Search results for: translocation
19 Phytoextraction of Some Heavy Metals from Artificially Polluted soil
Authors: Kareem Kalo Qassim, Hassan A. M. Mezori
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The bioaccumulation of heavy metals in the environment has become a matter of public interest because it persists in the soil longer than other components of the biosphere. Bioremediation has emerged as the ideal alternative environmentally friendly and ecological sound technology for removing pollutants from polluted sites. Phytoremediation is an attractive remediation technology that makes use of plants to remove contaminants from the environment. A pot experiment was conducted under lath house conditions to evaluate the ability of plants (H. Annuus, S. Bicolor, and Z. Mays) to phytoextract heavy metals from artificially polluted soils by different concentrations of Cadmium, Lead, and Copper (0, 100, 200, 200 + EDTA). The Seed germination was influenced by the presence of heavy metals and inhibition increased by increasing the heavy metals concentration. A significant difference was observed in the effect of lead and copper. Generally, the length of root, shoot, and intact plant was reduced by all the concentrations used in the experiments. The root system was affected more than the shoot system of the same plants. All heavy metals concentrations caused a reduction in the dry weight and chlorophyll content of all tested plant species; the reduction was increased by increasing the concentration of all heavy metals, especially when EDTA was added. The Bioaccumulation of heavy metals concentration of all the tested plants increased by increasing the concentration. The highest accumulation of cadmium was (81.77mg kg⁻¹), and copper was ( 65.07 mg kg⁻¹) in S. bicolor, while lead-in H. annuus was (60.74 mg kg⁻¹). The order of accumulation of heavy metals in all the tested plant species in the root system and the intact plant was as follows: H. annuus ˃ S. bicolor ˃ Z. mays and shoot system was: H. annuus ˃ Z. mays ˃ S. bicolor. The highest TF of cadmium was (0.41) in H. annuus; lead was (0.72) in Z. mays and S. bicolor, and copper was (0.44) in Z. mays. The tested plant species varied in their response to the heavy metals and the inhibition was concentration depended. In general, the roots system was more affected by heavy metals toxicity than the shoots system; the roots system accumulated more heavy metals in the roots than the shoots system. The addition of EDTA to the last concentration of heavy metals facilitated the availably and absorption of heavy metals from the polluted soil by all tested plant species.Keywords: phytoextyraction, phytoremediation, translocation, heavy metals, soil pollution
Procedia PDF Downloads 14818 Fastidious Enteric Pathogens in HIV
Authors: S. Pathak, R. Lazarus
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A 25-year-old male HIV patient (CD4 cells 20/µL and HIV viral load 14200000 copies/ml) with a past medical history of duodenal ulcer, pneumocystis carinii pneumonia, oesophageal candidiasis presented with fever and a seizure to hospital. The only recent travel had been a religious pilgrimage from Singapore to Malaysia 5 days prior; during the trip he sustained skin abrasions. The patient had recently started highly active antiretroviral therapy 2 months prior. Clinical examination was unremarkable other than a temperature of 38.8°C and perianal warts. Laboratory tests showed a leukocyte count 12.5x109 cells/L, haemoglobin 9.4 g/dL, normal biochemistry and a C-reactive protein 121 mg/L. CT head and MRI head were unremarkable and cerebrospinal fluid analysis performed after a delay (due to technical difficulties) of 11 days was unremarkable. Blood cultures (three sets) taken on admission showed Gram-negative rods in the anaerobic bottles only at the end of incubation with culture result confirmed by molecular sequencing showing Helicobacter cinaedi. The patient was treated empirically with ceftriaxone for seven days and this was converted to oral co-amoxiclav for a further seven days after the blood cultures became positive. A Transthoracic echocardiogram was unremarkable. The patient made a full recovery. Helicobacter cinaedi is a gram-negative anaerobic fastidious organism affecting patients with comorbidity. Infection may manifest as cellulitius, colitis or as in this case as bloodstream infection – the latter is often attributed to faeco-oral infection. Laboratory identification requires prolonged culture. Therapeutic options may be limited by resistance to macrolides and fluoroquinolones. The likely pathogen inoculation routes in the case described include gastrointestinal translocation due to proctitis at the site of perianal warts, or breach of the skin via abrasions occurring during the pilgrimage. Such organisms are increasing in prevalence as our patient population ages and patients have multiple comorbidities including HIV. It may be necessary in patients with unexplained fever to prolong incubation of sterile sites including blood in order to identify this unusual fastidious organism.Keywords: fastidious, Helicobacter cinaedi, HIV, immunocompromised
Procedia PDF Downloads 37917 Qualitative Modeling of Transforming Growth Factor Beta-Associated Biological Regulatory Network: Insight into Renal Fibrosis
Authors: Ayesha Waqar Khan, Mariam Altaf, Jamil Ahmad, Shaheen Shahzad
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Kidney fibrosis is an anticipated outcome of possibly all types of progressive chronic kidney disease (CKD). Epithelial-mesenchymal transition (EMT) signaling pathway is responsible for production of matrix-producing fibroblasts and myofibroblasts in diseased kidney. In this study, a discrete model of TGF-beta (transforming growth factor) and CTGF (connective tissue growth factor) was constructed using Rene Thomas formalism to investigate renal fibrosis turn over. The kinetic logic proposed by Rene Thomas is a renowned approach for modeling of Biological Regulatory Networks (BRNs). This modeling approach uses a set of constraints which represents the dynamics of the BRN thus analyzing the pathway and predicting critical trajectories that lead to a normal or diseased state. The molecular connection between TGF-beta, Smad 2/3 (transcription factor) phosphorylation and CTGF is modeled using GenoTech. The order of BRN is CTGF, TGF-B, and SMAD3 respectively. The predicted cycle depicts activation of TGF-B (TGF-β) via cleavage of its own pro-domain (0,1,0) and presentation to TGFR-II receptor phosphorylating SMAD3 (Smad2/3) in the state (0,1,1). Later TGF-B is turned off (0,0,1) thereby activating SMAD3 that further stimulates the expression of CTGF in the state (1,0,1) and itself turns off in (1,0,0). Elevated CTGF expression reactivates TGF-B (1,1,0) and the cycle continues. The predicted model has generated one cycle and two steady states. Cyclic behavior in this study represents the diseased state in which all three proteins contribute to renal fibrosis. The proposed model is in accordance with the experimental findings of the existing diseased state. Extended cycle results in enhanced CTGF expression through Smad2/3 and Smad4 translocation in the nucleus. The results suggest that the system converges towards organ fibrogenesis if CTGF remains constructively active along with Smad2/3 and Smad 4 that plays an important role in kidney fibrosis. Therefore, modeling regulatory pathways of kidney fibrosis will escort to the progress of therapeutic tools and real-world useful applications such as predictive and preventive medicine.Keywords: CTGF, renal fibrosis signaling pathway, system biology, qualitative modeling
Procedia PDF Downloads 17916 Phytoremediation of Heavy Metals by the Perennial Tussock Chrysopogon Zizanioides Grown on Zn and Cd Contaminated Soil Amended with Biochar
Authors: Dhritilekha Deka, Deepak Patwa, Ravi K., Archana M. Nair
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Bioaccumulation of heavy metal contaminants due to intense anthropogenic interference degrades the environment and ecosystem functions. Conventional physicochemical methods involve energy-intensive and costly methodologies. Phytoremediation, on the other hand, provides an efficient nature-based strategy for the reclamation of heavy metal-contaminated sites. However, the slow process and adaptation to high-concentration contaminant sequestration often limit the efficiency of the method. This necessitates natural amendments such as biochar to improve phytoextraction and stabilize the green cover. Biochar is a highly porous structure with high carbon sequestration potential and containing negatively charged functional groups that provide binding sites for the positively charged metals. This study aims to develop and determine the synergy between sugarcane bagasse biochar content and phytoremediation. A 60-day pot experiment using perennial tussock vetiver grass (Chrysopogon zizanioides) was conducted for different biochar contents of 1%, 2%, and 4% for the removal of cadmium and zinc. A concentration of 500 ppm is maintained for the amended and unamended control (CK) samples. The survival rates of the plants, biomass production, and leaf area index were measured for the plant growth characteristics. Results indicate a visible change in the plant growth and the heavy metal concentration with the biochar content. The bioconcentration factor (BCF) in the plant improved significantly for the 4% biochar content by 57% in comparison to the control CK treatment in Cd-treated soils. The Zn soils indicated the highest reduction in the metal concentration by 50% in the 2% amended samples and an increase in the BCF in all the amended samples. The translocation from the rhizosphere to the shoots was low but not dependent on the amendment content and varied for each contaminant type. The root-to-shoot ratio indicates higher values compared to the control samples. The enhanced tolerance capacities can be attributed to the nutrients released by the biochar in the soil. The study reveals the high potential of biochar as a phytoremediation amendment, but its effect is dependent on the soil and heavy metal and accumulator species.Keywords: phytoextraction, biochar, heavy metals, chrysopogon zizanioides, bioaccumulation factor
Procedia PDF Downloads 6515 Comparison of Home Ranges of Radio Collared Jaguars (Panthera onca L.) in the Dry Chaco and Wet Chaco of Paraguay
Authors: Juan Facetti, Rocky McBride, Karina Loup
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The Chaco Region of Paraguay is a key biodiverse area for the conservation of jaguars (Panthera onca), the largest feline of the Americas. It comprises five eco-regions, which holds important but decreasing populations of this species. The last decades, the expansion of soybean over the Atlantic Forest, forced the translocation of cattle-ranches towards the Chaco. Few studies of Jaguar's population densities in the American hemisphere were done until now. In the region, the specie is listed as vulnerable or threatened and more information is needed to implement any conservation policy. Among the factors that threaten the populations are land-use change, habitat fragmentation, prey depletion and illegal hunting. Two largest eco-regions were studied: the Wet Chaco and the Dry Chaco. From 2002 more than 20 jaguars were captured and fitted with GPS-collar. Data collected from 11 GPS-collars were processed, transformed numerically and finally converted into maps for analyzing. 8.092 locations were determined for four adult females (AF) and one adult male (AM) in the Wet Chaco, and one AF, one juvenile male (JM) and four AM in the Dry Chaco, during 1,867 days. GIS and kernel methodology were used to calculate daily distance of movement, home range-HR (95% isopleth), and core area (considered as 50% isopleth). In the Wet Chaco HR were 56 Km2 and 238 km2 for females and males respectively; while in the Dry Chaco HR were 685 Km2 and 844.5 km2 for females and males respectively, and 172 Km2 for a juvenile. Core areas of individual activity for each jaguar, were on average 11.5 Km2 and 33.55 km2 for AF and AM respectively in the Wet Chaco, while in the Dry Chaco were larger: 115 km2 for five AM and 225 Km2 for an AF and 32.4 Km2 for a JM. In both ecoregions, only one relevant overlap of HR of adults was reported. During the reproduction season, the HR (95% K) of one AM overlapped 49.83% with that of one AF. At the Wet Chaco, the maximum daily distance moved by an AF was 14.5 Km and 11.6 Km for the AM, while the Maximum Mean Daily Moved (MMDM) distance was 5.6 km for an AF and 3.1 km for an AM. At the Dry Chaco, the maximum daily distance for an AF was 61.7Km., 50.9Km for the AM and 6.6 Km for the JM, while the MMDM distance was 13.2 km for an AM and 8.4 km for an AF. This study confirmed that, as the invasion to jaguar habitat increased, it resulted in fragmented landscapes that influence spacing patterns of jaguars. Males used largest HR that of the smaller females and males covers largest distances that of the females. There appeared to be important spatial segregation between not only females but also males. It is likely that the larger areas used by males are partly caused by the sexual dimorphism in body size that entails differences in prey requirements. These could explain the larger distances travelled daily by males.Keywords: Chaco ecoregions, Jaguar, home range, Panthera onca, Paraguay
Procedia PDF Downloads 30214 Characterization of the Blood Microbiome in Rheumatoid Arthritis Patients Compared to Healthy Control Subjects Using V4 Region 16S rRNA Sequencing
Authors: D. Hammad, D. P. Tonge
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Rheumatoid arthritis (RA) is a disabling and common autoimmune disease during which the body's immune system attacks healthy tissues. This results in complicated and long-lasting actions being carried out by the immune system, which typically only occurs when the immune system encounters a foreign object. In the case of RA, the disease affects millions of people and causes joint inflammation, ultimately leading to the destruction of cartilage and bone. Interestingly, the disease mechanism still remains unclear. It is likely that RA occurs as a result of a complex interplay of genetic and environmental factors including an imbalance in the microorganism population inside our body. The human microbiome or microbiota is an extensive community of microorganisms in and on the bodies of animals, which comprises bacteria, fungi, viruses, and protozoa. Recently, the development of molecular techniques to characterize entire bacterial communities has renewed interest in the involvement of the microbiome in the development and progression of RA. We believe that an imbalance in some of the specific bacterial species in the gut, mouth and other sites may lead to atopobiosis; the translocation of these organisms into the blood, and that this may lead to changes in immune system status. The aim of this study was, therefore, to characterize the microbiome of RA serum samples in comparison to healthy control subjects using 16S rRNA gene amplification and sequencing. Serum samples were obtained from healthy control volunteers and from patients with RA both prior to, and following treatment. The bacterial community present in each sample was identified utilizing V4 region 16S rRNA amplification and sequencing. Bacterial identification, to the lowest taxonomic rank, was performed using a range of bioinformatics tools. Significantly, the proportions of the Lachnospiraceae, Ruminococcaceae, and Halmonadaceae families were significantly increased in the serum of RA patients compared with healthy control serum. Furthermore, the abundance of Bacteroides and Lachnospiraceae nk4a136_group, Lachnospiraceae_UGC-001, RuminococcaceaeUCG-014, Rumnococcus-1, and Shewanella was also raised in the serum of RA patients relative to healthy control serum. These data support the notion of a blood microbiome and reveal RA-associated changes that may have significant implications for biomarker development and may present much-needed opportunities for novel therapeutic development.Keywords: blood microbiome, gut and oral bacteria, Rheumatoid arthritis, 16S rRNA gene sequencing
Procedia PDF Downloads 13213 Microscopic Insights into Water Transport Through a Biomimetic Artificial Water Nano-Channels-Polyamide Membrane
Authors: Aziz Ghoufi, Ayman Kanaan
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Clean water is ubiquitous from drinking to agriculture and from energy supply to industrial manufacturing. Since the conventional water sources are becoming increasingly rare, the development of new technologies for water supply is crucial to address the world’s clean water needs in the 21st century. Desalination is in many regards the most promising approach to long-term water supply since it potentially delivers an unlimited source of fresh water. Seawater desalination using reverse osmosis (RO) membranes has become over the past decade a standard approach to produce fresh water. While this technology has proven to be efficient, it remains however relatively costly in terms of energy input due to the use of high-pressure pumps resulting of the low water permeation through polymeric RO membranes. Recently, water channels incorporated in lipidic and polymeric membranes were demonstrated to provide a selective water translocation that enables to break permeability- selectivity trade-off. Biomimetic Artificial Water channels (AWCs) are becoming highly attractive systems to achieve a selective transport of water. The first developed AWCs formed from imidazole quartet (I-quartet) embedded in lipidic membranes exhibited an ion selectivity higher than AQPs however associated with a lower water flow performance. Recently it has been conducted pioneer work in this field with the fabrication of the first AWC@Polyamide(PA) composite membrane with outstanding desalination performance. However, the microscopic desalination mechanism in play is still unknown and its understanding represents the shortest way for a long-term conception and design of AWC@PA composite membranes with better performance. In this work we gain an unprecedented fundamental understanding and rationalization of the nanostructuration of the AWC@PA membranes and the microscopic mechanism at the origin of their water transport performance from advanced molecular simulations. Using osmotic molecular dynamics simulations and a non-equilibrium method with water slab control, we demonstrate an increase in porosity near the AWC@PA interfaces, enhancing water transport without compromising the rejection rate. Indeed, the water transport pathways exhibit a single-file structure connected by hydrogen bonds. Finally, by comparing AWC@PA and PA membranes, we show that the difference in water flux aligns well with experimental results, validating the model used.Keywords: water desalination, biomimetic membranes, molecular simulation, nanochannels
Procedia PDF Downloads 1712 Biodegradation of Triclosan and Tetracycline in Sewage Sludge by Pleurotus Ostreatus Fungal Pellets
Authors: Ayda Maadani Mallak, Amir lakzian, Elham Khodaverdi, Gholam Hossein Haghnia
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The use of pharmaceuticals and personal care products such as antibiotics and antibacterials has been increased in recent years. Since the major part of consumed compounds remains unchanged in the wastewater treatment plant, they will easily find their way into the human food chain following the land use of sewage sludge (SS). Biological treatment of SS is one the most effective methods for expunging contaminants. White rot fungi, due to their ligninolytic enzymes, are extensively used to degrade organic compounds. Among all three different morphological forms and growth patterns of filamentous fungi (mycelia, clumps, and pellets), fungal pellet formation has been the subject of interest in industrial bioprocesses. Therefore this study was aimed to investigate the uptake of tetracycline (TC) and triclosan (TCS) by radish plant (Raphanus sativus) from soil amended with untreated and pretreated SS by P. ostreatus fungal pellets under greenhouse conditions. The experimental soil was amended with 1) Contaminated SS with TC at a concentration of 100 mgkg-1 and pretreated by fungal pellets, 2) Contaminated SS with TC at 100 mgkg-1 and untreated with fungal pellets, 3) Contaminated SS with TCS at a concentration of 50 mgkg-1 and pretreated by fungal pellets, 4) contaminated SS with TCS at 50 mgkg-1 and untreated with fungal pellets. An uncontaminated and untreated SS-amended soil also was considered as control treatment. An AB SCIEX 3200 QTRAP LC-MS/MS system was used in order to analyze the concentration of TC and TCS in plant tissues and soil medium. Results of this study revealed that the presence of TC and TCS in SS-amended soil decreased the radish biomass significantly. The reduction effect of TCS on dry biomass of shoot and root was 39 and 45% compared to controls, whereas for TC, the reduction percentage for shoot and root was 27 and 40.6%, respectively. However, fungal treatment of SS by P. ostreatus pellets reduced the negative effect of both compounds on plant biomass remarkably, as no significant difference was observed compared to control treatments. Pretreatment of SS with P. ostreatus also caused a significant reduction in translocation factor (concentration in shoot/root), especially for TC compound up to 32.3%, whereas this reduction for TCS was less (8%) compared to untreated SS. Generally, the results of this study confirmed the positive effect of using fungal pellets in SS amendment to decrease TC and TCS uptake by radish plants. In conclusion, P. ostreatus fungal pellets might provide future insights into bioaugmentation to remove antibiotics from environmental matrices.Keywords: antibiotic, fungal pellet, sewage sludge, white-rot fungi
Procedia PDF Downloads 15711 The Determination of Pb and Zn Phytoremediation Potential and Effect of Interaction between Cadmium and Zinc on Metabolism of Buckwheat (Fagopyrum Esculentum)
Authors: Nurdan Olguncelik Kaplan, Aysen Akay
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Nowadays soil pollution has become a global problem. External added polluters to the soil are destroying and changing the structure of the soil and the problems are becoming more complex and in this sense the correction of these problems is going to be harder and more costly. Cadmium has got a fast mobility in the soil and plant system because of that cadmium can interfere very easily to the human and animal food chain and in the same time this can be very dangerous. The cadmium which is absorbed and stored by the plants is causing to many metabolic changes of the plants like; protein synthesis, nitrogen and carbohydrate metabolism, enzyme (nitrate reductase) activation, photo and chlorophyll synthesis. The biological function of cadmium is not known over the plants and it is not a necessary element. The plant is generally taking in small amounts the cadmium and this element is competing with the zinc. Cadmium is causing root damages. Buckwheat (Fagopyrum esculentum) is an important nutraceutical because of its high content of flavonoids, minerals and vitamins, and their nutritionally balanced amino-acid composition. Buckwheat has relatively high biomass productivity, is adapted to many areas of the world, and can flourish in sterile fields; therefore buckwheat plants are widely used for the phytoremediation process.The aim of this study were to evaluate the phytoremediation capacity of the high-yielding plant Buckwheat (Fagopyrum esculentum) in soils contaminated with Cd and Zn. The soils were applied to differrent doses cd(0-12.5-25-50-100 mg Cd kg−1 soil in the form of 3CdSO4.8H2O ) and Zn (0-10-30 mg Zn kg−1 soil in the form of ZnSO4.7H2O) and incubated about 60 days. Later buckwheat seeds were sown and grown for three mounth under greenhouse conditions. The test plants were irrigated by using pure water after the planting process. Buckwheat seeds (Gunes and Aktas species) were taken from Bahri Dagdas International Agricultural Research. After harvest, Cd and Zn concentrations of plant biomass and grain, yield and translocation factors (TFs) for Cd and Cd were determined. Cadmium accumulation in biomass and grain significantly increased in dose-dependent manner. Long term field trials are required to further investigate the potential of buckwheat to reclaimed the soil. But this could be undertaken in conjunction with actual remediation schemes. However, the differences in element accumulation among the genotypes were affected more by the properties of genotypes than by the soil properties. Gunes genotype accumulated higher lead than Aktas genotypes.Keywords: buckwheat, cadmium, phytoremediation, zinc
Procedia PDF Downloads 41710 Grassland Development on Evacuated Sites for Wildlife Conservation in Satpura Tiger Reserve, India
Authors: Anjana Rajput, Sandeep Chouksey, Bhaskar Bhandari, Shimpi Chourasia
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Ecologically, grassland is any plant community dominated by grasses, whether they exist naturally or because of management practices. Most forest grasslands are anthropogenic and established plant communities planted for forage production, though some are established for soil and water conservation and wildlife habitat. In Satpura Tiger Reserve, Madhya Pradesh, India, most of the grasslands have been established on evacuated village sites. Total of 42 villages evacuated, and study was carried out in 23 sites to evaluate habitat improvement. Grasslands were classified into three categories, i.e., evacuated sites, established sites, and controlled sites. During the present study impact of various management interventions on grassland health was assessed. Grasslands assessment was done for its composition, status of palatable and non-palatable grasses, the status of herbs and legumes, status of weeds species, and carrying capacity of particular grassland. Presence of wild herbivore species in the grasslands with their abundance, availability of water resources was also assessed. Grassland productivity is dependent mainly on the biotic and abiotic components of the area, but management interventions may also play an important role in grassland composition and productivity. Variation in the status of palatable and non-palatable grasses, legumes, and weeds was recorded and found effected by management intervention practices. Overall in all the studied grasslands, the most dominant grasses recorded are Themeda quadrivalvis, Dichanthium annulatum, Ischaemum indicum, Oplismenus burmanii, Setaria pumilla, Cynodon dactylon, Heteropogon contortus, and Eragrostis tenella. Presence of wild herbivores, i.e., Chital, Sambar, Bison, Bluebull, Chinkara, Barking deer in the grassland area has been recorded through the installation of camera traps and estimated their abundance. Assessment of developed grasslands was done in terms of habitat suitability for Chital (Axis axis) and Sambar (Rusa unicolor). The parameters considered for suitability modeling are biotic and abiotic life requisite components existing in the area, i.e., density of grasses, density of legumes, availability of water, site elevation, site distance from human habitation. Findings of the present study would be useful for further grassland management and animal translocation programmes.Keywords: carrying capacity, dominant grasses, grassland, habitat suitability, management intervention, wild herbivore
Procedia PDF Downloads 1279 Role of Artificial Intelligence in Nano Proteomics
Authors: Mehrnaz Mostafavi
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Recent advances in single-molecule protein identification (ID) and quantification techniques are poised to revolutionize proteomics, enabling researchers to delve into single-cell proteomics and identify low-abundance proteins crucial for biomedical and clinical research. This paper introduces a different approach to single-molecule protein ID and quantification using tri-color amino acid tags and a plasmonic nanopore device. A comprehensive simulator incorporating various physical phenomena was designed to predict and model the device's behavior under diverse experimental conditions, providing insights into its feasibility and limitations. The study employs a whole-proteome single-molecule identification algorithm based on convolutional neural networks, achieving high accuracies (>90%), particularly in challenging conditions (95–97%). To address potential challenges in clinical samples, where post-translational modifications affecting labeling efficiency, the paper evaluates protein identification accuracy under partial labeling conditions. Solid-state nanopores, capable of processing tens of individual proteins per second, are explored as a platform for this method. Unlike techniques relying solely on ion-current measurements, this approach enables parallel readout using high-density nanopore arrays and multi-pixel single-photon sensors. Convolutional neural networks contribute to the method's versatility and robustness, simplifying calibration procedures and potentially allowing protein ID based on partial reads. The study also discusses the efficacy of the approach in real experimental conditions, resolving functionally similar proteins. The theoretical analysis, protein labeler program, finite difference time domain calculation of plasmonic fields, and simulation of nanopore-based optical sensing are detailed in the methods section. The study anticipates further exploration of temporal distributions of protein translocation dwell-times and the impact on convolutional neural network identification accuracy. Overall, the research presents a promising avenue for advancing single-molecule protein identification and quantification with broad applications in proteomics research. The contributions made in methodology, accuracy, robustness, and technological exploration collectively position this work at the forefront of transformative developments in the field.Keywords: nano proteomics, nanopore-based optical sensing, deep learning, artificial intelligence
Procedia PDF Downloads 958 Status and Management of Grape Stem Borer, Celosterna scrabrator with Soil Application of Chlorantraniliprole 0.4 gr
Authors: D. N. Kambrekar, S. B. Jagginavar, J. Aruna
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Grape stem borer, Celosterna scrabrator is an important production constraint in grapes in India. Hitherto this pest was a severe menace only on the aged and unmanaged fields but during the recent past it has also started damaging the newly established fields. In India, since Karnataka, Andra Pradesh, Tamil Nadu and Maharashtra are the major grape production states, the incidence of stem borer is also restricted and severe in these states. The grubs of the beetle bore in to the main stem and even the branches, which affect the translocation of nutrients to the areal parts of the plant. Since, the grubs bore inside the stem, the chewed material along with its excreta is discharged outside the holes and the frass is found on the ground just below the bored holes. The portion of vines above the damaged part has a sticky appearance. The leaves become pale yellow which looks like a deficiency of micronutrients. The leaves ultimately dry and drop down. The status of the incidence of the grape stem borer in different grape growing districts of Northern Karnataka was carried out during three years. In each taluka five locations were surveyed for the incidence of grape stem borer. Further, the experiment on management of stem borer was carried out in the grape gardens of Vijayapur districts under farmers field during three years. Stem borer infested plants that show live holes were selected per treatments and it was replicated three times. Live and dead holes observed during pre-treatment were closely monitored and only plants with live holes were selected and tagged. Different doses of chlorantraniliprole 0.4% GR were incorporated into the soil around the vine basins near root zone surrounded to trunk region by removing soils up to 5-10 cm with a peripheral distance of 1 to 1.5 feet from the main trunk where feeder roots are present. Irrigation was followed after application of insecticide for proper incorporation of the test chemical. The results indicated that there was sever to moderate incidence of the stem borer in all the grape growing districts of northern Karnataka. Maximum incidence was recorded in Belagavi (11 holes per vine) and minimum was in Gadag district (8.5 holes per vine). The investigations carried out to study the efficacy of chlorantraniliprole on grape stem borer for successive three years under farmers field indicated that chlorantraniliprole @ 15g/vine applied just near the active root zone of the plant followed by irrigation has successfully managed the pest. The insecticide has translocated to all the parts of the plants and thereby stopped the activity of the pest which has resulted in to better growth of the plant and higher berry yield compared to other treatments under investigation. Thus, chlorantraniliprole 0.4 GR @ 15g/vine can be effective means in managing the stem borer.Keywords: chlorantraniliprole, grape stem borer, Celosterna scrabrator, management
Procedia PDF Downloads 4427 Effects of Macro and Micro Nutrients on Growth and Yield Performances of Tomato (Lycopersicon esculentum MILL.)
Authors: K. M. S. Weerasinghe, A. H. K. Balasooriya, S. L. Ransingha, G. D. Krishantha, R. S. Brhakamanagae, L. C. Wijethilke
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Tomato (Lycopersicon esculentum Mill.) is a major horticultural crop with an estimated global production of over 120 million metric tons and ranks first as a processing crop. The average tomato productivity in Sri Lanka (11 metric tons/ha) is much lower than the world average (24 metric tons/ha).To meet the tomato demand for the increasing population the productivity has to be intensified through the agronomic-techniques. Nutrition is one of the main factors which govern the growth and yield of tomato and the main nutrient source soil affect the plant growth and quality of the produce. Continuous cropping, improper fertilizer usage etc., cause widespread nutrient deficiencies. Therefore synthetic fertilizers and organic manures were introduced to enhance plant growth and maximize the crop yields. In this study, effects of macro and micronutrient supplementations on improvement of growth and yield of tomato were investigated. Selected tomato variety is Maheshi and plants were grown in Regional Agricultural and Research Centre Makadura under the Department of Agriculture recommended (DOA) macro nutrients and various combination of Ontario recommended dosages of secondary and micro fertilizer supplementations. There were six treatments in this experiment and each treatment was replicated in three times and each replicate consisted of six plants. Other than the DOA recommendation, five combinations of Ontario recommended dosage of secondary and micronutrients for tomato were also used as treatments. The treatments were arranged in a Randomized Complete Block Design. All cultural practices were carried out according to the DOA recommendations. The mean data was subjected to the statistical analysis using SAS package and mean separation (Duncan’s Multiple Range test at 5% probability level) procedures. Secondary and micronutrients containing treatments significantly increased most of the growth parameters. Plant height, plant girth, number of leaves, leaf area index etc. Fruits harvested from pots amended with macro, secondary and micronutrients performed best in terms of total yield; yield quality; to pots amended with DOA recommended dosage of fertilizer for tomato. It could be due to the application of all essential macro and micro nutrients that rise in photosynthetic activity, efficient translocation and utilization of photosynthates causing rapid cell elongation and cell division in actively growing region of the plant leading to stimulation of growth and yield were caused. The experiment revealed and highlighted the requirements of essential macro, secondary and micro nutrient fertilizer supplementations for tomato farming. The study indicated that, macro and micro nutrient supplementation practices can influence growth and yield performances of tomato fruits and it is a promising approach to get potential tomato yields.Keywords: macro and micronutrients, tomato, SAS package, photosynthates
Procedia PDF Downloads 4746 Theoretical Study on the Visible-Light-Induced Radical Coupling Reactions Mediated by Charge Transfer Complex
Authors: Lishuang Ma
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Charge transfer (CT) complex, also known as Electron donor-acceptor (EDA) complex, has received attentions increasingly in the field of synthetic chemistry community, due to the CT complex can absorb the visible light through the intermolecular charge transfer excited states, various of catalyst-free photochemical transformations under mild visible-light conditions. However, a number of fundamental questions are still ambiguous, such as the origin of visible light absorption, the photochemical and photophysical properties of the CT complex, as well as the detailed mechanism of the radical coupling pathways mediated by CT complex. Since these are critical factors for target-specific design and synthesis of more new-type CT complexes. To this end, theoretical investigations were performed in our group to answer these questions based on multiconfigurational perturbation theory. The photo-induced fluoroalkylation reactions are mediated by CT complexes, which are formed by the association of an acceptor of perfluoroalkyl halides RF−X (X = Br, I) and a suitable donor molecule such as β-naphtholate anion, were chosen as a paradigm example in this work. First, spectrum simulations were carried out by both CASPT2//CASSCF/PCM and TD-DFT/PCM methods. The computational results showed that the broadening spectra in visible light range (360-550nm) of the CT complexes originate from the 1(σπ*) excitation, accompanied by an intermolecular electron transfer, which was also found closely related to the aggregate states of the donor and acceptor. Moreover, from charge translocation analysis, the CT complex that showed larger charge transfer in the round state would exhibit smaller charge transfer in excited stated of 1(σπ*), causing blue shift relatively. Then, the excited-state potential energy surface (PES) was calculated at CASPT2//CASSCF(12,10)/ PCM level of theory to explore the photophysical properties of the CT complexes. The photo-induced C-X (X=I, Br) bond cleavage was found to occur in the triplet state, which is accessible through a fast intersystem crossing (ISC) process that is controlled by the strong spin-orbit coupling resulting from the heavy iodine and bromine atoms. Importantly, this rapid fragmentation process can compete and suppress the backward electron transfer (BET) event, facilitating the subsequent effective photochemical transformations. Finally, the reaction pathways of the radical coupling were also inspected, which showed that the radical chain propagation pathway could easy to accomplish with a small energy barrier no more than 3.0 kcal/mol, which is the key factor that promote the efficiency of the photochemical reactions induced by CT complexes. In conclusion, theoretical investigations were performed to explore the photophysical and photochemical properties of the CT complexes, as well as the mechanism of radical coupling reactions mediated by CT complex. The computational results and findings in this work can provide some critical insights into mechanism-based design for more new-type EDA complexesKeywords: charge transfer complex, electron transfer, multiconfigurational perturbation theory, radical coupling
Procedia PDF Downloads 1435 Diagnosis, Treatment, and Prognosis in Cutaneous Anaplastic Lymphoma Kinase-Positive Anaplastic Large Cell Lymphoma: A Narrative Review Apropos of a Case
Authors: Laura Gleason, Sahithi Talasila, Lauren Banner, Ladan Afifi, Neda Nikbakht
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Primary cutaneous anaplastic large cell lymphoma (pcALCL) accounts for 9% of all cutaneous T-cell lymphomas. pcALCL is classically characterized as a solitary papulonodule that often enlarges, ulcerates, and can be locally destructive, but overall exhibits an indolent course with overall 5-year survival estimated to be 90%. Distinguishing pcALCL from systemic ALCL (sALCL) is essential as sALCL confers a poorer prognosis with average 5-year survival being 40-50%. Although extremely rare, there have been several cases of ALK-positive ALCL diagnosed on skin biopsy without evidence of systemic involvement, which poses several challenges in the classification, prognostication, treatment, and follow-up of these patients. Objectives: We present a case of cutaneous ALK-positive ALCL without evidence of systemic involvement, and a narrative review of the literature to further characterize that ALK-positive ALCL limited to the skin is a distinct variant with a unique presentation, history, and prognosis. A 30-year-old woman presented for evaluation of an erythematous-violaceous papule present on her right chest for two months. With the development of multifocal disease and persistent lymphadenopathy, a bone marrow biopsy and lymph node excisional biopsy were performed to assess for systemic disease. Both biopsies were unrevealing. The patient was counseled on pursuing systemic therapy consisting of Brentuximab, Cyclophosphamide, Doxorubicin, and Prednisone given the concern for sALCL. Apropos of the patient we searched for clinically evident, cutaneous ALK-positive ALCL cases, with and without systemic involvement, in the English literature. Risk factors, such as tumor location, number, size, ALK localization, ALK translocations, and recurrence, were evaluated in cases of cutaneous ALK-positive ALCL. The majority of patients with cutaneous ALK-positive ALCL did not progress to systemic disease. The majority of cases that progressed to systemic disease in adults had recurring skin lesions and cytoplasmic localization of ALK. ALK translocations did not influence disease progression. Mean time to disease progression was 16.7 months, and significant mortality (50%) was observed in those cases that progressed to systemic disease. Pediatric cases did not exhibit a trend similar to adult cases. In both the adult and pediatric cases, a subset of cutaneous-limited ALK-positive ALCL were treated with chemotherapy. All cases treated with chemotherapy did not progress to systemic disease. Apropos of an ALK-positive ALCL patient with clinical cutaneous limited disease in the histologic presence of systemic markers, we discussed the literature data, highlighting the crucial issues related to developing a clinical strategy to approach this rare subtype of ALCL. Physicians need to be aware of the overall spectrum of ALCL, including cutaneous limited disease, systemic disease, disease with NPM-ALK translocation, disease with ALK and EMA positivity, and disease with skin recurrence.Keywords: anaplastic large cell lymphoma, systemic, cutaneous, anaplastic lymphoma kinase, ALK, ALCL, sALCL, pcALCL, cALCL
Procedia PDF Downloads 834 Colocalization Analysis to Understand Yttrium Uptake in Saxifraga paniculata Using Complementary Imaging Technics
Authors: Till Fehlauer, Blanche Collin, Bernard Angeletti, Andrea Somogyi, Claire Lallemand, Perrine Chaurand, Cédric Dentant, Clement Levard, Jerome Rose
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Over the last decades, yttrium (Y) has gained importance in high-tech applications. It is an essential part of alloys and compounds used for lasers, displays, or cell phones, for example. Due to its chemical similarities with the lanthanides, Y is often considered a rare earth element (REE). Despite their increased usage, the environmental behavior of REEs remains poorly understood. Especially regarding their interactions with plants, many uncertainties exist. On the one hand, Y is known to have a negative effect on root development and germination, but on the other hand, it appears to promote plant growth at low concentrations. In order to understand these phenomena, a precise knowledge is necessary about how Y is absorbed by the plant and how it is handled once inside the organism. Contradictory studies exist, stating that due to a similar ionic radius, Y and the other REEs might be absorbed through Ca²⁺-channels, while others suspect that Y has a shared pathway with Al³⁺. In this study, laser ablation coupled ICP-MS, and synchrotron-based micro-X-ray fluorescence (µXRF, beamline Nanoscopium, SOLEIL, France) have been used in order to localize Y within the plant tissue and identify associated elements. The plant used in this study is Saxifraga paniculata, a rugged alpine plant that has shown an affinity for Y in previous studies (in prep.). Furthermore, Saxifraga paniculata performs guttation, which means that it possesses phloem sap secreting openings on the leaf surface that serve to regulate root pressure. These so-called hydathodes could provide special insights in elemental transport in plants. The plants have been grown on Y doped soil (500mg/kg DW) for four months. The results showed that Y was mainly concentrated in the roots of Saxifraga paniculata (260 ± 85mg/kg), and only a small amount was translocated to the leaves (10 ± 7.8mg/kg). µXRF analysis indicated that within the root transects, the majority of Y remained in the epidermis and hardly penetrated the stele. Laser ablation coupled ICP-MS confirmed this finding and showed a positive correlation in the roots between Y, Fe, Al, and to a lesser extent Ca. In the stem transect, Y was mainly detected in a hotspot of approximately 40µm in diameter situated in the endodermis area. Within the stem and especially in the hotspot, Y was highly colocalized with Al and Fe. Similar-sized Y hotspots have been detected in/on the leaves. All of them were strongly colocalized with Al and Fe, except for those situated within the hydathodes, which showed no colocalization with any of the measured elements. Accordingly, a relation between Y and Ca during root uptake remains possible, whereas a correlation to Fe and Al appears to be dominant in the aerial parts, suggesting common storage compartments, the formation of complexes, or a shared pathway during translocation.Keywords: laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS), Phytoaccumulation, Rare earth elements, Saxifraga paniculata, Synchrotron-based micro-X-ray fluorescence, Yttrium
Procedia PDF Downloads 1483 The Regulation of the Cancer Epigenetic Landscape Lies in the Realm of the Long Non-coding RNAs
Authors: Ricardo Alberto Chiong Zevallos, Eduardo Moraes Rego Reis
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Pancreatic adenocarcinoma (PDAC) patients have a less than 10% 5-year survival rate. PDAC has no defined diagnostic and prognostic biomarkers. Gemcitabine is the first-line drug in PDAC and several other cancers. Long non-coding RNAs (lncRNAs) contribute to the tumorigenesis and are potential biomarkers for PDAC. Although lncRNAs aren’t translated into proteins, they have important functions. LncRNAs can decoy or recruit proteins from the epigenetic machinery, act as microRNA sponges, participate in protein translocation through different cellular compartments, and even promote chemoresistance. The chromatin remodeling enzyme EZH2 is a histone methyltransferase that catalyzes the methylation of histone 3 at lysine 27, silencing local expression. EZH2 is ambivalent, it can also activate gene expression independently of its histone methyltransferase activity. EZH2 is overexpressed in several cancers and interacts with lncRNAs, being recruited to a specific locus. EZH2 can be recruited to activate an oncogene or silence a tumor suppressor. The lncRNAs misregulation in cancer can result in the differential recruitment of EZH2 and in a distinct epigenetic landscape, promoting chemoresistance. The relevance of the EZH2-lncRNAs interaction to chemoresistant PDAC was assessed by Real Time quantitative PCR (RT-qPCR) and RNA Immunoprecipitation (RIP) experiments with naïve and gemcitabine-resistant PDAC cells. The expression of several lncRNAs and EZH2 gene targets was evaluated contrasting naïve and resistant cells. Selection of candidate genes was made by bioinformatic analysis and literature curation. Indeed, the resistant cell line showed higher expression of chemoresistant-associated lncRNAs and protein coding genes. RIP detected lncRNAs interacting with EZH2 with varying intensity levels in the cell lines. During RIP, the nuclear fraction of the cells was incubated with an antibody for EZH2 and with magnetic beads. The RNA precipitated with the beads-antibody-EZH2 complex was isolated and reverse transcribed. The presence of candidate lncRNAs was detected by RT-qPCR, and the enrichment was calculated relative to INPUT (total lysate control sample collected before RIP). The enrichment levels varied across the several lncRNAs and cell lines. The EZH2-lncRNA interaction might be responsible for the regulation of chemoresistance-associated genes in multiple cancers. The relevance of the lncRNA-EZH2 interaction to PDAC was assessed by siRNA knockdown of a lncRNA, followed by the analysis of the EZH2 target expression by RT-qPCR. The chromatin immunoprecipitation (ChIP) of EZH2 and H3K27me3 followed by RT-qPCR with primers for EZH2 targets also assess the specificity of the EZH2 recruitment by the lncRNA. This is the first report of the interaction of EZH2 and lncRNAs HOTTIP and PVT1 in chemoresistant PDAC. HOTTIP and PVT1 were described as promoting chemoresistance in several cancers, but the role of EZH2 is not clarified. For the first time, the lncRNA LINC01133 was detected in a chemoresistant cancer. The interaction of EZH2 with LINC02577, LINC00920, LINC00941, and LINC01559 have never been reported in any context. The novel lncRNAs-EZH2 interactions regulate chemoresistant-associated genes in PDAC and might be relevant to other cancers. Therapies targeting EZH2 alone weren’t successful, and a combinatorial approach also targeting the lncRNAs interacting with it might be key to overcome chemoresistance in several cancers.Keywords: epigenetics, chemoresistance, long non-coding RNAs, pancreatic cancer, histone modification
Procedia PDF Downloads 962 A 3d Intestine-On-Chip Model Allows Colonization with Commensal Bacteria to Study Host-Microbiota Interaction
Authors: Michelle Maurer, Antonia Last, Mark S. Gresnigt, Bernhard Hube, Alexander S. Mosig
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The intestinal epithelium forms an essential barrier to prevent translocation of microorganisms, toxins or other potentially harmful molecules into the bloodstream. In particular, dendritic cells of the intestinal epithelium orchestrate an adapted response of immune tolerance to commensals and immune defense against invading pathogens. Systemic inflammation is typically associated with a dysregulation of this adapted immune response and is accompanied by a disruption of the epithelial and endothelial gut barrier which enables dissemination of pathogens within the human body. To understand the pathophysiological mechanisms underlying the inflammation-associated gut barrier breakdown, it is crucial to elucidate the complex interplay of the host and the intestinal microbiome. A microfluidically perfused three-dimensional intestine-on-chip model was established to emulate these processes in the presence of immune cells, commensal bacteria, and facultative pathogens. Multi-organ tissue flow (MOTiF) biochips made from polystyrene were used for microfluidic perfusion of the intestinal tissue model. The biochips are composed of two chambers separated by a microporous membrane. Each chamber is connected to inlet and outlet channels allowing independent perfusion of the individual channels and application of microfluidic shear stress. Human umbilical vein endothelial cells (HUVECs), monocyte-derived macrophages and intestinal epithelial cells (Caco-2) were assembled on the biochip membrane. Following 7 – 14 days of growth in the presence of physiological flow conditions, the epithelium was colonized with the commensal bacterium Lactobacillus rhamnosus, while the endothelium was perfused with peripheral blood mononuclear cells (PBMCs). Additionally, L. rhamnosus was co-cultivated with the opportunistic fungal pathogen Candida albicans. Within one week of perfusion, the epithelial cells formed self-organized and well-polarized villus- and crypt-like structures that resemble essential morphological characteristics of the human intestine. Dendritic cells were differentiated in the epithelial tissue that specifically responds to bacterial lipopolysaccharide (LPS) challenge. LPS is well-tolerated at the luminal epithelial side of the intestinal model without signs of tissue damage or induction of an inflammatory response, even in the presence of circulating PBMC at the endothelial lining. In contrast, LPS stimulation at the endothelial side of the intestinal model triggered the release of pro-inflammatory cytokines such as TNF, IL-1β, IL-6, and IL-8 via activation of macrophages residing in the endothelium. Perfusion of the endothelium with PBMCs led to an enhanced cytokine release. L. rhamnosus colonization of the model was tolerated in the immune competent tissue model and was demonstrated to reduce damage induced by C. albicans infection. A microfluidic intestine-on-chip model was developed to mimic a systemic infection with a dysregulated immune response under physiological conditions. The model facilitates the colonization of commensal bacteria and co-cultivation with facultative pathogenic microorganisms. Both, commensal bacteria alone and facultative pathogens controlled by commensals, are tolerated by the host and contribute to cell signaling. The human intestine-on-chip model represents a promising tool to mimic microphysiological conditions of the human intestine and paves the way for more detailed in vitro studies of host-microbiota interactions under physiologically relevant conditions.Keywords: host-microbiota interaction, immune tolerance, microfluidics, organ-on-chip
Procedia PDF Downloads 1301 Cardiac Hypertrophy in Diabetes; The Role of Factor Forkhead Box Class O-Regulation by O-GlcNAcylation
Authors: Mohammadjavad Sotoudeheian, Navid Farahmandian
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Cardiac hypertrophy arises in response to persistent increases in hemodynamic loads. In comparison, diabetic cardiomyopathy is defined by an abnormal myocardial changes without other cardiac-related risk factors. Pathological cardiac hypertrophy and myocardial remodeling are hallmarks of cardiovascular diseases and are risk factors for heart failure. The transcription factor forkhead box class O (FOXOs) can protect heart tissue by hostile oxidative stress and stimulating apoptosis and autophagy. FOXO proteins, as sensitive elements and mediators in response to environmental changes, have been revealed to prevent and inverse cardiac hypertrophy. FOXOs are inhibited by insulin and are critical mediators of insulin action. Insulin deficiency and uncontrolled diabetes lead to a catabolic state. FOXO1 acts downstream of the insulin-dependent pathways, which are dysregulated in diabetes. It regulates cardiomyocyte hypertrophy downstream of IGF1R/PI3K/Akt activation, which are critical regulators of cardiac hypertrophy. The complex network of signaling pathways comprising insulin/IGF-1 signaling, AMPK, JNK, and Sirtuins regulate the development of cardiovascular dysfunction by modulating the activity of FOXOs. Insulin receptors and IGF1R act via the PI3k/Akt and the MAPK/ERK pathways. Activation of Akt in response to insulin or IGF-1 induces phosphorylation of FOXOs. Increased protein synthesis induced by activation of the IGF-I/Akt/mTOR signaling pathway leads to hypertrophy. This pathway and the myostatin/Smad pathway are potent negative muscle development regulators. In cardiac muscle, insulin receptor substrates (IRS)-1 or IRS-2 activates the Akt signaling pathway and inactivate FOXO1. Under metabolic stress, p38 MAPK promotes degradation of IRS-1 and IRS-2 in cardiac myocytes and activates FOXO1, leading to cardiomyopathy. Sirt1 and FOXO1 interaction play an essential role in starvation-induced autophagy in cardiac metabolism. Inhibition of Angiotensin-II induced cardiomyocyte hypertrophy is associated with reduced FOXO1 acetylation and activation of Sirt1. The NF-κB, ERK, and FOXOs are de-acetylated by SIRT1. De-acetylation of FOXO1 induces the expression of genes involved in autophagy and stimulates autophagy flux. Therefore, under metabolic stress, FOXO1 can cause diabetic cardiomyopathy. The overexpression of FOXO1 leads to decreased cardiomyocyte size and suppresses cardiac hypertrophy through inhibition of the calcineurin–NFAT pathway. Diabetes mellitus is associated with elevation of O-GlcNAcylation. Some of its binding partners regulate the substrate selectivity of O-GlcNAc transferase (OGT). O-GlcNAcylation of essential contractile proteins may inhibit protein-protein interactions, reduce calcium sensitivity, and modulate contractile function. Uridine diphosphate (UDP)-GlcNAc is the obligatory substrate of OGT, which catalyzes a reversible post-translational protein modification. The increase of O-GlcNAcylation is accompanied by impaired cardiac hypertrophy in diabetic hearts. Inhibition of O-GlcNAcylation blocks activation of ERK1/2 and hypertrophic growth. O-GlcNAc modification on NFAT is required for its translocation from the cytosol to the nucleus, where NFAT stimulates the transcription of various hypertrophic genes. Inhibition of O-GlcNAcylation dampens NFAT-induced cardiac hypertrophic growth. Transcriptional activity of FOXO1 is enriched by improved O-GlcNAcylation upon high glucose stimulation or OGT overexpression. In diabetic conditions, the modification of FOXO1 by O-GlcNAc is promoted in cardiac troponin I and myosin light chain 2. Therefore targeting O-GlcNAcylation represents a potential therapeutic option to prevent hypertrophy in the diabetic heart.Keywords: diabetes, cardiac hypertrophy, O-GlcNAcylation, FOXO1, Akt, PI3K, AMPK, insulin
Procedia PDF Downloads 108