Configuration-driven Logging Deprecation Without Software Updates

Software that executes on client devices may generate log files that can be utilized for troubleshooting and debugging purposes. When generation of certain log data is no longer useful, logging is disabled. However, disabling logging requires the pushing of a binary update to client devices. In situations where some fraction of client devices does not receive updates, such unwanted logging continues, which is wasteful of device resources. This disclosure describes techniques to automatically disable logging on client devices without a binary update. The binary on a client device is configured to automatically stop logging after a certain date unless a configuration update is received that instructs it to do otherwise. If the device does not receive such an update, logging automatically stops on the date. If the device receives a configuration update to continue logging, logging is continued. Zombie logging is thus eliminated

Mechanistic Studies of Intracellular Delivery of Proteins by Cell-Penetrating Peptides in Cyanobacteria

Background: The plasma membrane plays an essential role in selective permeability, compartmentalization, osmotic balance, and cellular uptake. The characteristics and functions of cyanobacterial membranes have been extensively investigated in recent years. Cell-penetrating peptides (CPPs) are special nanocarriers that can overcome the plasma membrane barrier and enter cells directly, either alone or with associated cargoes. However, the cellular entry mechanisms of CPPs in cyanobacteria have not been studied. Results: In the present study, we determine CPP-mediated transduction efficiency and internalization mechanisms in cyanobacteria using a combination of biological and biophysical methods. We demonstrate that both Synechocystis sp. PCC 6803 and Synechococcus elongatus PCC 7942 strains of cyanobacteria possess red autofluorescence. Green fluorescent protein (GFP), either alone or noncovalently associated with a CPP comprised of nine arginine residues (R9/GFP complexes), entered cyanobacteria. The ATP-depleting inhibitor of classical endocytosis, N-ethylmaleimide (NEM), could block the spontaneous internalization of GFP, but not the transduction of R9/GFP complexes. Three specific inhibitors of macropinocytosis, cytochalasin D (CytD), 5-(N-ethyl-N-isopropyl)-amiloride (EIPA), and wortmannin, reduced the efficiency of R9/GFP complex transduction, indicating that entry of R9/GFP complexes involves macropinocytosis. Both the 1-(4,5-dimethylthiazol-2-yl)-3,5-diphenylformazan (MTT) and membrane leakage analyses confirmed that R9/GFP complexes were not toxic to the cyanobacteria, nor were the endocytic and macropinocytic inhibitors used in these studies. Conclusions: In summary, we have demonstrated that cyanobacteria use classical endocytosis and macropinocytosis to internalize exogenous GFP and CPP/GFP proteins, respectively. Moreover, the CPP-mediated delivery system is not toxic to cyanobacteria, and can be used to investigate biological processes at the cellular level in this species. These results suggest that both endocytic and macropinocytic pathways can be used for efficient internalization of regular protein and CPP-mediated protein delivery in cyanobacteria, respectively

Hypotoxic Fluorescent Nanoparticles Delivery by Cell-Penetrating Peptides in Multiple Organisms: From Prokaryotes to Mammalians Cells

Nanotechnology is the study of materials in the nanoscale. By its nature, nanotechnology is interdisciplinary. Nanotechnology has made a significant stride in recent two decades in various industries. Numerous nanomaterials are devised for biomedical applications which include intracellular tracking and labeling, gene detection and hybridization, tumor or tissue targeting, pharmaceutical therapies, pathogenic inhibiting, and medical instrument coating for disinfections. High photostability and quantum yield of fluorescent nanoparticles are ideal for long-term monitoring of molecular events in living organisms. Here, we discuss delivery of three fluorescent nanoparticles in A549 cells, rotifers, Gram-negative bacteria, Gram-positive bacteria, and archaea. As these nanoparticles cannot enter cells, arginine-rich cell-penetrating peptides (CPPs) were used to enhance their internalization at the cellular or organismal level. The 1-(4,5-dimethylthiazol-2-yl)-3,5-diphenylformazan (MTT) assay and sulforhodamine B (SRB) assay demonstrated that CPP complexed fluorescent nanoparticles did not produce lethal effect in all organisms tested. The discussion of these nanomaterials in this chapter intends to broaden our understanding of their biocompatibility in organisms of various hierarchical levels

Identification of a Short Cell-Penetrating Peptide from Bovine Lactoferricin for Intracellular Delivery of DNA in Human A549 Cells

Cell-penetrating peptides (CPPs) have been shown to deliver cargos, including protein, DNA, RNA, and nanomaterials, in fully active forms into live cells. Most of the CPP sequences in use today are based on non-native proteins that may be immunogenic. Here we demonstrate that the L5a CPP (RRWQW) from bovine lactoferricin (LFcin), stably and noncovalently complexed with plasmid DNA and prepared at an optimal nitrogen/phosphate ratio of 12, is able to efficiently enter into human lung cancer A549 cells. The L5a CPP delivered a plasmid containing the enhanced green fluorescent protein (EGFP) coding sequence that was subsequently expressed in cells, as revealed by real-time PCR and fluorescent microscopy at the mRNA and protein levels, respectively. Treatment with calcium chloride increased the level of gene expression, without affecting CPP-mediated transfection efficiency. Zeta-potential analysis revealed that positively electrostatic interactions of CPP/DNA complexes correlated with CPP-mediated transport. The L5a and L5a/DNA complexes were not cytotoxic. This biomimetic LFcin L5a represents one of the shortest effective CPPs and could be a promising lead peptide with less immunogenic for DNA delivery in gene therapy

Nogo-66 Receptor Prevents Raphespinal and Rubrospinal Axon Regeneration and Limits Functional Recovery from Spinal Cord Injury

AbstractAxon regeneration after injury to the adult mammalian CNS is limited in part by three inhibitory proteins in CNS myelin: Nogo-A, MAG, and OMgp. All three of these proteins bind to a Nogo-66 receptor (NgR) to inhibit axonal outgrowth in vitro. To explore the necessity of NgR for responses to myelin inhibitors and for restriction of axonal growth in the adult CNS, we generated ngr−/− mice. Mice lacking NgR are viable but display hypoactivity and motor impairment. DRG neurons lacking NgR do not bind Nogo-66, and their growth cones are not collapsed by Nogo-66. Recovery of motor function after dorsal hemisection or complete transection of the spinal cord is improved in the ngr−/− mice. While corticospinal fibers do not regenerate in mice lacking NgR, regeneration of some raphespinal and rubrospinal fibers does occur. Thus, NgR is partially responsible for limiting the regeneration of certain fiber systems in the adult CNS

Immunotherapeutic Approaches of Rheumatoid Arthritis and the Implication on Novel Interventions for Refractoriness

Rheumatoid arthritis is an autoimmune disorder involving the chronic inflammation of affected joints which lead to the distortion and eventually destruction of the articular tissues. Clinically, many therapeutic methods are being used for RA treatment. Non-steroidal anti-inflammatory drugs (NSAIDs), steroid, and disease-modifying anti-rheumatic drugs (DMARDs) are the three main categories of intervention approaches. Among which DMARDs, targeting mainly the release of pro-inflammatory cytokines, demonstrated high efficacy because of its direct drug action that alter the underlying disease mechanisms rather than simply to mediate symptoms relieve. However, the use of DMARDs also accompanying some unwanted adverse side effects, in particular, the development of refractoriness, which hampers the successful rate of treatment. In this chapter, the conventional RA drugs will be reviewed, focusing on the currently used and latest development of DMARDs. Novel methods that could improve RA pathogenesis will also be introduced. Because of the critical role of refractory RA, the progress of the disease to develop resistance to standard drug treatment will also be described. Finally, innovative RA therapeutic methods inspired by researches concerning the pathogenesis and contemporary treatments of RA will be discussed

Students of Color and COVID-19: Experiences, Coping Strategies, and Supports

The coronavirus disease of 2019, known as the COVID-19 pandemic, is a disaster event that posed significant physical, social, financial, and mental health risks to college students. Disproportionate experiences of stressors position students of color as a population particularly vulnerable to the negative impacts of COVID-19, thus, the current study assessed the impact of COVID-19 on undergraduate students of color in the United States. Students participated in semi-structured in-depth interviews about their experiences with stressors during the pandemic. Data were analyzed using conventional content analysis and revealed themes including (a) the pandemic’s impact on students; (b) basic needs as college students; (c) strategies used to cope with stressors; and (d) supports desired from institutions and faculty. Findings inform colleges and healthcare providers of the most salient concerns for students of color and the sources found helpful. As such, these findings may guide effective prevention and intervention strategies to minimize the effects of future disasters

Expression of executive control in situational context: Effects of facilitating versus restraining cues on snack food consumption

Objectives: To examine the effects of executive function (EF) on objectively measured high-calorie snack food consumption in 2 age groups and to explore the moderating influence of environmental cues. Methods: In Study 1, 43 older adults (Mage = 74.81) and in Study 2, 79 younger adults (Mage = 18.71) completed measures of EF and subsequently participated in a bogus taste-test paradigm wherein they were required to rate 3 highly appetitive (but high-calorie) snack foods on taste and texture. Grams of snack food consumed was measured covertly in the presence randomly assigned contextual cues (explicit semantic cues in Study 1; implicit visual cues in Study 2) that were facilitating or restraining in nature. Results: Findings indicated that in both age groups, stronger EF predicted lower consumption of snack foods across conditions, and the effects of EF were most pronounced in the presence of facilitating cues. Conclusions: Older and younger adults with weaker EF tend to consume more high-calorie snack food compared with their stronger EF counterparts. These tendencies appear to be especially amplified in the presence of facilitating cues

A novel auxiliary subunit critical to BK channel function in caenorhabditis elegans

The BK channel is a Ca(2+)- and voltage-gated potassium channel with many important physiological functions. To identify proteins important to its function in vivo, we screened for C. elegans mutants that suppressed a lethargic phenotype caused by expressing a gain-of-function (gf) isoform of the BK channel α-subunit SLO-1. BKIP-1, a small peptide with no significant homology to any previously characterized molecules was thus identified. BKIP-1 and SLO-1 showed similar expression and subcellular localization patterns, and appeared to interact physically through discrete domains. bkip-1 loss-of-function (lf) mutants phenocopied slo-1(lf) mutants in behavior and synaptic transmission, and suppressed the lethargy, egg-laying defect, and deficient neurotransmitter release caused by SLO-1(gf). In heterologous expression systems, BKIP-1 decreased the activation rate and shifted the conductance-voltage (G-V) relationship of SLO-1 in a Ca(2+)-dependent manner, and increased SLO-1 surface expression. Thus, BKIP-1 is a novel auxiliary subunit critical to SLO-1 function in vivo

Vasoplegic Shock treated with Methylene Blue complicated by Severe Serotonin Syndrome.

Introduction: Management of severe vasoplegic shock in overdose can be very challenging. We describe a case of severe refractory vasodilatory shock in poisoning where methylene blue (MB) was used with success. Case Report. A 70kg 15-year-old male presented 1.5 hours post ingestion of a large polypharmacy overdose of quetiapine slow release 1.5g, quetiapine immediate release 12g, desvenlafaxine slow release 5.6g, venlafaxine 1050mg, amlodipine 290mg, ramipril 100mg, fluoxetine 560mg, promethazine 500mg and an unknown amount of lithium. He developed severe vasoplegic shock that was resistant to maximal doses of noradrenaline and vasopressin. MB was administered 6.5 hour post ingestion. Within 1 hour there was an improvement in his haemodynamic status and reduction of catecholamine requirements. Twelve hours post ingestion, he developed severe serotonin syndrome that lasted 5 days as a result of interaction between MB, a reversible monoamine oxidase inhibitor, and the antidepressants taken in overdose. MB had a calculated half-life of 38 hours. Conclusion MB is a useful second or third line strategy for severe drug induced vasodilatory shock, and may be potentially life-saving. Conversely, physicians should be aware that it can interact with other drugs and cause life-threatening serotonin syndrome. Lower doses or shorter durations may be wise in patients at risk of this interaction