Polyherbal decoction modulates redox homeostasis during Malachite green induced metabolic stress in Saccharomyces cerevisiae

Malachite green (MG) is aquatic pollutant that induces oxidative stress when comes in contact with the living organisms. In Saccharomyces cerevisiae, MG produces intracellular reactive oxygen species (ROS) and these ROS disturb redox homeostasis and cellular functions leading to early cell death. Exogenous supply of natural antioxidants containing polyherbal decoction may play a crucial role in re-establishment of redox homeostasis by ensuring the cell survival. Exposure of MG to Saccharomyces cerevisiae resulted in a significant decrease (97.8%) in colony forming units (CFU). An Ayurvedic polyherbal formulation ‘Vayasthapana Rasayana’ (VR) which contains natural antioxidants from plants viz. Terminalia chebula, Clitoria ternatea, Boerhaavia diffusa, Centella asiatica, Phyllanthus emblica, Asparagus racemossus and Tinospora cordifolia at 1.0 mg/mL concentration could arrest the oxidative stress during MG exposure. Levels of ROS elevated up to 67.3% on MG exposure; while VR supplementation reduced it by 54.7%. MG induced cellular apoptosis in 38% and necrosis in 27% cells, while VR augmentation reduced it to 8%. Activities of antioxidant enzymes like catalase, superoxide dismutase and glutathione peroxidase in MG exposed cells were induced by 408, 144 and 140%, respectively, whereas, VR supplementation lowered the expressions to 102, 57 and 111%, respectively. Induction in caspases 3/7 activity was also found to be reduced by 65.39% after VR augmentation. Similarly, VR modulated activities of oxido-reductases like lignin peroxidase, laccase, NADH-DCIP reductase and MG reductase. VR supplementation also maintained the MG utilization potential of S. cerevisiae up to 20th exposure cycle which was otherwise arrested to 8th cycle. The treatment also decreased the ROS accumulation and nuclear damage, restoring the cell viability up to 94% and retained normal growth dynamics. Thus, VR supplementation could significantly decrease oxidative stress, enhance cell viability and ultimately protect the dying S. cerevisiae cells during MG exposure

Polyherbal decoction modulates redox homeostasis during Malachite green induced metabolic stress in Saccharomyces cerevisiae

17-26Malachite green (MG) is aquatic pollutant that induces oxidative stress when comes in contact with the living organisms. In Saccharomyces cerevisiae, MG produces intracellular reactive oxygen species (ROS) and these ROS disturb redox homeostasis and cellular functions leading to early cell death. Exogenous supply of natural antioxidants containing polyherbal decoction may play a crucial role in re-establishment of redox homeostasis by ensuring the cell survival. Exposure of MG to Saccharomyces cerevisiae resulted in a significant decrease (97.8%) in colony forming units (CFU). An Ayurvedic polyherbal formulation ‘Vayasthapana Rasayana’ (VR) which contains natural antioxidants from plants viz. Terminalia chebula, Clitoria ternatea, Boerhaavia diffusa, Centella asiatica, Phyllanthus emblica, Asparagus racemossus and Tinospora cordifolia at 1.0 mg/mL concentration could arrest the oxidative stress during MG exposure. Levels of ROS elevated up to 67.3% on MG exposure; while VR supplementation reduced it by 54.7%. MG induced cellular apoptosis in 38% and necrosis in 27% cells, while VR augmentation reduced it to 8%. Activities of antioxidant enzymes like catalase, superoxide dismutase and glutathione peroxidase in MG exposed cells were induced by 408, 144 and 140%, respectively, whereas, VR supplementation lowered the expressions to 102, 57 and 111%, respectively. Induction in caspases 3/7 activity was also found to be reduced by 65.39% after VR augmentation. Similarly, VR modulated activities of oxido-reductases like lignin peroxidase, laccase, NADH-DCIP reductase and MG reductase. VR supplementation also maintained the MG utilization potential of S. cerevisiae up to 20th exposure cycle which was otherwise arrested to 8th cycle. The treatment also decreased the ROS accumulation and nuclear damage, restoring the cell viability up to 94% and retained normal growth dynamics. Thus, VR supplementation could significantly decrease oxidative stress, enhance cell viability and ultimately protect the dying S. cerevisiae cells during MG exposure

Interactome Networks of FOSL1 and FOSL2 in Human Th17 Cells

Dysregulated function of Th17 cells has implications in immunodeficiencies and autoimmune disorders. Th17 cell differentiation is orchestrated by a complex network of transcription factors, including several members of the activator protein (AP-1) family. Among the latter, FOSL1 and FOSL2 modulate the effector functions of Th17 cells. However, the molecular mechanisms underlying these effects are unclear, owing to the poorly characterized protein interaction networks of FOSL factors. Here, we establish the first interactomes of FOSL1 and FOSL2 in human Th17 cells, using affinity purification–mass spectrometry analysis. In addition to the known JUN proteins, we identified several novel binding partners of FOSL1 and FOSL2. Gene ontology analysis found a significant fraction of these interactors to be associated with RNA-binding activity, which suggests new mechanistic links. Intriguingly, 29 proteins were found to share interactions with FOSL1 and FOSL2, and these included key regulators of Th17 fate. We further validated the binding partners identified in this study by using parallel reaction monitoring targeted mass spectrometry and other methods. Our study provides key insights into the interaction-based signaling mechanisms of FOSL proteins that potentially govern Th17 cell differentiation and associated pathologies. </p

Late Diagnosis of Oral Mucosal Melanoma: Case Report

Primary oral malignant melanoma usually presents as a dark brown or black lesion. It is a rare malignancy, accounting for less than 1% of all melanomas and 1.6% of all head and neck malignancies, thus forming up to 0.5% of all oral malignancies in the world literature. In general, the prognosis of oral melanoma is poor and worse than that of cutaneous melanoma. The preferred treatment is radical surgery alone or in combination with radiotherapy, chemotherapy, and immunotherapy and immunomodulatory agents. A case of Oral Malignant Melanoma is presented here which was undetected during the first visit to the hospital. When a simple oral surgical treatment was carried out in that region, it resulted in the appearance of a massive pigmented lesion which was histopathologically diagnosed as malignant melanoma. This paper is presented to re- emphasize the fact that any pigmented lesion in the oral cavity should be viewed with suspicion and proper investigation (biopsy) should be carried out to rule out any untoward experiences later

A comparative surface evaluation of orthodontic mini-implants before and after en masse retraction—A SEM study

OBJECTIVE: To evaluate the changes in surface morphology of two different types of mini-implants after clinical en masse retraction using scanning electron microscopy. MATERIAL AND METHODS: Fifty mini-implants of Dentos (Korea, Absoanchor, BH-1817-08) and Orlus (Korea, Yesanchor, C-1817) were inserted in patients in a split-mouth design who required en masse anterior retraction and absolute anchorage. Surface characteristics of mini-implants such as pitch (distance between consecutive threads), flank width (distance between root and crest), and taper were studied using scanning electron microscope (FEI nanosem450) before and after clinical use. RESULTS: Statistically significant difference (p value = 0.003) was found in a mean reduction of pitch dimension among the two groups with a mean difference of 25.000 μm. Also, a statistically significant difference was noted (p value = 0.001) in a mean reduction of flank width among Dentos implants as compared to Orlus implants. A statistically significant difference (p = 0.001) was seen in the mean reduction of taper dimension among Dentos group (0.0140 ± 0.02271) as compared to the Orlus group (0.0810 ± 0.05152). CONCLUSION: A marked reduction in surface morphology such as the pitch, flank width, and taper of both mini-implants after retrieval was observed. Dentos group of mini-implants displayed better dimensional stability post-retrieval as compared to the Orlus group of mini-implants. All the mini-screws showed milling defects in form of scratches on observation under scanning electron microscopy despite a smooth appearance to the naked eye

COMPREHENSIVE MOLECULAR STUDY REVEALS THE POTENTIAL ROLE OF CHEBULINIC ACID AND BOERAVINONE B TO ESTABLISH REDOX HOMEOSTASIS IN METABOLICALLY STRESSED CELLS

Objective: Our objective was to assess the multi-dimensional protective mechanism of Chebulinic acid (CA) and Boeravinone B (BB) against MG-induced metabolic stress and cytotoxicity. Methods: In this study, we have studied CA-and BB-mediated changes in molecular markers of highly dynamic mechanisms of mitochondrial disintegration, nuclear damage and cell death pathways associated with early ageing in MG-exposed Saccharomyces cerevisiae cells using biochemical assays, qRT-PCR and flow cytometry. Results: We found that CA and BB interventions during MG-induced stress in S. cerevisiae reduce the rate of extracellular nitrite production, protein carbonyl content, lipid peroxidation and in addition, photo components positively modulate the expression patterns of genes involved in different cell death pathways. Furthermore, CA and BB treatments to MG-stressed cells reduced the number of cells in late apoptosis by 13.4% and 28.3%, respectively. On top of that, CA and BB supplementation during MG-stress restored mitochondrial membrane potential (ΔΨm) by 63.0% and 62.5%, respectively. Conclusion: Based on the results of this study, it seems CA and BB phytotherapy protect against MG-induced cytotoxicity through their natural antioxidant properties by establishing redox homeostasis; thus, CA and BB defend the cell components from oxidative damage of different biomolecules and organelles, ultimately increase longevity

Photocatalytic One-Pot Conversion of Aldehydes to Esters and Degradation of Rhodamine B Dye Using Mesoporous Graphitic Carbon Nitride

Graphitic carbon nitride (g-C3N4) is a prominent semiconductor material for numerous photocatalytic applications. The mesoporous g-C3N4 (mpg-C3N4), a polymeric metal-free semiconductor proficient photocatalyst with triazine rings, is lightweight and has a high surface area. In this work, we elucidated the role of the mpg-C3N4 photocatalyst for the esterification of substituted benzaldehyde without any additives, a metal-free approach, and under visible light irradiation at room temperature. The mpg-C3N4 has been well characterized by various physicochemical techniques like X-ray diffraction, Fourier-transform infrared, UV–visible spectrophotometry, transmission electron microscopy (TEM), high-resolution TEM, elemental mapping, and the Brunauer–Emmett–Teller surface area. The catalyst characterization revealed that the as-prepared mpg-C3N4 material has a high surface area (97.39 m2/g), porosity, a medium band gap (2.83 eV), and a nanowire-like shape. The mpg-C3N4 properties enhanced the material’s photocatalytic activity toward the esterification of substituted benzaldehyde to afford a maximum of 35% conversion with 99% selectivity. A further extension of this work is explored for the organic dye degradation reaction, where it is observed that the materials are stable, sturdy, and reusable for more than eight cycles with a slight loss in activity. This research widens the versatile application of mpg-C3N4 with specific catalytic applications

Phytoextracts protect <i>Saccharomyces cerevisiae</i> from oxidative stress with simultaneous enhancement in bioremediation efficacy

469-478Bioremediation efficacies are highly affected by abiotic stresses imparted by a verity of pollutants due to generation of reactive oxygen species (ROS). These stressed cells can be treated using natural or synthetic antioxidants. Such an approach could prove beneficial to bioremediation agents as the exogenously added antioxidant compounds would scavenge the generated free radicals. This would definitely lead to increased longevity of the involved organism and carry out superior treatments. In present study, Malachite Green (MG) was found to exert oxidative stress on Saccharomyces cerevisiae through generation ROS. A 2 h exposure of MG though achieved 99% decolourization, the cells revealed a significant decrease (97.8%) in colony forming units (CFU) upon further subculture. Natural antioxidants from Centella asiatica, Phyllanthus emblica, Asperagus racemosus and Tinospora cordifolia extracts, however, restored the CFU with a loss of only 16-33%. The MG stressed cells indicated an increase in ROS by 6.7 fold which was reduced to near normal due to augmentation with plant extracts. MG damaged the nuclear material up to 90% and inclusion of phytoextracts protected the cells revealing only 0-7% nuclear damage. Induction in apoptosis (92%) and necrosis (23%) in MG exposed cells was noted, while plant extracts augmentation reduced apoptosis to 15-49% and necrosis to 10-16%. Activities of antioxidant enzymes such as superoxide dismutase, catalase and glutathione peroxidase were significantly decreased in phyto-augmented cells when compared to MG stressed cells. Dye degrading enzymes, namely lignin peroxidase, laccase, NADH-DCIP reductase and MG reductase were found to show induction in activities during MG utilization. Since antioxidants from plant extracts could protect the cells form oxidative stress, they were used to treat MG for 20 continuous decolourization cycles. Augmentation of C. asiatica, P. emblica, A. racemosus and T. cordifolia extracts at 20th decolourization cycle revealed 75, 79, 74 and 93% superior decolorization efficacies as compared to unaugmented cells. These natural antioxidants to protect bioremediation agents form oxidative stress, thus concluded to show enhanced treatment

NF-kappa B Signaling and IL-4 Signaling Regulate SATB1 Expression via Alternative Promoter Usage During Th2 Differentiation

SATB1 is a genome organizer protein that is expressed in a lineage specific manner in CD4(+) T-cells. SATB1 plays a crucial role in expression of multiple genes throughout the thymic development and peripheral differentiation of T cells. Although SATB1 function has been subjected to intense investigation, regulation of SATB1 gene expression remains poorly understood. Analysis of RNA-seq data revealed multiple transcription start sites at the upstream regulatory region of SATB1. We further demonstrated that SATB1 gene is expressed via alternative promoters during T-helper (Th) cell differentiation. The proximal promoter "P1" is used more by the naive and activated CD4(+) T-cells whereas the middle "P2" and the distal "P3" promoters are used at a significantly higher level by polarized T-helper cells. Cytokine and TCR signaling play crucial roles toward SATB1 alternative promoter usage. Under Th2 polarization conditions, transcription factor STAT6, which operates downstream of the cytokine signaling binds to the P2 and P3 promoters. Genetic perturbation by knockout and chemical inhibition of STAT6 activation resulted in the loss of P2 and P3 promoter activity. Moreover, chemical inhibition of activation of NF-KB, a transcription factor that operates downstream of the TCR signaling, also resulted in reduced P2 and P3 promoter usage. Furthermore, usage of the P1 promoter correlated with lower SATB1 protein expression whereas P2 and P3 promoter usage correlated with higher SATB1 protein expression. Thus, the promoter switch might play a crucial role in fine-tuning of SATB1 protein expression in a cell type specific manner

NF-κB signaling and IL-4 signaling regulate SATB1 expression via alternative promoter usage during Th2 differentiation

Abstract SATB1 is a genome organizer protein that is expressed in a lineage specific manner in CD4+ T-cells. SATB1 plays a crucial role in expression of multiple genes throughout the thymic development and peripheral differentiation of T cells. Although SATB1 function has been subjected to intense investigation, regulation of SATB1 gene expression remains poorly understood. Analysis of RNA-seq data revealed multiple transcription start sites at the upstream regulatory region of SATB1. We further demonstrated that SATB1 gene is expressed via alternative promoters during T-helper (Th) cell differentiation. The proximal promoter “P1” is used more by the naïve and activated CD4+ T-cells whereas the middle “P2” and the distal “P3” promoters are used at a significantly higher level by polarized T-helper cells. Cytokine and TCR signaling play crucial roles toward SATB1 alternative promoter usage. Under Th2 polarization conditions, transcription factor STAT6, which operates downstream of the cytokine signaling binds to the P2 and P3 promoters. Genetic perturbation by knockout and chemical inhibition of STAT6 activation resulted in the loss of P2 and P3 promoter activity. Moreover, chemical inhibition of activation of NF-κB, a transcription factor that operates downstream of the TCR signaling, also resulted in reduced P2 and P3 promoter usage. Furthermore, usage of the P1 promoter correlated with lower SATB1 protein expression whereas P2 and P3 promoter usage correlated with higher SATB1 protein expression. Thus, the promoter switch might play a crucial role in fine-tuning of SATB1 protein expression in a cell type specific manner