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This study investigates a novel microfluidic mixing technique that uses the resonant oscillation of coalescent droplets. During the vertical contact-separation process, solutes are initially separated as a result of the combined effects of diffusion and gravity. We show that the application of alternating current (AC) voltage to microelectrodes below the droplets causes a resonant oscillation, which enhances the even distribution of the solute. The difference in concentration between the top and bottom droplets exhibits frequency dependence and indicates the existence of a particular AC frequency that results in a homogeneous concentration. This frequency corresponds to the resonance frequency of the droplet oscillation that is determined using particle tracking velocimetry. To understand the mixing process, a phenomenological model based on the equilibrium between surface tension, viscosity, and electrostatic force was developed. This model accurately predicted the resonance frequency of droplet flow and was consistent with the experimental results. These results suggest that the resonant oscillation of droplets driven by AC voltage significantly enhances the diffusion of solutes, which is an effective approach to microfluid mixing., (© 2024. The Author(s).)

Cardiovascular and cerebrovascular diseases (CCVD) are prominent mortality causes in Japan, necessitating effective preventative measures, early diagnosis, and treatment to mitigate their impact. A diagnostic model was developed to identify patients with ischemic heart disease (IHD), stroke, or both, using specific health examination data. Lifestyle habits affecting CCVD development were analyzed using five causal inference methods. This study included 473,734 patients aged ≥ 40 years who underwent specific health examinations in Kanazawa, Japan between 2009 and 2018 to collect data on basic physical information, lifestyle habits, and laboratory parameters such as diabetes, lipid metabolism, renal function, and liver function. Four machine learning algorithms were used: Random Forest, Logistic regression, Light Gradient Boosting Machine, and eXtreme-Gradient-Boosting (XGBoost). The XGBoost model exhibited superior area under the curve (AUC), with mean values of 0.770 (± 0.003), 0.758 (± 0.003), and 0.845 (± 0.005) for stroke, IHD, and CCVD, respectively. The results of the five causal inference analyses were summarized, and lifestyle behavior changes were observed after the onset of CCVD. A causal relationship from 'reduced mastication' to 'weight gain' was found for all causal species theory methods. This prediction algorithm can screen for asymptomatic myocardial ischemia and stroke. By selecting high-risk patients suspected of having CCVD, resources can be used more efficiently for secondary testing., (© 2024. The Author(s).)

Implementing a signal-switching mechanism for the selective use of integrated sensors and actuators plays a crucial role in streamlining the functionality of miniaturized devices. Here, a liquid metal droplet (LMD)-based signal-switching mechanism is introduced to achieve such functionality. Pressure modulation with a 100-μm spatial resolution enabled precise control of the position of the LMDs within a channel. After integrating the channel with asymmetrically structured electrodes, the effect of the shuttle-like movement of LMD on the temporal changes in the overall capacitance was investigated. Consequently, analysis of the capacitive peaks revealed the directional movement of the LMDs, enabling estimation of the position of the LMDs without direct observation. In addition, we achieved successful signal extraction from the capacitive sensor that was linked to the activated electrodes, thereby enabling selective data retrieval. The proposed signal-switching mechanism method achieved a detection accuracy of ~0.1 pF. The sensor's ability to simultaneously detect the LMD position and generate a signal underscores its significant potential for multiplexing in multisensing systems, particularly in concealed environments, such as in vivo settings., Competing Interests: Competing interestsSatoshi Konishi is editorial board member of the journal, no other author has reported any competing interest., (© The Author(s) 2024.)

This study assessed the clinical performance of point-of-care testing (POCT) for quick cortisol assay (QCA) during adrenal vein sampling (AVS) using a newly invented portable quantitative assay instrument. An observational study was conducted prospectively at two centres in Japan. Forty-eight patients with primary aldosteronism considered for adrenalectomy were enrolled in this study and underwent AVS. Three basal adrenal vein samples from each adrenal vein and two from the inferior vena cava were collected sequentially. The cortisol concentration of adrenal vein samples was measured by routine method and QCA. A total of 338 adrenal vein samples were analysed from 250 sites to determine AVS success or failure. The distribution of turnaround time of the QCA for AVS success or failure followed a normal distribution with an average of 20.5 min. A positive correlation between the routine method and QCA was observed regarding cortisol concentration or selectivity index. No significant difference between the two methods was observed regarding the success rate of AVS. Using the routine method as a reference, the sensitivity and specificity of AVS success or failure were 99.1% (210/212) and 81.6% (31/38), respectively. Easy, quick, portable, and precise POCT-QCA demonstrated its compatibility with routine methods regarding clinical performance., (© 2023. The Author(s).)

The renin-angiotensin-aldosterone system (RAAS) is a regulatory mechanism of the endocrine system and is associated with various diseases, including hypertension and renal and cardiovascular diseases. The gut microbiota (GM) have been associated with various diseases, mainly in animal models. However, to our knowledge, no studies have examined the relationship between the RAAS and GM in humans. The present study aimed to assess the association between the systemic RAAS and GM genera and their causal relationships. The study participants were 377 members of the general population aged 40 years or older in Shika-machi, Japan. Plasma renin activity (PRA), plasma aldosterone concentration (PAC), aldosterone-renin ratio (ARR), and GM composition were analyzed using the 16S rRNA method. The participants were divided into high and low groups according to the PRA, PAC, and ARR values. U-tests, one-way analysis of covariance, and linear discriminant analysis of effect size were used to identify the important bacterial genera between the two groups, and binary classification modeling using Random Forest was used to calculate the importance of the features. The results showed that Blautia, Bacteroides, Akkermansia, and Bifidobacterium were associated with the RAAS parameters. Causal inference analysis using the linear non-Gaussian acyclic model revealed a causal effect of Blautia on PAC via SBP. These results strengthen the association between the systemic RAAS and GM in humans, and interventions targeting the GM may provide new preventive measures and treatments for hypertension and renal disease., (© 2023. The Author(s), under exclusive licence to The Japanese Society of Hypertension.)

Organs-on-chips using cultured cells have been developed and applied for evaluating in vitro biological phenomena. We previously reported an openable artificial intestinal tract system, as an in vitro model of the small intestine, for in vitro drug screening. The intestinal tract device could be transformed using an integrated artificial muscle actuator. An initial flat state was suitable for cell culture, and the transformed tubular structure was used as a fluidic channel for perfusion tests. The previously developed intestinal tract system could be used to evaluate drug absorption by cells through perfusion testing. This study presents an improved artificial intestinal tract system for analysis of drug permeation, in addition to absorption. Permeable filters were integrated into the intestinal tract device. Integration of additional filters into the design of the existing artificial muscle actuator was accomplished by considering device performance and available filter locations. Filter permeability was evaluated by perfusion testing. MDCK-II cells were cultured on the device and visually and electrically evaluated. The openable device, equipped with new functions for further pharmacokinetic analysis, could perform and evaluate drug disposition using cultured cells. We anticipate that the improved, openable organ-on-a-chip device system will contribute to advances in in vitro drug screening technology., (© 2023. The Author(s).)

In vertical contact control (VCC), a microdroplet array selectively contacts with an opposite microdroplet array. Generally, VCC is useful for the dispenser mechanism based on solute diffusion between microdroplet pairs. However, sedimentation due to gravity can cause an inhomogeneous distribution of solutes in microdroplets. Therefore, it is necessary to enhance solute diffusion to achieve the accurate dispensing of a large quantity of solute in the direction opposite to that of gravity. Herein, we applied a rotational magnetic field to the microrotors in microdroplets to enhance the solute diffusion in microdroplets. Driven by microrotors, the rotational flow can generate a homogeneous distribution of solutes in microdroplets. We analyzed the diffusion dynamics of solutes using a phenomenological model, and the results showed that the rotation of microrotors can increase the diffusion constant of solutes., (© 2023. The Author(s).)

Transportation of magnetized particles (MPs) against gravity is possible by applying a magnetic field to the particles. This transport phenomenon of MPs in microdroplets can be quantitatively assessed by determining the contribution of individual forces acting on the MPs. We studied the selective transportation of MPs in microdroplets. MPs in microdroplets were transported in the opposite direction to gravity when we applied an external magnetic field larger than a threshold value. We modulated the intensity of the external magnetic field and selectively manipulated the MPs. As a result, MPs were separated into different microdroplets based on their magnetic properties. Our quantitative investigation of transport dynamics shows that the threshold magnetic field depends only on the magnetic susceptibility and the density of MPs. This is a universal criterion for the selective transport of magnetized targets such as magnetized cells in microdroplets., (© 2023. The Author(s).)

Various soft actuators have been investigated to overcome the drawbacks of conventional solid machines and explore the applications of soft robotics. In particular, and because they are expected to be applicable in minimally invasive medicine because of their safety, soft inflatable microactuators using an actuation conversion mechanism from balloon inflation to bending motion have been proposed for high-output bending motion. These microactuators could be applied to create an operation space by safely moving organs and tissues; however, the conversion efficiency could be further improved. This study aimed to improve conversion efficiency by investigating the design of the conversion mechanism. The contact conditions between the inflated balloon and conversion film were examined to improve the contact area for force transmission, with the contact area dependent on the length of the contact arc between the balloon and force conversion mechanism and on the amount of balloon deformation. In addition, surface contact friction between the balloon and film, which affects actuator performance, was also investigated. The generated force of the improved device is 1.21 N at 80 kPa when it bends 10 mm, which is 2.2 times the generated force of the previous design. This improved soft inflatable microactuator is expected to assist in performing operations in a limited space, such as in endoscopic or laparoscopic operations., Competing Interests: Competing interestsThe authors declare no competing interests., (© The Author(s) 2023.)

The spatial resolution of typical sensor probes is sufficient for measuring the average electric properties of microelectrical devices, but they are unable to measure the distribution with a spatial precision. Liquid crystal droplets (LCDs) are promising candidate for visualizing the distribution. When voltage is applied, the LCDs show rotational and translational behaviors which depend on the location of LCDs within the devices. We demonstrate that by comparing the experimental and numerical results, the electric field and electrostatic energy distribution are visualized by rotating and transporting LCDs, with a spatial resolution of 10 µm and a detection accuracy of 5 µV/µm. In addition, we produced an array of LCDs by designing periodic modulation of the electrostatic energy density in the model device. These findings show that the LCDs serve as a periodic modulator of the refractive index as well as a sensor for the observation of electric properties of microelectrical devices., (© 2023. The Author(s).)

Palpation is widely used as the initial medical diagnosis. Integration of micro tactile sensors and artificial muscles enables a soft microfinger for active touch sensing using its bending actuation. Active touch sensing by pushing-in motion of microfinger enables to evaluate stiffness distribution on an elastic object. Due to its compactness, the microfinger can enter a narrow space, such as gastrointestinal and abdominal spaces in a body. However, a microfinger can only touch and sense limited points. We aim at efficient method for searching a stiffness singular part in an elastic object by the directional touch sensing of a microfinger. This study presents a microfinger for active touch sensing using bending and push-in actuation and proposes an algorithm utilizing directivity in touch sensing by a microfinger for efficient localization of the stiffness singular part in an object. A gelatin block structure with a small rigid ball was prepared and touch sensed by the microfinger. Consequently, the position of the buried rigid ball could be efficiently identified based on the proposed algorithm. This result implies that the proposed method has potential applications in endoscopic medical diagnosis, particularly in identifying tumor positions., (© 2022. The Author(s).)

Human-robot interaction technology has contributed to improving sociality for humanoid robots. At scales far from human scales, a microrobot can interact with an environment in a small world. Microsensors have been applied to measurement of forces by flying or walking insects. Meanwhile, most previous works focused on the measurement of the behavior of insects. Here, we propose microrobot-insect interactions by soft microfingers integrated with artificial muscle actuators and tactile sensors, which has been developed for a haptic teleoperation robot system. A soft pneumatic balloon actuator acts as the artificial muscle, and a flexible strain sensor using a liquid metal provides tactile sensing. Force interaction between a pill bug and the microfinger could be accomplished. The microfinger (12 mm × 3 mm × 490 μm) can move and touch an insect, and it can detect reaction force from an insect. The measured reaction force from the legs of a pill bug as a representative insect was less than 10 mN. This paper presents a microfinger as an end effector for the active sensing of reaction force from a small insect. We anticipate that our results will lead to further evaluation of small living things as well as technology development for human-environment interaction., (© 2022. The Author(s).)

Identifying the host genetic factors underlying severe COVID-19 is an emerging challenge 1-5 . Here we conducted a genome-wide association study (GWAS) involving 2,393 cases of COVID-19 in a cohort of Japanese individuals collected during the initial waves of the pandemic, with 3,289 unaffected controls. We identified a variant on chromosome 5 at 5q35 (rs60200309-A), close to the dedicator of cytokinesis 2 gene (DOCK2), which was associated with severe COVID-19 in patients less than 65 years of age. This risk allele was prevalent in East Asian individuals but rare in Europeans, highlighting the value of genome-wide association studies in non-European populations. RNA-sequencing analysis of 473 bulk peripheral blood samples identified decreased expression of DOCK2 associated with the risk allele in these younger patients. DOCK2 expression was suppressed in patients with severe cases of COVID-19. Single-cell RNA-sequencing analysis (n = 61 individuals) identified cell-type-specific downregulation of DOCK2 and a COVID-19-specific decreasing effect of the risk allele on DOCK2 expression in non-classical monocytes. Immunohistochemistry of lung specimens from patients with severe COVID-19 pneumonia showed suppressed DOCK2 expression. Moreover, inhibition of DOCK2 function with CPYPP increased the severity of pneumonia in a Syrian hamster model of SARS-CoV-2 infection, characterized by weight loss, lung oedema, enhanced viral loads, impaired macrophage recruitment and dysregulated type I interferon responses. We conclude that DOCK2 has an important role in the host immune response to SARS-CoV-2 infection and the development of severe COVID-19, and could be further explored as a potential biomarker and/or therapeutic target., (© 2022. The Author(s).)

We proposed the detection mechanism of the micro-levels of temperature gradient in a micro-electromechanical system using the unidirectional rotation of cholesteric-liquid crystal (Ch-LC) droplets. Ch-LC droplets in the presence of an isotropic phase subjected to a heat flux rotate with a speed proportional to the magnitude of the temperature gradient. We further quantified the temperature gradient-to-torque conversion efficiency to apply the thermomechanical cross-correlation to the detection of temperature gradient. Then, we observed the rotational behavior of Ch-LC droplets after introducing them onto model devices containing patterned Au thin-film electrodes. Direct electric current applied to these Au electrodes results in unidirectional rotation of the Ch-LC droplets in response to heat flux generated from the Au electrodes. By evaluating the possible temperature gradient detection resolution using Ch-LC droplet rotation, we show that Ch-LC droplets can achieve both high spatial resolution (~ 10 µm) and high detection accuracy (~ 0.1 mK/µm)., (© 2022. The Author(s).)

We aimed to determine the survival and staging benefit of limited pelvic lymph node dissection (PLND) during radical prostatectomy (RP) in high-risk prostate cancer (PC) patients treated with neoadjuvant chemohormonal therapy. We retrospectively analyzed 516 patients with high-risk localized PC (< cT4N0M0) who received neoadjuvant androgen-deprivation therapy plus estramustine phosphate followed by RP between January 2010 and March 2020. Since we stopped limited PLND for such patients in October 2015, we compared the surgical outcomes and biochemical recurrence-free survival (BCR-FS) between the limited-PLND group (before October 2015, n = 283) and the non-PLND group (after November 2015, n = 233). The rate of node metastases in the limited-PLND group were 0.8% (2/283). Operation time was significantly longer (176 vs. 162 min) and the rate of surgical complications were much higher (all grades; 19 vs. 6%, grade ≥ 3; 3 vs. 0%) in the limited-PLND group. The inverse probability of treatment weighting-Cox analysis revealed limited PLND had no significant impact on BCR-FS (hazard ratio, 1.44; P = 0.469). Limited PLND during RP after neoadjuvant chemohormonal therapy showed quite low rate of positive nodes, higher rate of complications, and no significant impact on BCR-FS., (© 2022. The Author(s).)

Multiple sensors and actuators integrated in a small space, especially an elongated thin structure, require equivalent number of signal lines between microdevices, but there is limited space for signal wires. Thus, we propose a mechanism using a single microchannel where a liquid metal droplet moves and shuttles. A shuttling droplet switches multiple terminals of signal lines along a microchannel based on a traditional switching mechanism using a liquid metal droplet. Electrically conductive gallium alloy liquid metals (Galinstan) can flow in a microchannel due to their fluidity. The terminals consist of opposing electrode pairs in a microchannel. A change in a variable impedance connected to a terminal as a pseudo sensor can be read when a droplet flows in and connects electrode pairs. This paper presents switching and addressing objective terminals of chromium electrodes by a shuttling conductive droplet (500 µm in diameter and 10 mm long) in a microchannel (500 µm in diameter and 100 mm long). A demonstrated simple mechanism enables communication between multiple microdevices along a microchannel. We anticipate wide application of proposed mechanism toward a multiplexer, especially in microfluidic devices because of the advantages of utilizing microchannels as common microstructures for both microdevices and signal lines., (© 2022. The Author(s).)

The discriminative power of CURB-65 for mortality in community-acquired pneumonia (CAP) is suspected to decrease with age. However, a useful prognostic prediction model for older patients with CAP has not been established. This study aimed to develop and validate a new scoring system for predicting mortality in older patients with CAP. We recruited two prospective cohorts including patients aged ≥ 65 years and hospitalized with CAP. In the derivation (n = 872) and validation cohorts (n = 1,158), the average age was 82.0 and 80.6 years and the 30-day mortality rate was 7.6% (n = 66) and 7.4% (n = 86), respectively. A new scoring system was developed based on factors associated with 30-day mortality, identified by multivariate analysis in the derivation cohort. This scoring system named CHUBA comprised five variables: confusion, hypoxemia (SpO 2  ≤ 90% or PaO 2  ≤ 60 mmHg), blood urea nitrogen ≥ 30 mg/dL, bedridden state, and serum albumin level ≤ 3.0 g/dL. With regard to 30-day mortality, the area under the receiver operating characteristic curve for CURB-65 and CHUBA was 0.672 (95% confidence interval, 0.607-0.732) and 0.809 (95% confidence interval, 0.751-0.856; P < 0.001), respectively. The effectiveness of CHUBA was statistically confirmed in the external validation cohort. In conclusion, a simpler novel scoring system, CHUBA, was established for predicting mortality in older patients with CAP., (© 2021. The Author(s).)

The evaluation of surgical damage is challenging because of the lack of specific biomarkers. Total cell-free DNA (cfDNA) levels have been reported to increase with external trauma and may be a biomarker for tissue damage. To investigate the utility of perioperative total cfDNA levels in evaluating surgical damage in urological surgeries. This multicenter, prospective, observational study included 196 patients scheduled for urological surgeries between September 2020 and July 2021. The primary outcome was the change in total cfDNA levels before and after urological surgery. The secondary outcome was the effect of surgical type on total cfDNA ratio before and after urological surgery. The postoperative median total cfDNA level of the 196 patients was significantly increased 2.5-fold compared to the preoperative level (185.2 ng/mL vs. 406.7 ng/mL, P < 0.001). The median total cfDNA before/after ratio was greater than four-fold for kidney transplantation, open cystectomy, and open adrenalectomy. The ratio was less than two-fold for laparoscopic adrenalectomy and robot-assisted radical prostatectomy. Major surgery showed a significant postoperative increase in total cfDNA levels, while minor surgery did not. Total cfDNA levels increased 2.5-fold after urological surgery and it can be used as an acute-phase biomarker for surgical damage., (© 2021. The Author(s).)

Skeletal muscle mass is critical for good quality of life. Mesenchymal stem cells (MSCs) are multipotent stem cells distributed across various tissues. They are characterized by the capacity to secrete growth factors and differentiate into skeletal muscle cells. These capabilities suggest that MSCs might be beneficial for muscle growth. Nevertheless, little is known regarding the effects on muscle protein anabolic and catabolic systems of intramuscular injection of MSCs into skeletal muscle. Therefore, in the present study, we measured changes in mechanistic target of rapamycin complex 1 (mTORC1) signaling, the ubiquitin-proteasome system, and autophagy-lysosome system-related factors after a single intramuscular injection of MSCs with green fluorescence protein (GFP) into mouse muscles. The intramuscularly-injected MSCs were retained in the gastrocnemius muscle for 7 days after the injection, indicated by detection of GFP and expression of platelet-derived growth factor receptor-alpha. The injection of MSCs increased the expression of satellite cell-related genes, activated mTORC1 signaling and muscle protein synthesis, and increased protein ubiquitination and autophagosome formation (indicated by the expression of microtubule-associated protein 1 light chain 3-II). These results suggest that the intramuscular injection of MSCs activated muscle anabolic and catabolic systems and accelerated muscle protein turnover., (© 2021. The Author(s).)

Methodological advances in on-chip technology enable high-throughput drug screening, such as droplet-array sandwiching technology. Droplet-array sandwiching technology involves upper and lower substrates with a droplet-array designed for a one-step process. This technology is, however, limited to batch manipulation of the droplet-array. Here, we propose a method for selective control of individual droplets, which allows different conditions for individual droplet pairs. Electrowetting-on-dielectric (EWOD) technology is introduced to control the height of the droplets so that the contact between droplet-pairs can be individually controlled. Circular patterns 4 mm in diameter composed of electrodes for EWOD and hydrophilic-hydrophobic patterns for droplet formation 4 μl in volume were developed. We demonstrate the selective control of the droplet height by EWOD for an applied voltage up to 160 V and selective control of the contact and transport of substances. Presented results will provide useful method for advanced drug screening, including cell-based screening.

Gallium alloy liquid metals (Galinstan) possessing fluidity, electric conductivity, and low toxicity are attractive for use in flexible devices and microfluidic devices. However, the oxide skin of Galinstan in the atmosphere adheres to the microchannel surface, preventing the transportation of Galinstan in the channel. To tackle the problem of the adhesion of Galinstan to microchannel, we introduced liquid with Galinstan into a channel with a diameter of 1000 μm. Then, we found that the cylindrical shape of the channel enabled smooth transportation of Galinstan independently of both the liquid and the channel material. The liquid introduced with Galinstan not only prevents adhesion but also improves the spatial controllability of Galinstan in the channel. We can control the position of Galinstan with 100 μm resolution using highly viscous (> 10 cSt) liquid. In addition, we combined the microchannel with patterned electrodes, fabricating a serially arranged capacitive device. The local capacitance detected by the patterned electrodes changed by more than 6% via the smooth transportation of Galinstan. The analysis results based on an equivalent circuit quantitatively agree with our experimental results. We can modulate the serially arranged capacitors using the smooth transportation of Galinstan in the channel.

This paper presents progress in the automation of cell and tissue systems and attempts toward the in situ feedback control of organs-on-a-chip. Our study aims to achieve feedback control of a cell and tissue system by a personal computer (PC), whereas most studies on organs-on-a-chip focus on the automation of status monitoring. The implemented system is composed of subsystems including automated culture, stimulation, and monitoring. The monitoring function provides imaging as well as sampling and dispensing in combination with an external analyzer. Individual subsystems can be combined accordingly. First, monitoring of skeletal muscle (SM) and adipose tissues using this system was demonstrated. The highlight of this paper is the application of the system to the feedback control of the lipid droplet (LD) size, where biochemical stimulation using insulin and adrenaline is controlled by a PC according to the obtained LD imaging data. In this study, the system demonstrated its function of maintaining the desired size of LDs. Our results expand the possibility of PC-controllable cell and tissue systems by addressing the challenge of feedback control of organs-on-a-chip. The PC-controllable cell and tissue systems will contribute to living systems-on-a-chip based on homeostasis phenomena involving interactions between organs or tissues.

It has been suggested that aldosterone breakthrough during treatment with a type 1 angiotensin II receptor (AT1R) blocker (ARB) may be an important risk factor for the progression of renal and cardiovascular disease. We examined whether the direct renin inhibitor, aliskiren caused aldosterone breakthrough in angiotensin II (Ang II)-dependent hypertensive mice. The effect of combination therapy with aliskiren and eplerenone was compared with that of therapy using renin-angiotensin system (RAS) blockade. Tsukuba hypertensive mice were treated for 12 weeks with aliskiren (30 mg/kg/day, i.p), candesartan (5 mg/kg/day, p.o), eplerenone (100 mg/kg/day, p.o) aliskiren and candesartan, aliskiren and eplerenone or candesartan and eplerenone. Blood pressure, urinary aldosterone and angiotensinogen (AGTN) excretion; plasma endothelin-1 concentration; kidney weight; urinary albumin excretion (UAE); glomerular injury; and renal messenger RNA (mRNA) levels for transforming growth factor (TGF)-β1, plasminogen activator inhibitor (PAI)-1, angiotensin-converting enzyme (ACE) and AT1R were measured. Combination therapy with aliskiren and candesartan caused a further decrease in blood pressure (p < 0.05) compared with either agent alone. Urinary aldosterone excretion was decreased significantly by 4 weeks of treatment with aliskiren or candesartan (p < 0.05). However, it was increased again by treatment with candesartan or aliskiren for 12 weeks. Combination therapy with aliskiren and eplerenone significantly decreased UAE, the glomerulosclerosis index, and PAI-1 and TGF-β1 mRNA levels compared with all other therapies (p < 0.05). Treatment with aliskiren decreased urinary aldosterone excretion at 4 weeks and increased it at 12 weeks. Combination therapy with a direct renin inhibitor and a mineralocorticoid receptor blocker may be effective for the prevention of renal injury in Ang II-dependent hypertension.

The improvement of soft inflatable microactuators using an actuation conversion mechanism is presented in terms of high-output generation; a bending inflatable microactuator with the conversion mechanism is designed to generate high-output bending motion. The designed microactuator consists of a pneumatic balloon on a base film and a conversion film over the balloon and ribs on the backside of the base film. A conversion film converts the inflating motion of a pneumatic balloon into a bending motion. The fabricated microactuator with a pneumatic balloon of 13 mm in diameter is 16 mm × 40 mm × 850 μm. A 25 μm thick polyimide film is used as a conversion film over the pneumatic balloon because polyimide film is both non-stretchable and flexible. An array of Si ribs (15 mm × 40 mm × 400 μm) is integrated on the backside of the base film. Analysis of the microactuators with and without the conversion mechanism indicates that the output performance is improved with the addition of the conversion mechanism, as designed. As a result, the microactuator with the conversion film generates a maximum force of 1.72 N at 80 kPa, whereas the microactuator without the conversion film generates a maximum force of 0.15 N at 40 kPa. The improved microactuator can provide 4.2 mN/mm 3 as the force density. In addition to fundamental characterization, the performance characteristics of the actuators are examined by combining the fundamental results.

The integration of a flexible temperature sensor with a soft microactuator (a pneumatic balloon actuator) for a functional microfinger is presented herein. A sensor integrated with a microactuator can actively approach a target for contact detection when a distance exists from the target or when the target moves. This paper presents a microfinger with temperature sensing functionality. Moreover, thermocouples, which detect temperature based on the Seebeck effect, are designed for use as flexible temperature sensors. Thermocouples are formed by a pair of dissimilar metals or alloys, such as copper and constantan. Thin-film metals or alloys are patterned and integrated in the microfinger. Two typical thermocouples (K-type and T-type) are designed in this study. A 2.0 mm × 2.0 mm sensing area is designed on the microfinger (3.0 mm × 12 mm × 400 μm). Characterization indicates that the output voltage of the sensor is proportional to temperature, as designed. It is important to guarantee the performance of the sensor against actuation effects. Therefore, in addition to the fundamental characterization of the temperature sensors, the effect of bending deformation on the characteristics of the temperature sensors is examined with a repeated bending test consisting of 1000 cycles.

Microactuators are the most distinctive and challenging microdevices among micro electromechanical systems (MEMS) relative to microsensors or electronic circuits. Soft and flexible microactuators have been achieved by introducing polymers as structural materials in addition to conventional materials. Expanding the application of MEMS to the biomedical field requires particular features, such as softness and small devices. It is important to address small and fragile biological objects while satisfying the demand for minimally invasive medicine. Both MEMS and biomedical applications require three-dimensional microstructures for higher-order functions. In general, microactuators are limited to simple motions such as bending. Our group has developed an openable artificial small intestinal tract system. An array of pneumatic balloon actuators (PBAs) transforms a flat structure into a tube structure representing the small intestine. Coordination of the bending motions of the PBAs enables the formation of a three-dimensional tube structure. Each PBA is 400 μm × 1800 μm × 100 μm. The diameter of the tube structure is 1 mm. Additional higher-order functions of the artificial small intestine, such as peristaltic motion, are currently of interest for us. This paper describes the morphological transformation of a soft microstructure and further potential possibilities of coordinated motions of soft microactuators.

This paper describes a pneumatic balloon actuator (PBA) composed of polydimethylsiloxane (PDMS) for cellular aggregate manipulation. We evaluated the ability of the microdevice to manipulate a tiny and sensitive cellular aggregate without causing serious damage. We used human mesenchymal stem cells (hMSCs) for the cellular aggregate. We describe the design, fabrication, characterization and operation of the soft microfingers to pinch and release a spherical hMSC aggregate ( φ 200 μm), and we employed a PBA to serve as an artificial muscle to drive the microfingers. A design of the microfingers in terms of dimensions, generated force and contact conditions was accomplished. The designed dimensions of a single finger were 560 μm×900 μm. In summary, we demonstrate the utility of the surface modification of a fingertip for pinching and releasing a cellular aggregate and describe a manipulation system that was constructed to drive and control the microfingers. The implemented manipulation system, which is composed of microfingers and a positioning mechanism, was tested and verified in a series of operations., Competing Interests: The authors declare no conflict of interest.

Silicon is an important nutrient for the optimal growth and sustainable production of rice. Rice accumulates up to 10% silicon in the shoot, and this high accumulation is required to protect the plant from multiple abiotic and biotic stresses. A gene, Lsi1, that encodes a silicon influx transporter has been identified in rice. Here we describe a previously uncharacterized gene, low silicon rice 2 (Lsi2), which has no similarity to Lsi1. This gene is constitutively expressed in the roots. The protein encoded by this gene is localized, like Lsi1, on the plasma membrane of cells in both the exodermis and the endodermis, but in contrast to Lsi1, which is localized on the distal side, Lsi2 is localized on the proximal side of the same cells. Expression of Lsi2 in Xenopus oocytes did not result in influx transport activity for silicon, but preloading of the oocytes with silicon resulted in a release of silicon, indicating that Lsi2 is a silicon efflux transporter. The identification of this silicon transporter revealed a unique mechanism of nutrient transport in plants: having an influx transporter on one side and an efflux transporter on the other side of the cell to permit the effective transcellular transport of the nutrients.

Silicon is beneficial to plant growth and helps plants to overcome abiotic and biotic stresses by preventing lodging (falling over) and increasing resistance to pests and diseases, as well as other stresses. Silicon is essential for high and sustainable production of rice, but the molecular mechanism responsible for the uptake of silicon is unknown. Here we describe the Low silicon rice 1 (Lsi1) gene, which controls silicon accumulation in rice, a typical silicon-accumulating plant. This gene belongs to the aquaporin family and is constitutively expressed in the roots. Lsi1 is localized on the plasma membrane of the distal side of both exodermis and endodermis cells, where casparian strips are located. Suppression of Lsi1 expression resulted in reduced silicon uptake. Furthermore, expression of Lsi1 in Xenopus oocytes showed transport activity for silicon only. The identification of a silicon transporter provides both an insight into the silicon uptake system in plants, and a new strategy for producing crops with high resistance to multiple stresses by genetic modification of the root's silicon uptake capacity.

Metabotropic gamma-aminobutyric acid type B (GABAB) and glutamate receptors (mGluRs) are postsynaptically co-expressed at cerebellar parallel fiber (PF)-Purkinje cell (PC) excitatory synapses, but their functional interactions are unclear. We found that mGluR1 agonist-induced currents and [Ca2+]i increases in PCs were enhanced following co-activation of GABAB receptors. A GABAB antagonist and a G-protein uncoupler suppressed these effects. Low-concentration baclofen, a GABAB agonist, augmented mGluR1-mediated excitatory synaptic current produced by stimulating PFs. These results indicate that postsynaptic GABAB receptors functionally interact with mGluR1 and enhance mGluR1-mediated excitatory transmission at PF-PC synapses. The interaction between the two types of metabotropic receptors provides a likely mechanism for regulating cerebellar synaptic plasticity.

A single neurotransmitter elicits diverse physiological responses through activation of multiple receptor subtypes and/or heterosynaptic interactions involving distinct synaptic targets. We found that a typical excitatory transmitter released from the climbing fiber (CF) in the cerebellar cortex not only excited Purkinje cells directly but also presynaptically inhibited GABAergic transmission from interneurons converging on the same Purkinje cells. Both homosynaptic and heterosynaptic actions of the CF transmitter (possibly glutamate) were mediated by activation of AMPA receptors. Dual AMPA receptor-mediated functions of excitation and disinhibition may ensure transmission of cerebellar CF signals controlling sensorimotor coordination.

The Wisconsin Card Sorting Test, which probes the ability to shift attention from one category of stimulus attributes to another (shifting cognitive sets), is the most common paradigm used to detect human frontal lobe pathology. However, the exact relationship of this card test to prefrontal function and the precise anatomical localization of the cognitive shifts involved are controversial. By isolating shift-related signals using the temporal resolution of functional magnetic resonance imaging, we reproducibly found transient activation of the posterior part of the bilateral inferior frontal sulci. This activation was larger as the number of dimensions (relevant stimulus attributes that had to be recognized) were increased. These results suggest that the inferior frontal areas play an essential role in the flexible shifting of cognitive sets.

Recent biochemical and immunohistochemical studies have shown that the opioid peptides, enkephalins, occur in nerve terminals and cell bodies in mammalian sympathetic ganglia1-3. Opiates and enkephalins are thought to inhibit synaptic transmission in the peripheral nervous tissues as well as in the central nervous system4-12. The mechanisms of the opiate actions, however, are not entirely clear; both pre- and postsynaptic sites of action have been proposed7-9,11,12. As acetylcholine is known to be the major neurotransmitter in the autonomic ganglia and as the mechanism of synaptic transmission is well clarified13, analysis of the peptide action could be more easily but equally usefully carried out in the peripheral synapses than in central synapses. We now report that enkephalins presynaptically inhibit cholinergic transmission in sympathetic ganglia.