tRF-29-79 regulates lung adenocarcinoma progression through mediating glutamine transporter SLC1A5.

In recent decades, considerable evidence has emerged indicating the involvement of tRNA-derived fragments (tRFs) in cancer progression through various mechanisms. However, the biological effects and mechanisms of tRFs in lung adenocarcinoma (LUAD) remain unclear. In this study, we screen out tRF-29-79, a 5'-tRF derived from tRNAGlyGCC, through profiling the tRF expressions in three pairs of LUAD tissues. We show that tRF-29-79 is down-regulated in LUAD and down-regulation of tRF-29-79 is associated with poorer prognosis. In vivo and in vitro assay reveal that tRF-29-79 inhibits proliferation, migration, and invasion of LUAD cells. Mechanistically, we discovered that tRF-29-79 interacts with the RNA-binding protein PTBP1 and facilitates the transportation of PTBP1 from nucleus to cytoplasm, which regulates alternative splicing in the 3' untranslated region (UTR) of SLC1A5 pre-mRNA. Given that SLC1A5 is a core transporter of glutamine, we proved that tRF-29-79 mediate glutamine metabolism of LUAD through affecting the stability of SLC1A5 mRNA, thus exerts its anticancer function. In summary, our findings uncover the novel mechanism that tRF-29-79 participates in glutamine metabolism through interacting with PTBP1 and regulating alternative splicing in the 3' UTR of SLC1A5 pre-mRNA.

Rapid Room-Temperature Aerosol Dehydration Versus Spray Drying: A Novel Paradigm in Biopharmaceutical Drying Technologies.

To ensure the high quality of biopharmaceutical products, it is imperative to implement specialized unit operations that effectively safeguard the structural integrity of large molecules. While lyophilization has long been a reliable process, spray drying has recently garnered attention for its particle engineering capabilities for the pulmonary route of administration. However, maintaining the integrity of biologics during spray drying remains a challenge. To address this issue, we explored a novel dehydration system based on aerosol-assisted room-temperature drying of biological formulations recently developed at Princeton University, called Rapid Room-Temperature Aerosol Dehydration. We compared the quality attributes of the bulk powder of biopharmaceutical products manufactured using this drying technology with that of traditional spray drying. For all the fragment antigen-binding formulations tested, in terms of protein degradation and aerosol performance, we were able to achieve a better product quality using this drying technology compared to the spray drying technique. We also highlight areas for improvement in future prototypes and prospective commercial versions of the system. Overall, the offered dehydration system holds potential for improving the quality and diversity of biopharmaceutical products and may pave the way for more efficient and effective production methods in the biopharma industry.

Flows of a nonequilibrated aqueous two-phase system in a microchannel.

Liquid-liquid phase separation is a rich and dynamic process, which recently has gained new interest, especially in biology and for material synthesis. In this work, we experimentally show that co-flow of a nonequilibrated aqueous two-phase system within a planar flow-focusing microfluidic device results in a three-dimensional flow, as the two nonequilibrated solutions move downstream along the length of the microchannel. After the system reaches steady-state, invasion fronts from the outer stream are formed along the top and bottom walls of the microfluidic device. The invasion fronts advance towards the center of the channel, until they merge. We first show by tuning the concentration of polymer species within the system that the formation of these fronts is due to liquid-liquid phase separation. Moreover, the rate of invasion from the outer stream increases with increasing polymer concentrations in the streams. We hypothesize the invasion front formation and growth is driven by Marangoni flow induced by the polymer concentration gradient along the width of the channel, as the system is undergoing phase separation. In addition, we show how at various downstream positions the system reaches its steady-state configuration once the two fluid streams flow side-by-side in the channel.

Controlling extrudate volume fraction through poroelastic extrusion of entangled looped fibers.

When a suspension of spherical or near-spherical particles passes through a constriction the particle volume fraction either remains the same or decreases. In contrast to these particulate suspensions, here we observe that an entangled fiber suspension increases its volume fraction up to 14-fold after passing through a constriction. We attribute this response to the entanglements among the fibers that allows the network to move faster than the liquid. By changing the fiber geometry, we find that the entanglements originate from interlocking shapes or high fiber flexibility. A quantitative poroelastic model is used to explain the increase in velocity and extrudate volume fraction. These results provide a new strategy to use fiber volume fraction, flexibility, and shape to tune soft material properties, e.g., suspension concentration and porosity, during delivery, as occurs in healthcare, three-dimensional printing, and material repair.

Conformal single cell hydrogel coating with electrically induced tip streaming of an AC cone.

Encapsulation of single cells in a thin hydrogel provides a more precise control of stem cell niches and better molecular transport. Despite the recent advances in microfluidic technologies to allow encapsulation of single cells, existing methods rely on special crosslinking agents that are pre-coated on the cell surface and subject to the variation of the cell membrane, which limits their widespread adoption. This work reports a high-throughput single-cell encapsulation method based on the "tip streaming" mode of alternating current (AC) electrospray, with encapsulation efficiencies over 80% after tuned centrifugation. Dripping with multiple cells is curtailed due to gating by the sharp conic meniscus of the tip streaming mode that only allows one cell to be ejected at a time. Moreover, the method can be universally applied to both natural and synthetic hydrogels, as well as various cell types, including human multipotent mesenchymal stromal cells (hMSCs). Encapsulated hMSCs maintain good cell viability over an extended culture period and exhibit robust differentiation potential into osteoblasts and adipocytes. Collectively, electrically induced tip streaming enables high-throughput encapsulation of single cells with high efficiency and universality, which is applicable for various applications in cell therapy, pharmacokinetic studies, and regenerative medicine.

Chemically Triggered Coalescence and Reactivity of Droplet Fibers.

We describe the role of functional polymer surfactants in the construction and triggered collapse of droplet-based fibers and the use of these macroscopic supracolloidal structures for reagent compartmentalization. Copolymer surfactants containing both zwitterionic and tertiary amine pendent groups were synthesized for stabilization of oil-in-water droplets, in which the self-adherent properties of the selected zwitterions impart interdroplet adherence, while the amine groups provide access to pH-triggered coalescence. Macroscopic fibers, obtained by droplet extrusion, were prepared with reagents embedded in spatially distinct components of the fibers. Upon acidification of the continuous aqueous phase, protonation of the polymer surfactants increases their hydrophilicity and causes rapid fiber disruption and collapse. Cross-linked versions of these supracolloidal fibers were stable upon acidification and appeared to direct interdroplet passage of encapsulants along the fiber length. Overall, these functional, responsive emulsions provide a strategy to impart on-demand chemical reactivity to soft materials structures that benefits from the interfacial chemistry of the system.

Slowing down DNA translocation through solid-state nanopores by edge-field leakage.

Solid-state nanopores allow high-throughput single-molecule detection but identifying and even registering all translocating small molecules remain key challenges due to their high translocation speeds. We show here the same electric field that drives the molecules into the pore can be redirected to selectively pin and delay their transport. A thin high-permittivity dielectric coating on bullet-shaped polymer nanopores permits electric field leakage at the pore tip to produce a voltage-dependent surface field on the entry side that can reversibly edge-pin molecules. This mechanism renders molecular entry an activated process with sensitive exponential dependence on the bias voltage and molecular rigidity. This sensitivity allows us to selectively prolong the translocation time of short single-stranded DNA molecules by up to 5 orders of magnitude, to as long as minutes, allowing discrimination against their double-stranded duplexes with 97% confidence.

Deep Learning for Hemorrhagic Lesion Detection and Segmentation on Brain CT Images.

Stroke is an acute cerebral vascular disease that is likely to cause long-term disabilities and death. Immediate emergency care with accurate diagnosis of computed tomographic (CT) images is crucial for dealing with a hemorrhagic stroke. However, due to the high variability of a stroke's location, contrast, and shape, it is challenging and time-consuming even for experienced radiologists to locate them. In this paper, we propose a U-net based deep learning framework to automatically detect and segment hemorrhage strokes in CT brain images. The input of the network is built by concatenating the flipped image with the original CT slice which introduces symmetry constraints of the brain images into the proposed model. This enhances the contrast between hemorrhagic area and normal brain tissue. Various Deep Learning topologies are compared by varying the layers, batch normalization, dilation rates, and pre-train models. This could increase the respective filed and preserves more information on lesion characteristics. Besides, the adversarial training is also adopted in the proposed network to improve the accuracy of the segmentation. The proposed model is trained and evaluated on two different datasets, which achieve the competitive performance with human experts with the highest location accuracy 0.9859 for detection, 0.8033 Dice score, and 0.6919 IoU for segmentation. The results demonstrate the effectiveness, robustness, and advantages of the proposed deep learning model in automatically hemorrhage lesion diagnosis, which make it possible to be a clinical decision support tool in stroke diagnosis.

Regime Map and Triple Point in Selective Withdrawal.

Entrainment in selective withdrawal occurs when both the top and bottom phases are withdrawn through a capillary tube oriented perpendicular to a flat gravitationally separated liquid-liquid interface. The tube introduces two distinct features to the conditions for fluid entrainment. First, the ratio of the two phases being withdrawn is affected by the region of influence of the flow upstream of the tube's orifice. Second, a minimum withdrawal flow rate must be reached for entrainment regardless of the distance between the interface and the tube. We show that these phenomena can be understood based on the Reynolds number that governs the external flow field around the capillary tube and the capillary number that regulates the effect of the viscosity and capillarity.

Is laparoscopic lavage safe in purulent diverticulitis versus colonic resection? A systematic review and meta-analysis.

BACKGROUND: Diverticulitis is one of the most common gastrointestinal diseases in western population. Colonic resection is recommended by international guidelines as a routinely used technique for purulent diverticulitis. Laparoscopic lavage was introduced as a non-resection alternative. The studies available so far have shown contradictory results. This meta-analysis aims to compare laparoscopic lavage versus colonic resection in patients with Hinchey Ⅲ-Ⅳ diverticulitis. METHODS: We did a systematic review of articles published before March 20, 2019, with no language restriction by searching PubMed, Cochrane library, EMBASE databases, clinicaltrials.gov, and Google Scholar databases. We included all RCTs and cohort studies comparing outcomes between patients with Hinchey Ⅲ-Ⅳ diverticulitis undergoing laparoscopic lavage versus colonic resection. Important outcomes were mortality, complications, length of stay, readmission and reoperation rates. We combined data to assess the outcomes using DerSimonian and Laird random-effects model. RESULTS: A total of 569 patients with diverticulitis of which more than 80% were Hinchey Ⅲ were enrolled from 3 RCTs and 5 cohort studies. Laparoscopic lavage was associated with shorter operative time (WMD -78.9, 95%CI -100.58 to -57.11, P < 0.0001) and total postoperative hospital stay (WMD -7.62, 95%CI -11.60 to -3.63, P = 0.0002) but a higher rate of intra-abdominal abscess (OR 2.69, 95%CI 1.39 to 5.21, P = 0.0032) and secondary peritonitis (OR 5.30, 95%CI 1.91 to 14.73, P = 0.0014). CONCLUSION: Laparoscopic lavage for patients with Hinchey Ⅲ to Ⅳ diverticulitis does provide similar mortality, shorter operative time and hospital stay. However, the evidence so far suggests that it might be inadequate for sepsis control and may result in more unplanned reoperations. Further studies are needed to standardize the formal indication for laparoscopic lavage.

Immersed AC electrospray (iACE) for monodispersed aqueous droplet generation.

We report a new immersed alternating current (AC) electrospray droplet generation method that can generate monodispersed water-in-oil droplets, with diameters ranging from 5 µm to 150 µm, in a stationary oil phase. This method offers high through-put, easy size tuning, and droplets with a viscous aqueous phase at high ionic strengths (raw physiological samples). Yet, it does not require coordinated flows of the dispersed/continuous phases or even a microfluidic chip. The design relies on a small constant back pressure (less than 0.1 atm) to drive the water phase through a nozzle (glass micropipette) and a non-isotropic AC electric Maxwell pressure to eject it into the oil phase. Undesirable field-induced discharge and nanojet formation at the tip are suppressed with a biocompatible polymer, polyethylene oxide. Its viscoelastic property favors the monodispersed dripping mechanism, with a distinct neck forming at the capillary tip before pinch-off, such that the tip dimension is the only controlling length scale. Consecutive droplets are connected by a whipping filament that disperses the drops away from the high-field nozzle to prevent electro-coalescence. A scaling theory is developed to correlate the droplet size with the applied pressure, the most important tuning parameter, and to determine the optimum frequency. The potential applications of this technology to biological systems are demonstrated with a digital loop-mediated isothermal amplification experiment, with little damage to the nucleic acids and other biomolecules, but with easy adaptive tuning for the optimum droplet number for accurate quantification.

Concentration-Gradient Stabilization with Segregated Counter- and Co-Ion Paths: A Quasistationary Depletion Front for Robust Molecular Isolation or Concentration.

We study the spatiotemporal dynamics of a microfluidic system with a nonselective microfluidic channel gated by an ion-selective membrane which separates the ion flux paths of cations and anions. To preserve electroneutrality, the ionic concentration in the system is shown to converge to a specific inhomogeneous distribution with robust constant current fluxes. A circuit scaling theory that collapses measured asymptotic currents verifies that this is a generic and robust mechanism insensitive to channel geometry, ion selectivity, and electrolyte ionic strength. This first temporally stationary but spatially inhomogeneous depletion front can be used for modulating ionic current and for isotachophoretic isolation of low-mobility molecules and exosomes on small diagnostic chips for various medical applications that require robust high-throughput and integrated platforms.

Universal Scaling of Robust Thermal Hot Spot and Ionic Current Enhancement by Focused Ohmic Heating in a Conic Nanopore.

A stable nanoscale thermal hot spot, with temperature approaching 100 °C, is shown to be sustained by localized Ohmic heating of a focused electric field at the tip of a slender conic nanopore. The self-similar (length-independent) conic geometry allows us to match the singular heat source at the tip to the singular radial heat loss from the slender cone to obtain a self-similar steady temperature profile along the cone and the resulting ionic current conductance enhancement due to viscosity reduction. The universal scaling, which depends only on a single dimensionless parameter Z, collapses the measured conductance data and computed temperature profiles in ion-track conic nanopores and conic nanopipettes. The collapsed numerical data reveal universal values for the hot-spot location and temperature in an aqueous electrolyte.

Future microfluidic and nanofluidic modular platforms for nucleic acid liquid biopsy in precision medicine.

Nucleic acid biomarkers have enormous potential in non-invasive diagnostics and disease management. In medical research and in the near future in the clinics, there is a great demand for accurate miRNA, mRNA, and ctDNA identification and profiling. They may lead to screening of early stage cancer that is not detectable by tissue biopsy or imaging. Moreover, because their cost is low and they are non-invasive, they can become a regular screening test during annual checkups or allow a dynamic treatment program that adjusts its drug and dosage frequently. We briefly review a few existing viral and endogenous RNA assays that have been approved by the Federal Drug Administration. These tests are based on the main nucleic acid detection technologies, namely, quantitative reverse transcription polymerase chain reaction (PCR), microarrays, and next-generation sequencing. Several of the challenges that these three technologies still face regarding the quantitative measurement of a panel of nucleic acids are outlined. Finally, we review a cluster of microfluidic technologies from our group with potential for point-of-care nucleic acid quantification without nucleic acid amplification, designed to overcome specific limitations of current technologies. We suggest that integration of these technologies in a modular design can offer a low-cost, robust, and yet sensitive/selective platform for a variety of precision medicine applications.