Critical Control Element Research Articles

“I just cannot take medication for the rest of my life”: Non adherence Towards Diabetes Treatment

Diabetes is a rapidly growing health challenge and potential epidemic across the low-and-middle-income countries. In primary health care settings, the management of diabetes is mainly by providing hypoglycemic drugs and lifestyle change. The level of adherence and compliance to the therapy is a critical element in glycaemic control. In developed countries, adherence to long-term therapies in the general population is around 50% and is much lower in developing countries. A specific trend of non compliance and non adherence towards medication was seen in patients coming to a rural primary health care facility in India. Several beliefs regarding non adherence were identified in such patients which are - Beliefs about chronic diseases in general, beliefs about long term drug intake, behavior towards diabetes medication, peers observations and switching between different treatments. There is a need to identify specific reasons for non compliance and/or non adherence so that more focused behavioral change strategies can be made for patients of chronic diseases.

Global sensitivity analysis of a generator-absorber heat exchange (GAX) system’s thermal performance with a hybrid energy source: An approach using artificial intelligence models

• Temperature, heat and flow measurements of an experimental GAX cycle. • Evaluation of artificial intelligence (AI) techniques for multivariate modeling. • Artificial neural networks represent the GAX system with the best accuracy. • AI model coupled to the PAWN method for a global sensitivity analysis of GAX cycle. • COP is affected the most by the temperature measured at the generator inlet. Generator-absorber heat exchange (GAX) systems represent a promising alternative to substitute environmentally harmful refrigeration devices based on conventional vapor compression, as long as a proper analysis of thermal performance and the complex interactions of heat transfer that occur into GAX cycle is taken in consideration. In this research, a cooling process based on a GAX system that uses ammonia-water working fluid and a hybrid source (natural gas-solar) is studied to analyze the variables that affect the system’s thermal performance. The work’s novelty is the hybridization between artificial intelligence (AI) modeling and the global sensitivity analysis (GSA) developed with the PAWN method. Experimental data was obtained from a system with a cooling capacity of 10.5 kW (3 Ton), designed to work at heat source temperatures of 200 °C. The measured variables were the temperatures at generator, heat at evaporator, and working fluid volumetric flow. Three AI techniques (artificial neural networks, genetic programming, and support vector machines) were evaluated for modeling the thermodynamic cycle. Results obtained from the PAWN method applied to the artificial neural network, since it was the best AI model, indicates that the operational parameters with a greater impact in the system’s performance are the inlet temperature at the generator (30.7 %) and the heat measured at the evaporator for NH 3 (27.4 %), for the first output COP N H 3 . For the second output COP H 2 O , the inlet temperature at the generator (32.5 %) and the and heat measured at the evaporator for H 2 O (26.7 %), have a greater impact for such output. The proposed IA-GSA methodology contributes to the development of operational decision-making related to instrumentation, operation performance, and corrective and/or preventive maintenance actions of GAX systems. The developed thermal performance model has potential for implementation in embedded systems (smart sensors) as a critical element in control and optimization strategies to improve the performance of these cycles.

Online detection of residual stress near the gate using cavity pressure for injection molding

Abstract Residual stress negatively affects the quality of optical products and is a difficult problem to solve. Regulating process parameters is a critical element in quality control. Therefore, the online monitoring of critical process variables is necessary for stabilizing product quality and reducing manufacturing cost. This study proposes a method for the online monitoring of residual stress near the gate of a mold using multiple pressure sensors to measure process variables and verifies the feasibility of online monitoring. The study found that residual stress near the gate could not be accurately measured with only a single sensor because measurements were susceptible to interference with process parameters. When a single sensor was used to measure process variables, correlation coefficients of between 0.48 and 0.59 were obtained, and when multiple sensors were used, correlation coefficients of between 0.80 and 0.92 were obtained, which indicated strong correlation. Therefore, the online monitoring of residual stress near the gate is feasible. This monitoring method can provide a new concept of online monitoring technology for the injection molding of optical products.

INITIAL EVIDENCE OF THE IMPACT OF COVID-19 ON GOVERNMENTAL FINANCIAL PERFORMANCE AND RISK MANAGEMENT

ABSTRACT Risk assessment, disclosure, and management have become one of most critical internal control elements for all types of organizations, including governments. In this paper, we performed initial investigation of how COVID-19, an acute pandemic with unprecedented severity, has impacted the operations of top US governments during the fiscal year of 2020, and analyzed the related management implications for state and local governments. Keywords Covid-19, risk assessment, financial disclosure, local governments

Lysosome activates AKT inducing cancer and metastasis

Hyperactivated lysosome causes cancer and induces metastasis or cancer relapse. Such activation occurs during excessive, intense, and protracted oxidative burst in the lysosome. The burst induces the formation of the constitutively active (permanently active) AKT locus generating cancer complexity and robustness. Such condition has the tendency to persist by stabilized intense signaling inducing upregulation of cell function and metabolic setup at the higher level. Most intense activator of the lysosome is the fungus Aspergillus fumigatus, which activates the AKT, a critical element in lysosome control, inducing cancer development, metastatic progression, or cancer relapse. Targeting the AKT active site of hydrogen network, by redox balance change or hydrogen balance change or muon-catalyzed fusion or laser-induced fusion with anti- A. fumigatus medication converts active AKT locus into inactive element, inducing disappearance of malignant phenotype.

Anesthesia Medication Handling Needs a New Vision.

Anesthesia Medication Handling Needs a New Vision.

Alkaline hydrolysis to remove potentially infectious viral RNA contaminants from DNA.

BackgroundDiagnostics and research of high-consequence animal disease agents is often limited to laboratories with a high level of biosecurity that restrict the transport of biological material. Often, sharing of DNA with external partners is needed to support diagnostics, forensics, or research. Even in the absence of virus, RNA from positive-sense single stranded RNA (+ssRNA) viruses that may contaminate otherwise purified DNA preparations continues to pose a threat due to its potential to be infectious via direct translation to yield viral proteins. While the risk of animal infection or accidental reconstitution and release of a virus from RNA is very low, the high impact of an animal disease event associated with the accidental release of some + ssRNA viruses, such as classical swine fever or foot-and-mouth disease viruses, necessitates the precaution of having procedures to ensure the complete inactivation of viruses and + ssRNA viral genomes. RNA and DNA are differentially susceptible to enzymatic degradations; however, such procedures are susceptible to unintended DNA damage and/or failure due to enzyme or cofactor instabilities. Therefore, we describe the development and verification of a robust and simple chemical and physical method to selectively degrade RNA from purified DNA preparations. The procedure employs incubation of DNA in 0.25 N sodium hydroxide at 65 °C for 1 h followed by neutralization and boiling for 10 min to hydrolyze contaminating RNA and inactivate animal disease viruses from DNA preparations. Additional critical quality control elements include use of a synthetic control RNA (SCR) and an SCR-specific real-time RT-PCR to track effectiveness of the procedure in a parallel treated control sample, and a pH check of reagents to ensure proper neutralization of alkaline conditions.ResultsThe new procedure reduced intact RNA beyond the limit of detection by realtime RT-PCR and inactivated viruses by in vitro culture infectivity assays.ConclusionsTreated DNA, while denatured, remains suitable for most common molecular biology procedures including PCR, transformation of E. coli, and molecular sequencing. The procedure ensures not only the inactivation of a variety of viruses but also the degradation through hydrolysis of potentially contaminating infectious + ssRNA viral genomes.

A Selective C−H Deprotonation Strategy to Access Functionalized Arynes by Using Hypervalent Iodine

AbstractDescribed here is an efficient method to access highly functionalized arynes from unsymmetrical aryl(mesityl)iodonium tosylate salts. The iodonium salts are prepared in a single pot from either commercially available aryl iodides or arylboronic acids. The aryne intermediates are generated by ortho‐C−H deprotonation of aryl(mesityl)iodonium salt with a commercially available amide base and trapped in a cycloaddition reaction with furan in moderate to good yields. Coupling partners for the aryne intermediates beyond furan are also described, including benzyl azide and alicyclic amine nucleophiles. The regio‐ and chemoselectivity of this reaction is discussed and evidence for the spectator aryl ligand of the iodonium salt as a critical control element in selectivity is presented.

A Selective C-H Deprotonation Strategy to Access Functionalized Arynes by Using Hypervalent Iodine.

Described here is an efficient method to access highly functionalized arynes from unsymmetrical aryl(mesityl)iodonium tosylate salts. The iodonium salts are prepared in a single pot from either commercially available aryl iodides or arylboronic acids. The aryne intermediates are generated by ortho-C-H deprotonation of aryl(mesityl)iodonium salt with a commercially available amide base and trapped in a cycloaddition reaction with furan in moderate to good yields. Coupling partners for the aryne intermediates beyond furan are also described, including benzyl azide and alicyclic amine nucleophiles. The regio- and chemoselectivity of this reaction is discussed and evidence for the spectator aryl ligand of the iodonium salt as a critical control element in selectivity is presented.

HCF1 and OCT2 Cooperate with EBNA1 To Enhance OriP-Dependent Transcription and Episome Maintenance of Latent Epstein-Barr Virus.

ABSTRACTEpstein-Barr virus (EBV) establishes latent infections as multicopy episomes with complex patterns of viral gene transcription and chromatin structure. The EBV origin of plasmid replication (OriP) has been implicated as a critical control element for viral transcription, as well as viral DNA replication and episome maintenance. Here, we examine cellular factors that bind OriP and regulate histone modification, transcription regulation, and episome maintenance. We found that OriP is enriched for histone H3 lysine 4 (H3K4) methylation in multiple cell types and latency types. Host cell factor 1 (HCF1), a component of the mixed-lineage leukemia (MLL) histone methyltransferase complex, and transcription factor OCT2 (octamer-binding transcription factor 2) bound cooperatively with EBNA1 (Epstein-Barr virus nuclear antigen 1) at OriP. Depletion of OCT2 or HCF1 deregulated latency transcription and histone modifications at OriP, as well as the OriP-regulated latency type-dependent C promoter (Cp) and Q promoter (Qp). HCF1 depletion led to a loss of histone H3K4me3 (trimethylation of histone H3 at lysine 4) and H3 acetylation at Cp in type III latency and Qp in type I latency, as well as an increase in heterochromatic H3K9me3 at these sites. HCF1 depletion resulted in the loss of EBV episomes from Burkitt's lymphoma cells with type I latency and reactivation from lymphoblastoid cells (LCLs) with type III latency. These findings indicate that HCF1 and OCT2 function at OriP to regulate viral transcription, histone modifications, and episome maintenance. As HCF1 is best known for its function in herpes simplex virus 1 (HSV-1) immediate early gene transcription, our findings suggest that EBV latency transcription shares unexpected features with HSV gene regulation.IMPORTANCE EBV latency is associated with several human cancers. Viral latent cycle gene expression is regulated by the epigenetic control of the OriP enhancer region. Here, we show that cellular factors OCT2 and HCF1 bind OriP in association with EBNA1 to maintain elevated histone H3K4me3 and transcriptional enhancer function. HCF1 is known as a transcriptional coactivator of herpes simplex virus (HSV) immediate early (IE) transcription, suggesting that OriP enhancer shares aspects of HSV IE transcription control.

Translation Initiation Factor eIF4E Is a Target for Tumor Cell Radiosensitization

A core component in the cellular response to radiation occurs at the level of translational control of gene expression. Because a critical element in translation control is the availability of the initiation factor eIF4E, which selectively enhances the cap-dependent translation of mRNAs, we investigated a regulatory role for eIF4E in cellular radiosensitivity. eIF4E silencing enhanced the radiosensitivity of tumor cell lines but not normal cells. Similarly, pharmacologic inhibition of eIF4E with ribavirin also enhanced tumor cell radiosensitivity. eIF4E attenuation did not affect cell-cycle phase distribution or radiation-induced apoptosis, but it delayed the dispersion of radiation-induced γH2AX foci and increased the frequency of radiation-induced mitotic catastrophe. Radiation did not affect 4E-BP1 phosphorylation or cap-complex formation but it increased eIF4E binding to more than 1,000 unique transcripts including many implicated in DNA replication, recombination, and repair. Taken together, our findings suggest that eIF4E represents a logical therapeutic target to increase tumor cell radiosensitivity.

Nuclear Transmutations in HFIR’s Beryllium Reflector and Their Impact on Reactor Operation and Reflector Disposal

The High Flux Isotope Reactor (HFIR) located at the Oak Ridge National Laboratory utilizes a large annular beryllium reflector that is subdivided into three concentric regions and encompasses the compact reactor core. Nuclear transmutations caused by neutron activation occur in the beryllium reflector regions, which leads to unwanted neutron-absorbing and radiation-emitting isotopes. During the past year, two topics related to the HFIR beryllium reflector were reviewed. The first topic included studying the neutron poison (3He and 6Li) buildup in the reflector regions and its effect on beginning-of-cycle reactivity. A new methodology was developed to predict the reactivity impact and estimated symmetrical critical control element positions as a function of outage time between cycles due to 3He buildup and was shown to be in better agreement with actual symmetrical critical control element position data than the current methodology. The second topic included studying the composition of the beryllium reflector regions at discharge and during postdischarge decay to assess the viability of transporting, storing, and ultimately disposing of the reflector regions currently stored in the spent-fuel pool. The postirradiation curie inventories were used to determine whether, for disposal purposes, the reflector regions are discharged as transuranic (TRU) waste or become TRU waste during the decay period and to determine the nuclear hazard category, which may affect the controls invoked for transportation and temporary storage. Two of the reflector regions were determined to be TRU waste at discharge, and the other region was determined to become TRU waste <2 yr after being discharged due to irradiation of the initial uranium impurity content (0.0044 wt% uranium). It was also concluded that all three of the reflector regions could be classified as nuclear hazard category 3 (potential for localized consequences only).

Slowly Produced MicroRNAs Control Protein Levels

Proteins are the primary agents of function in biological systems, and their levels are critical control elements, reflecting the interplay between transcription, translation, and protein degradation. Here, we consider the role of microRNAs (miRNAs) in the post-transcriptional regulation of protein synthesis. To determine their impact on protein concentration, we constructed a mechanistic model consisting of four state variables and nine kinetic parameters that account for transcript sequestration and degradation via miRNA-mRNA complex formation. Our dynamical model predicts that, even when present in low copy number, miRNAs can exert potent effects on protein concentration. Sensitivity analysis of the steady-state solution indicates that miRNA synthesis commonly acts to fine-tune protein concentrations. However, the same analysis shows that for a small subset of miRNA-mRNA pairs characterized by slowly produced miRNAs, the miRNA synthesis rate is the dominant control element. Our model equations provide a tool to evaluate the importance of particular miRNAs on their target proteins and promote the development of miRNA-based therapies that target proteins associated with cancer, inflammation, and metabolic disorders.

Simulation of the Unloading Pressure and Recovery Pressure of Internal Unloading Valve for Emulsion Pump Station

Automatic unloading valve is the critical pressure control element of emulsion pump station, the unloading pressure and the recovery pressure after unloading of the pump are two important technical parameters of the emulsion pump station. The working performance of the unloading valve can be improved by research and simulation of the two parameters, making clear the state of the unloading valve for emulsion pump station when working, calculating the electromagnetic thrust when the pilot valve and main valve core act as well as the response speed of the unloading pressure, checking and correcting the original spring force. The variation range of pump station pressure can be controlled within ±0.5MPa through the effective control on the unloading pressure and recovery pressure of the automatic unloading valve for emulsion pump station, which ensures the stability, reliability and sensitivity of the pump station when working. The improvement of the automatic unloading valve technology of internal unloading type is adaptable to the development needs of high-yield and high-efficiency modern mine for high pressure and super larger flow pump station.

Factor binding sites within the antisilencer/enhancer region of the myelin proteolipid protein gene are important for transcriptional activity in cultured oligodendroglial cells

The first intron of the myelin proteolipid protein (Plp) gene contains a positive regulatory region known as the antisilencer/enhancer (ASE) that is located between nucleotides 1083 and 1177 of the intron and is sufficient to restrict high‐level expression of the gene to oligodendrocytes in mice. To identify critical transcriptional control elements within the ASE, we have created a series of twelve non‐overlapping 8 bp linker scan mutations across the ASE region. These mutant ASE fragments were inserted in both orientations into a Plp‐lacZ fusion gene containing a shortened form of intron 1 that lacked the ASE. Relative transcriptional activities of these PLP‐lacZ constructs were assessed indirectly by measuring the β‐galactosidase activities in extracts of transiently transfected oligodendroglial (N20.1) cells. Five of the mutants with linkers inserted near the center of the ASE demonstrated reduced activity (10–50% of wild‐type levels). This effect was relatively independent of the orientation of the ASE inserted into the Plp‐lacZ constructs, consistent with the function of this region as a transcriptional enhancer. LS mutations at these same positions were previously shown to interfere with the binding of factors present in nuclear extracts of N20.1 cells, suggesting that these factors may be important in the oligodendrocyte‐specific regulation of the Plp gene. Supported by NIH grant P20 RR‐16460.

Field evaluation of yield effects on the U.S.A. heirloom sweetpotato cultivars infected by Sweet potato leaf curl virus

The incidence of Sweet potato leaf curl virus (SPLCV), a Begomovirus, on sweetpotato, Ipomoea batatas (L.) Lam. (Convolvulaceae), in South Carolina, U.S.A. has increased rapidly in recent years. This is likely due to the use of infected propagating materials and the increasing population of its vector, the sweetpotato whitefly, Bemisia tabaci (Gennadius). In this study, we demonstrated in field experiments that SPLCV infection reduced the yields of most heirloom sweetpotato lines relative to the yields of non-infected plants. Real-time polymerase chain reaction (PCR) technology specific to several common sweetpotato viruses was used to determine the virus infection status in 69 selected accessions of heirloom sweetpotato lines. Meristem tip culture technology was used to regenerate virus-free plants from these materials. To ensure the virus-free status, each mericlone was evaluated using real-time PCR and graft bio-indexing on the indicator species, Ipomoea setosa Ker Gawl. Mericlones of 27 cultivars were found to be free of the viruses. The 27 cultivars were included in a field test to determine the effect of SPLCV infection on yield. Yields of virus-free plants of the cultivars ranged from 10 to 80% greater than the yields of SPLCV-infected plants. However, the yield differences between virus-free and infected plants were diminished in the second year of the field experiment due to a rapid re-infection by SPLCV. These results demonstrate the importance of using certified, virus-tested seed roots or cuttings. The rapid re-infection of the virus-tested sweetpotato plants with SPLCV observed in these studies suggests that management of the whitefly population should be a critical element in control of this important virus.

A Critical Control Element for Interleukin-4 Memory Expression in T Helper Lymphocytes

Naive T helper (Th) lymphocytes are induced to express the il4 (interleukin-4) gene by simultaneous signaling through the T cell receptor and the interleukin (IL)-4 receptor. Upon restimulation with antigen, such preactivated Th lymphocytes can reexpress the il4 gene independent of IL-4 receptor signaling. This memory for expression of the il4 gene depends on epigenetic modification of the il4 gene locus and an increased expression of GATA-3, the key transcription factor for Th2 differentiation. Here, we have identified a phylogenetically conserved sequence, the conserved intronic regulatory element, in the first intron of the il4 gene containing a tandem GATA-3 binding site. We show that GATA-3 binds to this sequence in a position- and orientation-dependent manner, in vitro and in vivo. DNA demethylation and histone acetylation of this region occurs early and selectively in differentiating, IL-4-secreting Th2 lymphocytes. Deletion of the conserved element by replacement of the first exon and part of the first intron of the il4 gene with gfp leads to a defect in the establishment of memory for expression of IL-4, in that reexpression of IL-4 still requires costimulation by exogenous IL-4. The conserved intronic regulatory element thus links the initial epigenetic modification of the il4 gene to GATA-3 and serves as a genetic control element for memory expression of IL-4.

GABA receptors containing the alpha5 subunit mediate the trace effect in aversive and appetitive conditioning and extinction of conditioned fear.

A reduction in alpha5 subunit-containing gamma-aminobutyric acid (GABA)A receptors has been reported to enhance some forms of learning in mutant mouse models. This effect has been attributed to impaired alpha5 GABAA receptor-mediated inhibitory modulation in the hippocampus. The introduction of a point mutation (H105R) in the alpha5 subunit is associated with a specific reduction of alpha5 subunit-containing GABAA receptors in the hippocampus. The present study examined the modulation of associative learning and the extinction of conditioned response in these animals. The strength of classical conditioning can be weakened when a trace interval is interposed between the conditioned stimulus and unconditioned stimulus. Here we report that this 'trace effect' in classical conditioning was absent in the mutant mice--they were insensitive to the imposition of a 20-s trace interval. This effect of the mutation was most clearly in the female mice using an aversive conditioning paradigm, and in the male mice using an appetitive conditioning paradigm. These gender-specific phenotypes were accompanied by a resistance to extinction of conditioned fear response in the mutant mice that was apparent in both genders. Our results identify neuronal inhibition in the hippocampus mediated via alpha5 GABAA receptors as a critical control element in the regulation of the acquisition and expression of associative memory.

Identification of a critical control element directing expression of the muscle-specific transcription factor MRF4 in the mouse embryo

Skeletal muscle development in the vertebrate embryo critically depends on the myogenic regulatory factors (MRFs) including MRF4 and Myf5. Both genes exhibit distinct expression patterns during mouse embryogenesis, although they are genetically closely linked with multiple regulatory elements dispersed throughout the common gene locus. MRF4 has a biphasic expression profile, first in somites and later in foetal skeletal muscles. Here, we demonstrate by transgenic analysis that elements within a 7.5-kb promoter fragment of the MRF4 gene are sufficient to drive the embryonic wave of expression very similar to the endogenous gene in somites of mouse embryos. In contrast, a 3-kb fragment of the proximal promoter fails to support expression in the myotome, suggesting that essential cis-acting elements are located between −7.5 and −3 kb upstream of MRF4. Further analysis of this sequence delimits an essential region between −6.6 and −5.6 kb that together with the 3-kb promoter fragment directs transgene expression in the epaxial myotome of all somites during the appropriate developmental period. These data provide evidence that the partly overlapping expression patterns of Mrf4 and Myf5 in somites are controlled by distinct regulatory elements. We also show that 11.4 kb sequence upstream of MRF4, including the promoter and the somitic control region identified in this study, is not sufficient to elicit target specificity towards the strong Myf5 (−58/−48 kb) enhancer, suggesting that additional yet unidentified elements are necessary to convey promoter selectivity and protect the MRF4 gene from this enhancer.

Altered Balance of Half-reactions in p-Hydroxybenzoate Hydroxylase Caused by Substituting the 2′-Carbon of FAD with Fluorine

Apo-p-hydroxybenzoate hydroxylase was reconstituted using 2'-fluoro-2'-deoxy-arabino-FAD, a synthetic flavin in which the hydroxyl of the 2'-center of the ribityl chain was replaced with fluorine in an inverted configuration. The absorbance spectral changes caused by the binding of either p-hydroxybenzoate (pOHB) or 2,4-dihydroxybenzoate (2,4-diOHB) indicated that the isoalloxazine of the artificial flavin adopts the more solvent-exposed "out" conformation rather than the partially buried "in" conformation near the aromatic substrate. In contrast, the flavin of the natural enzyme adopts the in conformation when pOHB is bound. Much of the behavior of the artificial enzyme can be rationalized in light of the preference of the flavin for the out conformation, including the weaker binding of pOHB, the tighter binding of 2,4-diOHB, and the slower reactions involved in the hydroxylation of pOHB and 2,4-diOHB. Particularly noteworthy is the enhancement of the reduction of the flavin by NADPH when pOHB is bound to the active site, consistent with the recent finding that the reaction occurs when the flavin adopts the out conformation (Palfey, B. A., Moran, G. R., Entsch, B., Ballou, D. P., and Massey, V. (1999) Biochemistry 38, 1153-1158). Thus, whereas the change that induces the out conformation is detrimental to the oxidative half-reaction, it improves the reductive half-reaction, showing that the control of the flavin position in p-hydroxybenzoate hydroxylase represents a compromise between the conflicting needs of two chemically disparate half-reactions, and demonstrating that the 2'-hydroxyl of FAD can serve as a critical control element in flavoenzyme catalysis.