Efficient intracellular delivery of proteins is needed to fully realize the potential of protein therapeutics. Current methods of protein delivery commonly suffer from low tolerance for serum, poor ...endosomal escape and limited in vivo efficacy. Here we report that common cationic lipid nucleic acid transfection reagents can potently deliver proteins that are fused to negatively supercharged proteins, that contain natural anionic domains or that natively bind to anionic nucleic acids. This approach mediates the potent delivery of nM concentrations of Cre recombinase, TALE- and Cas9-based transcription activators, and Cas9:sgRNA nuclease complexes into cultured human cells in media containing 10% serum. Delivery of unmodified Cas9:sgRNA complexes resulted in up to 80% genome modification with substantially higher specificity compared to DNA transfection. This approach also mediated efficient delivery of Cre recombinase and Cas9:sgRNA complexes into the mouse inner ear in vivo, achieving 90% Cre-mediated recombination and 20% Cas9-mediated genome modification in hair cells.
Immune-checkpoint inhibitors (ICIs), including anti-cytotoxic T lymphocyte antigen 4 (CTLA-4), anti-programmed cell death 1 (PD-1) and anti-programmed cell death 1 ligand 1 (PD-L1) antibodies, are ...arguably the most important development in cancer therapy over the past decade. The indications for these agents continue to expand across malignancies and disease settings, thus reshaping many of the previous standard-of-care approaches and bringing new hope to patients. One of the costs of these advances is the emergence of a new spectrum of immune-related adverse events (irAEs), which are often distinctly different from the classical chemotherapy-related toxicities. Owing to the growing use of ICIs in oncology, clinicians will increasingly be confronted with common but also rare irAEs; hence, awareness needs to be raised regarding the clinical presentation, diagnosis and management of these toxicities. In this Review, we provide an overview of the various types of irAEs that have emerged to date. We discuss the epidemiology of these events and their kinetics, risk factors, subtypes and pathophysiology, as well as new insights regarding screening and surveillance strategies. We also highlight the most important aspects of the management of irAEs.
The incidence of primary cutaneous melanoma continues to increase each year. Melanoma accounts for the majority of skin cancer–related deaths, but treatment is usually curative following early ...detection of disease. In this American Academy of Dermatology clinical practice guideline, updated treatment recommendations are provided for patients with primary cutaneous melanoma (American Joint Committee on Cancer stages 0-IIC and pathologic stage III by virtue of a positive sentinel lymph node biopsy). Biopsy techniques for a lesion that is clinically suggestive of melanoma are reviewed, as are recommendations for the histopathologic interpretation of cutaneous melanoma. The use of laboratory, molecular, and imaging tests is examined in the initial work-up of patients with newly diagnosed melanoma and for follow-up of asymptomatic patients. With regard to treatment of primary cutaneous melanoma, recommendations for surgical margins and the concepts of staged excision (including Mohs micrographic surgery) and nonsurgical treatments for melanoma in situ, lentigo maligna type (including topical imiquimod and radiation therapy), are updated. The role of sentinel lymph node biopsy as a staging technique for cutaneous melanoma is described, with recommendations for its use in clinical practice. Finally, current data regarding pregnancy and melanoma, genetic testing for familial melanoma, and management of dermatologic toxicities related to novel targeted agents and immunotherapies for patients with advanced disease are summarized.
Immune checkpoint inhibitors are reshaping the prognosis of many cancer and are progressively becoming the standard of care in the treatment of many tumour types. Immunotherapy is bringing new hope ...to patients, but also a whole new spectrum of toxicities for healthcare practitioners to manage. Oncologists and specialists involved in the pluridisciplinary management of patients with cancer are increasingly confronted with the therapeutic challenge of treating patients with severe and refractory immune-related adverse events. In this Personal View, we summarise the therapeutic strategies that have been used to manage such toxicities resulting from immune checkpoint inhibitor treatment. On the basis of current knowledge about their pathogenesis, we discuss the use of new biological and non-biological immunosuppressive drugs to treat severe and steroid refractory immune-related adverse events. Depending on the immune infiltrate type that is predominant, we propose a treatment algorithm for personalised management that goes beyond typical corticosteroid use. We propose a so-called shut-off strategy that aims at inhibiting key inflammatory components involved in the pathophysiological processes of immune-related adverse events, and limits potential adverse effects of drug immunosuppression on tumour response. This approach develops on current guidelines and challenges the step-by-step increase approach to drug immunosuppression.
Why has the United States assumed so extensive and costly a role in world affairs over the last hundred years? The two most common answers to this question are "because it could" and "because it had ...to." Neither answer will do, according to this challenging re-assessment of the way that America came to assume its global role. The country's vast economic resources gave it the capacity to exercise great influence abroad, but Americans were long reluctant to meet the costs of wielding that power. Neither the country's safety from foreign attack nor its economic well-being required the achievement of ambitious foreign policy objectives.
InA Sense of Power, John A. Thompson takes a long view of America's dramatic rise as a world power, from the late nineteenth century into the post-World War II era. How, and more importantly why, has America come to play such a dominant role in world affairs? There is, he argues, no simple answer. Thompson challenges conventional explanations of America's involvement in World War I and World War II, seeing neither the requirements of national security nor economic interests as determining. He shows how American leaders from Wilson to Truman developed an ever more capacious understanding of the national interest, and why by the 1940s most Americans came to support the price tag, in blood and treasure, attached to strenuous efforts to shape the world. The beliefs and emotions that led them to do so reflected distinctive aspects of U.S. culture, not least the strength of ties to Europe. Consciousness of the nation's unique power fostered feelings of responsibility, entitlement, and aspiration among the people and leaders of the United States.
This original analysis challenges some widely held beliefs about the determinants of United States foreign policy and will bring new insight to contemporary debates about whether the nation should-or must-play so active a part in world politics.
Why has the United States assumed so extensive and costly a role in world affairs over the last hundred years? The two most common answers to this question are "because it could" and "because it had to." Neither answer will do, according to this challenging re-assessment of the way that America came to assume its global role. The country's vast economic resources gave it the capacity to exercise great influence abroad, but Americans were long reluctant to meet the costs of wielding that power. Neither the country's safety from foreign attack nor its economic well-being required the achievement of ambitious foreign policy objectives.InA Sense of Power, John A. Thompson takes a long view of America's dramatic rise as a world power, from the late nineteenth century into the post-World War II era. How, and more importantlywhy, has America come to play such a dominant role in world affairs? There is, he argues, no simple answer. Thompson challenges conventional explanations of America's involvement in World War I and World War II, seeing neither the requirements of national security nor economic interests as determining. He shows how American leaders from Wilson to Truman developed an ever more capacious understanding of the national interest, and why by the 1940s most Americans came to support the price tag, in blood and treasure, attached to strenuous efforts to shape the world. The beliefs and emotions that led them to do so reflected distinctive aspects of U.S. culture, not least the strength of ties to Europe. Consciousness of the nation's unique power fostered feelings of responsibility, entitlement, and aspiration among the people and leaders of the United States.This original analysis challenges some widely held beliefs about the determinants of United States foreign policy and will bring new insight to contemporary debates about whether the nation should-or must-play so active a part in world politics.
Fatigue and its management in the workplace Caldwell, John A.; Caldwell, J. Lynn; Thompson, Lauren A. ...
Neuroscience & biobehavioral reviews/Neuroscience and biobehavioral reviews,
01/2019, Volume:
96, Issue:
C
Journal Article
Peer reviewed
Open access
•Fatigue, or workplace sleepiness, is a consequence of modern industrial society.•Excessive sleepiness in the workplace and on highways is a serious safety hazard.•The greatest cause of fatigue is ...insufficient or disrupted sleep.•Validated sleep-enhancement and alertness-management strategies can mitigate fatigue.
Fatigue and workplace sleepiness are consequences of modern industrial society. Fatigue is a complex biological phenomenon that occurs as a function of time awake, time-of-day, workload, health, and off-duty lifestyle. Fatigue is a function of two major biological factors – the homeostatic drive for sleep and circadian rhythm of sleepiness. The greatest cause of fatigue is insufficient or disrupted sleep. Excessive sleepiness in the workplace and on highways is a serious safety hazard, and insufficient or disrupted sleep results in numerous accidents and adverse mental and physical health outcomes. Evidence-based strategies that promote better sleep and optimize work/rest schedules can mitigate the impact of fatigue and sleep loss. Proper nap and sleep scheduling, work breaks, modeling and monitoring tools, fatigue detection technologies, and pharmacological countermeasures can be implemented at home and/or in the workplace to reduce performance and safety hazards. Education about obtaining adequate sleep, the dangers of fatigue in terms of both health and cognitive consequences, and the availability of scientifically-proven sleep-enhancement and alertness-management strategies is essential.
Directly modulating the activity of genome-editing proteins has the potential to increase their specificity by reducing activity following target locus modification. We developed Cas9 nucleases that ...are activated by the presence of a cell-permeable small molecule by inserting an evolved 4-hydroxytamoxifen-responsive intein at specific positions in Cas9. In human cells, conditionally active Cas9s modify target genomic sites with up to 25-fold higher specificity than wild-type Cas9.
We provide evidence that the obliquities of stars with close-in giant planets were initially nearly random, and that the low obliquities that are often observed are a consequence of star-planet tidal ...interactions. The evidence is based on 14 new measurements of the Rossiter-McLaughlin effect (for the systems HAT-P-6, HAT-P-7, HAT-P-16, HAT-P-24, HAT-P-32, HAT-P-34, WASP-12, WASP-16, WASP-18, WASP-19, WASP-26, WASP-31, G1 436, and Kepler-8), as well as a critical review of previous observations. The low-obliquity (well-aligned) systems are those for which the expected tidal timescale is short, and likewise the high-obliquity (misaligned and retrograde) systems are those for which the expected timescale is long. At face value, this finding indicates that the origin of hot Jupiters involves dynamical interactions like planet-planet interactions or the Kozai effect that tilt their orbits rather than inspiraling due to interaction with a protoplanetary disk. We discuss the status of this hypothesis and the observations that are needed for a more definitive conclusion.
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