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1.
Lab Chip ; 24(13): 3243-3251, 2024 Jun 25.
Artigo em Inglês | MEDLINE | ID: mdl-38836406

RESUMO

The parathyroid gland is an endocrine organ that plays a crucial role in regulating calcium levels in blood serum through the secretion of parathyroid hormone (PTH). Hypoparathyroidism is a chronic disease that can occur due to parathyroid defects, but due to the difficulty of creating animal models of this disease or obtaining human normal parathyroid cells, the evaluation of parathyroid functionality for drug development is limited. Although parathyroid-like cells that secrete PTH have recently been reported, their functionality may be overestimated using traditional culture methods that lack in vivo similarities, particularly vascularization. To overcome these limitations, we obtained parathyroid organoids from tonsil-derived mesenchymal stem cells (TMSCs) and fabricated a parathyroid-on-a-chip, capable of simulating PTH secretion based on calcium concentration. This chip exhibited differences in PTH secretion according to calcium concentration and secreted PTH within the range of normal serum levels. In addition, branches of organoids, which are difficult to observe in animal models, were observed in this chip. This could serve as a guideline for successful engraftment in implantation therapies in the future.


Assuntos
Cálcio , Dispositivos Lab-On-A-Chip , Células-Tronco Mesenquimais , Glândulas Paratireoides , Hormônio Paratireóideo , Hormônio Paratireóideo/metabolismo , Cálcio/metabolismo , Humanos , Glândulas Paratireoides/metabolismo , Glândulas Paratireoides/citologia , Células-Tronco Mesenquimais/metabolismo , Células-Tronco Mesenquimais/citologia , Organoides/metabolismo , Organoides/citologia , Células Cultivadas
2.
J Colloid Interface Sci ; 670: 729-741, 2024 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-38788440

RESUMO

This study explores a strategy to mitigate capacity fading in secondary batteries, which is primarily attributed to side reactions caused by residual Li impurities (LiOH or Li2CO3) on the surface of Ni-rich LiNi0.8Co0.1Mn0.1O2 (NCM811) layered cathode materials. By applying a 1.5 wt% Co3(PO4)2 coating, we successfully formed a thin and stable LiF cathode-electrolyte interface (CEI) layer, which resulted in decreased battery resistance and enhanced diffusion of Li+ ions within the electrolyte. This coating significantly improved the interface stability of NCM811, leading to superior battery performance. Specifically, the discharge capacity of uncoated NCM811 was 190 mA h g-1 at a charge of 4.3 V and a rate of 0.1C, whereas the 1.5Co3(PO4)2/NCM811 exhibited an increased capacity of 213 mA h g-1. Furthermore, the Co3(PO4)2 coating effectively reduced the levels of LiOH and Li2CO3 on the NCM811 surface to only 0.1 %, thereby minimizing adverse side reactions with the electrolyte salt (LiPF6), cation mixing between Ni2+ and Li+, and defects at the NCM811 interface. As a result, battery lifespan was significantly extended. This study presents a robust approach for enhancing battery stability and performance by efficiently reducing residual Li+ ions on the surface of NCM811 through strategic Co3(PO4)2 coating.

3.
Small Methods ; : e2400284, 2024 Apr 23.
Artigo em Inglês | MEDLINE | ID: mdl-38651527

RESUMO

Perovskite materials that aren't stable during the oxygen evolution reaction (OER) are unsuitable for anion-exchange membrane water electrolyzers (AEMWE). But through manipulating their electronic structures, their performance can further increase. Among the first-row transition metals, nickel and iron are widely recognized as prominent electrocatalysts; thus, the researchers are looking into how combining them can improve the OER. Recent research has actively explored the design and study of heterostructures in this field, showcasing the dynamic exploration of innovative catalyst configurations. In this study, a heterostructure is used to manipulate the electronic structure of LaNiO3 (LNO) to improve both OER properties and durability. Through adsorbing iron onto the LNO (LNO@Fe) as γ iron oxyhydroxide (γ-FeOOH), the binding energy of nickel in the LNO exhibited negative shifts, inferring nickel movement toward the metallic state. Consequently, the electrochemical properties of LNO@Fe are further improved. LNO@Fe showed excellent performance (1.98 A cm-2, 1 m KOH, 50 °C at 1.85 V) with 84.1% cell efficiency in AEMWE single cells, demonstrating great improvement relative to LNO. The degradation for the 850 h durability analysis of LNO@Fe is ≈68 mV kh-1, which is ≈58 times less than that of LNO.

4.
Magn Reson Med ; 91(6): 2483-2497, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38342983

RESUMO

PURPOSE: We introduced a novel reconstruction network, jointly unrolled cross-domain optimization-based spatio-temporal reconstruction network (JUST-Net), aimed at accelerating 3D multi-echo gradient-echo (mGRE) data acquisition and improving the quality of resulting myelin water imaging (MWI) maps. METHOD: An unrolled cross-domain spatio-temporal reconstruction network was designed. The main idea is to combine frequency and spatio-temporal image feature representations and to sequentially implement convolution layers in both domains. The k-space subnetwork utilizes shared information from adjacent frames, whereas the image subnetwork applies separate convolutions in both spatial and temporal dimensions. The proposed reconstruction network was evaluated for both retrospectively and prospectively accelerated acquisition. Furthermore, it was assessed in simulation studies and real-world cases with k-space corruptions to evaluate its potential for motion artifact reduction. RESULTS: The proposed JUST-Net enabled highly reproducible and accelerated 3D mGRE acquisition for whole-brain MWI, reducing the acquisition time from fully sampled 15:23 to 2:22 min within a 3-min reconstruction time. The normalized root mean squared error of the reconstructed mGRE images increased by less than 4.0%, and the correlation coefficients for MWI showed a value of over 0.68 when compared to the fully sampled reference. Additionally, the proposed method demonstrated a mitigating effect on both simulated and clinical motion-corrupted cases. CONCLUSION: The proposed JUST-Net has demonstrated the capability to achieve high acceleration factors for 3D mGRE-based MWI, which is expected to facilitate widespread clinical applications of MWI.


Assuntos
Bainha de Mielina , Água , Imageamento por Ressonância Magnética/métodos , Estudos Retrospectivos , Imageamento Tridimensional/métodos , Processamento de Imagem Assistida por Computador/métodos
5.
Stem Cells Int ; 2023: 6845571, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-38020205

RESUMO

Sensorineural hearing loss is very difficult to treat. Currently, one of the techniques used for hearing rehabilitation is a cochlear implant that can transform sound into electrical signals instead of inner ear hair cells. However, the prognosis remains very poor if sufficient auditory nerve cells are not secured. In this study, the effect of mouse embryonic stem cells (mESC) and photobiomodulation (PBM) combined treatment on auditory function and auditory nerve cells in a secondary neuropathy animal model was investigated. To confirm the engraftment of stem cells in vitro, cochlear explants were treated with kanamycin (KM) to mimic nerve damage and then cocultured with GFP-mESC. GFP-mESCs were observed to have attached and integrated into the explanted samples. An animal model for secondary neurodegeneration was achieved by KM treatment and was treated by a combination therapy of GFP-mESC and NIR-PBM at 8 weeks of KM treatment. Hearing recovery by functional testing using auditory brain stem response (ABR) and eABR was measured as well as morphological changes and epifluorescence analysis were conducted after 2 weeks of combination therapy. KM treatment elevated the hearing threshold at 70-80 dB and even after the combination treatment with GFP-mESC and PBM was applied, the auditory function was not restored. In addition, the stem cells transplanted into cochlea has exponentially increased due to PBM treatment although did not produce any malignancy. This study confirmed that the combined treatment with mESC and PBM could not improve hearing or increase the response of the auditory nerve. Nevertheless, it is noteworthy in this study that the cells are distributed in most cochlear tissues and the proliferation of stem cells was very active in animals irradiated with PBM compared to other groups wherein the stem cells had disappeared immediately after transplantation or existed for only a short period of time.

7.
Int J Mol Sci ; 24(18)2023 Sep 06.
Artigo em Inglês | MEDLINE | ID: mdl-37762066

RESUMO

Hepatocellular Carcinoma (HCC) continues to pose a substantial global health challenge due to its high incidence and limited therapeutic options. In recent years, the Janus Kinase (JAK) and Signal Transducer and Activator of Transcription (STAT) pathway has emerged as a critical signaling cascade in HCC pathogenesis. The review commences with an overview of the JAK/STAT pathway, delving into the dynamic interplay between the JAK/STAT pathway and its numerous upstream activators, such as cytokines and growth factors enriched in pathogenic livers afflicted with chronic inflammation and cirrhosis. This paper also elucidates how the persistent activation of JAK/STAT signaling leads to diverse oncogenic processes during hepatocarcinogenesis, including uncontrolled cell proliferation, evasion of apoptosis, and immune escape. In the context of therapeutic implications, this review summarizes recent advancements in targeting the JAK/STAT pathway for HCC treatment. Preclinical and clinical studies investigating inhibitors and modulators of JAK/STAT signaling are discussed, highlighting their potential in suppressing the deadly disease. The insights presented herein underscore the necessity for continued research into targeting the JAK/STAT signaling pathway as a promising avenue for HCC therapy.


Assuntos
Carcinoma Hepatocelular , Neoplasias Hepáticas , Humanos , Transdução de Sinais , Carcinoma Hepatocelular/metabolismo , Janus Quinases/metabolismo , Neoplasias Hepáticas/metabolismo , Fatores de Transcrição STAT/metabolismo
8.
Nat Metab ; 5(9): 1506-1525, 2023 09.
Artigo em Inglês | MEDLINE | ID: mdl-37653043

RESUMO

The lateral hypothalamic area (LHA) regulates food intake and energy balance. Although LHA neurons innervate adipose tissues, the identity of neurons that regulate fat is undefined. Here we show that GABRA5-positive neurons in LHA (GABRA5LHA) polysynaptically project to brown and white adipose tissues in the periphery. GABRA5LHA are a distinct subpopulation of GABAergic neurons and show decreased pacemaker firing in diet-induced obesity mouse models in males. Chemogenetic inhibition of GABRA5LHA suppresses fat thermogenesis and increases weight gain, whereas gene silencing of GABRA5 in LHA decreases weight gain. In the diet-induced obesity mouse model, GABRA5LHA are tonically inhibited by nearby reactive astrocytes releasing GABA, which is synthesized by monoamine oxidase B (Maob). Gene silencing of astrocytic Maob in LHA facilitates fat thermogenesis and reduces weight gain significantly without affecting food intake, which is recapitulated by administration of a Maob inhibitor, KDS2010. We propose that firing of GABRA5LHA suppresses fat accumulation and selective inhibition of astrocytic GABA is a molecular target for treating obesity.


Assuntos
Astrócitos , Obesidade , Masculino , Animais , Camundongos , Aumento de Peso , Neurônios , Modelos Animais de Doenças , Monoaminoxidase , Ácido gama-Aminobutírico
9.
Comput Biol Med ; 163: 107229, 2023 09.
Artigo em Inglês | MEDLINE | ID: mdl-37413852

RESUMO

Osteoarthritis knee can be restored by total knee arthroplasty (TKA). Imageless TKA requires several anatomical points to construct a reference coordinate system to measure bone resections and implant placement. Inaccuracies in the definition of the coordinate system lead to malalignment and failure of the implant. While the surgical transepicondylar axis (sTEA) is a reliable anatomical axis to define the lateromedial axis for the femoral coordinate system (FCS), the presence of the collateral ligaments and deterioration of the medial sulcus (MS) make the registration of sTEA a challenging task. In this work, sTEA is assigned using the articular surfaces of the femoral condyles, independent of the lateral epicondyle (LE) and MS. A single 3D arc is marked on each condyle, which is transformed into a 2D arc to get the best-fit curve according to the profile of condyles. The turning point of each best-fit curve, when transformed back to 3D, defines an axis parallel to sTEA. The condyles-based sTEA is measured experimentally on a 3D-printed bone using an Optitrack tracking setup. Using the proposed method, the angle between the aTEA, sTEA, and Whiteside's line was (3.77, 0.55, and 92.72)°, respectively. The proposed method provides the same level of accuracy and improves the anatomical points registration efficiency, as there is no need to register the LE or MS.


Assuntos
Artroplastia do Joelho , Osteoartrite do Joelho , Humanos , Artroplastia do Joelho/métodos , Articulação do Joelho/diagnóstico por imagem , Articulação do Joelho/cirurgia , Fêmur/diagnóstico por imagem , Fêmur/cirurgia , Computadores
10.
Front Genet ; 14: 1150822, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37388940

RESUMO

Background: Fabry disease (FD) is a rare X-linked lysosomal storage disease caused by mutations in the GLA gene that encodes α-galactosidase A (α-GAL). Clinical phenotypes tend to vary in monozygotic female twins because mutations are located on the X-chromosome, whereas similar phenotypes are found in male monozygotic twins. Here we report the case of male monozygotic twins with FD presenting with distinguishable renal phenotypes. Case: A 49-year-old male patient who visited the hospital with proteinuria 14 years prior was readmitted for the same issue. His monozygotic twin brother had started hemodialysis 6 months prior due to renal failure of unknown origin. The patient's renal function was within the normal range, while his spot urine protein-to-creatinine ratio was 557 mg/g. Echocardiography revealed left ventricular hypertrophy (LVH). The findings of a renal biopsy were consistent with FD. Genetic testing identified a c.656T>C mutation in the GLA gene, and α-GAL activity was significantly decreased. Genetic screening of his family clarified that his mother, older sister, twin brother, and his daughter had the same genetic mutations. The patient received enzyme replacement therapy 34 times. Subsequently, migalastat was initiated that continues today. Renal function and proteinuria remain stable, and the LVH has mildly improved. Conclusion: This is the first case of male monozygotic twins expressing different progressions of FD. Our findings demonstrate the possibility that environmental or epigenetic factors may critically influence genotype-phenotype discordance.

11.
PLoS One ; 18(5): e0284626, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37216352

RESUMO

Noise exposure can destroy the synaptic connections between hair cells and auditory nerve fibers without damaging the hair cells, and this synaptic loss could contribute to difficult hearing in noisy environments. In this study, we investigated whether delivering lithium chloride to the round-window can regenerate synaptic loss of cochlea after acoustic overexposure. Our rat animal model of noise-induced cochlear synaptopathy caused about 50% loss of synapses in the cochlear basal region without damaging hair cells. We locally delivered a single treatment of poloxamer 407 (vehicle) containing lithium chloride (either 1 mM or 2 mM) to the round-window niche 24 hours after noise exposure. Controls included animals exposed to noise who received only the vehicle. Auditory brainstem responses were measured 3 days, 1 week, and 2 weeks post-exposure treatment, and cochleas were harvested 1 week and 2 weeks post-exposure treatment for histological analysis. As documented by confocal microscopy of immunostained ribbon synapses, local delivery of 2 mM lithium chloride produced synaptic regeneration coupled with corresponding functional recovery, as seen in the suprathreshold amplitude of auditory brainstem response wave 1. Western blot analyses revealed that 2 mM lithium chloride suppressed N-methyl-D-aspartate (NMDA) receptor expression 7 days after noise-exposure. Thus, round-window delivery of lithium chloride using poloxamer 407 reduces cochlear synaptic loss after acoustic overexposure by inhibiting NMDA receptor activity in rat model.


Assuntos
Perda Auditiva Provocada por Ruído , Receptores de N-Metil-D-Aspartato , Ratos , Animais , Receptores de N-Metil-D-Aspartato/metabolismo , Cloreto de Lítio , Perda Auditiva Provocada por Ruído/etiologia , Poloxâmero , Limiar Auditivo/fisiologia , Cóclea/patologia , Sinapses/metabolismo , Potenciais Evocados Auditivos do Tronco Encefálico/fisiologia
12.
Med Phys ; 50(3): 1660-1669, 2023 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-36585806

RESUMO

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.


Assuntos
Encéfalo , Imageamento por Ressonância Magnética , Imageamento por Ressonância Magnética/métodos , Algoritmos , Tomografia/métodos , Condutividade Elétrica , Imagens de Fantasmas , Processamento de Imagem Assistida por Computador/métodos
13.
Stem Cell Res ; 65: 102970, 2022 12.
Artigo em Inglês | MEDLINE | ID: mdl-36399926

RESUMO

Cytochrome P450 (CYP) reaction phenotyping has become crucial for predicting drug reactions and side effects. Single nucleotide polymorphisms (SNPs) in CYP genes alter drug metabolism capacity and cause unexpected drug-related reactions. Here, we established two human induced pluripotent stem cell (hiPSC) lines with pharmacologically important SNPs in CYP2D6 in conjunction with CYP2C19 or CYP3A5 genes. These hiPSC lines can serve as valuable resources for expanding our understanding of the relationships between genotypes and drug reactions.


Assuntos
Citocromo P-450 CYP2C19 , Citocromo P-450 CYP2D6 , Citocromo P-450 CYP3A , Células-Tronco Pluripotentes Induzidas , Preparações Farmacêuticas , Humanos , Citocromo P-450 CYP2C19/genética , Citocromo P-450 CYP2D6/genética , Citocromo P-450 CYP3A/genética , Células-Tronco Pluripotentes Induzidas/enzimologia , Linhagem Celular , Polimorfismo de Nucleotídeo Único , Preparações Farmacêuticas/metabolismo
14.
Med Phys ; 49(9): 5929-5942, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-35678751

RESUMO

PURPOSE: To enable acceleration in 3D multi-echo gradient echo (mGRE) acquisition for myelin water imaging (MWI) by combining joint parallel imaging (JPI) and joint deep learning (JDL). METHODS: We implemented a multistep reconstruction process using both advanced parallel imaging and deep learning network which can utilize joint spatiotemporal components between the multi-echo images to further accelerate 3D mGRE acquisition for MWI. In the first step, JPI was performed to estimate missing k-space lines. Next, JDL was implemented to reduce residual artifacts and produce high-fidelity reconstruction by using variable splitting optimization consisting of spatiotemporal denoiser block, data consistency block, and weighted average block. The proposed method was evaluated for MWI with 2D Cartesian uniform under-sampling for each echo, enabling scan times of up to approximately 2 min for 2 mm × 2 mm × 2 mm $2\ {\rm mm} \times 2\ {\rm mm} \times 2\ {\rm mm}$ 3D coverage. RESULTS: The proposed method showed acceptable MWI quality with improved quantitative values compared to both JPI and JDL methods individually. The improved performance of the proposed method was demonstrated by the low normalized mean-square error and high-frequency error norm values of the reconstruction with high similarity to the fully sampled MWI. CONCLUSION: Joint spatiotemporal reconstruction approach by combining JPI and JDL can achieve high acceleration factors for 3D mGRE-based MWI.


Assuntos
Imageamento por Ressonância Magnética , Bainha de Mielina , Encéfalo , Processamento de Imagem Assistida por Computador/métodos , Imageamento Tridimensional , Imageamento por Ressonância Magnética/métodos , Tomografia Computadorizada por Raios X , Água
15.
Glia ; 70(10): 1799-1825, 2022 10.
Artigo em Inglês | MEDLINE | ID: mdl-35713516

RESUMO

The organ of Corti, located in the cochlea in the inner ear, is one of the major sensory organs involved in hearing. The organ of Corti consists of hair cells, glia-like supporting cells, and the cochlear nerve, which work in harmony to receive sound from the outer ear and transmit auditory signals to the cochlear nucleus in the auditory ascending pathway. In this process, maintenance of the endocochlear potential, with a high potassium gradient and clearance of electrolytes and biochemicals in the inner ear, is critical for normal sound transduction. There is an emerging need for a thorough understanding of each cell type involved in this process to understand the sophisticated mechanisms of the organ of Corti. Hair cells have long been thought to be active, playing a primary role in the cochlea in actively detecting and transmitting signals. In contrast, supporting cells are thought to be silent and function to support hair cells. However, growing lines of evidence regarding the membrane proteins that mediate ionic movement in supporting cells have demonstrated that supporting cells are not silent, but actively play important roles in normal signal transduction. In this review, we summarize studies that characterize diverse membrane proteins according to the supporting cell subtypes involved in cochlear physiology and hearing. This review contributes to a better understanding of supporting cell functions and facilitates the development of potential therapeutic tools for hearing loss.


Assuntos
Proteínas de Membrana , Órgão Espiral , Cóclea/fisiologia , Audição/fisiologia , Neuroglia , Órgão Espiral/fisiologia
16.
Mater Today Bio ; 14: 100241, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-35313446

RESUMO

Redox activity is known to regulate migration, invasion, metastasis, proliferation, and vascularization of cancer. Because cancer is heterogeneous, the role of redox activity in different cancers and cancer-related processes vary widely. In this study, water soluble, Tween 80-coated polyaniline (TPAni) nanoparticles were synthesized and used as nano-agents for sensing the redox activities of various cancer cells. To identify the relationship between the redox activity and the aggressiveness of cancer cells, two different cancer cell lines, derived from the same tissue but different with regards to aggressiveness, were selected for study. First, the cancer cell lines were incubated with TPAni nanoparticles, and an absorbance ratio obtained from the cell culture media was used as a colorimetric indicator of the redox activities of the cells. Simultaneously, hydrophobically modified filter papers coated with silver nanosnowflakes (SNSF) were used as sensing substrates for surface enhanced Raman scattering (SERS). SERS spectra obtained from varying concentrations of rhodamine 6G were used to confirm the detection limit of the SNSF-based SERS substrate. Cell culture media containing TPAni nanoparticles were treated with the SNSF-containing SERS substrates to examine the redox activities of the various cancer cell lines.The redox activities of cancer cell lines were confirmed by absorbance spectral analysis, and these redox activities were better identified via an SERS analysis method. A SNSF-containing SERS substrate, fabricated from SNSF and filter paper, was used to sense redox activity in cancer cell lines and to further identify correlations between redox activity and cancer cell line aggressiveness, as indicated by the use of EpCAM as a biomarker. Finally, potential of â€‹in vivo â€‹redox activity sensing was also confirmed.

17.
Biosensors (Basel) ; 12(2)2022 Feb 14.
Artigo em Inglês | MEDLINE | ID: mdl-35200378

RESUMO

Droplet-based microfluidics has been widely used as a potent high-throughput platform due to various advantages, such as a small volume of reagent consumption, massive production of droplets, fast reaction time, and independent control of each droplet. Therefore, droplet microfluidic systems demand the reliable generation of droplets with precise and effective control over their size and distribution, which is critically important for various applications in the fields of chemical analysis, material synthesis, lab-on-a-chip, cell research, diagnostic test, and so on. In this study, we propose a microfluidic device with a high-aspect-ratio (HAR) channel, which has a parallelogram cross-section, for generating monodisperse droplets. The HAR channel was fabricated using simple and cheap MEMS processes, such as photolithography, anisotropic wet etching, and PDMS molding, without expensive equipment. In addition, the parallelogram cross-section channel structure, regarded as a difficult shape to implement in previous fabrication methods, was easily formed by the self-alignment between the silicon channel and the PDMS mold, both of which were created from a single crystal silicon through an anisotropic etching process. We investigated the effects of the cross-sectional shape (parallelogram vs. rectangle) and height-to-width ratio of microfluidic channels on the size and uniformity of generated droplets. Using the developed HAR channel with the parallelogram cross-section, we successfully obtained smaller monodisperse droplets for a wider range of flow rates, compared with a previously reported HAR channel with a rectangular cross-section.


Assuntos
Técnicas Analíticas Microfluídicas , Microfluídica , Dispositivos Lab-On-A-Chip , Técnicas Analíticas Microfluídicas/instrumentação , Microfluídica/métodos , Silício/química
18.
J Biol Chem ; 298(3): 101626, 2022 03.
Artigo em Inglês | MEDLINE | ID: mdl-35074425

RESUMO

The bacterial second messenger bis-(3'-5')-cyclic diguanylate monophosphate (c-di-GMP) controls various cellular processes, including motility, toxin production, and biofilm formation. c-di-GMP is enzymatically synthesized by GGDEF domain-containing diguanylate cyclases and degraded by HD-GYP domain-containing phosphodiesterases (PDEs) to 2 GMP or by EAL domain-containing PDE-As to 5'-phosphoguanylyl-(3',5')-guanosine (pGpG). Since excess pGpG feedback inhibits PDE-A activity and thereby can lead to the uncontrolled accumulation of c-di-GMP, a PDE that degrades pGpG to 2 GMP (PDE-B) has been presumed to exist. To date, the only enzyme known to hydrolyze pGpG is oligoribonuclease Orn, which degrades all kinds of oligoribonucleotides. Here, we identified a pGpG-specific PDE, which we named PggH, using biochemical approaches in the gram-negative bacteria Vibrio cholerae. Biochemical experiments revealed that PggH exhibited specific PDE activity only toward pGpG, thus differing from the previously reported Orn. Furthermore, the high-resolution structure of PggH revealed the basis for its PDE activity and narrow substrate specificity. Finally, we propose that PggH could modulate the activities of PDE-As and the intracellular concentration of c-di-GMP, resulting in phenotypic changes including in biofilm formation.


Assuntos
GMP Cíclico/análogos & derivados , Diester Fosfórico Hidrolases , Vibrio cholerae , Proteínas de Bactérias/metabolismo , Biofilmes , GMP Cíclico/metabolismo , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/metabolismo , Diester Fosfórico Hidrolases/metabolismo , Transdução de Sinais , Especificidade por Substrato , Vibrio cholerae/enzimologia , Vibrio cholerae/metabolismo
19.
Anal Chem ; 93(45): 14996-15004, 2021 11 16.
Artigo em Inglês | MEDLINE | ID: mdl-34736319

RESUMO

The conventional tissue biopsy method yields isolated snapshots of a narrow region. Therefore, it cannot facilitate comprehensive disease characterization and monitoring. Recently, the detection of tumor-derived components in body fluids─a practice known as liquid biopsy─has attracted increased attention from the biochemical research and clinical application viewpoints. In this vein, surface-enhanced Raman scattering (SERS) has been identified as one of the most powerful liquid-biopsy analysis techniques, owing to its high sensitivity and specificity. Moreover, it affords high-capacity spectral multiplexing for simultaneous target detection and a unique ability to obtain intrinsic biomolecule-fingerprint spectra. This paper presents the fabrication of silver nanosnowflakes (SNSFs) using the polyol method and their subsequent dropping onto a hydrophobic filter paper. The SERS substrate, which comprises the SNSFs and hydrophobic filter paper, facilitates the simultaneous detection of creatinine and cortisol in human sweat using a hand-held Raman spectrometer. The proposed SERS system affords Raman spectrometry to be performed on small sample volumes (2 µL) to identify the normal and at-risk creatinine and cortisol groups.


Assuntos
Nanopartículas Metálicas , Prata , Creatinina , Humanos , Hidrocortisona , Análise Espectral Raman , Suor
20.
J Anim Sci Technol ; 63(5): 1086-1097, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34796349

RESUMO

This study examined the impact of using total mixed ration (TMR) and concentrate on feed intake, daily gain, carcass yield grade, and carcass quality grade of Hanwoo steers and its subsequent economic efficiency. Thirty six 7-month-old Hanwoo steers were assigned to one of the four treatment groups, and each group was divided into three repeated pens, with each repeated pen comprising three steers. The treatment groups were: 1) separate feeding with commercial concentrate and forage (namely, SCF) for the entire experimental period; 2) TMR feeding for a growing period followed by SCF for the early and late fattening period (namely, TMRGSCF); 3) TMR feeding for growing and an early fattening period followed by SCF for the late fattening period (namely, TMREFSCF); and 4) TMR feeding for the entire experimental period (namely, TMRW). The results showed that the SCF treatment had significantly (p < 0.05) higher feed intake during the growing period than other treatments. In contrast, the total feed intake had little difference during early and late fattening as well as the whole period regardless of feeding strategies. Daily gain showed no difference during the growing period. However, it was significantly higher in SCF and TMREFSCF treatments for the early and late fattening period, respectively (p < 0.05). The daily gain during the total raising period is in the order of TMREFSCF > TMRGSCF > SCF > TMRW. Carcass characteristics, including carcass weight, loin eye muscle area, and carcass yield grade, did not significantly differ among different treatments. However, TMRW treatment, wherein TMR was fed for a long time, showed that the cold carcass weight was less compared with other treatments, but carcass yield grade was higher with thinner backfat. Backfat thickness was in the order of SCF > TMRGSCF > TMREFSCF > TMRW, showing that the thickness reduced with longer TMR feeding (p < 0.05). TMRGSCF, which numerically had a higher carcass quality grade, showed higher economic efficiency, whereas SCF showed low economic efficiency. In conclusion, it was more feasible to apply TMR strategy in the growing and early fattening period and then SCF for the early or late fattening period to improve carcass yield, quality grade, and economic efficiency.

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