). Reports suggest hepatic de novo lipogenesis (DNL) as an important contributor to and regulator of carbohydrate-induced hepatic lipid accumulation in NAFLD. The mechanisms responsible for the increase in hepatic DNL due to overconsumption of carbohydrate diet are less than clear; however, literatures suggest high carbohydrate diet to activate the lipogenic transcription factor carbohydrate response element-binding protein (ChREBP), which further transcribes genes involved in DNL. Here, we provide an evidence of an unknown link between nuclear factor kappa-light chain enhancer of activated B cells (NF-κB) activation and increased DNL. Our data indicates high carbohydrate diet to enforce nuclear shuttling of hepatic NF-κB p65 and repress transcript levels of sorcin, a cytosolic interacting partner of ChREBP. Reduced sorcin levels, further prompted ChREBP nuclear translocation, leading to enhanced DNL and intrahepatic lipid accumulation both in vivo and in vitro. We further report that pharmacological inhibition of NF-κB abrogated high carbohydrate diet-mediated sorcin repression and thereby prevented ChREBP nuclear translocation and this, in turn, attenuated hepatic lipid accumulation both in in vitro and in vivo. Additionally, sorcin knockdown blunted the lipid-lowering ability of the NF-κB inhibitor in vitro. Together, these data suggest a heretofore unknown role for NF-κB in regulating ChREBP nuclear localization and activation, in response to high carbohydrate diet, for further explorations in lines of NAFLD therapeutics.


Basic Helix-Loop-Helix Leucine Zipper Transcription Factors/metabolism , Cell Nucleus/drug effects , Dietary Carbohydrates/pharmacology , Lipogenesis/drug effects , Liver/metabolism , Transcription Factor RelA/metabolism , Active Transport, Cell Nucleus/drug effects , Cell Nucleus/metabolism , Hep G2 Cells , Humans
6.
Environ Monit Assess ; 192(4): 221, 2020 Mar 07.
Article En | MEDLINE | ID: mdl-32146574

The deterioration of water quality of river Ganga is a huge concern for Govt. of India. Apart from various pollution sources, the religious and ritualistic activities also have a good share in deteriorating Ganga water quality. Thus, the aim of the present study was to evaluate the changes in physico-chemical properties, microbial diversity and role of bacteriophages in controlling bacterial population of Ganga water during mass ritualistic bathing on the occasion of Maha-Kumbh in 2013. The BOD, COD, hardness, TDS and level of various ions significantly increased, while DO decreased in Ganga water during Maha-Kumbh. Ganga water was more affluent in trace elements than Yamuna and their levels further increased during Maha-Kumbh, which was correlated with decreased level of trace elements in the sediment. The bacterial diversity and evenness were increased and correlated with the number of devotees taking a dip at various events. Despite enormous increase in bacterial diversity during mass ritualistic bathing, the core bacterial species found in pre-Kumbh Ganga water were present in all the samples taken during Kumbh and post-Kumbh. In addition, the alteration in bacterial population during mass bathing was well under 2 log units which can be considered negligible. The study of bacteriophages at different bathing events revealed that Ganga was richer with the presence of bacteriophages in comparison with Yamuna against seven common bacteria found during the Maha-Kumbh. These bacteriophages have played a role in controlling bacterial growth and thus preventing putrefaction of Ganga water. Further, the abundance of trace elements in Ganga water might also be a reason for suppression of bacterial growth. Thus, the current study showed that Ganga has characteristic water quality in terms of physico-chemical property and microbial diversity that might have a role in the reported self-cleansing property of Ganga; however, the increased pollution load has surpassed its self-cleansing properties. Since water has been celebrated in all cultures, the outcome of the current study will not only be useful for the policy maker of cleaning and conservation of Ganga but also for restoration of other polluted rivers all over the world.


Environmental Monitoring , Water Pollutants, Chemical , Water Quality , India , Rivers
7.
Int J Nanomedicine ; 14: 9837-9854, 2019.
Article En | MEDLINE | ID: mdl-31849472

INTRODUCTION: An ideal wound dressing material needs to be predisposed with desirable attributes like anti-infective effect, skin hydration balance, adequate porosity and elasticity, high mechanical strength, low wound surface adherence, and enhanced tissue regeneration capability. In this work, we have synthesized hydrogel-based wound patches having antibacterial silver nanoparticles and antioxidant epigallocatechin gallate (EGCG) and showed fast wound closure through their synergistic interaction without any inherent toxicity. METHODS AND RESULTS: Wound patches were synthesized from modified guar gum polymer and assessed to determine accelerated wound healing. The modified polymer beget chemical-free in-situ synthesis of monodispersed silver NPs (~12 nm), an antimicrobial agent, besides lending ionic surface charges. EGCG impregnated during ionotropic gelation process amplified the efficacy of wound patches that possess apt tensile strength, porosity, and swellability for absorbing wound exudates. Further, in vitro studies endorsed them as non-cytotoxic and the post agent effect following exposure to the patch showed an unbiased response to E coli K12 and B. subtilis. In vivo study using sub-cutaneous wounds in Wistar rats validated its accelerated healing properties when compared to a commercially available wound dressing material (skin graft; Neuskin-F®) through better wound contraction, promoted collagen deposition and enhanced vascularization of wound region by modulating growth factors and inflammatory cytokines. CONCLUSION: Synthesized wound patches showed all the desired attributes of a clinically effective dressing material and the results were validated in various in vitro and in vivo assays.


Bandages , Catechin/analogs & derivatives , Metal Nanoparticles/chemistry , Silver/pharmacology , Wound Healing/drug effects , Animals , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/pharmacology , Antioxidants/pharmacology , Bacillus subtilis/drug effects , Catechin/chemistry , Catechin/pharmacology , Escherichia coli/drug effects , Galactans/chemistry , Hydrogels/chemistry , Mannans/chemistry , Metal Nanoparticles/therapeutic use , Plant Gums/chemistry , Polymers/chemistry , Rats, Wistar , Silver/chemistry , Tensile Strength
8.
Neurotoxicology ; 74: 184-195, 2019 09.
Article En | MEDLINE | ID: mdl-31330156

Proteomic analysis was carried out in substantia nigra (SNi) and hippocampus (Hi) isolated from rat offspring born to mothers exposed to lindane (orally; 0.25 mg/kg) from gestation day 5 (GD5) to GD 21 and subsequently rechallenged (orally; 2.5 mg/kg X 21 days) at adulthood (12 weeks). 2D gel electrophoresis revealed no significant differences in the expression of proteins in brain regions isolated from prenatally exposed offspring at adulthood. Significantly greater magnitude of alterations was observed in the expression of proteins related to mitochondrial and energy metabolism, ubiquitin-proteasome pathway, structural and axonal growth leading to increased oxidative stress in Hi and SNi isolated from rechallenged offspring when compared to control offspring treated postnatally with lindane. Western blotting and DNA laddering showed a greater magnitude of increase in apoptosis in the Hi and SNi of rechallenged offspring. Ultrastructural analysis demonstrated disrupted mitochondrial integrity, synaptic disruption and necrotic structures in the brain region of rechallenged offspring. Neurobehavioral studies also demonstrated a greater magnitude of alterations in cognitive and motor functions in rechallenged rats. The data suggest that prenatal exposure of lindane induces persistent molecular changes in the nervous system of offspring which are unmasked leading to neurodegeneration following rechallenge at adulthood.


Hexachlorocyclohexane/toxicity , Insecticides/toxicity , Neurotoxicity Syndromes/pathology , Prenatal Exposure Delayed Effects , Proteomics/methods , Animals , Axons/drug effects , Behavior, Animal/drug effects , Brain Chemistry/drug effects , Energy Metabolism/drug effects , Female , Hippocampus/drug effects , Hippocampus/growth & development , Male , Mitochondria/drug effects , Mitochondria/metabolism , Nerve Tissue Proteins/biosynthesis , Nerve Tissue Proteins/genetics , Neurotoxicity Syndromes/psychology , Oxidative Stress/drug effects , Pregnancy , Proteasome Endopeptidase Complex/drug effects , Rats , Rats, Wistar , Substantia Nigra/drug effects , Substantia Nigra/growth & development
9.
FEBS Lett ; 593(21): 3084-3097, 2019 11.
Article En | MEDLINE | ID: mdl-31309533

Dysregulated hepatic de novo lipogenesis contributes to the pathogenesis of nonalcoholic fatty liver disease in both humans and rodents. Clinical evidence suggests fatty liver to have a positive correlation with serum lead (Pb2+ ) levels. However, an exact mechanism of Pb2+ -induced fatty liver progression is still unknown. Here, we show that exposure to Pb2+ regulates ChREBP-dependent hepatic lipogenesis. Presence of Pb2+ ions within the hepatocytes reduces transcript and protein levels of sorcin, a cytosolic adaptor partner of ChREBP. Adenovirus-mediated overexpression of sorcin in Pb2+ exposed hepatocytes and an in vivo mouse model ameliorates liver steatosis and hepatotoxicity. Hereby, we present Pb2+ exposure to be a lethal disruptor of lipid metabolism in hepatocytes and highlight sorcin as a novel therapeutic target against Pb2+ -induced hepatic dyslipidemia.


Basic Helix-Loop-Helix Leucine Zipper Transcription Factors/genetics , Calcium-Binding Proteins/genetics , Calcium-Binding Proteins/metabolism , Lead/toxicity , Non-alcoholic Fatty Liver Disease/genetics , Transcriptional Activation/drug effects , Animals , Disease Models, Animal , Down Syndrome , Gene Expression Regulation/drug effects , Hep G2 Cells , Humans , Lipogenesis/drug effects , Male , Mice , Non-alcoholic Fatty Liver Disease/chemically induced , Non-alcoholic Fatty Liver Disease/metabolism , Up-Regulation
10.
Bioresour Technol ; 285: 121314, 2019 Aug.
Article En | MEDLINE | ID: mdl-30992159

Biosurfactants from the yeast strains Candida albicans SC5314 and Candida glabrata CBS138 were isolated and characterized. Surface tension of the cell-free broth was reduced from 72 N/m to 42 N/m and 55 N/m respectively. The biosurfactants showed emulsifying ability as the indices against castor oil were determined to be 51% and 53% for C. albicans and C. glabrata respectively and were found stable between pH 2 and 10, temperature 4-120 °C and salt concentration 2-14%. The partially purified surfactants were identified as sophorolipid using Fourier transform infrared spectroscopy. Isolated sophorolipid showed antibacterial properties against pathogenic bacteria and generated reactive oxygen species in Bacillus subtilis and Escherichia coli. Flow cytometric analysis revealed that 60 mg/L of C. glabrata biosurfactant killed 65.8% B. subtilis and 4% E. coli. The data here obtained indicates applications of biosurfactant focusing mainly as antimicrobial and therapeutic perspectives.


Anti-Bacterial Agents , Candida , Emulsifying Agents , Escherichia coli , Surface-Active Agents
11.
Bioresour Technol ; 272: 19-25, 2019 Jan.
Article En | MEDLINE | ID: mdl-30296609

Rhamnolipid produced from a Lysinibacillus sphaericus IITR51 was characterized and its ability for dissolution of hydrophobic pesticides were evaluated. L. sphaericus produced 1.6 g/L of an anionic biosurfactant that reduced surface tension from 72 N/m to 52 N/m with 48% emulsification index. The biosurfactant was found stable over a wide range of pH (4.0-10.0), temperature (4-100 °C), salt concentration (2-14%) and was identified as rhamnolipid. At the concentration of 90 mg/L rhamnolipid showed enhanced dissolution of α-, ß-endosulfan, and γ-hexachlorocyclohexane up to 7.2, 2.9, and 1.8 folds, respectively. The bacterium utilized benzoic acid, chlorobenzene, 3- and 4-chlorobenzoic acid as sole source of carbon and was found resistant to arsenic, lead and cadmium. Furthermore, the isolated biosurfactant showed antimicrobial activities against different pathogenic bacteria. The results obtained indicate the usefulness of rhamnolipid for enhanced dissolution and thereby increasing the bioavailability.


Bacillus/chemistry , Glycolipids/chemistry , Pesticides/chemistry , Hydrophobic and Hydrophilic Interactions , Solubility , Surface Tension , Temperature
12.
J Neurochem ; 149(5): 679-698, 2019 06.
Article En | MEDLINE | ID: mdl-30311190

The anti-diabetic drug and peroxisome proliferator-activated receptor-gamma (PPARγ) agonist, rosiglitazone, alters astrocyte activation; however, its mechanism remains less-known. We hypothesized participation of epidermal growth factor receptor (EGFR), known to control astrocyte reactivity. We first detected that rosiglitazone promoted glial fibrillary acidic protein (GFAP) expression in primary astrocytes as well as the mouse cerebral cortex, associated with increased EGFR activation. Screening for EGFR ligands revealed a rosiglitazone-mediated increase of heparin-binding epidermal growth factor (HB-EGF) in astrocytes, resulting in HB-EGF release into culture medium and mouse cerebrospinal fluid too. Treatment with HB-EGF-siRNA and EGFR inhibitors showed that the rosiglitazone-induced HB-EGF and p-EFGR were interdependent, which participated in GFAP increase. Interestingly, we observed that rosiglitazone could induce cellular and secreted-HB-EGF in neurons also, contributing toward the activated EGFR-induced GFAP in astrocytes. Probing whether these effects of rosiglitazone were PPARγ-linked, revealed potential PPARγ-responsive elements within HB-EGF gene. Moreover, gel-shift, site-directed mutagenesis, chromatin-immunoprecipitation and luciferase-reporter assays demonstrated a PPARγ-dependent HB-EGF transactivation. Subsequently, we examined effects of rosiglitazone in a high-fat diet-fed diabetes mouse model, and supporting observations in the normal cortical cells, identified a rosiglitazone-induced GFAP, astrocyte and neuronal HB-EGF and secreted-HB-EGF in the cerebral cortex of diabetic mice. Moreover, assessing relevance of increased HB-EGF and GFAP revealed an anti-apoptotic role of rosiglitazone in the cerebral cortex, supported by a GFAP-siRNA as well as HB-EGF-siRNA-mediated increase in cleaved-caspase 3 and 9 levels in the rosiglitazone-treated astrocyte-neuron coculture. Overall, our study indicates that rosiglitazone may protect the brain, via a PPARγ-dependent HB-EGF/EGFR signaling and increased GFAP.


Astrocytes/drug effects , Hypoglycemic Agents/pharmacology , Neurons/drug effects , Rosiglitazone/pharmacology , Signal Transduction/drug effects , Animals , Apoptosis/drug effects , Astrocytes/metabolism , Brain/drug effects , Brain/metabolism , Diabetes Mellitus, Experimental/metabolism , Diabetes Mellitus, Type 2/metabolism , Diet, High-Fat , Glial Fibrillary Acidic Protein/biosynthesis , Heparin-binding EGF-like Growth Factor/biosynthesis , Hypoglycemic Agents/adverse effects , Mice , Neurons/metabolism , PPAR gamma/drug effects , PPAR gamma/metabolism , Up-Regulation
15.
Anal Chem ; 89(19): 10343-10352, 2017 10 03.
Article En | MEDLINE | ID: mdl-28825960

Development of a highly photostable, renal clearable, and nontoxic new NIR probe (CyG) for precise quantification of albumin in different biofluids and liver targeted in vivo albumin visualization is demonstrated. CyG's inherent property to interact selectively with albumin among different biomolecules in intracellular environment with high degree of sensitivity helps CyG in targeted liver imaging. In addition to its long excitation/emission wavelengths (λex = 740 nm, λem = 804 nm), which are much above the biological tissue opaque window (400-700 nm) ensuring better photon penetration, diminished tissue autofluorescence and high contrasts, its molecular mass and size are far below the renal cutoff and hence, CyG qualifies as imaging material for clinical studies. We anticipate that CyG will provide new strategies to overcome the pitfall of present day albumin detection methods as well as accelerate the detection process at relatively lower costs without compromising the accuracy of detection. Moreover, the renal excretion kinetic and intrahepatic albumin binding affinity of CyG can further be used to differentiate between fatty liver from healthy liver in an experimentally arrived mouse model using noninvasive technique.


Albumins/analysis , Body Fluids/metabolism , Fluorescent Dyes/chemistry , Microscopy, Confocal , Albumins/metabolism , Animals , Fluorescent Dyes/chemical synthesis , Hep G2 Cells , Humans , Liver/diagnostic imaging , Liver/metabolism , Mice , Mice, Inbred BALB C , Non-alcoholic Fatty Liver Disease/diagnosis , Non-alcoholic Fatty Liver Disease/diagnostic imaging , RAW 264.7 Cells , Spectroscopy, Near-Infrared
16.
PLoS One ; 12(3): e0173381, 2017.
Article En | MEDLINE | ID: mdl-28278249

Bioethanol is an environment friendly and renewable source of energy produced by the fermentation of agricultural raw material by a variety of microorganisms including yeast. Obtaining yeast strains that are tolerant to stresses like high levels of ethanol and high temperature is highly desirable as it reduces cost and increases yield during bioethanol production. Here, we report that heterologous expression of C-5 Sterol desaturase (FvC5SD)-an ergosterol biosynthesis enzyme from an edible mushroom Flammulina velutipes in fission yeast, not only imparts increased thermotolerance but also tolerance towards high ethanol concentration and low pH. This tolerance could be attributed to an increase of ≈1.5 fold in the level of ergosterol and oleic acid (C-18 unsaturated fatty acid) as analysed by gas chromatography- mass spectrometry. FvC5SD is a membrane localized iron binding enzyme that introduces double bond at C-5 position into the Δ7-sterol substrates to yield Δ5, 7- sterols as products. In F. velutipes, FvC5SD transcript was observed to be upregulated by ≈5 fold under low pH condition and by ≈ 9 folds and ≈5 fold at 40°C and 4°C respectively when compared to normal growth temperature of 23°C. Besides, susceptibility to cell wall inhibiting drugs like Congo red and Calcoflour white was also found to increase in FvC5SD expressing S. pombe strain. Alteration in membrane sterol and fatty acid composition could also lead to increase in susceptibility to cell wall inhibiting drugs. Thus, this study has immense industrial application and can be employed to ensure competitiveness of fermentation process.


Ethanol/pharmacology , Flammulina/enzymology , Oxidoreductases/genetics , Schizosaccharomyces/drug effects , Schizosaccharomyces/genetics , Thermotolerance/genetics , Cell Membrane/drug effects , Cell Membrane/metabolism , Cell Wall/drug effects , Ergosterol/metabolism , Flammulina/genetics , Gene Expression , Hydrogen-Ion Concentration , Oleic Acid/metabolism , Oxidoreductases/metabolism , Promoter Regions, Genetic/genetics , Protein Transport , Schizosaccharomyces/metabolism , Schizosaccharomyces/physiology , Temperature
17.
J Neurochem ; 140(1): 96-113, 2017 01.
Article En | MEDLINE | ID: mdl-27787894

The astrocyte marker, glial fibrillary acidic protein (GFAP), has essential functions in the brain, but may trigger astroglial scarring when expressed in excess. Docosahexaenoic acid (DHA) is an n-3 fatty acid that is protective during brain development. However, the effect of DHA on GFAP levels of developing brain remains unexplored. Here, we detected that treating developing rats with DHA-enriched fish-oil caused dose-dependent GFAP augmentation. We investigated the mechanism promoting GFAP, hypothesizing the participation of fatty acid-binding protein-7 (FABP7), known to bind DHA. We identified that DHA stimulated FABP7 expression in astrocytes, and FABP7-silencing suppressed DHA-induced GFAP, indicating FABP7-mediated GFAP increase. Further investigation proved FABP7 expression to be phosphatidylinositide 3-kinases (PI3K)/AKT and nuclear receptor peroxisome proliferator-activated receptor-gamma (PPARγ)-dependent. We found that PI3K/AKT activated PPARγ that triggered FABP7 expression via PPARγ-responsive elements within its gene. Towards identifying FABP7-downstream pathways, we considered our previous report that demonstrated cyclin-dependent kinase-5 (CDK5)-PPARγ-protein-protein complex to suppress GFAP. We found that the DHA-induced FABP7 underwent protein-protein interaction with PPARγ, which impeded CDK5-PPARγ formation. Hence, it appeared that enhanced FABP7-PPARγ in lieu of CDK5-PPARγ resulted in increased GFAP. PI3K/AKT not only stimulated formation of FABP7-PPARγ protein-protein complex, but also up-regulated a FABP7-independent MAP-kinase-phosphatase-3 pathway that inactivated CDK5 and hence attenuated CDK5-PPARγ. Overall, our data reveal that via the proximal PI3K/AKT, DHA induces FABP7-PPARγ, through genomic and non-genomic mechanisms, and MAP-kinase-phosphatase-3 that converged at attenuated CDK5-PPARγ and therefore, enhanced GFAP. Accordingly, our study demonstrates a DHA-mediated astroglial hyperactivation, pointing toward a probable injurious role of DHA in brain development.


Astrocytes/metabolism , Docosahexaenoic Acids/pharmacology , Dual Specificity Phosphatase 6/biosynthesis , Fatty Acid-Binding Protein 7/biosynthesis , Glial Fibrillary Acidic Protein/biosynthesis , Oncogene Protein v-akt/biosynthesis , PPAR gamma/biosynthesis , Animals , Astrocytes/drug effects , Brain/drug effects , Brain/growth & development , Brain/metabolism , Cells, Cultured , Dose-Response Relationship, Drug , Female , Male , Protein Binding/physiology , Rats , Rats, Wistar , Up-Regulation/drug effects , Up-Regulation/physiology
18.
Theor Appl Genet ; 129(9): 1639-55, 2016 Sep.
Article En | MEDLINE | ID: mdl-27381849

KEY MESSAGE: New advances in crop genetic engineering can significantly pace up the development of genetically improved varieties with enhanced yield, nutrition and tolerance to biotic and abiotic stresses. Genetically modified (GM) crops can act as powerful complement to the crops produced by laborious and time consuming conventional breeding methods to meet the worldwide demand for quality foods. GM crops can help fight malnutrition due to enhanced yield, nutritional quality and increased resistance to various biotic and abiotic stresses. However, several biosafety issues and public concerns are associated with cultivation of GM crops developed by transgenesis, i.e., introduction of genes from distantly related organism. To meet these concerns, researchers have developed alternative concepts of cisgenesis and intragenesis which involve transformation of plants with genetic material derived from the species itself or from closely related species capable of sexual hybridization, respectively. Recombinase technology aimed at site-specific integration of transgene can help to overcome limitations of traditional genetic engineering methods based on random integration of multiple copy of transgene into plant genome leading to gene silencing and unpredictable expression pattern. Besides, recently developed technology of genome editing using engineered nucleases, permit the modification or mutation of genes of interest without involving foreign DNA, and as a result, plants developed with this technology might be considered as non-transgenic genetically altered plants. This would open the doors for the development and commercialization of transgenic plants with superior phenotypes even in countries where GM crops are poorly accepted. This review is an attempt to summarize various past achievements of GM technology in crop improvement, recent progress and new advances in the field to develop improved varieties aimed for better consumer acceptance.


Crops, Agricultural/genetics , Genetic Engineering/methods , Plants, Genetically Modified/genetics , Disease Resistance , Food Storage , Nutritive Value , Plant Diseases , Stress, Physiological , Transgenes
19.
Sci Rep ; 6: 25340, 2016 05 06.
Article En | MEDLINE | ID: mdl-27150014

Plant often responds to fungal pathogens by expressing a group of proteins known as pathogenesis-related proteins (PRs). The expression of PR is mediated through pathogen-induced signal-transduction pathways that are fine-tuned by phytohormones such as methyl jasmonate (MeJA). Here, we report functional characterization of an Ocimum basilicum PR5 family member (ObTLP1) that was identified from a MeJA-responsive expression sequence tag collection. ObTLP1 encodes a 226 amino acid polypeptide that showed sequence and structural similarities with a sweet-tasting protein thaumatin of Thaumatococcus danielli and also with a stress-responsive protein osmotin of Nicotiana tabacum. The expression of ObTLP1 in O. basilicum was found to be organ-preferential under unstressed condition, and responsive to biotic and abiotic stresses, and multiple phytohormone elicitations. Bacterially-expressed recombinant ObTLP1 inhibited mycelial growth of the phytopathogenic fungi, Scleretonia sclerotiorum and Botrytis cinerea; thereby, suggesting its antifungal activity. Ectopic expression of ObTLP1 in Arabidopsis led to enhanced tolerance to S. sclerotiorum and B. cinerea infections, and also to dehydration and salt stress. Moreover, induced expression of the defense marker genes suggested up-regulation of the defense-response pathways in ObTLP1-expressing Arabidopsis upon fungal challenge. Thus, ObTLP1 might be useful for providing tolerance to the fungal pathogens and abiotic stresses in crops.


Arabidopsis/genetics , Ocimum basilicum/metabolism , Plant Proteins/genetics , Plants, Genetically Modified/microbiology , Acetates/pharmacology , Arabidopsis/microbiology , Cyclopentanes/pharmacology , Disease Resistance , Fungi/growth & development , Ocimum basilicum/genetics , Oxylipins/pharmacology , Plant Growth Regulators/pharmacology , Plant Proteins/chemistry , Plant Proteins/metabolism , Stress, Physiological
20.
Genome Announc ; 4(2)2016 Mar 03.
Article En | MEDLINE | ID: mdl-26941147

Here, we report the 4.16-Mb draft genome sequence of an indole-degrading bacterium, Acinetobacter baumannii IITR88, isolated from the Bhagirathi river in India. A total of 4,069 coding regions (CDSs), 3 rRNAs, and 52 tRNAs were predicted. Genes for the degradation of indoles, phenylacetaldehyde, anthranilate, and several other aromatic compounds were present.

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