Inhibition of Autophagy Aggravates Arachis hypogaea L. Skin Extracts-Induced Apoptosis in Cancer Cells.

The skin of Arachis hypogaea L. (peanut or groundnut) is a rich source of polyphenols, which have been shown to exhibit a wider spectrum of noteworthy biological activities, including anticancer effects. However, the anticancer activity of peanut skin extracts against melanoma and colorectal cancer (CRC) cells remains elusive. In this study, we systematically investigated the cytotoxic, antiproliferative, pro-apoptotic, and anti-migration effects of peanut skin ethanolic extract and its fractions on melanoma and CRC cells. Cell viability results showed that the ethyl acetate fraction (AHE) of peanut skin ethanolic crude extract and one of the methanolic fractions (AHE-2) from ethyl acetate extraction exhibited the highest cytotoxicity against melanoma and CRC cells but not in nonmalignant human skin fibroblasts. AHE and AHE-2 effectively modulated the cell cycle-related proteins, including the suppression of cyclin-dependent kinase 4 (CDK4), cyclin-dependent kinase 6 (CDK6), phosphorylation of Retinoblastoma (p-Rb), E2F1, Cyclin A, and activation of tumor suppressor p53, which was associated with cell cycle arrest and paralleled their antiproliferative efficacies. AHE and AHE-2 could also induce caspase-dependent apoptosis and inhibit migration activities in melanoma and CRC cells. Moreover, it is noteworthy that autophagy, manifested by microtubule-associated protein light chain 3B (LC3B) conversion and the aggregation of GFP-LC3, was detected after AHE and AHE-2 treatment and provided protective responses in cancer cells. Significantly, inhibition of autophagy enhanced AHE- and AHE-2-induced cytotoxicity and apoptosis. Together, these findings not only elucidate the anticancer potential of peanut skin extracts against melanoma and CRC cells but also provide a new insight into autophagy implicated in peanut skin extracts-induced cancer cell death.

Emergence and dissemination of multidrug-resistant Escherichia coli ST8346 coharboring blaNDM-5 and blaOXA-181 in Southern Taiwan, 2017-2021.

BACKGROUND: Enterobacterales carrying blaNDM represent an emerging challenge in treating infectious diseases. In this study, we aimed to investigate the characteristics of blaNDM-producing Enterobacterales from three hospitals in southern Taiwan. METHODS: Enterobacterales strains that were nonsusceptible to more than one carbapenem (ertapenem, meropenem, imipenem, or doripenem) were collected from hospitalized patients. Molecular typing for New Delhi metallo-ß-lactamase (NDM) and antibiotic susceptibility tests were performed, followed by multilocus sequence typing (MLST), pulsed-field gel electrophoresis (PFGE), and plasmid analysis by PCR-based replicon typing. RESULTS: A total of 1311 carbapenem-nonsusceptible Enterobacterales were isolated from 2017 to 2021. blaNDM-encoding genes were detected in 108 isolates, with 53 (49.1%) harboring blaNDM-1 and 55 (50.9%) harboring blaNDM-5. The rate of blaNDM-1 detection among isolates decreased to 2% in 2021. However, the rate of E. coli harboring blaNDM-5 increased from 1% to 12% of total isolates during the study period. Of 47 NDM-5-positive E. coli isolates, 44 (93.6%) were ST8346 with high genetic relatedness. E. coli ST8346 isolates showed high-level resistance to both carbapenems and aminoglycosides. Most (38 out of 47, 80.9%) blaNDM-5-harboring E. coli isolates co-harbored blaOXA-181. We analyzed the regions harboring blaNDM-5 in E. coli ST8346 via PCR amplification. blaNDM-5 and blaOXA-181 were located on two separate plasmids, IncF and IncX3, respectively. CONCLUSION: The dissemination of E. coli ST8346 caused an increase in blaNDM-5 and blaOXA-181 co-harboring Enterobacterales in southern Taiwan, which show high-level resistance to both carbapenems and aminoglycosides. We identified a distinct IncF plasmid encoding blaNDM-5 that has the potential for rapid spread and needs further surveillance.

Antimicrobial Susceptibility of E. coli Isolates from Intra-Abdominal Infections in the Asia-Pacific Region: Trends in Ciprofloxacin, Ceftriaxone, Cefepime, and Piperacillin/Tazobactam Susceptibility.

Purpose: To investigate the antibiotic susceptibility of Escherichia coli isolates in patients diagnosed with intra-abdominal infections (IAIs) in the Asia-Pacific region. Patients and Methods: This study was conducted at 50 medical hospitals across 9 countries/regions as part of the Study for Monitoring Antimicrobial Resistance Trends (SMART) surveillance program from 2014 to 2018. Nonduplicate isolates of aerobic and facultative gram-negative bacilli were collected and processed for further antimicrobial susceptibility testing. Results: A total of 10,709 isolates were collected, with E. coli (n=4737, 44.2%) being the leading pathogen causing IAIs, followed by Klebsiella pneumoniae (n=2429, 22.7%) and Pseudomonas aeruginosa (n=931, 8.7%). Community-associated (CA) E. coli isolates generally exhibited higher susceptibility rates for most antibiotics than hospital-associated (HA) isolates. In countries/regions other than Hong Kong, South Korea, and Singapore, HA isolates displayed lower susceptibility rates for multiple classes (≥4) of antibiotics. Among the commonly used antibiotics in IAIs, the overall susceptibility rate for ciprofloxacin was low, with an average of 41.3%. Ceftriaxone susceptibility rates in all selected countries were below 80% starting in 2018, ranging from 23.3% to 75.8%. The cefepime susceptibility rates varied across regions, with consistently reduced susceptibility ranging from 45.5% to 57.8% in India, Thailand, and Vietnam. Piperacillin/tazobactam demonstrated effectiveness against E. coli isolates in almost all countries except India, with a downward trend observed in the Philippines and Taiwan. Carbapenems remained effective against more than 90% of E. coli isolates, except in India. Conclusion: Prudent use of fluoroquinolones and ceftriaxone is advised when treating both CA and HA IAIs in the Asia-Pacific region. The low susceptibility rate of cefepime in India, Thailand, and Vietnam needs careful consideration in its administration. Moreover, the increase in nonsusceptibility to piperacillin/tazobactam in the Philippines and Taiwan poses a potential risk that should be closely monitored.

In vitro activities of antimicrobial combinations against planktonic and biofilm forms of Stenotrophomonas maltophilia.

Objectives: To investigate the in vitro activity of antibiotic combinations against Stenotrophomonas maltophilia isolates and their associated biofilms. Methods: Thirty-two S. maltophilia clinical isolates with at least twenty-five different pulsotypes were tested. The antibacterial activity of various antibiotic combinations against seven randomly selected planktonic and biofilm-embedded S. maltophilia strains with strong biofilm formation was assessed using broth methods. Extraction of bacterial genomic DNA and PCR detection of antibiotic resistance and biofilm-related genes were also performed. Results: The susceptibility rates of levofloxacin (LVX), fosfomycin (FOS), tigecycline (TGC) and sulfamethoxazole-trimethoprim (SXT) against 32 S. maltophilia isolates were 56.3, 71.9, 71.9 and 90.6%, respectively. Twenty-eight isolates were detected with strong biofilm formation. Antibiotic combinations, including aztreonam-clavulanic (ATM-CLA) with LVX, ceftazidime-avibactam (CZA) with LVX and SXT with TGC, exhibited potent inhibitory activity against these isolates with strong biofilm formation. The antibiotic resistance phenotype might not be fully caused by the common antibiotic-resistance or biofilm-formation gene. Conclusion: S. maltophilia remained resistant to most antibiotics, including LVX and ß-lactam/ß-lactamases; however, TGC, FOS and SXT still exhibited potent activity. Although all tested S. maltophilia isolates exhibited moderate-to-strong biofilm formation, combination therapies, especially ATM-CLA with LVX, CZA with LVX and SXT with TGC, exhibited a higher inhibitory activity for these isolates.

Laparoendoscopic Single-Site Inguinal Herniorrhaphy: Experience of a Single Institute.

Background: Minimally invasive techniques for inguinal herniorrhaphy have focused on developing the laparoendoscopic single-site (LESS) procedure to improve cosmesis. Outcomes of total extraperitoneal (TEP) herniorrhaphy vary considerably because of being performed by different surgeons. We aimed to evaluate the perioperative characteristics and outcomes of patients undergoing the LESS-TEP approach for inguinal herniorrhaphy and to determine its overall safety and effectiveness. Methods: Data of 233 patients who underwent 288 laparoendoscopic single-site total extraperitoneal approach (LESS-TEP) herniorrhaphies at Kaohsiung Chang Gung Memorial Hospital between January 2014 and July 2021 were reviewed retrospectively. We reviewed the experiences and results of LESS-TEP herniorrhaphy performed by a single surgeon (CHC) using homemade glove access and standard laparoscopic instruments with a 50 cm long 30° telescope. Results: Among 233 patients, 178 patients had unilateral hernias and 55 patients had bilateral hernias. About 32% (n = 57) of patients in the unilateral group and 29% (n = 16) of patients in the bilateral group were obese (body mass index ≥ 25). The mean operative time was 66 min for the unilateral group and 100 min for the bilateral group. Postoperative complications occurred in 27 (11%) cases, which were minor morbidities except for one mesh infection. Three (1.2%) cases were converted to open surgery. Comparison of the variables between obese and non-obese patients found no significant differences in operative times or postoperative complications. Conclusion: LESS-TEP herniorrhaphy is a safe and feasible operation with excellent cosmetic results and a low rate of complication, even in obese patients. Further large-scale prospective controlled studies and long-term analyses are needed to confirm these results.

Targeting Triple Negative Breast Cancer Stem Cells by Heat Shock Protein 70 Inhibitors.

Triple negative breast cancer (TNBC) is considered the most aggressive breast cancer with high relapse rates and poor prognosis. Although great advances in the development of cancer therapy have been witnessed over the past decade, the treatment options for TNBC remain limited. In this study, we investigated the effect and potential underlying mechanism of the Hsp70 inhibitors, compound 1 and compound 6, on breast cancer stem cells (BCSCs) in TNBC cells. Our results showed that compound 1 and 6 exhibited potent tumor suppressive effects on cell viability and proliferation, and effectively inhibited BCSC expansion in TNBC cells. Reminiscent with the effect of Hsp70 inhibitors, Hsp70 knockdown effectively suppressed mammosphere formation and the expressions of BCSCs surface markers. Mechanistically, evidence showed that the Hsp70 inhibitors inhibited BCSCs by down-regulating ß-catenin in TNBC cells. Moreover, we used the Hsp70 inhibitors treated TNBC cells and a stable Hsp70 knockdown clone of MDA-MB-231 cells to demonstrate the in vivo efficacy of Hsp70 inhibition in suppressing tumorigenesis and xenograft tumor growth. Together, these findings suggest the potential role of Hsp70 as a target for TNBC therapy and foster new therapeutic strategies to eliminate BCSCs by targeting Hsp70.

Development of minimal physiologically-based pharmacokinetic-pharmacodynamic models for characterizing cellular kinetics of CAR T cells following local deliveries in mice.

Chimeric antigen receptor (CAR) T cell therapies have revolutionized the treatment of hematologic malignancies and have potentials for solid tumor treatment. To overcome limited CAR T cell infiltration to solid tumors, local delivery of CAR T cells is a practical strategy that has shown promising therapeutic outcome and safety profile in the clinic. It is of great interest to understand the impact of dosing routes on CAR T cell distribution, subsequent proliferation and tumor killing in a quantitative manner to identify key factors that contribute to CAR T efficacy and safety. In this study, we established mouse minimal physiologically-based pharmacokinetic (mPBPK) models combined with pharmacodynamic (PD) components to delineate CAR T cell distribution, proliferation, tumor growth, and tumor cell killing in the cases of pleural and liver tumors. The pleural tumor model reasonably captured published CAR T cellular kinetic and tumor growth profiles in mice. The mPBPK-PD simulation of a liver tumor mouse model showed a substantial increase in initial tumor infiltration and earlier CAR T cell proliferation with local hepatic artery delivery compared to portal vein and intravenous (i.v.) injections whereas portal vein injection showed little difference from i.v. administration, suggesting the importance of having the injection site close to tumor for maximal effect of non-systemic administration. Blood flow rate in the liver tumor was found to be a sensitive parameter for cellular kinetics and efficacy, indicating a potential role of tumor vascularization in the efficacy of CAR T cell therapies.

Universal Plasma Jet for Droplet Manipulation on a PDMS Surface towards Wall-Less Scaffolds.

Droplet manipulation is important in the fields of engineering, biology, chemistry, and medicine. Many techniques, such as electrowetting and magnetic actuation, have been developed for droplet manipulation. However, the fabrication of the manipulation platform often takes a long time and requires well-trained skills. Here we proposed a novel method that can directly generate and manipulate droplets on a polymeric surface using a universal plasma jet. One of its greatest advantages is that the jet can tremendously reduce the time for the platform fabrication while it can still perform stable droplet manipulation with controllable droplet size and motion. There are two steps for the proposed method. First, the universal plasma jet is set in plasma mode for modifying the manipulation path for droplets. Second, the jet is switched to air-jet mode for droplet generation and manipulation. The jetted air separates and pushes droplets along the plasma-treated path for droplet generation and manipulation. According to the experimental results, the size of the droplet can be controlled by the treatment time in the first step, i.e., a shorter treatment time of plasma results in a smaller size of the droplet, and vice versa. The largest and the smallest sizes of the generated droplets in the results are about 6 µL and 0.1 µL, respectively. Infrared spectra of absorption on the PDMS surfaces with and without the plasma treatment are investigated by Fourier-transform infrared spectroscopy. Tests of generating and mixing two droplets on a PDMS surface are successfully achieved. The aging effect of plasma treatment for the proposed method is also discussed. The proposed method provides a simple, fast, and low-cost way to generate and manipulate droplets on a polymeric surface. The method is expected to be applied to droplet-based cell culture by manipulating droplets encapsulating living cells and towards wall-less scaffolds on a polymeric surface.

Neural Network for Enhancing Microscopic Resolution Based on Images from Scanning Electron Microscope.

In this paper, an artificial neural network is applied for enhancing the resolution of images from an optical microscope based on a network trained with the images acquired from a scanning electron microscope. The resolution of microscopic images is important in various fields, especially for microfluidics because the measurements, such as the dimension of channels and cells, largely rely on visual information. The proposed method is experimentally validated with microfluidic structure. The images of structural edges from the optical microscope are blurred due to optical effects while the images from the scanning electron microscope are sharp and clear. Intensity profiles perpendicular to the edges and the corresponding edge positions determined by the scanning electron microscope images are plugged in a neural network as the input features and the output target, respectively. According to the results, the blurry edges of the microstructure in optical images can be successfully enhanced. The average error between the predicted channel position and ground truth is around 328 nanometers. The effects of the feature length are discussed. The proposed method is expected to significantly contribute to microfluidic applications, such as on-chip cell evaluation.

Grasp to See-Object Classification Using Flexion Glove with Support Vector Machine.

This paper proposes an object classification method using a flexion glove and machine learning. The classification is performed based on the information obtained from a single grasp on a target object. The flexion glove is developed with five flex sensors mounted on five finger sleeves, and is used for measuring the flexion of individual fingers while grasping an object. Flexion signals are divided into three phases, and they are the phases of picking, holding and releasing, respectively. Grasping features are extracted from the phase of holding for training the support vector machine. Two sets of objects are prepared for the classification test. One is printed-object set and the other is daily-life object set. The printed-object set is for investigating the patterns of grasping with specified shape and size, while the daily-life object set includes nine objects randomly chosen from daily life for demonstrating that the proposed method can be used to identify a wide range of objects. According to the results, the accuracy of the classifications are achieved 95.56% and 88.89% for the sets of printed objects and daily-life objects, respectively. A flexion glove which can perform object classification is successfully developed in this work and is aimed at potential grasp-to-see applications, such as visual impairment aid and recognition in dark space.

On-Chip Micro Mixer Driven by Elastic Wall with Virtual Actuator.

In this paper, we propose an on-chip micromixer driven by an elastic wall with a virtual actuator. The on-chip micro mixer is composed of a circular chamber surrounded by a ring-shaped channel under isolation with an elastic wall. When vibrational pressure is put on the driving channel by an actuator, the volume of the circular chamber changes through the deformation of the elastic wall, as if there exists a virtual actuator near the wall. As a result, the liquid in the circular chamber is pushed out and pulled through the neck channel. This action creates a swirling flow in the circular chamber while maintaining isolation from the driving channel. Through experiments, we confirmed the swirling flow under an isolated environment using an air-based valve. The advantage of this approach is that the micromixer can be designed with a single layer having a simple mechanism.

Push/Pull Inequality Based High-Speed On-Chip Mixer Enhanced by Wettability.

In this paper, a high-speed on-chip mixer using two effects is proposed, i.e., push/pull inequality and wettability. Push/pull inequality and wettability are effective for generating a rotational fluid motion in the chamber and for enhancing the rotational speed by reducing the viscous loss between the liquid and channel wall, respectively. An on-chip mixer is composed of three components, a microfluidic channel for making the main fluid flow, a circular chamber connected to the channel for generating a rotational flow, and an actuator connected at the end of the channel allowing a push/pull motion to be applied to the liquid in the main channel. The flow patterns in the chamber under push/pull motions are nonreversible for each motion and, as a result, produce one-directional torque to the fluid in the circular chamber. This nonreversible motion is called push/pull inequality and eventually creates a swirling flow in the chamber. Using hydrophilic treatments, we executed the experiment with a straight channel and a circular chamber to clarify the mixing characteristics at different flow speeds. According to the results, it is confirmed that the swirling velocity under appropriately tuned wettability is 100 times faster than that without tuning.

A Quantitative Systems Pharmacology Model of T Cell Engager Applied to Solid Tumor.

Cancer immunotherapy has recently drawn remarkable attention as promising results in the clinic have shown its ability to improve the overall survival, and T cells are considered to be one of the primary effectors for cancer immunotherapy. Enhanced and restored T cell tumoricidal activity has shown great potential for killing cancer cells. Bispecific T cell engagers (TCEs) are a growing class of molecules that are designed to bind two different antigens on the surface of T cells and cancer cells to bring them in close proximity and selectively activate effector T cells to kill target cancer cells. New T cell engagers are being investigated for the treatment of solid tumors. The activity of newly developed T cell engagers showed a strong correlation with tumor target antigen expression. However, the correlation between tumor-associated antigen expression and overall response of cancer patients is poorly understood. In this study, we used a well-calibrated quantitative systems pharmacology (QSP) model extended to bispecific T cell engagers to explore their efficacy and identify potential biomarkers. In principle, patient-specific response can be predicted through this model according to each patient's individual characteristics. This extended QSP model has been calibrated with available experimental data and provides predictions of patients' response to TCE treatment.

The role of intravesical prostatic protrusion in the evaluation of overactive bladder in male patients with LUTS.

PURPOSE: To evaluate the association of intravesical prostatic protrusion (IPP) and overactive bladder (OAB) in male patients with lower urinary tract symptoms (LUTS). IPP has been suggested to correlate with storage symptoms in addition to bladder outlet obstruction. METHODS: This was an open-labeled, single-center, prospective study involving 128 men older than 40 years presenting with LUTS. We analyzed the relationship of IPP with age, prostate volume, uroflowmetry, post-void residual urine volume (PVR), International Prostate Symptom Score (IPSS), urgency severity scale (USS), and OAB symptom score (OABSS). The patients with an urgency score of ≥ 2 (OABSS question 2) and sum score of ≥ 3 were considered to have OAB. IPP was measured in the mid-sagittal section using transrectal ultrasound. The degree of IPP was classified as grade 1 (≤ 5 mm), grade 2 (> 5-10 mm), and grade 3 (> 10 mm). RESULTS: The mean age of the patients was 64.9 ± 9.2 years, and 101 patients were diagnosed with OAB (79%). Mean IPPs were 2.4 ± 1.4 mm (grade 1, n = 77), 7.6 ± 1.4 mm (grade 2, n = 27), and 14.8 ± 4.4 mm (grade 3, n = 24). IPP was positively correlated with age, prostate size, PSA, PVR, and OABSS nocturia subscore, but not correlated with the presence or severity of OAB. Areas under the receiver-operating characteristic (ROC) curves for the diagnosis of OAB were 0.807 and 0.604 for IPSS-storage subscore and IPP, respectively. CONCLUSION: IPP is not a good predictor of OAB in men presenting with LUTS. However, grade 3 IPP indicates higher frequency of nocturia.

Experimental Study on Microfluidic Mixing with Different Zigzag Angles.

This paper presents experimental investigations of passive mixing in a microfluidic channel with different zigzag angles. Zigzag channel is commonly used for microfluidic mixing because it does not need an additional control unit and can be easily implemented in a lab-on-a-chip system. In this work, microfluidic channels with six different zigzag angles, from θ = 0° to θ = 75°, are tested under ten different flow rates corresponding to Reynolds number from 0.309 to 309. Two colored liquids are mixed with the zigzag channels and mixing performance is evaluated based on the color of the pixels on the region of interest from captured images. According to the results, we found that the mixing performance is almost independent of the zigzag angle in the low-speed regime where its Reynolds number is less than 4. The mixing became very much depending on the zigzag angle in the high-speed regime where its Reynolds number is greater than 100. Microfluidic mixing is needed for Lab-on-a-chip applications in both low flow speed, such as medium perfusion for cell culture, and high flow speed, such as high-speed sensing on a point-of-care device. This work is aimed to provide practical information on zigzag mixing for chip design and applications.

On-Chip Cell Incubator for Simultaneous Observation of Culture with and without Periodic Hydrostatic Pressure.

This paper proposes a microfluidic device which can perform simultaneous observation on cell growth with and without applying periodic hydrostatic pressure (Yokoyama et al. Sci. Rep. 2017, 7, 427). The device is called on-chip cell incubator. It is known that culture with periodic hydrostatic pressure benefits the elasticity of a cultured cell sheet based on the results in previous studies, but how the cells respond to such a stimulus during the culture is not yet clear. In this work, we focused on cell behavior under periodic hydrostatic pressure from the moment of cell seeding. The key advantage of the proposed device is that we can compare the results with and without periodic hydrostatic pressure while all other conditions were kept the same. According to the results, we found that cell sizes under periodic hydrostatic pressure increase faster than those under atmospheric pressure, and furthermore, a frequency-dependent fluctuation of cell size was found using Fourier analysis.

On-chip density mixer enhanced by air chamber.

This paper proposes an on-chip density mixer that can achieve even density in a target chamber with a swirling flow enhanced by an air chamber. The system is composed of a main channel, a target chamber where two liquids with different densities are included, an isolated air chamber, and an external vibration pump driven by a piezo actuator at the entrance of the main channel. The air chamber is expected to amplify the vibration owing to structure softening. The amplification would be more pronounced at the resonance frequencies of the structure. We developed the system and conducted experiments. We showed that the swirling motion in the target chamber with an air chamber is stronger than that without an air chamber. We also confirmed that the time resulting in even density is shorter when the pump is driven at a resonance frequency. An air-based virtual valve is introduced for maintaining a constant density in the target chamber.

Virtual vortex gear: Unique flow patterns driven by microfluidic inertia leading to pinpoint injection.

An interesting phenomenon that vortices are sequentially generated on a microfluidic chip is investigated in this paper. The direction of every two adjacent vortices is opposite to each other, like a set of gears, and thus is named virtual vortex gear (VVG). Both experiments and computational simulations were conducted in order to make clear the mechanism of VVG. The experimental results show that only the flow from a particular point would form vortices and enter the target chamber. A technique of inverse mapping is proposed based on the phenomenon and it demonstrates that only a pinpoint injection is sufficient to control the contents of a microfluidic chamber. VVG can significantly reduce the volume of chemical usage in biological research and has potential for other on-chip applications, such as mixing and valving.

Constrained Adherable Area of Nanotopographic Surfaces Promotes Cell Migration through the Regulation of Focal Adhesion via Focal Adhesion Kinase/Rac1 Activation.

Cell migration is crucial in physiological and pathological processes such as embryonic development and wound healing; such migration is strongly guided by the surrounding nanostructured extracellular matrix. Previous studies have extensively studied the cell migration on anisotropic nanotopographic surfaces; however, only a few studies have reported cell migration on isotropic nanotopographic surfaces. We herein, for the first time, propose a novel concept of adherable area on cell migration using isotropic nanopore surfaces with sufficient nanopore depth by adopting a high aspect ratio. As the pore size of the nanopore surface was controlled to 200, 300, and 400 nm in a fixed center-to-center distance of 480 nm, it produced 86, 68, and 36% of adherable area, respectively, on the fabricated surface. A meticulous investigation of the cell migration in response to changes in the constrained adherable area of the nanotopographic surface showed 1.4-, 1.5-, and 1.6-fold increase in migration speeds and a 1.4-, 2-, and 2.5-fold decrease in the number of focal adhesions as the adherable area was decreased to 86, 68, and 36%, respectively. Furthermore, a strong activation of FAK/Rac1 signaling was observed to be involved in the promoted cell migration. These results suggest that the reduced adherable area promotes cell migration through decreasing the FA formation, which in turn upregulates FAK/Rac1 activation. The findings in this study can be utilized to control the cell migration behaviors, which is a powerful tool in the research fields involving cell migration such as promoting wound healing and tissue repair.

LED-CT Scan for pH Distribution on a Cross-Section of Cell Culture Medium.

In cell culture, the pH of the culture medium is one of the most important conditions. However, the culture medium may have non-uniform pH distribution due to activities of cells and changes in the environment. Although it is possible to measure the pH distribution with an existing pH meter using distributed electrodes, the method involves direct contact with the medium and would greatly increase the risk of contamination. Here in this paper, we propose a computed tomography (CT) scan for measuring pH distribution using the color change of phenol red with a light-emitting diode (LED) light source. Using the principle of CT scan, we can measure pH distribution without contacting culture medium, and thus, decrease the risk of contamination. We have developed the device with a LED, an array of photo receivers and a rotation mechanism. The system is firstly calibrated with different shapes of wooden objects that do not pass light, we succeeded in obtaining their 3D topographies. The system was also used for measuring a culture medium with two different pH values, it was possible to obtain a pH distribution that clearly shows the boundary.