Behavior of Complex Knots in Single DNA Molecules

We used optical tweezers to tie individual DNA molecules in knots. Although these knots become highly localized under tension, they remain surprisingly mobile and undergo thermal diffusion with classical random walk statistics. The diffusion constants of knots with different complexities correlate with theoretical calculations of knot sizes. We show that this correlation can be explained by a simple hydrodynamical model of "self-reptation" of the knot along a polymer

The Rate of Osmotic Downshock Determines the Survival Probability of Bacterial Mechanosensitive Channel Mutants

Mechanosensitive (MS) channels allow cells to sense and respond to environmental changes. In bacteria, these channels are believed to protect against an osmotic shock. The physiological function of these channels has been characterized primarily by a standardized assay, where aliquots of batch-cultured cells are rapidly pipetted into a hypotonic medium. Under this method, it has been inferred many types of MS channels (MscS homologs in Escherichia coli) demonstrate limited effectiveness against shock, typically rescuing less than 10% of the cells when expressed at native levels. We introduce a single-cell-based assay which allows us to control how fast the osmolarity changes, over time scales ranging from a fraction of a second to several minutes. We find that the protection provided by MS channels depends strongly on the rate of osmotic change, revealing that, under a slow enough osmotic drop, MscS homologs can lead to survival rates comparable to those found in wild-type strains. Further, after the osmotic downshift, we observe multiple death phenotypes, which are inconsistent with the prevailing paradigm of how cells lyse. Both of these findings require a reevaluation of our basic understanding of the physiology of MS channels

A trajectory approach to two-state kinetics of single particles on sculpted energy landscapes

We study the trajectories of a single colloidal particle as it hops between two energy wells A and B, which are sculpted using adjacent optical traps by controlling their respective power levels and separation. Whereas the dynamical behaviors of such systems are often treated by master-equation methods that focus on particles as actors, we analyze them here instead using a trajectory-based variational method called Maximum Caliber, which utilizes a dynamical partition function. We show that the Caliber strategy accurately predicts the full dynamics that we observe in the experiments: from the observed averages, it predicts second and third moments and covariances, with no free parameters. The covariances are the dynamical equivalents of Maxwell-like equilibrium reciprocal relations and Onsager-like dynamical relations. In short, this work describes an experimental model system for exploring full trajectory distributions in one-particle two-state systems, and it validates the Caliber approach as a useful way to understand trajectory-based dynamical distribution functions in this system.Comment: 4 pages, 4 figures, submitted to Physical Review Letter

Engineering Cooperativity in Biomotor-Protein Assemblies

A biosynthetic approach was developed to control and probe cooperativity in multiunit biomotor assemblies by linking molecular motors to artificial protein scaffolds. This approach provides precise control over spatial and elastic coupling between motors. Cooperative interactions between monomeric kinesin-1 motors attached to protein scaffolds enhance hydrolysis activity and microtubule gliding velocity. However, these interactions are not influenced by changes in the elastic properties of the scaffold, distinguishing multimotor transport from that powered by unorganized monomeric motors. These results highlight the role of supramolecular architecture in determining mechanisms of collective transport

Psycho-oncology in Korea: Past, present and future

Background: Psycho-oncology in Korea was introduced among the circle of consultation-liaison psychiatrists, in the 1990s. For almost 25 years, the field has been developing at a steady pace as the psychosocial needs of patients with cancer continue to increase. In this study, we review the history of psycho-oncology in Korea, in a chronological order, within the domains of clinical practice, research activity, training, and public policy. Main body: Before the 1990s, patients with cancer with psychiatric comorbidities were usually taken care of by consultation-liaison psychiatrists in general hospitals. In 1993, psycho-oncology was first introduced by psychiatrists. Psychologists, nurses, and social workers have also been increasingly involved in providing psychosocial care for patients with cancer. Professionals from various disciplines began to communicate, and agreed to found the Korean Psycho-Oncology Study Group (KPOSG) in 2006, the first academic society in this field. In 2009, National Cancer Center published the Recommendations for Distress Management in Patients with Cancer, which are consensus-based guidelines for Korean patients. In 2014, the KPOSG was dissolved and absorbed into a new organization, the Korean Psycho-Oncology Society (KPOS). It functions as a center of development of psycho-oncology, publishing official journals, and hosting annual conferences. There are many challenges, including, low awareness of psycho-oncology, presence of undertreated psychiatric disorders in patients with cancer, shortage of well-trained psycho-oncologists, stigma, and suicide risk. It is important to improve the cancer care system to the extent that psycho-oncology is integrated with mainstream oncology. Considering the socio-cultural characteristics of Korean cancer care, a Korean model of distress management is being prepared by the KPOS. Conclusion: This article provides an overview of the development, current issues, and future challenges of psycho-oncology in Korea. Through its long journey to overcome the many barriers and stigmas of cancer and mental illnesses, psycho-oncology is now acknowledged as an essential part of integrated supportive care in cancer. Active research and international cooperation can gradually shape the Korean model of distress management.ope

Controlling Organization and Forces in Active Matter Through Optically-Defined Boundaries

Living systems are capable of locomotion, reconfiguration, and replication. To perform these tasks, cells spatiotemporally coordinate the interactions of force-generating, "active" molecules that create and manipulate non-equilibrium structures and force fields that span up to millimeter length scales [1-3]. Experimental active matter systems of biological or synthetic molecules are capable of spontaneously organizing into structures [4,5] and generating global flows [6-9]. However, these experimental systems lack the spatiotemporal control found in cells, limiting their utility for studying non-equilibrium phenomena and bioinspired engineering. Here, we uncover non-equilibrium phenomena and principles by optically controlling structures and fluid flow in an engineered system of active biomolecules. Our engineered system consists of purified microtubules and light-activatable motor proteins that crosslink and organize microtubules into distinct structures upon illumination. We develop basic operations, defined as sets of light patterns, to create, move, and merge microtubule structures. By composing these basic operations, we are able to create microtubule networks that span several hundred microns in length and contract at speeds up to an order of magnitude faster than the speed of an individual motor. We manipulate these contractile networks to generate and sculpt persistent fluid flows. The principles of boundary-mediated control we uncover may be used to study emergent cellular structures and forces and to develop programmable active matter devices

Connecting the dots between mechanosensitive channel abundance, osmotic shock, and survival at single-cell resolution

Rapid changes in extracellular osmolarity are one of many insults microbial cells face on a daily basis. To protect against such shocks, Escherichia coli and other microbes express several types of transmembrane channels that open and close in response to changes in membrane tension. In E. coli, one of the most abundant channels is the mechanosensitive channel of large conductance (MscL). While this channel has been heavily characterized through structural methods, electrophysiology, and theoretical modeling, our understanding of its physiological role in preventing cell death by alleviating high membrane tension remains tenuous. In this work, we examine the contribution of MscL alone to cell survival after osmotic shock at single-cell resolution using quantitative fluorescence microscopy. We conducted these experiments in an E. coli strain which is lacking all mechanosensitive channel genes save for MscL, whose expression was tuned across 3 orders of magnitude through modifications of the Shine-Dalgarno sequence. While theoretical models suggest that only a few MscL channels would be needed to alleviate even large changes in osmotic pressure, we find that between 500 and 700 channels per cell are needed to convey upwards of 80% survival. This number agrees with the average MscL copy number measured in wild-type E. coli cells through proteomic studies and quantitative Western blotting. Furthermore, we observed zero survival events in cells with fewer than ∼100 channels per cell. This work opens new questions concerning the contribution of other mechanosensitive channels to survival, as well as regulation of their activity

Persistent fluid flows defined by active matter boundaries

Biological systems achieve precise control over ambient fluids through the self-organization of active protein structures including flagella, cilia, and cytoskeletal networks. In active structures individual proteins consume chemical energy to generate force and motion at molecular length scales. Self-organization of protein components enables the control and modulation of fluid flow fields on micron scales. The physical principles underlying the organization and control of active-matter driven fluid flows are poorly understood. Here, we apply an optically-controlled active-matter system composed of microtubule filaments and light-switchable kinesin motor proteins to analyze the emergence of persistent flow fields in a model active matter system. Using light, we form contractile microtubule networks of varying shape. We analyze the fluid flow fields generated by a wide range of microtubule network geometries and explain the resulting flow fields within a unified theoretical framework. We specifically demonstrate that the geometry of microtubule flux at the boundary of contracting microtubule networks predicts the steady-state fluid flow fields across polygonal network geometries through finite-element simulations. Our work provides a foundation for programming microscopic fluid-flows with controllable active matter and could enable the engineering of versatile and dynamic microfluidic devices