Master Sensor Research Articles

DYNAMIC PRESSURE MONITORING USING A WIRELESS SENSOR NETWORK

This study introduces the build out and implementation of a wireless sensor network that was used to monitor dynamic pressure of water flowing inside a pipe against leak. At 5 chosen points along a prototype water pipeline, pressure sensors (HK1100C) were used to collect pressure data and its voltage data equivalent. Five locally designed and constructed field sensor nodes made up of pressure sensors, Ardiuno boards containing an 8-bit ATmega328P microcontrollers, GSM SIM800L modules, and power sources were used to process and analyze the pressure data such that when the pressure data falls below 9psi, 9.5psi, 10psi, 10.5psi, and 11psi respectively at the 5 chosen points, the information is processed by the Ardiuno boards containing an 8-bit Atmega328P microcontrollers and then passed on to the GSM SIM800L modules of respective field sensor nodes which then transmit these information as radio waves to the master sensor node of the network that has an LCD JHD162 module and a buzzer.. The received radio waves were converted back into electrical signals by the transceiver contained in the GSM SIM800L module of the master sensor node and used to power the LCD and buzzer to indicate ‘low pressure’ and sound an alarm simultaneously. The location of the pressure drop is identified and the cause rectified by trained personnel. SIM cards of chosen GSM networks were inserted into the various GSM SIM800L modules contained in the sensor nodes (fields and master nodes) for communication. Also, an experiential evaluation of the relationship between leaks and pressure variations was carried out at one of the chosen 5 points along the prototype water pipeline.

Local residual random forest classifier for strain-based damage detection and localization in aerospace sandwich structures

To ensure the structural integrity of large aerospace structures during operation, structural health monitoring is a major challenge. The monitoring can be performed by distributed strain measurements using strain gauges or fiber optical sensors. In this work, an advanced local residual classifier for strain-based damage detection and localization is introduced. The key principle is that a change in the relationship between a strain sensor and its neighbors indicates the presence of damage. After defining a sensor grid with sensor locations and their orientation, the relationship can be obtained from numerical simulations of the healthy structure. Here, local regression models are estimated between each master sensor and its neighboring sensors. Then, residuals of the predicted and measured strains are evaluated using a random forest classifier. The evaluation of the residuals has the advantage that the method is independent of the load level, as well as the fact that it is independent of certain environmental influences that are uniformly distributed over the entire structure. In addition to the numerical healthy strains, synthetically generated damage data are used for training the classifier. The synthetic data are obtained by statistical modifications of the healthy strains. This procedure avoids time-consuming and expensive damage simulations. The health monitoring approach is applied to a glass fiber reinforced polymer sandwich structure, imitating an aircraft spoiler, with a hole in the face layer considered as damage. The validation is performed by numerical finite element simulations as well as physical experiments under random loading conditions. The results demonstrate the high potential of the presented approach for strain-based structural health monitoring in composite sandwich structures.

Minimum weight clustered dominating tree problem

We discuss minimum weight clustered dominating trees that find applications in the wireless sensor network design based on a clustered independent set structure. A cluster consists of a master sensor and the sensors belonging to its sensing radius. Masters collect, filter, and transmit the sensed data to a central sensor responsible for processing all sensed information. The data sent from a cluster to the central sensor follow a unique path. It alternates between a master and a bridge node, in this order. A bridge allows data communication between two neighboring clusters. The larger the distance between masters and bridges, the higher the energy consumption for data transmission. To reduce energy consumption and increase the network lifetime, we investigate a clustered tree structure of minimum total link distances. We propose hop- and flow-based models, introduce valid inequalities for them, and discuss five exponential families of cuts when embedded into the branch-and-cut framework of the CPLEX solver. Our models benefit properly of the CPLEX Benders’ decomposition. We also highlight the differences between the topology of the clustered tree of minimum cost and the one of the (non-clustered) minimum dominating tree of the corresponding instances in terms of cost and number of solution nodes.

Growing at the right time: interconnecting the TOR pathway with photoperiod and circadian regulation.

Plants can adjust their growth to specific times of the day and season. Different photoperiods result in distinct growth patterns, which correlate with specific carbon-partitioning strategies in source (leaves) and sink (roots) organs. Therefore, external cues such as light, day length, and temperature need to be integrated with intracellular processes controlling overall carbon availability and anabolism. The target of rapamycin (TOR) pathway is a signalling hub where environmental signals, circadian information, and metabolic processes converge to regulate plant growth. TOR complex mutants display altered patterns of root growth and starch levels. Moreover, depletion of TOR or reduction in cellular energy levels affect the pace of the clock by extending the period length, suggesting that this pathway could participate in circadian metabolic entrainment. However, this seems to be a mutual interaction, since the TOR pathway components are also under circadian regulation. These results strengthen the role of this signalling pathway as a master sensor of metabolic status, integrating day length and circadian cues to control anabolic processes in the cell, thus promoting plant growth and development. Expanding this knowledge from Arabidopsis thaliana to crops will improve our understanding of the molecular links connecting environmental perception and growth regulation under field conditions.

The Nitrate Transporter MtNPF6.8 Is a Master Sensor of Nitrate Signal in the Primary Root Tip of Medicago truncatula.

Nitrate is not only an essential nutrient for plants, but also a signal involved in plant development. We have previously shown in the model legume Medicago truncatula, that the nitrate signal, which restricts primary root growth, is mediated by MtNPF6.8, a nitrate transporter. Nitrate signal also induces changes in reactive oxygen species accumulation in the root tip due to changes in cell wall peroxidase (PODs) activity. Thus, it was interesting to determine the importance of the role of MtNPF6.8 in the regulation of the root growth by nitrate and identify the POD isoforms responsible for the changes in POD activity. For this purpose, we compared in M. truncatula a npf6.8 mutant and nitrate insensitive line deficient in MtNPF6.8 and the corresponding wild and sensitive genotype for their transcriptomic and proteomic responses to nitrate. Interestingly, only 13 transcripts and no protein were differently accumulated in the primary root tip of the npf6.8-3 mutant line in response to nitrate. The sensitivity of the primary root tip to nitrate appeared therefore to be strongly linked to the integrity of MtNPF6.8 which acts as a master mediator of the nitrate signal involved in the control of the root system architecture. In parallel, 7,259 and 493 genes responded, respectively, at the level of transcripts or proteins in the wild type, 196 genes being identified by both their transcript and protein. By focusing on these 196 genes, a concordance of expression was observed for most of them with 143 genes being up-regulated and 51 being down-regulated at the two gene expression levels. Their ontology analysis uncovered a high enrichment in POD genes, allowing the identification of POD candidates involved in the changes in POD activity previously observed in response to nitrate.

Elevated Bloom’s Taxonomy; Complete Learning through Sensors

India is known for Learning with appropriate methodologies. Vedas, Upanishads are the world treasure of knowledge. Ancient Guru’s used to teach students, with applicative skills, where at the end of the course students are fully equipped with knowledge and wisdom towards execution. Though lot of universities has been designing curriculum, with an objective to meet the real life applications, yet the efficiency of most of the graduates are not meeting, barring graduates of few institutes. Bloom’s Taxonomy describes about Learning concepts like Remembering, Understanding, Applying, Analyzing, Evaluating, Creating in ‘Cognitive’ domain., Physical movements, coordination of human body with muscle movement in ‘Psycho motor’ domain., as well as feel the pain or joy in ‘Affective’ domain. Blooms Taxonomy is widely accepted by most of the Universities across the globe, and is successful upto some extent only, as lot more is required in learning process. In current era, the overall efficiency of most of the graduates is not meeting employer expectations / real life applications, as 20% of overall learning is only taking place with the sensorization of approximately two of the human body sensors only out of five sensors governed by master sensor the mind. Blooms Taxonomy couldn’t stress on essential learning through sensors and there is not much emphasis given about learning in physiological point of view through human body sensors, therefore the current methodology of learning enunciates about Elevated Bloom’s Taxonomy, where complete learning takes place through sensorization of Human Body Sensors including ear, nose, eye, mouth, skin and mind through seven stages of learning. The results of sensorized learning of 7 different batches of students proved to be phenomenal. It is also hoped that, sensorized learning will give confidence to students / learners to earn while learning.

548 - Cognitive aging and cognitive reserve

IntroductionThe aging processes are accelerating in all regions of the world. The involvement of older people in production and social processes determines the need to maintain a high level of social and psychological adaptation, despite the progressive pathology of the brain caused by its aging. This increases the relevance of research related to the study of biological reserves of the brain and psychological and social mechanisms of human adaptation in late adulthood. The risk of developing cognitive disorders is not fatal. According to some observations, even in the hippocampal type of UKR, despite the content of amyloid in the brain, the functional and social activity of the elderly remains high. Prospective studies show that people with high cognitive reserve have a lower risk of developing dementia. Cognitive reserve is the brain’s resistance to damage. Cognitive reserve is the ability of the brain to cope with the consequences of damage caused by external influences, brain stroke, chronic brain ischemia, neurodegenerative diseases, and age-related changes. Cognitive reserve is the brain’s ability to functionally compensate for and minimize clinical manifestations of cognitive impairment. The mechanisms of cognitive reserve in normal and Alzheimer’s disease are different. In healthy older adults, a higher cognitive reserve correlates with larger brain sizes and effective strategies for performing cognitive tasks. In the early stages of Alzheimer’s disease and Alzheimer’s disease, the size of the brain decreases. But high brain activity helps preserve cognitive resources. Excessive brain activity in dementia is a compensatory mechanism. This is confirmed by the results of functional magnetic resonance imaging of the brain. Of course, the degree of brain atrophy is a predisposing factor for dementia, but it is not a mandatory factor for cognitive decline. So, the symptoms of dementia do not appear until you have crossed the critical border of damage to the brain substance. Progressive brain atrophy underlies the clinical manifestations of dementia in neurodegenerative diseases, but the correlation between the degree of brain damage and cognitive impairment is not linear.Research materials and methodsAn observational 10-year longitudinal study was conducted. In 2006, moderate cognitive impairment was found in 66 patients. The group of patients included 49 women and 49 men. Their average age in 2006 was 59.3±5.2 years. In 2006, the severity of cognitive decline was 26.2±1.9 points on the MMSE scale. This corresponds to indicators of moderate cognitive impairment. Research methods: clinical and psychopathological, psychometric, statistical. Questionnaire “Loss and acquisition of personal resources” (N. Vodopyanova, M. Stein), MMSE scale.Research resultIn 2006, amyloid was detected in the spinal fluid of all patients selected for the study group. If a patient developed dementia, they were given specific therapy. The dynamics of cognitive functions in patients was different. Mild dementia was formed in 53% of patients. Moderate dementia was formed in 10.6% of patients. Moderate cognitive impairment (pre-dementia) persisted in 36.4% of patients. Hereditary burden of dementia in patients with moderate dementia was detected 2 times more often. Back in 2006, we identified the leading sensory systems of patients. The master sensor system determines the modality of the main information flow. This is the most important part of the information that a person interacts with. This is the basis of interaction with reality. This is the basis of cognitive functions. Correlations of age-specific lesions of the corresponding sensory systems with the severity of cognitive decline were found in patients with the corresponding sensory modality (Spearman’s Correlation Coefficient-r, p<0.05): presbyacoussis – auditory r=0.667, presbyopia-visual r=0.705. The influence of psychosocial factors on the condition of patients was studied. In dementia, significant history of psychotrauma was found in 35.7%. Moderate stress was detected in the group of patients with moderate cognitive impairment in 33.3%. Moderate stress was detected in the group of patients with dementia in 83.3%. Stress of loss of life meaning was detected more often in patients with dementia 76.7%. It is important not only what stresses a person endures, but how they can cope with them. Dementia patients were statistically more likely to have unproductive coping strategies that did not help them cope adequately with stress.ConclusionsThe concept of cognitive reserve suggests possible causes of heterogeneity in the dynamics of cognitive decline in the initial stages of atrophic-degenerative brain diseases: biological causes and psychosocial causes. The concept of cognitive reserve helps to study and develop individual programs for the prevention of severe cognitive disorders.

AMPK: Potential Therapeutic Target for Vascular Calcification.

Vascular calcification (VC) is an urgent worldwide health issue with no available medical treatment. It is an active cell-driven process by osteogenic differentiation of vascular cells with complex mechanisms. The AMP-activated protein kinase (AMPK) serves as the master sensor of cellular energy status. Accumulating evidence reveals the vital role of AMPK in VC progression. AMPK is involved in VC in various ways, including inhibiting runt-related transcription factor 2 signaling pathways, triggering autophagy, attenuating endoplasmic reticulum stress and dynamic-related protein 1-mediated mitochondrial fission, and activating endothelial nitric oxide synthase. AMPK activators, like metformin, are associated with reduced calcification deposits in certain groups of patients, indicating that AMPK is a potential therapeutic target for VC.

Physiology of Energy Intake in the Weight-Reduced State.

Physiological adaptations to intentional weight loss can facilitate weight regain. This review summarizes emerging findings on hypothalamic and brainstem circuitry in the regulation of body weight and identifies promising areas for research to improve therapeutic interventions for sustainable weight loss. There is good evidence that body weight is actively regulated in a homeostatic fashion similar to other physiological parameters. However, the defended level of body weight is not fixed but rather depends on environmental conditions and genetic background in an allostatic fashion. In an environment with plenty of easily available energy-dense food and low levels of physical activity, prone individuals develop obesity. In a majority of individuals with obesity, body weight is strongly defended through counterregulatory mechanisms, such as hunger and hypometabolism, making weight loss challenging. Among the options for treatment or prevention of obesity, those directly changing the defended body weight would appear to be the most effective ones. There is strong evidence that the mediobasal hypothalamus is a master sensor of the metabolic state and an integrator of effector actions responsible for the defense of adequate body weight. However, other brain areas, such as the brainstem and limbic system, are also increasingly implicated in body weight defense mechanisms and may thus be additional targets for successful therapies.

Minimizing Redundant Sensing Data Transmissions in Energy-Harvesting Sensor Networks via Exploring Spatial Data Correlations

Energy harvesting rates of sensors in renewable (e.g., solar energy) wireless sensor networks are not only lower than their energy consumption rates but also temporally varying. Existing studies exploited spatial data correlations among sensors to reduce their energy consumptions, where the data correlations mean that the sensing data of nearby sensors have high similarities. They assumed that the sensing data of nearby sensors are very likely to highly correlated. They adopted a coarse-grained spatial-correlation model, in which sensors are partitioned into different clusters such that the sensors in the same cluster have high data similarities with each other. Then, only the sensor with the maximum residual energy in each cluster sends its sensing data, while the other sensors do not. We, however, notice that the data similarities among nearby sensors in real sensor networks may vary significantly, i.e., ranging from very similar to not similar at all. Since the existing algorithms require that the sensors in the same cluster have high data similarities with each other, the sensors in a network may be partitioned into many clusters and each cluster consists of only a few sensors, where two nearby sensors belong to two different clusters if the sensing data of the two sensors are not highly correlated. Therefore, in the existing studies, many sensors have to send all their data as there are many clusters. Unlike the existing studies, in this article, we first propose a fine-grained spatial correlation model, in which sensors are partitioned into only a few clusters and each cluster consists of many sensors. Then, each cluster master sensor sends all its data to the sink, while the majority of other sensors in the cluster transmit only their nonredundant data, thereby significantly saving sensor energy consumptions. We formulate a novel sensor clustering problem under the proposed model, which is to partition sensors into different clusters and choose a representative sensor for each cluster such that the amount of suppressed redundant data transmissions is maximized. We propose a randomized (0.5-ε)-approximation algorithm for the clustering problem, where E is a given constant with 0 <; ε ≤ 0.5. To further reduce sensor energy consumption, we consider temporal data correlations, where the sensing data by a sensor in a short period are likely to be highly correlated. We investigate a data utility maximization problem that allocates sensor data rates and routing so that the accumulative utility of both spatially and temporally correlated data received by the sink is maximized. We devise a near-optimal algorithm for the problem. We finally evaluate the performance of the proposed algorithms through experiments. the experimental results show that the proposed algorithms are very promising.

AMPK-dependent activation of the Cyclin Y/CDK16 complex controls autophagy

The AMP-activated protein kinase (AMPK) is a master sensor of the cellular energy status that is crucial for the adaptive response to limited energy availability. AMPK is implicated in the regulation of many cellular processes, including autophagy. However, the precise mechanisms by which AMPK controls these processes and the identities of relevant substrates are not fully understood. Using protein microarrays, we identify Cyclin Y as an AMPK substrate that is phosphorylated at Serine 326 (S326) both in vitro and in cells. Phosphorylation of Cyclin Y at S326 promotes its interaction with the Cyclin-dependent kinase 16 (CDK16), thereby stimulating its catalytic activity. When expressed in cells, Cyclin Y/CDK16 is sufficient to promote autophagy. Moreover, Cyclin Y/CDK16 is necessary for efficient AMPK-dependent activation of autophagy. This functional interaction is mediated by AMPK phosphorylating S326 of Cyclin Y. Collectively, we define Cyclin Y/CDK16 as downstream effector of AMPK for inducing autophagy.

Unique IL-13R\u03b12/STAT3 mediated IL-13 regulation detected in lung conventional dendritic cells, 24\u2009h post viral vector vaccination

This study demonstrates that 24 h following viral vector-based vaccination IL-13Rα2 functions as a master sensor on conventional dendritic cells (cDCs), abetted by high protein stability coupled with minimal mRNA expression, to rapidly regulate DC mediated IL-13 responses at the lung mucosae, unlike IL-13Rα1. Under low IL-13, IL-13Rα2 performs as a primary signalling receptor, whilst under high IL-13, acts to sequester IL-13 to maintain homeostasis, both in a STAT3-dependent manner. Likewise, we show that viral vector-derived IL-13 levels at the vaccination site can induce differential STAT3/STAT6 paradigms in lung cDC, that can get regulated collaboratively or independently by TGF-β1 and IFN-γ. Specifically, low IL-13 responses associated with recombinant Fowlpox virus (rFPV) is regulated by early IL-13Rα2, correlated with STAT3/TGF-β1 expression. Whilst, high IL-13 responses, associated with recombinant Modified Vaccinia Ankara (rMVA) is regulated in an IL-13Rα1/STAT6 dependent manner associated with IFN-γR expression bias. Different viral vaccine vectors have previously been shown to induce unique adaptive immune outcomes. Taken together current observations suggest that IL-13Rα2-driven STAT3/STAT6 equilibrium at the cDC level may play an important role in governing the efficacy of vector-based vaccines. These new insights have high potential to be exploited to improve recombinant viral vector-based vaccine design, according to the pathogen of interest and/or therapies against IL-13 associated disease conditions.

Metabolic Regulation of Hippocampal Neuroprogenitor Apoptosis After Irradiation.

The tumor suppressor p53 is an important regulator of cell fate response after DNA damage. Cell fate response following metabolic stresses has also been linked to p53-dependent pathways. In this study, we asked if 5'-adenosine monophosphate-activated protein kinase (AMPK), the master sensor of cellular energy balance, played a role in p53-dependent apoptosis of neural progenitor cells (NPCs) in the hippocampus after irradiation. Adult mice with targeted disruption of p53 or prkaa2 (gene that encodes AMPKα) in the brain were used to determine the role of p53 and AMPK, respectively, in radiation-induced apoptosis of NPCs in the hippocampus. The p53-dependent apoptosis of NPCs was associated with an increase in phospho-AMPK expression in the dentate gyrus at 8 hours after irradiation. Activation of AMPK was seen in granule neurons and subgranular NPCs. Compared with wildtype mice, apoptosis of NPCs was significantly attenuated in AMPK deficient (nestinCre: prkaa2fl/fl) mice after irradiation. AMPK deficiency did not however alter p53 activation in NPCs after irradiation. We conclude that AMPK may regulate apoptosis of hippocampal NPCs after irradiation. These findings suggest that cellular metabolism may play a role in determining cell fate response such as apoptosis after DNA damage in NPCs.

CSB affected on the sensitivity of lung cancer cells to platinum-based drugs through the global decrease of let-7 and miR-29

BackgroundTranscription-coupled nucleotide excision repair (TC-NER) plays a prominent role in the removal of DNA adducts induced by platinum-based chemotherapy reagents. Cockayne syndrome protein B (CSB), the master sensor of TCR, is also involved in the platinum resistant. Let-7 and miR-29 binding sites are highly conserved in the proximal 3′UTR of CSB.MethodsWe conducted immunohistochemisty to examine the expression of CSB in NSCLC. To determine whether let-7 family and miR-29 family directly interact with the putative target sites in the 3′UTR of CSB, we used luciferase reporter gene analysis. To detect the sensitivity of non-small cell lung cancer (NSCLC) cells to platinum-based drugs, CCK analysis and apoptosis analysis were performed.ResultsWe found that let-7 and miR-29 negatively regulate the expression of CSB by directly targeting to the 3′UTR of CSB. The endogenous CSB expression could be suppressed by let-7 and miR-29 in lung cancer cells. The suppression of CSB activity by endogenous let-7 and miR-29 can be robustly reversed by their sponges. Down-regulation of CSB induced apoptosis and increased the sensitivity of NSCLC cells to cisplatin and carboplatin drugs. Let-7 and miR-29 directly effect on cisplatin and carboplatin sensitivity in NSCLC.ConclusionsIn conclusion, the platinum-based drug resistant of lung cancer cells may involve in the regulation of let-7 and miR-29 to CSB.

Inhibition of miRNA-27b enhances neurogenesis via AMPK activation in a mouse ischemic stroke model.

Stroke is a leading cause of death and disability, but treatment options remain limited. Recent studies have suggested that cerebral ischemia‐induced neurogenesis plays a vital role in post‐stroke repair. Overactivation of AMP‐activated protein kinase (AMPK), a master sensor of energy balance, has been reported to exacerbate neuron apoptosis, but the role of chronic AMPK stimulus in post‐stroke recovery remains unclear. MicroRNAs have emerged as regulators of neurogenesis and have been reported to be involved in neurological function. In this study, we verified that miR‐27b directly targets AMPK and inhibits AMPK expression. In cultured neural stem cells, miR‐27b inhibitor improved proliferation and differentiation via the AMPK signaling pathway, but did not have an obvious effect on cell viability under oxygen and glucose deprivation conditions. In a mouse middle cerebral artery occlusion model, administration of miR‐27b inhibitor significantly enhanced behavioral function recovery and spatial memory. Up‐regulation of neurogenesis was observed both in the subventricular zone and in the hippocampal dentate gyrus. Collectively, our data suggest that miR‐27b inhibition promotes recovery after ischemic stroke by regulating AMPK activity. These findings may facilitate the development of novel therapeutic strategies for stroke.

Master–slave structure‐based capacitor voltage measuring technique for hybrid modular multilevel converters

The measurement of the capacitor voltage of each sub-module (SM) is a crucial part of achieving robust control of modular multilevel converters (MMCs). This study proposes a master-slave structure-based capacitor voltage measuring technique (MS-MT) for hybrid MMCs, in which the SM capacitor voltages are calculated using nine processes at the corresponding sampling instants according to the changing rule of the arm AC-side voltage and SM operation states. In MS-MT, the master sensor is used to measure the overall arm voltage while the slave sensor, which measures the full bridge, is used additionally to improve accuracy. Furthermore, MS-MT performs well even under pre-charging and DC short-circuit fault conditions, for the reason that a single SM short-circuit fault can be detected during above two conditions by comparing the voltages measured by the two sensors. Compared with the direct measuring technique, which has low measurement errors, and the overall measuring technique, which requires less hardware cost, MS-MT inherits both advantages. Finally, extensive simulation and experimental results validate the effectiveness of the proposed MS-MT.

Abstract 5147: Lactate transporters are a potential therapeutic target for LKB1-deficient lung cancers

Abstract The serine/threonine kinase LKB1 is mutated and inactivated in 20-30% of lung adenocarcinoma representing the second most commonly altered tumor suppressor in non-small cell lung cancer (NSCLC), and to date there are no effective targeting strategies for tumors bearing this mutation. Therefore, new therapeutic approaches are urgently needed for LKB1-deficient tumors. LKB1 activates AMPK, the master sensor of cellular energy and because of this, many of the best known functions of LKB1 are attributed to its ability to control metabolic alterations. Our laboratory and others have demonstrated that loss of LKB1 can promote enhanced glucose metabolism and a shift from aerobic to anaerobic respiration. A result of enhanced glycolysis is an elevated lactate production even under normoxic conditions. It has been reported that lactate-mediated extracellular acidification is a key factor mediating tumor cell invasion and metastasis. We found that LKB1-deficient NSCLC tumors significantly upregulate lactate transporters and, because these tumors exhibit an immunologically inert phenotype, we propose that inhibition of lactate transporters could represent a rational strategy to inhibit tumor growth and enhance immune responses in LKB1 mutant NSCLC. To characterize lactate transporter expression, a panel of NSCLC cell lines was stably transduced to overexpress LKB1 and with shRNA targeting LKB1. We analyzed gene expression of lactate transporters in TCGA dataset of lung adenocarcinoma (LUAD). We validated these results by qPCR and western blot analysis of expression levels of MCT1, MCT4 and MCT14 in human LKB1-intact and deficient cells and in LKB1 KO murine NSCLC cell lines that were generated using CRISPR/Cas9 in a KRASG12D mutant background. To study lactate transporter expression in tumors in vivo, we generated syngeneic NSCLC mouse models via s.c. injection of LKB1-intact and KO murine cells in immunocompetent mice and analyzed the protein expression levels of MCT4 in these tumor samples. The analysis of TCGA LUAD dataset revealed a significant upregulation of lactate transporter SLC16A14 (MCT14) gene expression levels in LKB1-deficient tumors compared with LKB1 wild-type (p&amp;lt;0.001). In vitro, NSCLC LKB1-deficient cells (A549, H460 and H2030) showed increased RNA and protein expression of MCT4 and MCT14 compared to cells where a copy of LKB1 was introduced. In vivo, KRASG12D mutant LKB1 KO tumors from syngeneic mouse models significantly showed an upregulation of MCT4 protein expression compared with KRASG12D mutant LKB1 wild type tumors (p&amp;lt;0.0001). In conclusion, LKB1-deficient NSCLC showed higher levels of glycolysis and significantly elevated expression of lactate transporters in vitro and in vivo compared to LKB1-wild type NSCLC. Additional studies are ongoing to determine the efficacy of lactate transporter inhibition in LKB1-mutant NSCLC. Citation Format: Irene Guijarro, Alissa Poteete, Pan Tong, Jing Wang, John Heymach. Lactate transporters are a potential therapeutic target for LKB1-deficient lung cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5147.

Inhibition of the mTOR Pathway Exerts Cardioprotective Effects Partly through Autophagy in CLP Rats.

Background Sepsis-induced myocardial dysfunction is a severe clinical problem. Recent studies have indicated that autophagy and myocardial energy depletion play a major role in myocardial dysfunction during sepsis, a mechanistic target of rapamycin (mTOR) as a master sensor of energy status and autophagy mediator; however, there are little data describing its role during sepsis in the heart. Methods Cecal ligation and puncture (CLP) or sham operation (SHAM) was performed in rats. After treatment, pathological changes were determined by H&E staining, cardiac functions by echocardiography, the distribution of microtubule-associated protein light chain 3 (LC-3) type II and hypoxia-inducible factor 1α (HIF-1a) by immunohistochemical staining, and autophagic vacuoles by transmission electron microscopy. Moreover, the mTOR signaling pathway and LC3II, p62, and HIF-1a expression were measured by western blotting. Results Rapamycin alleviated the pathological damage of myocardial tissue, attenuated cardiac dysfunction (left ventricular ejection fraction (LVEF), p < 0.05; fractional shortening (FS), p < 0.05), and reduced HIF-1a expression (p < 0.05). Expectedly, rapamycin decreased the activity of the mTOR pathway in both sham-operated rats (p < 0.0001) and CLP rats (p < 0.01). Interestingly, we also found inhibition of the mTOR pathway in CLP rats compared with sham-operated rats; phosphorylation of both mTOR (p < 0.001) and pS6K1 (p < 0.01) was significantly suppressed following CLP challenge. Furthermore, autophagic processes were elevated by CLP; the ratio of LC3II/LC3I (p < 0.05) was increased while p62 expression (p < 0.001) was decreased significantly; there were also more autophagic vacuoles in CLP rats; and rapamycin could further elevate the autophagic processes compared with CLP rats (LC3II/LC3I, p < 0.05; P62, p < 0.05). Conclusion Inhibition of the mTOR pathway has cardioprotective effects on myocardial dysfunction during sepsis induced by CLP, which is partly mediated through autophagy.

Concurrent regulation of LKB1 and CaMKK2 in the activation of AMPK in castrate-resistant prostate cancer by a well-defined polyherbal mixture with anticancer properties

BackgroundZyflamend, a blend of herbal extracts, effectively inhibits tumor growth using preclinical models of castrate-resistant prostate cancer mediated in part by 5′-adenosine monophosphate-activated protein kinase (AMPK), a master energy sensor of the cell. Clinically, treatment with Zyflamend and/or metformin (activators of AMPK) had benefits in castrate-resistant prostate cancer patients who no longer responded to treatment. Two predominant upstream kinases are known to activate AMPK: liver kinase B1 (LKB1), a tumor suppressor, and calcium-calmodulin kinase kinase-2 (CaMKK2), a tumor promotor over-expressed in many cancers. The objective was to interrogate how Zyflamend activates AMPK by determining the roles of LKB1 and CaMKK2.MethodsAMPK activation was determined in CWR22Rv1 cells treated with a variety of inhibitors of LKB1 and CaMKK2 in the presence and absence of Zyflamend, and in LKB1-null HeLa cells that constitutively express CaMKK2, following transfection with wild type LKB1 or catalytically-dead mutants. Upstream regulation by Zyflamend of LKB1 and CaMKK2 was investigated targeting protein kinase C-zeta (PKCζ) and death-associated protein kinase (DAPK), respectively.ResultsZyflamend’s activation of AMPK appears to be LKB1 dependent, while simultaneously inhibiting CaMKK2 activity. Zyflamend failed to rescue the activation of AMPK in the presence of pharmacological and molecular inhibitors of LKB1, an effect not observed in the presence of inhibitors of CaMKK2. Using LKB1-null and catalytically-dead LKB1-transfected HeLa cells that constitutively express CaMKK2, ionomycin (activator of CaMKK2) increased phosphorylation of AMPK, but Zyflamend only had an effect in cells transfected with wild type LKB1. Zyflamend appears to inhibit CaMKK2 by DAPK-mediated phosphorylation of CaMKK2 at Ser511, an effect prevented by a DAPK inhibitor. Alternatively, Zyflamend mediates LKB1 activation via increased phosphorylation of PKCζ, where it induced translocation of PKCζ and LKB1 to their respective active compartments in HeLa cells following treatment. Altering the catalytic activity of LKB1 did not alter this translocation.DiscussionZyflamend’s activation of AMPK is mediated by LKB1, possibly via PKCζ, but independent of CaMKK2 by a mechanism that appears to involve DAPK.ConclusionsTherefore, this is the first evidence that natural products simultaneously and antithetically regulate upstream kinases, known to be involved in cancer, via the activation of AMPK.

AMPK Regulation of Glucose, Lipid and Protein Metabolism: Mechanisms and Nutritional Significance

AMP-activated protein kianse (AMPK) is a master sensor of cellular energy levels and a crucial regulator of nutrient metabolism such as the synthesis of fatty acids, glucose and protein as well as their oxidation to CO2 and water . Thus, AMPK signaling has important implications for fat deposition and glucose homeostasis in animals and humans. Much experimental and clinical evidence show that AMPK is a key therapeutic target for the prevention of diseases such as obesity, diabetes, cancer, inflammation and cardiac dysfunction. In this review we highlight recent advances on the upstream and downstream targets of AMPK, as well as the specific mechanisms whereby AMPK regulates digestive functions and chronic energy balance in animals and humans.