Split T7 switch-mediated cell-free protein synthesis system for detecting target nucleic acids.

Cell-free protein synthesis (CFPS) reactions can be used to detect nucleic acids. However, most CFPS systems rely on a toehold switch and exhibit the following critical limitations: (i) off-target signals due to leaky translation in the absence of target nucleic acids, (ii) a suboptimal detection limit of approximately 30 nM without pre-amplification, and (iii) labor-intensive screening processes due to sequence constraints for the target nucleic acids. To overcome these shortcomings, we developed a new split T7 switch-mediated CFPS system in which the split T7 promoter was applied to a three-way junction structure to selectively initiate transcription-translation only in the presence of target nucleic acids. Both fluorescence and colorimetric detection systems were constructed by employing different reporter proteins. Notably, we introduced the self-complementation of split fluorescent proteins to streamline preparation of the proposed system, enabling versatile applications. Operation of this one-pot approach under isothermal conditions enabled the detection of target nucleic acids at concentrations as low as 10 pM, representing more than a thousand times improvement over previous toehold switch-based approaches. Furthermore, the proposed system demonstrated high specificity in detecting target nucleic acids and compatibility with various reporter proteins encoded in the expression region. By eliminating issues associated with the previous toehold switch system, our split T7 switch-mediated CFPS system could become a core platform for detecting various target nucleic acids.

Identification, Characterization, and Electronic Structures of Interconvertible Cobalt-Oxygen TAML Intermediates.

The reaction of Li[(TAML)CoIII]·3H2O (TAML = tetraamido macrocyclic tetraanionic ligand) with iodosylbenzene at 253 K in acetone in the presence of redox-innocent metal ions (Sc(OTf)3 and Y(OTf)3) or triflic acid affords a blue species 1, which is converted reversibly to a green species 2 upon cooling to 193 K. The electronic structures of 1 and 2 have been determined by combining advanced spectroscopic techniques (X-band electron paramagnetic resonance (EPR), electron nuclear double resonance (ENDOR), X-ray absorption spectroscopy/extended X-ray absorption fine structure (XAS/EXAFS), and magnetic circular dichroism (MCD)) with ab initio theoretical studies. Complex 1 is best represented as an S = 1/2 [(Sol)(TAML•+)CoIII---OH(LA)]- species (LA = Lewis/Brønsted acid and Sol = solvent), where an S = 1 Co(III) center is antiferromagnetically coupled to S = 1/2 TAML•+, which represents a one-electron oxidized TAML ligand. In contrast, complex 2, also with an S = 1/2 ground state, is found to be multiconfigurational with contributions of both the resonance forms [(H-TAML)CoIV═O(LA)]- and [(H-TAML•+)CoIII═O(LA)]-; H-TAML and H-TAML•+ represent the protonated forms of TAML and TAML•+ ligands, respectively. Thus, the interconversion of 1 and 2 is associated with a LA-associated tautomerization event, whereby H+ shifts from the terminal -OH group to TAML•+ with the concomitant formation of a terminal cobalt-oxo species possessing both singlet (SCo = 0) Co(III) and doublet (SCo = 1/2) Co(IV) characters. The reactivities of 1 and 2 at different temperatures have been investigated in oxygen atom transfer (OAT) and hydrogen atom transfer (HAT) reactions to compare the activation enthalpies and entropies of 1 and 2.

Ynamide and Azaalleneyl. Acid-Base Promoted Chelotropic and Spin-State Rearrangements in a Versatile Heterocumulene [(Ad)NCC(tBu)].

We introduce the heterocumulene ligand [(Ad)NCC(tBu)]- (Ad=1-adamantyl (C10H15), tBu=tert-butyl, (C4H9)), which can adopt two forms, the azaalleneyl and ynamide. This ligand platform can undergo a reversible chelotropic shift using Brønsted acid-base chemistry, which promotes an unprecedented spin-state change of the [VIII] ion. These unique scaffolds are prepared via addition of 1-adamantyl isonitrile (C≡NAd) across the alkylidyne in complexes [(BDI)V≡CtBu(OTf)] (A) (BDI-=ArNC(CH3)CHC(CH3)NAr), Ar=2,6-iPr2C6H3) and [(dBDI)V≡CtBu(OEt2)] (B) (dBDI2-=ArNC(CH3)CHC(CH2)NAr). Complex A reacts with C≡NAd, to generate the high-spin [VIII] complex with a κ1-N-ynamide ligand, [(BDI)V{κ1-N-(Ad)NCC(tBu)}(OTf)] (1). Conversely, B reacts with C≡NAd to generate a low-spin [VIII] diamagnetic complex having a chelated κ2-C,N-azaalleneyl ligand, [(dBDI)V{κ2-N,C-(Ad)NCC(tBu)}] (2). Theoretical studies have been applied to better understand the mechanism of formation of 2 and the electronic reconfiguration upon structural rearrangement by the alteration of ligand denticity between 1 and 2.

Analysis of production efficiency of cloned transgenic Yucatan miniature pigs according to recipient breeds with embryo transfer conditions.

The purpose of this study was to compare the efficiency of the production of cloned transgenic Yucatan miniature pigs (YMPs) using two recipient breeds, i.e., YMPs and domestic pigs (DPs), under various embryo transfer conditions. We initially assessed the in vitro developmental competence of embryos obtained via somatic cell nuclear transfer (SCNT) from three different transgenic donor cells. No difference was observed among the three groups regarding developmental competence. Furthermore, the cloning efficiency remained consistent among the three groups after the transfer of the SCNT embryos to each surrogate mother. Subsequently, to compare the efficiency of the production of cloned transgenic YMPs between the two recipient breeds using varying parameters, including ovulation status (preovulation and postovulation), duration of in vitro culture (IVC) (incubated within 24 h and 24-48 h), and the number of transferred SCNT embryos (less than and more than 300), we assessed the pregnancy rates, delivery rates, mean offspring counts, and cloning efficiency. Regarding the ovulation status, YMPs exhibited higher pregnancy rates, delivery rates, and cloning efficiency compared with DPs in both statuses. Moreover, the pregnancy rates, delivery rates, and cloning efficiency were affected by the ovulation status in DPs, but not in YMPs. The comparison of IVC duration between groups revealed that YMPs had higher pregnancy rates vs. DPs in both conditions. SCNT embryos cultured for 24-48 h in YMPs yielded higher delivery rates and cloning efficiency compared with those cultured for less than 24 h in DPs. Finally, the analysis based on the number of transferred SCNT embryos showed that both the pregnancy and delivery rates were higher in YMPs vs. DPs. However, the highest average number of offspring was obtained when more than 300 SCNT embryos were transferred into DPs, whereas the cloning efficiency was higher in YMPs vs. DPs. Our results suggest that YMPs are more suitable recipients than are DPs under various conditions for the production of cloned transgenic YMPs.

An open-source, 3D printed inkjet DNA synthesizer.

Synthetic oligonucleotides have become a fundamental tool in a wide range of biological fields, including synthetic biology, biosensing, and DNA storage. Reliable access to equipment for synthesizing high-density oligonucleotides in the laboratory ensures research security and the freedom of research expansion. In this study, we introduced the Open-Source Inkjet DNA Synthesizer (OpenIDS), an open-source inkjet-based microarray synthesizer that offers ease of construction, rapid deployment, and flexible scalability. Utilizing 3D printing, Arduino, and Raspberry Pi, this newly designed synthesizer achieved robust stability with an industrial inkjet printhead. OpenIDS maintains low production costs and is therefore suitable for self-fabrication and optimization in academic laboratories. Moreover, even non-experts can create and control the synthesizer with a high degree of freedom for structural modifications. Users can easily add printheads or alter the design of the microarray substrate according to their research needs. To validate its performance, we synthesized oligonucleotides on 144 spots on a 15 × 25-mm silicon wafer filled with controlled pore glass. The synthesized oligonucleotides were analyzed using urea polyacrylamide gel electrophoresis.

Problem Solving MRI to Reduce False-Positive Biopsy Related to Breast US: Conductivity vs. DWI vs. Abbreviated Contrast-Enhanced MRI.

BACKGROUND: While breast ultrasound (US) is a useful tool for diagnosing breast masses, it can entail false-positive biopsy results because of some overlapping features between benign and malignant breast masses and subjective interpretation. PURPOSE: To evaluate the performance of conductivity imaging for reducing false-positive biopsy results related to breast US, as compared to diffusion-weighted imaging (DWI) and abbreviated MRI consisting of one pre- and one post-contrast T1-weighted imaging. STUDY TYPE: Prospective. SUBJECTS: Seventy-nine women (median age, 44 years) with 86 Breast Imaging Reporting and Data System (BI-RADS) category 4 masses as detected by breast US. FIELD STRENGTH/SEQUENCE: 3-T, T2-weighted turbo spin echo sequence, DWI, and abbreviated contrast-enhanced MRI (T1-weighted gradient echo sequence). ASSESSMENT: US-guided biopsy (reference standard) was obtained on the same day as MRI. The maximum and mean conductivity parameters from whole and single regions of interest (ROIs) were measured. Apparent diffusion coefficient (ADC) values were obtained from an area with the lowest signal within a lesion on the ADC map. The performance of conductivity, ADC, and abbreviated MRI for reducing false-positive biopsies was evaluated using the following criteria: lowest conductivity and highest ADC values among malignant breast lesions and BI-RADS categories 2 or 3 on abbreviated MRI. STATISTICAL TESTS: One conductivity parameter with the maximum area under the curve (AUC) from receiver operating characteristics was selected. A P-value <0.05 was considered statistically significant. RESULTS: US-guided biopsy revealed 65 benign lesions and 21 malignant lesions. The mean conductivity parameter of the single ROI method was selected (AUC = 0.74). Considering conductivity (≤0.10 S/m), ADC (≥1.60 × 10-3 mm2 /sec), and BI-RADS categories 2 or 3 reduced false-positive biopsies by 23% (15 of 65), 38% (25 of 65), and 43% (28 of 65), respectively, without missing malignant lesions. DATA CONCLUSION: Conductivity imaging may show lower performance than DWI and abbreviated MRI in reducing unnecessary biopsies. EVIDENCE LEVEL: 2 TECHNICAL EFFICACY: Stage 2.

Deep neural network-based phase calibration in integrated optical phased arrays.

Calibrating the phase in integrated optical phased arrays (OPAs) is a crucial procedure for addressing phase errors and achieving the desired beamforming results. In this paper, we introduce a novel phase calibration methodology based on a deep neural network (DNN) architecture to enhance beamforming in integrated OPAs. Our methodology focuses on precise phase control, individually tailored to each of the 64 OPA channels, incorporating electro-optic phase shifters. To effectively handle the inherent complexity arising from the numerous voltage set combinations required for phase control across the 64 channels, we employ a tandem network architecture, further optimizing it through selective data sorting and hyperparameter tuning. To validate the effectiveness of the trained DNN model, we compared its performance with 20 reference beams obtained through the hill climbing algorithm. Despite an average intensity reduction of 0.84 dB in the peak values of the beams compared to the reference beams, our experimental results demonstrate substantial agreements between the DNN-predicted beams and the reference beams, accompanied by a slight decrease of 0.06 dB in the side-mode-suppression-ratio. These results underscore the practical effectiveness of the DNN model in OPA beamforming, highlighting its potential in scenarios that necessitate the intelligent and time-efficient calibration of multiple beams.

Data-driven electrical conductivity brain imaging using 3 T MRI.

Magnetic resonance electrical properties tomography (MR-EPT) is a non-invasive measurement technique that derives the electrical properties (EPs, e.g., conductivity or permittivity) of tissues in the radiofrequency range (64 MHz for 1.5 T and 128 MHz for 3 T MR systems). Clinical studies have shown the potential of tissue conductivity as a biomarker. To date, model-based conductivity reconstructions rely on numerical assumptions and approximations, leading to inaccuracies in the reconstructed maps. To address such limitations, we propose an artificial neural network (ANN)-based non-linear conductivity estimator trained on simulated data for conductivity brain imaging. Network training was performed on 201 synthesized T2-weighted spin-echo (SE) data obtained from the finite-difference time-domain (FDTD) electromagnetic (EM) simulation. The dataset was composed of an approximated T2-w SE magnitude and transceive phase information. The proposed method was tested three in-silico and in-vivo on two volunteers and three patients' data. For comparison purposes, various conventional phase-based EPT reconstruction methods were used that ignore B 1 + magnitude information, such as Savitzky-Golay kernel combined with Gaussian filter (S-G Kernel), phase-based convection-reaction EPT (cr-EPT), magnitude-weighted polynomial-fitting phase-based EPT (Poly-Fit), and integral-based phase-based EPT (Integral-based). From the in-silico experiments, quantitative analysis showed that the proposed method provides more accurate and improved quality (e.g., high structural preservation) conductivity maps compared to conventional reconstruction methods. Representatively, in the healthy brain in-silico phantom experiment, the proposed method yielded mean conductivity values of 1.97 ± 0.20 S/m for CSF, 0.33 ± 0.04 S/m for WM, and 0.52 ± 0.08 S/m for GM, which were closer to the ground-truth conductivity (2.00, 0.30, 0.50 S/m) than the integral-based method (2.56 ± 2.31, 0.39 ± 0.12, 0.68 ± 0.33 S/m). In-vivo ANN-based conductivity reconstructions were also of improved quality compared to conventional reconstructions and demonstrated network generalizability and robustness to in-vivo data and pathologies. The reported in-vivo brain conductivity values were in agreement with literatures. In addition, the proposed method was observed for various SNR levels (SNR levels = 10, 20, 40, and 58) and repeatability conditions (the eight acquisitions with the number of signal averages = 1). The preliminary investigations on brain tumor patient datasets suggest that the network trained on simulated dataset can generalize to unforeseen in-vivo pathologies, thus demonstrating its potential for clinical applications.

Effects of extracellular vesicles derived from steroids-primed oviductal epithelial cells on porcine in vitro embryonic development.

Extracellular vesicles (EVs) play an active role in regulating different physiological events, however, endocrine control of EVs cargo contents remain poorly understood. In this study, we aimed to isolate EVs from the porcine oviductal epithelial cells (POECs) that were primed with steroid hormones including estradiol (E2) and progesterone (P4), mimicking the in vivo conditions of the reproductive cycle and studied their effects on in vitro produced embryonic development. For this purpose, POECs were treated either with 0 concentration (control) or two different combinations of E2 and P4 including 50 pg/mL E2 + 0.5 ng/mL P4 (group H1), and 10 pg/mL E2 + 35 ng/mL P4 (group H2). Embryos were prepared after in vitro maturation either by parthenogenetic activation or somatic cell nuclear transfer (SCNT) technique. Treating parthenogenetic embryo with EVs, led a significantly higher rate of the blastocyst formation in the group supplemented with each EVs, compared to the control group. In addition, TUNEL assay and gene expression level analysis revealed that apoptosis was significantly reduced in the H2 EVs group. Furthermore, EVs from hormone-primed POECs improved the formation rate of porcine SCNT embryos compared to the control group. While in each EVs supplemented group (control EVs, H1 EVs, H2 EVs), the expression of cell reprogramming-related genes in cloned embryos showed a tendency of increase, the effect was stronger in H1 EVs and H2 EVs. In conclusion, EVs derived from POECs cultured in hormonal conditions simulating the in vivo environment had a positive effect on porcine blastocysts formation, which will likely facilitate in the production of cloned embryos.

Correlation between standardized uptake value of 18F-FDG PET/CT and conductivity with pathologic prognostic factors in breast cancer.

We investigated the correlation between standardized uptake value (SUV) of 18F-fluorodeoxyglucose (FDG) positron emission tomography (PET)/computed tomography (CT) and conductivity parameters in breast cancer and explored the feasibility of conductivity as an imaging biomarker. Both SUV and conductivity have the potential to reflect the tumors' heterogeneous characteristics, but their correlations have not been investigated until now. Forty four women diagnosed with breast cancer who underwent breast MRI and 18F-FDG PET/CT at the time of diagnosis were included. Among them, 17 women received neoadjuvant chemotherapy followed by surgery and 27 women underwent upfront surgery. For conductivity parameters, maximum and mean values of the tumor region-of-interests were examined. For SUV parameters, SUVmax, SUVmean, and SUVpeak of the tumor region-of-interests were examined. Correlations between conductivity and SUV were evaluated, and among them, the highest correlation was observed between mean conductivity and SUVpeak (Spearman's correlation coefficient = 0.381). In a subgroup analysis for 27 women with upfront surgery, tumors with lymphovascular invasion (LVI) showed higher mean conductivity than those without LVI (median: 0.49 S/m vs 0.06 S/m, p < 0.001). In conclusion, our study shows a low positive correlation between SUVpeak and mean conductivity in breast cancer. Furthermore, conductivity showed a potential to noninvasively predict LVI status.

Trogocytic molting of T cell microvilli upregulates T cell receptor surface expression and promotes clonal expansion.

Although T cell activation is known to involve the internalization of the T cell antigen receptor (TCR), much less is known regarding the release of TCRs following T cell interaction with cognate antigen-presenting cells. In this study, we examine the physiological mechanisms underlying TCR release following T cell activation. We show that T cell activation results in the shedding of TCRs in T cell microvilli, which involves a combined process of trogocytosis and enzymatic vesiculation, leading to the loss of membrane TCRs and microvilli-associated proteins and lipids. Surprisingly, unlike TCR internalization, this event results in the rapid upregulation of surface TCR expression and metabolic reprogramming of cholesterol and fatty acid synthesis to support cell division and survival. These results demonstrate that TCRs are lost through trogocytic 'molting' following T cell activation and highlight this mechanism as an important regulator of clonal expansion.

Peli3 ablation ameliorates acetaminophen-induced liver injury through inhibition of GSK3ß phosphorylation and mitochondrial translocation.

The signaling pathways governing acetaminophen (APAP)-induced liver injury have been extensively studied. However, little is known about the ubiquitin-modifying enzymes needed for the regulation of APAP-induced liver injury. Here, we examined whether the Pellino3 protein, which has E3 ligase activity, is needed for APAP-induced liver injury and subsequently explored its molecular mechanism. Whole-body Peli3-/- knockout (KO) and adenovirus-mediated Peli3 knockdown (KD) mice showed reduced levels of centrilobular cell death, infiltration of immune cells, and biomarkers of liver injury, such as alanine aminotransferase (ALT) and aspartate aminotransferase (AST), upon APAP treatment compared to wild-type (WT) mice. Peli3 deficiency in primary hepatocytes decreased mitochondrial and lysosomal damage and reduced the mitochondrial reactive oxygen species (ROS) levels. In addition, the levels of phosphorylation at serine 9 in the cytoplasm and mitochondrial translocation of GSK3ß were decreased in primary hepatocytes obtained from Peli3-/- KO mice, and these reductions were accompanied by decreases in JNK phosphorylation and mitochondrial translocation. Pellino3 bound more strongly to GSK3ß compared with JNK1 and JNK2 and induced the lysine 63 (K63)-mediated polyubiquitination of GSK3ß. In rescue experiments, the ectopic expression of wild-type Pellino3 in Peli3-/- KO hepatocytes restored the mitochondrial translocation of GSK3ß, but this restoration was not obtained with expression of a catalytically inactive mutant of Pellino3. These findings are the first to suggest a mechanistic link between Pellino3 and APAP-induced liver injury through the modulation of GSK3ß polyubiquitination.

Demonstration of high-accuracy 3D imaging using a Si optical phased array with a tunable radiator.

Precise imaging in three-dimension (3D) is an essential technique for solid-state light detection and ranging (LiDAR). Among various solid-state LiDAR technologies, silicon (Si) optical phased array (OPA)-based LiDAR has the significant advantage of robust 3D imaging due to its high scanning speed, low power consumption, and compactness. Numerous techniques employing a Si OPA have utilized two-dimensional arrays or wavelength tuning for longitudinal scanning but the operation of those systems is restricted by additional requirements. Here, we demonstrate high-accuracy 3D imaging using a Si OPA with a tunable radiator. As we adapted a time-of-flight approach for distance measurement, we have developed an optical pulse modulator that allows a ranging accuracy of less than 2 cm. The implemented Si OPA is composed of an input grating coupler, multimode interferometers, electro-optic p-i-n phase shifters, and thermo-optic n-i-n tunable radiators. With this system, it is possible to attain a wide beam steering range of 45° in a transversal angle with a 0.7° divergence angle, and 10° in a longitudinal angle with a 0.6° divergence angle can be achieved using Si OPA. The character toy model was successfully imaged in three dimensions with a range resolution of 2 cm using the Si OPA. The further improvement of each component of the Si OPA will allow even more accurate 3D imaging over a longer distance.

dDrop-Chip: disposable film-chip microfluidic device for real-time droplet feedback control.

A cost-effective, simple to use, and automated technique that can provide real-time feedback control for droplet generation is required to obtain droplets with high-throughput, stability, and uniformity. This study introduces a disposable droplet generation microfluidic device (dDrop-Chip) that can simultaneously control both droplet size and production rate in real time. The dDrop-Chip consists of a reusable sensing substrate and a disposable microchannel that can be assembled using vacuum pressure. It also integrates a droplet detector and a flow sensor on-chip, enabling real-time measurement and feedback control of droplet size and sample flow rate. The dDrop-Chip has the additional advantage of being disposable, which can prevent chemical and biological contamination, due to low manufacturing cost by the film-chip technique. We demonstrate benefits of the dDrop-Chip by controlling droplet size at a fixed sample flow rate and the production rate at a fixed droplet size using real-time feedback control. The experimental results show that the dDrop-Chip consistently generates monodisperse droplets with a length of 219.36 ± 0.08 µm (CV 0.036%) at a production rate of 32.38 ± 0.48 Hz using the feedback control, while without feedback control, there is a significant deviation in droplet length (224.18 ± 6.69 µm, CV 2.98%) and production rate (33.94 ± 1.72 Hz) despite the use of identical devices. Therefore, the dDrop-Chip is a reliable, cost-effective, and automated technique for generating droplets of controlled size and production rate in real time, making it suitable for various droplet-based applications.

Fractional Charge Density Functional Theory and Its Application to the Electro-inductive Effect.

We employed the chemical potential equalization principle to demonstrate that fractional electrons are involved in the electro-inductive effect as well as the vibrational Stark effect. By the chemical potential model, we were able to deduce that the frontier molecular orbitals of immobilized molecules can provide valuable insight into these effects. To further understand and quantify these findings, we introduced fractional charge density functional theory (FC-DFT), a canonical ensemble approach for open systems. This method allows for the calculation of electronic energies, nuclear gradients, and the Hessian matrix of fractional electronic systems. To correct the spurious delocalization error commonly found in approximate density functionals for small systems, we imposed the Perdew-Parr-Levy-Balduz (PPLB) condition through linear interpolation of two adjacent integer points (LI-FC-DFT). Although this approach is relatively simple in terms of molecular modeling, the results obtained through LI-FC-DFT calculations predict the same trend seen in experimental reactivity and the frequency change of immobilized molecules.

Inverse design of an on-chip optical response predictor enabled by a deep neural network.

We proposed inverse-designed nanophotonic waveguide devices which have the desired optical responses in the wide band of 1450-1650 nm. The proposed devices have an ultra-compact size of just 1.5 µm × 3.0 µm and are designed on a silicon-on-insulator (SOI) waveguide platform. Individual nano-pixels with dimensions of 150 nm × 150 nm were made of either silicon or silicon dioxide, and the materials for the 200 total cells were determined using a trained deep neural network. While training the two networks, the hyperparameter optimization method was applied to make the training process efficient. We then fabricated the proposed devices using a CMOS-compatible fabrication process, and experimentally verified the fabricated device performance.

The Role of the Redox Non-Innocent Hydroxyl Ligand in the Activation of O2 Performed by [Ni(H)(OH)].

The cationic complex [Ni(H)(OH)]+ was previously found to activate dioxygen and methane in gas phase under single collision conditions. These remarkable reactivities were thought to originate from a non-classical electronic structure, where the Ni-center adopts a Ni(II), instead of the classically expected Ni(III) oxidation state by formally accepting an electron from the hydroxo ligand, which formally becomes a hydroxyl radical in the process. Such radicaloid oxygen moieties are envisioned to easily react with otherwise inert substrates, mimicking familiar reactivities of free radicals. In this study, the reductive activation of dioxygen by [Ni(H)(OH)]+ to afford the hydroperoxo species was investigated using coupled cluster, multireference ab initio and density functional theory calculations. Orbital and wave function analyses indicate that O2 binding tranforms the aforementioned non-classical electronic structure to a classical Ni(III)-hydroxyl system, before O2 reduction takes place. Remarkably, we found no evidence for a direct involvement of the radicaloid hydroxyl in the reaction with O2 , as is often assumed. The function of the redox non-innocent character of the activator complex is to protect the reactive electronic structure until the complex engages O2 , upon which a dramatic electronic reorganization releases internal energy and drives the chemical reaction to completion.

Technical note: Multi-receiver combination method for phase-based electrical property tomography of the breast.

BACKGROUND: Phase-based electrical property tomography (EPT) is a technique that allows conductivity reconstruction with only phase of the B1 field under the assumption that the magnitude of the B1 fields are homogeneous. The more this assumption is violated, the less accurate the reconstructed conductivity. Thus, a method that ensures homogeneity of | B 1 - | $| {{\rm{B}}_1^ - } |$ field is important for breast image using multi-receiver coil. PURPOSE: To develop a method for multi-receiver combination for phase-based EPT usable for breast EPT imaging in the clinic. METHODS: Theory of the proposed method is presented. To validate the proposed method, the phantom and in-vivo experiments were conducted. Conductivity images were reconstructed using the transceive phase of the combined image and results were compared with another combination method. RESULTS: The proposed method's conductivity results were more stable than those of the previous method when | B 1 + | $| {{\rm{B}}_1^ + } |$ was not homogeneous and when the homogeneous contrast region was small. The phantom and in-vivo results indicate that the proposed method produces improved conductivity images than the previous method. The proposed combination method also increased the conductivity contrast between benign and cancerous tissues. CONCLUSION: The proposed method produced more stable multi-receiver combination for phase-based EPT of the breast in a clinical environment.

Performance Assessment of Carbon Dioxide Sequestration in Cement Composites with a Granulation Technique.

The cement industry emits a significant amount of carbon dioxide (CO2). Therefore, the cement industry should recycle the emitted CO2. However, sequestration by carbonation in cement composites absorbs a very small amount of CO2. Therefore, a direct way of achieving this is to improve the absorption performance of CO2 in cement composites. In this study, to improve absorption, unlike in existing studies, a granulation technique was applied, and the material used was calcium hydroxide (CH). In addition, granulated CH was coated to prevent a reaction during the curing of cement paste. The coated CH granule (CCHG) was applied to 5% of the cement weight as an additive material, and the specimens were cured for 91 days to wait for the coating of CCHG to fully phase-change. The experiment of CO2 absorption showed an unexpected result, where the use of blast furnace slag (BFS) and fly ash (FA) had a negative effect on CO2 sequestration. This was because BFS and FA had a filler effect in the cement matrix, and the filler effect caused the blocking of the path of CO2. In addition, BFS and FA are well-known pozzolanic materials; the pozzolan reaction caused a reduction in the amount of CH because the pozzolan reaction consumed the CH to produce a calcium silicate hydrate. Therefore, the pozzolan reaction also had a negative effect on the CO2 sequestration performance combined with the filler effect. The CO2 sequestration efficiency was decreased between ordinary cement paste and BFS-applied specimens by 45.45%. In addition, compared to cases of ordinary cement paste and FA-applied specimens, the CO2 sequestration performance was decreased by 63.64%. Comprehensively, CO2 sequestration performance depends on the porosity and amount of CH.

An Improved In Vitro Blood-Brain Barrier Model for the Evaluation of Drug Permeability Using Transwell with Shear Stress.

The development of drugs targeting the central nervous system (CNS) is challenging because of the presence of the Blood-Brain barrier (BBB). Developing physiologically relevant in vitro BBB models for evaluating drug permeability and predicting the activity of drug candidates is crucial. The transwell model is one of the most widely used in vitro BBB models. However, this model has limitations in mimicking in vivo conditions, particularly in the absence of shear stress. This study aimed to overcome the limitations of the transwell model using immortalized human endothelial cells (hCMEC/D3) by developing a novel dish design for an orbital shaker, providing shear stress. During optimization, we assessed cell layer integrity using trans-endothelial electrical resistance measurements and the % diffusion of lucifer yellow. The efflux transporter activity and mRNA expression of junctional proteins (claudin-5, occludin, and VE-cadherin) in the newly optimized model were verified. Additionally, the permeability of 14 compounds was evaluated and compared with published in vivo data. The cell-layer integrity was substantially increased using the newly designed annular shaking-dish model. The results demonstrate that our model provided robust conditions for evaluating the permeability of CNS drug candidates, potentially improving the reliability of in vitro BBB models in drug development.