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Dielectrophoresis (DEP) manipulation combined with micro-electric impedance spectroscopy (µEIS) presents a sophisticated approach for cellular analysis and dielectric characterization. While conventional cell analysis techniques rely on complex labeling methods with inherent limitations, integrating DEP and µEIS offers non-invasive, label-free cellular characterization with enhanced sensitivity. This study presents an innovative dual-mode DEP platform incorporating both levitation (LEV DEP ) and rotational (ROT DEP ) forces, integrated with high-precision impedance measurement capabilities on one chip, enabling simultaneous Cell controlling and manipulation and dielectric signature extraction within a single microfluidic device. The fabricated and developed microfluidic platform demonstrated exceptional particle discrimination through the dual mode, with distinct responses for both particle populations. Under F l E V . D E P 10.4 μ m 2.01 MHz showed a 63.4% magnitude increase, while F l E V . D E P 24.9 μ m , particles exhibited a higher 81.2% increase at the same force, yielding a 2.48 × enhancement in discrimination ratio compared to no-DEP conditions. ROT DEP at 110 kHz induced even more pronounced differences, with F R O T . D E P 10.4 μ m showing a 120% magnitude increase (phase patterns: −24.501° to −34.363°) and F R O T . D E P 24.9 μ m µm particles demonstrating a 145% increase (phase patterns: −31.267° to −42.891°), achieving a 3.16 × discrimination ratio enhancement. The impedance spectrum revealed distinct frequency-dependent signatures, with ROT DEP showing superior mid-frequency discrimination (10.4 µm: 1.9370× 10 4 Ω vs 24.9 µm: 2.0542× 10 4 Ω at 110 kHz) and LEV DEP optimizing high-frequency characterization (10.4 µm: 1.6677× 10 4 Ω vs 24.9 µm: 1.5849× 10 4 Ω at 2.01 MHz). These signatures demonstrate the platform’s comprehensive particle characterization capabilities through complementary DEP forces. The dual-mode approach enhanced discrimination ratios by 2.48 × under L e v . f o r c e and 3.16 × under L E V . f o r c e at selected characteristic frequency range compared to NonDEPforce conditions. Comprehensive impedance analysis through frequency spectrum (10 kHz—2.01 MHz) revealed unique frequency-dependent cell signatures, R O T . f o r c e demonstrating superior mid-frequency discrimination (magnitude differences of 1.9370 × 10 4 Ω vs 2.0542 × 10 4 Ω at 110 kHz) and LEV DEP optimizing high-frequency characterization (1.6677 × 10 4 Ω vs 1.5849 × 10 4 Ω at 2.01 MHz). Impedance dielectric analysis conducted over the 10 kHz to 2.01 MHz frequency range demonstrated frequency-dependent characteristics for each selected cell population. ROT DEP enhanced the discrimination in the mid-frequency range (110 kHz), with 10.4 µm particles presenting impedance magnitudes of 1.9370 × 10 4 Ω, while 24.9 µm particles displayed 2.0542 × 10 4 Ω, yielding a distinct separation ratio of 1.06 × . In the high-frequency domain (2.01 MHz), LEV DEP optimized particle characterization revealed that 10.4 µm particles exhibited a resistance of 1.6677 × 10 4 Ω. In contrast, 24.9 µm particles showed a resistance of 1.5849 × 10 4 Ω, resulting in a separation ratio of 1.05 × . The dual-mode approach markedly improved discrimination capabilities, with LEV DEP demonstrating a 2.48 × enhancement and ROT DEP exhibiting a 3.16 × increase in separation ratios relative to no-DEP conditions. This proposed dual-force implementation exhibited notable efficacy in designated frequency ranges: ROT DEP excelled in mid-frequency discrimination, achieving magnitude differences of 11.72 × 10 3 Ω between particle populations, whereas LEV DEP optimized high-frequency characterization with differences of 8.28 × 10 3 Ω, facilitating comprehensive particle discrimination through complementary DEP forces. This study establishes a novel microfluidic platform integrating dual-mode DEP manipulation with high-sensitivity dielectric features impedance detection, achieving a 163.1% enhancement in signal-to-noise SNR ratio compared to the conventional impedance mode. The proposed system demonstrates exceptional particle discrimination capabilities, with LEV DEP achieving a 63.4% and 81.2% magnitude increase for 10.4 µm and 24.9 µm particles, respectively, at 2.01 MHz. In comparison, ROT DEP induced more pronounced increases of 120% and 145% at 110 kHz. The proposed system significantly improved discrimination ratios (2.48 × under LEV DEP and 3.16 × under ROT DEP ) relative to no-DEP conditions, identifying clear phase behavior patterns for both particle populations. Impedance analysis over the 10 kHz to 2.01 MHz frequency range identified distinct frequency-dependent characteristics. ROT DEP exhibited enhanced mid-frequency discrimination, measuring 1.9370 × 10 4 Ω compared to 2.0542 × 10 4 Ω, while LEV DEP provided optimized high-frequency characterization, with values of 1.6677 × 10 4 Ω versus 1.5849 × 10 4 Ω. This system, which is label-free and non-invasive, facilitates cellular dielectric analysis with improved throughput and measurement precision. It provides substantial benefits for biological research, medical diagnostics, and drug analysis and development through its dual-force implementation and extensive impedance characterization capabilities.

The Ecuadorian Chocó is a global conservation hotspot with high levels of biodiversity and endemism but is threatened by extensive deforestation. Here, we report the discovery of an additional locality for the Critically Endangered poison-dart frog Leucostethus bilsa (Amphibia, Dendrobatidae) in a privately protected forest reserve in Esmeraldas, Ecuador. This may represent a distinct IUCN location for the species, thereby downgrading its IUCN threat level to Endangered. This finding increases the extent of occurrence for L. bilsa from 0.9 to 4.78 km2, and the area of occupancy from 8.0 to 16.0 km2, demonstrating the importance of small forest reserves for the conservation of forest-dependent amphibians. Further surveys are warranted to determine whether the species is present in additional forest fragments in the region, the degree of connectivity, if any, between individuals inhabiting different fragments, and whether the species’ population is increasing, stable, or in decline.

Rice leaf morphology is an essential agronomic trait to develop drought-tolerant genotypes for adequate and stable crop production in drought-prone areas. Here, rolled leaf mutant plants were acquired by CRISPR/Cas9-based mutagenesis of Semi-rolled leaf1,2 (SRL1 and SRL2) genes, and isobaric tags for relative and absolute quantification (iTRAQ) based proteomic analysis was performed to analyze the subsequent proteomic regulation events. Homozygous mutants exhibit decreased chlorophyll content, transpiration rate, stomatal conductance, vascular bundles (VB), stomatal number, and agronomic traits with increased panicle number and bulliform cells (BCs). Under drought stress, mutant plants displayed lower malondialdehyde (MDA) content while higher survival rate, abscisic acid (ABA) content, superoxide dismutase (SOD), catalase (CAT) activities, and grain filling percentage compare with their wild type (WT). Proteomic results revealed that 270 proteins were significantly downregulated, and 107 proteins were upregulated in the mutant line compared with WT. Proteins related to lateral organ boundaries’ (LOB) domain (LBD) were downregulated, whereas abiotic stress-responsive proteins were upregulated in the CRISPR mutant. LBD proteins (Q5KQR7, Q6K713, Q7XGL4, Q8LQH4), probable indole-3-acetic acid-amido synthetase (Q60EJ6), putative auxin transporter-like protein 4 (Q53JG7), Monoculm 1 (Q84MM9) and AP2 (Apetala2) domain-containing protein (Q10A97) were found to be hub-proteins. The hybrids developed from mutant restorers showed a semi-rolled leaf phenotype with increased panicle number, grain number per panicle, and yield per plant. Our findings reveal the intrinsic value of genome editing and expand the knowledge about the network of proteins for leaf rolling and drought avoidance in rice.

The preservation of image details in the defogging process is still one key challenge in the field of deep learning. The network uses the generation of confrontation loss and cyclic consistency loss to ensure that the generated defog image is similar to the original image, but it cannot retain the details of the image. To this end, we propose a detail enhanced image CycleGAN to retain the detail information during the process of defogging. Firstly, the algorithm uses the CycleGAN network as the basic framework and combines the U-Net network’s idea with this framework to extract visual information features in different spaces of the image in multiple parallel branches, and it introduces Dep residual blocks to learn deeper feature information. Secondly, a multi-head attention mechanism is introduced in the generator to strengthen the expressive ability of features and balance the deviation produced by the same attention mechanism. Finally, experiments are carried out on the public data set D-Hazy. Compared with the CycleGAN network, the network structure of this paper improves the SSIM and PSNR of the image dehazing effect by 12.2% and 8.1% compared with the network and can retain image dehazing details.

Purpose We have developed and reported on the usefulness of single and double endplates penetrating screw trajectories (SEPST/DEPST), which penetrate the endplates of the vertebrae to increase the fixation strength in patients with diffuse idiopathic skeletal hyperostosis (DISH). The purpose of this study is to compare the peak insertion torque (PIT) and pullout force (POF) of traditional trajectory (TT) and SEPST/DEPST using cadaveric spine. Methods The eighth thoracic vertebra to the fifth lumbar vertebra from six formalin-fixed adult human cadavers (two males, four females; age at death 85.7 ± 8.8 years) were used and PIT and POF were evaluated in the following three studies. Firstly, a comparison among TT/SEPST/DEPST. Secondly, a comparison of reinsertion of screws with DEPST (re-DEPST) in vertebrae inserted with TT and removed, assuming reoperation. Lastly, a comparison between the endplate thread screw (ETS) which has a smaller pitch and the conventional cancellous thread screw (CTS) was made. Results SEPST/DEPST had a 32–70% higher PIT and a 45–82% higher POF than TT, and DEPST was about 1.4 times higher than SEPST. Assuming reoperation, the results show that fixation with re-DEPST provided almost the same fixation strength as initial TT insertion. ETS, which was considered useful for DEPS, showed a relatively higher fixation strength than CTS, however, the difference was not significant. Conclusion The present study using cadaver demonstrated the strong fixation of SEPST/DEPST and its usefulness for revision surgery, however, there were no significant differences in the mechanical properties between ETS and CTS.

Currently, cell separation occurs almost exclusively by density gradient methods and by fluorescence- and magnetic-activated cell sorting (FACS/MACS). These variously suffer from lack of specificity, high cell loss, use of labels, and high capital/operating cost. We present a dielectrophoresis (DEP)-based cell-separation method, using 3D electrodes on a low-cost disposable chip; one cell type is allowed to pass through the chip whereas the other is retained and subsequently recovered. The method advances usability and throughput of DEP separation by orders of magnitude in throughput, efficiency, purity, recovery (cells arriving in the correct output fraction), cell losses (those which are unaccounted for at the end of the separation), and cost. The system was evaluated using three example separations: live and dead yeast; human cancer cells/red blood cells; and rodent fibroblasts/red blood cells. A single-pass protocol can enrich cells with cell recovery of up to 91.3% at over 300,000 cells per second with >3% cell loss. A two-pass protocol can process 300,000,000 cells in under 30 min, with cell recovery of up to 96.4% and cell losses below 5%, an effective processing rate >160,000 cells per second. A three-step protocol is shown to be effective for removal of 99.1% of RBCs spiked with 1%cancer cells while maintaining a processing rate of ∼170,000 cells per second. Furthermore, the self-contained and low-cost nature of the separator device means that it has potential application in low-contamination applications such as cell therapies, where good manufacturing practice compatibility is of paramount importance.

Particulate matter in the air exacerbates airway inflammation (AI) in asthma; moreover, prenatal exposure to concentrated urban air particles (CAPs) and diesel exhaust particles (DEPs) predisposes the offspring to asthma and worsens the resolution of AI in response to allergens. We previously tested the hypothesis that such exposure impairs the pathways of specialized proresolving mediators that are critical for resolution and found declined Lipoxin A4 (LxA4) and Resolvin E2 (RvE2) levels in the “at-risk” pups of exposed mothers. Here, we hypothesized that supplementation with synthetic LxA4 or RvE2 via the airway can ameliorate AI after allergen exposure, which has not been tested in models with environmental toxicant triggers. BALB/c newborns with an asthma predisposition resultant from prenatal exposure to CAPs and DEPs were treated once daily for 3 days with 750 ng/mouse of LxA4 or 300 ng/mouse of RvE2 through intranasal instillation, and they were tested with the intentionally low-dose ovalbumin protocol that elicits asthma in the offspring of particle-exposed mothers but not control mothers, mimicking the enigmatic maternal transmission of asthma seen in humans. LxA4 and RvE2 ameliorated the asthma phenotype and improved AI resolution, which was seen as declining airway eosinophilia, lung tissue infiltration, and proallergic cytokine levels.

Background Disordered eating behaviors (DEBs) may lead to full blown eating disorders. Both type 1 diabetes mellitus (T1DM) and celiac disease (CD) have been linked to DEBs. Objective To compare the presence of DEBs between adolescents and young adults with a dual diagnosis of T1DM and CD, and individuals with only one of the diagnoses. Methods Individuals with a dual diagnosis of T1DM and CD (“T1DM + CD group” n = 39), with a diagnosis of T1DM only (“T1DM group” n = 97) and with a diagnosis of CD only (“CD group” n = 267) filled the Eating Attitude Test‐26 (EAT‐26) questionnaire. Those with T1DM completed in addition to the Diabetes Eating Problem Survey‐Revised (DEPS‐R). Results The study population comprised of 403 individuals, of whom 65% were females. There were no statistically significant differences among the groups in distribution of sex, age, hemoglobin A1c (HbA1c) levels, age of disease diagnosis and duration. The prevalence of DEBs in the T1DM + CD group was 3‐fold higher (26.0%) than in the T1DM (8.2%) and CD (8.2%) groups (P = .003). This trend was observed for both females and males. Multivariate analysis demonstrated that the T1DM + CD group had an increased risk for DEBs (odds ratio, OR: 4.7, 95% confidence interval, CI: 1.9‐11.2, P = .001) after adjustment for age, sex, and body mass index. Additionally, being female, older and overweight increased the risk for DEBs. HbA1c values were not associated with an increased DEBs rate. Conclusions Individuals with the dual diagnoses of T1DM and CD have an increased likelihood to develop DEBs compared to those with only one of these diagnoses.

Background During inhalation, airborne particles such as particulate matter ≤ 2.5 μm (PM 2.5 ), can deposit and accumulate on the alveolar epithelial tissue. In vivo studies have shown that fractions of PM 2.5 can cross the alveolar epithelium to blood circulation, reaching secondary organs beyond the lungs. However, approaches to quantify the translocation of particles across the alveolar epithelium in vivo and in vitro are still not well established. In this study, methods to assess the translocation of standard diesel exhaust particles (DEPs) across permeable polyethylene terephthalate (PET) inserts at 0.4, 1, and 3 μm pore sizes were first optimized with transmission electron microscopy (TEM), ultraviolet-visible spectroscopy (UV-VIS), and lock-in thermography (LIT), which were then applied to study the translocation of DEPs across human alveolar epithelial type II (A549) cells. A549 cells that grew on the membrane (pore size: 3 μm) in inserts were exposed to DEPs at different concentrations from 0 to 80 µg.mL − 1 ( 0 to 44 µg.cm − 2 ) for 24 h. After exposure, the basal fraction was collected and then analyzed by combining qualitative (TEM) and quantitative (UV-VIS and LIT) techniques to assess the translocated fraction of the DEPs across the alveolar epithelium in vitro. Results We could detect the translocated fraction of DEPs across the PET membranes with 3 μm pore sizes and without cells by TEM analysis, and determine the percentage of translocation at approximatively 37% by UV-VIS (LOD: 1.92 µg.mL − 1 ) and 75% by LIT (LOD: 0.20 µg.cm − 2 ). In the presence of cells, the percentage of DEPs translocation across the alveolar tissue was determined around 1% at 20 and 40 µg.mL − 1 (11 and 22 µg.cm − 2 ), and no particles were detected at higher and lower concentrations. Interestingly, simultaneous exposure of A549 cells to DEPs and EDTA can increase the translocation of DEPs in the basal fraction. Conclusion We propose a combination of analytical techniques to assess the translocation of DEPs across lung tissues. Our results reveal a low percentage of translocation of DEPs across alveolar epithelial tissue in vitro and they correspond to in vivo findings. The combination approach can be applied to any traffic-generated particles, thus enabling us to understand their involvement in public health.

Background The epididymis is a highly regionalized tubular organ possesses vectorial functions of sperm concentration, maturation, transport, and storage. The epididymis-expressed genes and proteins are characterized by regional and developmental dependent pattern. However, a systematic and comprehensive insight into the postnatal development dependent changes in gene and protein expressions of porcine epididymis is still lacking. Here, the RNA and protein of epididymis of Duroc pigs at different postnatal development stages were extracted by using commercial RNeasy Midi kit and extraction buffer (7 M Urea, 2 M thiourea, 3% CHAPS, and 1 mM PMSF) combined with sonication, respectively, which were further subjected to transcriptomic and proteomic profiling. Results Transcriptome analysis indicated that 198 and 163 differentially expressed genes (DEGs) were continuously up-regulated and down-regulated along with postnatal development stage changes, respectively. Most of the up-regulated DEGs linked to functions of endoplasmic reticulum and lysosome, while the down-regulated DEGs mainly related to molecular process of extracellular matrix. Moreover, the following key genes INSIG1 , PGRMC1 , NPC2 , GBA , MMP2 , MMP14 , SFRP1 , ELN , WNT-2 , COL3A1 , and SPARC were highlighted. A total of 49 differentially expressed proteins (DEPs) corresponding to postnatal development stages changes were uncovered by the proteome analysis. Several key proteins ACSL3 and ACADM, VDAC1 and VDAC2, and KNG1, SERPINB1, C3, and TF implicated in fatty acid metabolism, voltage-gated ion channel assembly, and apoptotic and immune processes were emphasized. In the integrative network, the key genes and proteins formed different clusters and showed strong interactions. Additionally, NPC2 , COL3A1 , C3, and VDAC1 are located at the hub position in each cluster. Conclusions The identified postnatal development dependent genes and proteins in the present study will pave the way for shedding light on the molecular basis of porcine epididymis functions and are useful for further studies on the specific regulation mechanisms responsible for epididymal sperm maturation.