Flow control in a laminate capillary-driven microfluidic device.

Capillary-driven microfluidic devices are of significant interest for on-site analysis because they do not require external pumps and can be made from inexpensive materials. Among capillary-driven devices, those made from paper and polyester film are among the most common and have been used in a wide array of applications. However, since capillary forces are the only driving force, flow is difficult to control, and passive flow control methods such as changing the geometry must be used to accomplish various analytical applications. This study presents several new flow control methods that can be utilized in a laminate capillary-driven microfluidic device to increase available functionality. First, we introduce push and burst valve systems that can stop and start flow. These valves can stop flow for >30 min and be opened by either pressing the channel or inflowing other fluids to the valve region. Next, we propose flow control methods for Y-shaped channels that enable more functions. In one example, we demonstrate the ability to accurately control concentration to create laminar, gradient, and fully mixed flows. In a second example, flow velocity in the main channel is controlled by adjusting the length of the inlet channel. In addition, the flow velocity is constant as the inlet length increases. Finally, the flow velocity in the Y-shaped device as a function of channel height and fluid properties such as viscosity and surface tension was examined. As in previous studies on capillary-driven channels, the flow rate was affected by each parameter. The fluidic control tools presented here will enable new designs and functions for low cost point of need assays across a variety of fields.

Development of a Paper-Based Viscometer for Blood Plasma Using Colorimetric Analysis.

The viscosity of biofluids can be used to acquire meaningful medical information on the conditions of a patient but has seldom been utilized in clinical practices owing to cumbersome measurement procedures and the need for large sample volumes. We present a colorimetric method to measure the viscosity of blood plasma using a paper-based viscometer developed in this study specifically for clinical diagnosis. The proposed analytical device consists of multilayered papers with fluid-loading, -mixing, and -measuring regions, and it can be fabricated readily in a simple manner using three-layered paper channels and tape. Moreover, the colorimetric analysis enables viscosity estimations by analyzing a single optical image. To validate the device performance, we measured the viscosities of fluids such as glycerin aqueous solutions, bovine-serum-albumin solution, dimethyl sulfoxide, and blood plasma. We found that the measured viscosities were in good agreement with the reference values. Finally, we developed a simple smartphone application for the viscosity measurements that helped enhance the convenience and utility of the paper-based viscometer while maintaining the measurement accuracy.

Consolidation and maturation of the orthopaedic medical device market between 1999 and 2015.

Orthopaedic surgeons often require highly specialized medical devices, implants, and equipment, which are usually offered by several vendors/companies. This study assesses long-term market trends for orthopaedic medical device companies and examines various implications for healthcare cost. Using S&P Capital IQ, a Wall Street database, financial data were gathered on orthopaedic device companies, ranked by worldwide sales, from 1999 to 2015. Annual sales were aggregated to calculate market share and compounded annual growth rates (CAGRs). Overall, the global orthopaedic device market grew at 12.0% CAGR from 1999 to 2008, before slowing to 2.8% from 2009 to 2015. Between 1999 and 2015, the top 5 companies increased total market share from 52.8 to 62.2%. The orthopaedic device market is not only consolidating under a few dominant players, but also growing at a decreasing rate, both of which signal a maturing industry. These trends are likely to shape patient care and healthcare costs in orthopaedic surgery in years to come.

Stem cell industry update: 2012 to 2016 reveals accelerated investment, but market capitalization and earnings lag.

Treatments based on stem cells have long been heralded for their potential to drive the future of regenerative medicine and have inspired increasing medical and business interest. The stem cell therapy market has been expanding since 2012, but earnings and profitability still lag the broader health care sector (compounded annual growth rate in annual financing of 31.5% versus 13.4%, respectively). On the basis of historical financial data, approximately $23 billion has been invested in stem cell companies since 1994, with more than 80% of this raised from 2011 through 2016. This reflects a marked acceleration in capital investment, as companies began late-stage clinical trials, initiate partnerships or are acquired by large pharmaceutical companies. All of these data reflect a field that is emerging from infancy, which will demand more time and capital to mature. This update is relevant to researchers, clinicians and investors who wish to quantify the potential in this field.

Size-Controlled Fabrication of Polyaniline Microfibers Based on 3D Hydrodynamic Focusing Approach.

Owing to the relatively high conductivity and unique redox behavior, polyaniline (PANI) has been one of the most technologically promising conducting polymers. Although various methodologies have been developed, fabrication of PANI microfibers has been a challenging task owing to the poor solubility in most organic solvents. By taking advantage of a microfluidic technology and organic soluble acid labile t-Boc-protected PANI (t-Boc-PANI) as the conducting polymer precursor, fabrication of PANI microfibers in a size-controlled manner is possible. Introduction of a THF solution containing t-Boc-PANI, and dodecylbenzenesulfonic acid (DBSA) as a core flow, and water as a sheath flow into a microfluidic channel with a 3D hydrodynamic focusing effect results in crystallization of the polymer fiber. By changing the flow rate, linear PANI microfibers that range from 16.2 to 39.4 µm in diameter are readily obtained.

Fragmented mitochondrial genomes of the rat lice, Polyplax asiatica and Polyplax spinulosa: intra-genus variation in fragmentation pattern and a possible link between the extent of fragmentation and the length of life cycle.

BACKGROUND: Blood-sucking lice (suborder Anoplura) parasitize eutherian mammals with 67% of the 540 described species found on rodents. The five species of blood-sucking lice that infest humans and pigs have fragmented mitochondrial genomes and differ substantially in the extent of fragmentation. To understand whether, or not, any life-history factors are linked to such variation, we sequenced the mt genomes of Polyplax asiatica and Polyplax spinulosa, collected from the greater bandicoot rat, Bandicota indica, and the Asian house rat, Rattus tanezumi, respectively. RESULTS: We identified all of the 37 mitochondrial genes common to animals in Polyplax asiatica and Polyplax spinulosa. The mitochondrial genes of these two rat lice are on 11 circular minichromosomes; each minichromosome is 2-4 kb long and has 2-7 genes. The two rat lice share the same pattern for the distribution of the protein-coding genes and ribosomal RNA genes over the minichromosomes, but differ in the pattern for the distribution of 8 of the 22 transfer RNA genes. The mitochondrial genomes of the Polyplax rat lice have 3.4 genes, on average, on each minichromosome and, thus, are less fragmented than those of the human lice (2.1 and 2.4 genes per minichromosome), but are more fragmented than those of the pig lice (4.1 genes per minichromosome). CONCLUSIONS: Our results revealed distinct patterns of mitochondrial genome fragmentation within the genus Polyplax and, furthermore, indicated a possible inverse link between the extent of mitochondrial genome fragmentation and the length of life cycle of the blood-sucking lice.

Fragmented mitochondrial genomes are present in both major clades of the blood-sucking lice (suborder Anoplura): evidence from two Hoplopleura rodent lice (family Hoplopleuridae).

BACKGROUND: The suborder Anoplura contains 540 species of blood-sucking lice that parasitize over 840 species of eutherian mammals. Fragmented mitochondrial (mt) genomes have been found in the lice of humans, pigs, horses and rats from four families: Pediculidae, Pthiridae, Haematopinidae and Polyplacidae. These lice, eight species in total, are from the same major clade of the Anoplura. The mt genomes of these lice consist of 9-20 minichromosomes; each minichromosome is 1.5-4 kb in size and has 1-8 genes. To understand mt genome fragmentation in the other major clade of the Anoplura, we sequenced the mt genomes of two species of rodent lice in the genus Hoplopleura (family Hoplopleuridae). RESULTS: We identified 28 mt genes on 10 minichromosomes in the mouse louse, Ho. akanezumi; each minichromosome is 1.7-2.7 kb long and has 1-6 genes. We identified 34 mt genes on 11 minichromosomes in the rat louse, Ho. kitti; each minichromosome is 1.8-2.8 kb long and has 1-5 genes. Ho. akanezumi also has a chimeric minichromosome with parts of two rRNA genes and a full-length tRNA gene for tyrosine. These two rodent lice share the same pattern for the distribution of all of the protein-coding and rRNA genes but differ in tRNA gene content and gene arrangement in four minichromosomes. Like the four genera of blood-sucking lice that have been investigated in previous studies, the Hoplopleura species have four minichromosomes that are only found in this genus. CONCLUSIONS: Our results indicate that fragmented mt genomes were present in the most recent common ancestor of the two major clades of the blood-sucking lice, which lived ~75 million years ago. Intra-genus variation in the pattern of mt genome fragmentation is common in the blood-sucking lice (suborder Anoplura) and genus-specific minichromosomes are potential synapomorphies. Future studies should expand into more species, genera and families of blood-sucking lice to explore further the phylogenetic utility of the novel features associated with fragmented mt genomes.

Transient freezing behavior in photophobic responses of Euglena gracilis investigated in a microfluidic device.

We found that the transient freezing behavior in photophobic responses of Euglena gracilis is a good indicator of the metabolic status of the cells. The transient blue light photophobic responses of E. gracilis cells were investigated on-chip using a new measurement, 'trace momentum' (TM), to evaluate their swimming activity quantitatively in real time. When blue light of intensity >30 mW cm(-2) was repeatedly switched on and off, a large negative spike in the TM was observed at the onset of the 'blue-light-off' phase. Single-cell trace analysis at a blue light intensity of 40 mW cm(-2) showed that 48% (on average, n = 15) of tumbling Euglena cells ceased activity ('freezing') for 2-30 s at the onset of blue-light-off before commencing forward motion in a straight line (termed 'straightforward swimming'), while 45% smoothly commenced straightforward swimming without delay. The proportion of freezing Euglena cells depended on the blue light intensity (only 20% at 20 mW cm(-2)). When the cells were stimulated by four blue light pulses at the higher intensity, without pre-exposure, the transient freezing behavior was more prominent but, on repeating the stimuli after an 80 min interval in red light, the same cells did not freeze. This shows that the metabolism of the cells had changed to anti-freezing during the interval. The relationship between the interval time with/without light irradiation and the blue light adaptation was elucidated experimentally. The origin of the freezing behavior is considered to be a shortage of a metabolic substance that promotes smooth switching of flagellum movement from in situ rotation mode to a straightforward swimming mode.

Correct Closure of the Left Atrial Appendage Reduces Stagnant Blood Flow and the Risk of Thrombus Formation: A Proof-of-Concept Experimental Study Using 4D Flow Magnetic Resonance Imaging.

OBJECTIVE: The study was conducted to investigate the effect of correct occlusion of the left atrial appendage (LAA) on intracardiac blood flow and thrombus formation in patients with atrial fibrillation (AF) using four-dimensional (4D) flow magnetic resonance imaging (MRI) and three-dimensional (3D)-printed phantoms. MATERIALS AND METHODS: Three life-sized 3D-printed left atrium (LA) phantoms, including a pre-occlusion (i.e., before the occlusion procedure) model and correctly and incorrectly occluded post-procedural models, were constructed based on cardiac computed tomography images from an 86-year-old male with long-standing persistent AF. A custom-made closed-loop flow circuit was set up, and pulsatile simulated pulmonary venous flow was delivered by a pump. 4D flow MRI was performed using a 3T scanner, and the images were analyzed using MATLAB-based software (R2020b; Mathworks). Flow metrics associated with blood stasis and thrombogenicity, such as the volume of stasis defined by the velocity threshold (|V̅| < 3 cm/s), surface-and-time-averaged wall shear stress (WSS), and endothelial cell activation potential (ECAP), were analyzed and compared among the three LA phantom models. RESULTS: Different spatial distributions, orientations, and magnitudes of LA flow were directly visualized within the three LA phantoms using 4D flow MRI. The time-averaged volume and its ratio to the corresponding entire volume of LA flow stasis were consistently reduced in the correctly occluded model (70.82 mL and 39.0%, respectively), followed by the incorrectly occluded (73.17 mL and 39.0%, respectively) and pre-occlusion (79.11 mL and 39.7%, respectively) models. The surface-and-time-averaged WSS and ECAP were also lowest in the correctly occluded model (0.048 Pa and 4.004 Pa-1 , respectively), followed by the incorrectly occluded (0.059 Pa and 4.792 Pa-1 , respectively) and pre-occlusion (0.072 Pa and 5.861 Pa-1 , respectively) models. CONCLUSION: These findings suggest that a correctly occluded LAA leads to the greatest reduction in LA flow stasis and thrombogenicity, presenting a tentative procedural goal to maximize clinical benefits in patients with AF.

Size-controlled fabrication of polydiacetylene-embedded microfibers on a microfluidic chip.

A microfluidic technique was employed to fabricate polydiacetylene (PDA)-embedded hydrogel microfibers. By taking advantage of calcium ion-induced insoluble hydrogel formation, supramolecularly assembled diacetylene (DA)-surfactant complexes were successfully immobilized in the calcium alginate fibers. Thus, instantaneous microfiber formation was observed when the core flow of DA supramolecules-containing alginate solution met the sheath flow of calcium ions. UV irradiation of the resulting fibers afforded blue colored PDAs, and the formation of a conjugated polymer was confirmed by heat-induced phase transition and by Raman spectroscopy. By adjusting the core and sheath flow rates, PDA-embedded hydrogel fibers of various sizes were obtained.

Temporal Evolution of the Gravitaxis of Euglena gracilis from a Single Cell.

Gravitaxis is one of the most important issues in the growth of microalgae in the water column; it determines how easily cells receive sunlight with a comfortable intensity that is below the damaging threshold. We quantitatively investigated and analyzed the gravitaxis and cell multiplication of Euglena gracilis using vertically placed microchambers containing a single cell. A temporal change in gravitaxis and cell multiplication was observed after transferring the cells to fresh culture medium for 9 days. We performed 29 individual experiments with 2.5 mm × 2.5 mm × 0.1 mm square microchambers and found that the cells showed positive, negative, and moderate gravitaxis in 8, 7, and 14 cases, respectively, after transferring to fresh culture medium. A common trend was observed for the temporal change in gravitaxis for the eight initially positive gravitaxis cases. The cells with initially positive gravitaxis showed a higher rate of cell multiplication than those with initially negative gravitaxis. We also discussed the gravitaxis mechanism of E. gracilis from the observed trend of gravitaxis change and swimming traces. In addition, bioconvection in a larger and thicker chamber was investigated at a millimeter scale and visualized.

Regeneration of the Eyespot and Flagellum in Euglena gracilis during Cell Division.

Cell division of unicellular microalgae is a fascinating process of proliferation, at which whole organelles are regenerated and distributed to two new lives. We performed dynamic live cell imaging of Euglena gracilis using optical microscopy to elucidate the mechanisms involved in the regulation of the eyespot and flagellum during cell division and distribution of the organelles into the two daughter cells. Single cells of the wild type (WT) and colorless SM-ZK cells were confined in a microfluidic device, and the appearance of the eyespot (stigma) and emergent flagellum was tracked in sequential video-recorded images obtained by automatic cell tracking and focusing. We examined 12 SM-ZK and 10 WT cells and deduced that the eyespot diminished in size and disappeared at an early stage of cell division and remained undetected for 26-97 min (62 min on average, 22 min in deviation). Subsequently, two small eyespots appeared and were distributed into the two daughter cells. Additionally, the emergent flagellum gradually shortened to zero-length, and two flagella emerged from the anterior ends of the daughter cells. Our observation revealed that the eyespot and flagellum of E. gracilis are degraded once in the cell division, and the carotenoids in the eyespot are also decomposed. Subsequently, the two eyespots/flagella are regenerated for distribution into daughter cells. As a logical conclusion, the two daughter cells generated from a single cell division possess the equivalent organelles and each E. gracilis cell has eternal or non-finite life span. The two newly regenerated eyespot and flagellum grow at different rates and mature at different timings in the two daughter cells, resulting in diverse cell characteristics in E. gracilis.

Size-controlled fabrication of supramolecular vesicles for the construction of conjugated polymer sensors with enhanced optical properties.

Polymerizable supramolecular monomer vesicles are readily fabricated by employing a hydrodynamic focusing method on a microfluidic chip. The polymerized diacetylenene nanovesicles, generated using the microfluidic method, display an improved fluorescence property compared to those prepared by employing a conventional bulk method. The flexibility of the vesicle size control by manipulating the flow conditions is another significant feature of the new microfluidic approach.

U.S. Healthcare Insurance Market Concentration from 2001 to 2016: Increased Growth in Direct Written Premiums and Overall Decreased Market Consolidation.

With the establishment of state-based health insurance marketplaces, how U.S. health insurers are responding to market pressures and influencing premiums have represented important questions. We made novel use of the Standard and Poor's (S&P) Financial, a Wall Street financial dataset platform, to analyze trends in market capitalization and total direct written premiums (DWPs) from 2001 to 2016 of the top 5, 10, and 25 health insurance companies. Our results indicate that the market concentration of publicly traded companies has remained relatively stable over the past decade. The top 5, 10, and 25 health insurance companies were 43.5%, 57.5%, and 78.6% of the total market share in 2001 and 39.4%, 52.9%, and 72.8% in 2016, respectively. DWPs have grown nearly four-fold from $177 billion to $631 billion at a compounded annual rate of 8.8%, consistent with overall healthcare sector growth. Aggregating state-specific data, the overall U.S. health insurance market has become slightly less consolidated over recent years, as measured using the population-weighted Herfindahl-Hirschman index, a measure for market concentration, falling from 3,817 to 2,174 during this time period. As health insurance costs place a growing burden on American families, additional efforts are needed to study the impact on choice, quality, access, cost, and value to patients and providers from evolving health insurance markets.

A thermoresponsive fluorogenic conjugated polymer for a temperature sensor in microfluidic devices.

A new, microfluidic temperature sensor system, based on a thermoresponsive conjugated polymer supramolecule, has been developed. By generating blue-phase polydiacetylene (PDA) sensor droplets using hydrodynamic instability, we were able to monitor the variation of fluorescent intensity of the droplets with respect to the flow temperature.

Novel and facile criterion to assess the accuracy of WSS estimation by 4D flow MRI.

Four-dimensional flow magnetic resonance imaging (4D flow MRI) is a versatile tool to obtain hemodynamic information and anatomic information simultaneously. The wall shear stress (WSS), a force exerted on a vessel wall in parallel, is one of the hemodynamic parameters available with 4D flow MRI and is thought to play an important role in clinical applications such as assessing the development of atherosclerosis. Nevertheless, the accuracy of WSS obtained with 4D flow MRI is rarely evaluated or reported in literature, especially in the in vivo studies. We propose a novel and facile criterion called Reynolds resolution to assess the accuracy of WSS estimation in 4D flow MRI studies. Reynolds resolution consists of a spatial resolution, encoding velocity, kinematic viscosity of a working fluid, and signal-to-noise ratio, which are readily accessible information in 4D flow MRI measurements. We explored the relationship between Reynolds resolution and the WSS error. To include diverse and extensive cases, we measured three circular tubing flows with a diameter of 40, 8, and 2 mm. The 40 mm tubing flow was measured by 3 Tesla (T) human MR scanner with a knee coil and spatial resolution of 0.5 mm. The 8 and 2 mm tubing flows were both measured by 4.7 T MR scanner, but the scans were performed with a conventional birdcage coil (8 mm tubing) and a custom-made solenoid coil (2 mm tubing), respectively. The spatial resolution was varied from 0.2, 0.4 or 0.8 mm for the 8 mm tubing flow, but was fixed at 0.090 mm for 2 mm tubing flow. In addition, the near-wall velocity gradient, required to be determined prior to the WSS, was calculated using two methods; these included assuming a linear velocity profile or quadratic velocity profile near wall. The accuracy of WSS obtained using each method and tubing flow was evaluated against the theoretical WSS value. As a result, we found that Reynolds resolution is in logarithmic relation to the WSS error.

Patient-specific Hemodynamics of Severe Carotid Artery Stenosis Before and After Endarterectomy Examined by 4D Flow MRI.

Carotid endarterectomy (CEA) influences the carotid endoluminal anatomy, which results in hemodynamic changes before and after surgery. We investigated the hemodynamics of severe carotid artery stenosis before and after conventional endarterectomy with/without patch repair. An in vitro experiment utilizing carotid phantoms, which underwent a procedure that emulated CEA with/without the patch repair, was performed with a high-spatiotemporal resolution using 4D flow MRI. We evaluated an abnormal region of carotids, which consists of the normalized time-averaged wall shear stress (NTA|WSS|) and the oscillatory shear index (OSI), to account for continuous high-shear regions (high NTA|WSS| and low OSI) and chaotic low-shear regions, i.e., stenosis-prone regions (low NTA|WSS| and high OSI). The use of normalized hemodynamic parameters (e.g., NTA|WSS|) allowed comparison of diverse cases with different conditions of hemodynamics and vessel geometry. We observed that the stenosis-prone regions of the carotids with patches were noticeably larger than the corresponding regions in no-patch carotids. A large recirculating flow zone found in the stenosis-prone region of the internal carotid artery (ICA) of the postoperative carotids with patches partially blocks the flow path into ICA, and consequently the flow rate was not recovered after surgery unlike an expectation.

Water-repellent Hybrid Nanowire and Micro-scale Denticle Structures on Flexible Substrates of Effective Air Retention.

The air retention capability of a superhydrophobic surface plays the crucial role of drag reduction in an aqueous environment. Here, fabrication of water-repellent hybrid structural surfaces by synthesizing superhydrophobic nanowires with a high aspect ratio on micro-scale denticle structures to improve their air holding capacity in water is reported. The hybrid structure is realized by carrying out polymer molding of denticle structures on flexible substrates, hydrothermal growth of nanowires, and subsequent ultra-thin film coating. This technique is readily applicable to large areas, and the fabricated substrates are attachable onto curved surfaces. Our engineered, super water-repellent hybrid structures are found to effectively maintain air bubbles on their surfaces in a highly shear flow condition with a wall shear stress of up to 33.4 Pa, due to the combined effects of the micro-scale denticle structure, which reduces flow resistance, and the superhydrophobic, high-aspect-ratio nanowire structure, which enhances the capillary force to maintain the air bubbles. Our results show the importance of developing superhydrophobic structures of improved air retention capability.

Estimation of the thermocapillary force and its applications to precise droplet control on a microfluidic chip.

Droplet control through the use of light-induced thermocapillary effects has recently garnered attention due to its non-intrusive and multifunctional nature. An important issue in droplet control is the estimation of the thermocapillary force. The purpose of the present study is to estimate the thermocapillary force and propose empirical equations between the force and simply measurable key parameters such as droplet diameter and power of heat source. In addition, we aim to shift the droplet trajectory and develop an on-demand droplet routing system based on the estimation of the thermocapillary force. We illuminated a continuous phase with a 532 nm laser beam to minimize possible damage or property changes to target molecules contained within droplets. A mixture of light-absorbing material and oleic acid was used for the continuous phase fluid, while deionized water (DI water) was used for the dispersed phase fluid. We proposed empirical equations to estimate the thermocapillary force, which was then applied to precise droplet shifting and routing. We found that the shifting distance was linearly proportional to the thermocapillary force, and that an on-demand droplet routing system resulted in a success rate greater than 95%.

The Mitochondrial Genome of the Guanaco Louse, Microthoracius praelongiceps: Insights into the Ancestral Mitochondrial Karyotype of Sucking Lice (Anoplura, Insecta).

Fragmented mitochondrial (mt) genomes have been reported in 11 species of sucking lice (suborder Anoplura) that infest humans, chimpanzees, pigs, horses, and rodents. There is substantial variation among these lice in mt karyotype: the number of minichromosomes of a species ranges from 9 to 20; the number of genes in a minichromosome ranges from 1 to 8; gene arrangement in a minichromosome differs between species, even in the same genus. We sequenced the mt genome of the guanaco louse, Microthoracius praelongiceps, to help establish the ancestral mt karyotype for sucking lice and understand how fragmented mt genomes evolved. The guanaco louse has 12 mt minichromosomes; each minichromosome has 2-5 genes and a non-coding region. The guanaco louse shares many features with rodent lice in mt karyotype, more than with other sucking lice. The guanaco louse, however, is more closely related phylogenetically to human lice, chimpanzee lice, pig lice, and horse lice than to rodent lice. By parsimony analysis of shared features in mt karyotype, we infer that the most recent common ancestor of sucking lice, which lived ∼75 Ma, had 11 minichromosomes; each minichromosome had 1-6 genes and a non-coding region. As sucking lice diverged, split of mt minichromosomes occurred many times in the lineages leading to the lice of humans, chimpanzees, and rodents whereas merger of minichromosomes occurred in the lineage leading to the lice of pigs and horses. Together, splits and mergers of minichromosomes created a very complex and dynamic mt genome organization in the sucking lice.