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Scientific Reports Jun 2021Mechanical forces are pervasive in the inflammatory site where dendritic cells (DCs) are activated to migrate into draining lymph nodes. For example, fluid shear stress...
Mechanical forces are pervasive in the inflammatory site where dendritic cells (DCs) are activated to migrate into draining lymph nodes. For example, fluid shear stress modulates the movement patterns of DCs, including directness and forward migration indices (FMIs), without chemokine effects. However, little is known about the effects of biomechanical forces on the activation of DCs. Accordingly, here we fabricated a microfluidics system to assess how biomechanical forces affect the migration and activity of DCs during inflammation. Based on the structure of edema, we proposed and experimentally analyzed a novel concept for a microchip model that mimicked such vascular architecture. The intensity of shear stress generated in our engineered chip was found as 0.2-0.6 dyne/cm by computational simulation; this value corresponded to inflammation in tissues. In this platform, the directness and FMIs of DCs were significantly increased, whereas the migration velocity of DCs was not altered by shear stress, indicating that mechanical stimuli influenced DC migration. Moreover, DCs with shear stress showed increased expression of the DC activation markers MHC class I and CD86 compared with DCs under static conditions. Taken together, these data suggest that the biomechanical forces are important to regulate the migration and activity of DCs.
Topics: Animals; B7-2 Antigen; Biomechanical Phenomena; Bone Marrow; Bone Marrow Cells; Cell Movement; Cell Separation; Chemokines; Computer Simulation; Dendritic Cells; Edema; Flow Cytometry; Inflammation; Lymph Nodes; Mice; Mice, Inbred C57BL; Microfluidics; Shear Strength; Stress, Mechanical
PubMed: 34103554
DOI: 10.1038/s41598-021-91117-2 -
Optimizing Binding Site Spacing in Fluidic Self-Assembly for Enhanced Microchip Integration Density.Micromachines Feb 2024This manuscript presents a comprehensive study on the assembly of microchips using fluidic self-assembly (FSA) technology, with a focus on optimizing the spacing between...
This manuscript presents a comprehensive study on the assembly of microchips using fluidic self-assembly (FSA) technology, with a focus on optimizing the spacing between binding sites to improve yield and assembly. Through a series of experiments, we explored the assembly of microchips on substrates with varying binding site spacings, revealing the impact of spacing on the rate of undesired chip assembly across multiple sites. Our findings indicate a significant reduction in incorrect assembly rates as the spacing increases beyond a critical threshold of 140 μm. This study delves into the mechanics of chip alignment within the fluid medium, hypothesizing that the extent of the alloy's grip on the chips at different spacings influences assembly outcomes. By analyzing cases of undesired assembly, we identified the relationship between binding site spacing and the area of chip contact, demonstrating a decrease in the combined left and right areas of chips as the spacing increases. The results highlight a critical spacing threshold, which, when optimized, could significantly enhance the efficiency and precision of microchip assembly processes using FSA technology. This research contributes to the field of microcomponent assembly, offering insights into achieving higher integration densities and precision in applications, such as microLED displays and augmented reality (AR) devices.
PubMed: 38542547
DOI: 10.3390/mi15030300 -
Advanced Science (Weinheim,... Jan 2023Most multiplex nucleic acids detection methods require numerous reagents and high-priced instruments. The emerging clustered regularly interspaced short palindromic...
Most multiplex nucleic acids detection methods require numerous reagents and high-priced instruments. The emerging clustered regularly interspaced short palindromic repeats (CRISPR)/Cas has been regarded as a promising point-of-care (POC) strategy for nucleic acids detection. However, how to achieve CRISPR/Cas multiplex biosensing remains a challenge. Here, an affordable means termed CRISPR-RDB (CRISPR-based reverse dot blot) for multiplex target detection in parallel, which possesses the advantages of high sensitivity and specificity, cost-effectiveness, instrument-free, ease to use, and visualization is reported. CRISPR-RDB integrates the trans-cleavage activity of CRISPR-Cas12a with a commercial RDB technique. It utilizes different Cas12a-crRNA complexes to separately identify multiple targets in one sample and converts targeted information into colorimetric signals on a piece of accessible nylon membrane that attaches corresponding specific-oligonucleotide probes. It has demonstrated that the versatility of CRISPR-RDB by constructing a four-channel system to simultaneously detect influenza A, influenza B, respiratory syncytial virus, and SARS-CoV-2. With a simple modification of crRNAs, the CRISPR-RDB can be modified to detect human papillomavirus, saving two-thirds of the time compared to a commercial PCR-RDB kit. Further, a user-friendly microchip system for convenient use, as well as a smartphone app for signal interpretation, is engineered. CRISPR-RDB represents a desirable option for multiplexed biosensing and on-site diagnosis.
Topics: Humans; COVID-19; CRISPR-Cas Systems; Influenza, Human; Nylons; SARS-CoV-2; RNA, Guide, CRISPR-Cas Systems
PubMed: 36442853
DOI: 10.1002/advs.202204689 -
Blood Advances Nov 2020Our previous real-time imaging studies directly demonstrated the spatiotemporal regulation of clot formation and lysis by activated platelets. In addition to their...
Our previous real-time imaging studies directly demonstrated the spatiotemporal regulation of clot formation and lysis by activated platelets. In addition to their procoagulant functions, platelets enhanced profibrinolytic potential by augmenting the accumulation of tissue-type plasminogen activator (tPA) and plasminogen, in vivo in a murine microthrombus model, and in vitro in a platelet-containing plasma clot model. To clarify the role of thrombin-activatable fibrinolysis inhibitor (TAFI), which regulates coagulation-dependent anti-fibrinolytic potential, we analyzed tPA-induced clot lysis times in platelet-containing plasma. Platelets prolonged clot lysis times in a concentration-dependent manner, which were successfully abolished by a thrombomodulin-neutralizing antibody or an activated TAFI inhibitor (TAFIaI). The results obtained using TAFI- or factor XIII-deficient plasma suggested that TAFI in plasma, but not in platelets, was essential for this prolongation, though its cross-linkage with fibrin was not necessary. Confocal laser scanning microscopy revealed that fluorescence-labeled plasminogen accumulated on activated platelet surfaces and propagated to the periphery, similar to the propagation of fibrinolysis. Plasminogen accumulation and propagation were both enhanced by TAFIaI, but only accumulation was enhanced by thrombomodulin-neutralizing antibody. Labeled TAFI also accumulated on both fibrin fibers and activated platelet surfaces, which were Lys-binding-site-dependent and Lys-binding-site-independent, respectively. Finally, TAFIaI significantly prolonged the occlusion times of tPA-containing whole blood in a microchip-based flow chamber system, suggesting that TAFI attenuated the tPA-dependent prolongation of clot formation under flow. Thus, activated platelet surfaces are targeted by plasma TAFI, to attenuate plasminogen accumulation and fibrinolysis, which may contribute to thrombogenicity under flow.
Topics: Animals; Blood Coagulation; Blood Platelets; Carboxypeptidase B2; Fibrin Clot Lysis Time; Fibrinolysis; Mice
PubMed: 33166409
DOI: 10.1182/bloodadvances.2020002923 -
Journal of Pharmaceutical Analysis Aug 2020Researches on detection of human papillomavirus (HPV) high-risk samples were carried out by polymerase chain reaction (PCR) coupled with microchip electrophoresis (MCE)....
Researches on detection of human papillomavirus (HPV) high-risk samples were carried out by polymerase chain reaction (PCR) coupled with microchip electrophoresis (MCE). Herein, we introduced a simple, rapid, automated method for detecting high-risk samples HPV16 and HPV18. In this research, general primers were initially selected to obtain sufficient detectable yield by PCR to verify feasibility of MCM method for HPV detection, then type-specific primers were further used to evaluate the specificity of MCE method. The results indicated MCE method was capable of specifically detecting high-risk HPV16 and HPV18, and also enabled simultaneous detection of multiplex samples. This MCE method described here has been successfully applied to HPV detection and displayed excellent reliability demonstrating by sequencing results. The inherent capability of MCE facilitated HPV detection conducted in a small chip with automated, high throughput, massive parallelized analysis. We envision that MCE method will definitely pave a way for clinical diagnosis, and even on-site screening of cervical cancer.
PubMed: 32923006
DOI: 10.1016/j.jpha.2020.04.003 -
"State of the Mewnion": Practices of Feral Cat Care and Advocacy Organizations in the United States.Frontiers in Veterinary Science 2021Over the last several decades, feral cats have moved from the fringes to the mainstream in animal welfare and sheltering. Although many best practice guidelines have...
Over the last several decades, feral cats have moved from the fringes to the mainstream in animal welfare and sheltering. Although many best practice guidelines have been published by national non-profits and veterinary bodies, little is known about how groups “in the trenches” actually operate. Our study sought to address that gap through an online survey of feral cat care and advocacy organizations based in the United States. Advertised as “The State of the Mewnion,” its topics included a range of issues spanning non-profit administration, public health, caretaking and trapping, adoptions of friendly kittens and cats, veterinary medical procedures and policies, data collection and program efficacy metrics, research engagement and interest, and relationships with wildlife advocates and animal control agencies. Respondents from 567 organizations participated, making this the largest and most comprehensive study on this topic to date. Respondents came primarily from grassroots organizations. A majority reported no paid employees (74.6%), served 499 or fewer feral cats per year (75.0%), engaged between 1 and 9 active volunteers (54.9%), and did not operate a brick and mortar facility (63.7%). Some of our findings demonstrate a shared community of practice, including the common use of a minimum weight of 2.0 pounds for spay/neuter eligibility, left side ear tip removals to indicate sterilization, recovery holding times after surgery commonly reported as 1 night for male cats and 1 or 2 nights for females, requiring or recommending to adopters of socialized kittens/cats that they be kept indoor-only, and less than a quarter still engaging in routine testing of cats for FIV and FeLV. Our survey also reveals areas for improvement, such as most organizations lacking a declared goal with a measurable value and a time frame, only sometimes scanning cats for microchips, and about a third not using a standardized injection site for vaccines. This study paints the clearest picture yet available of what constitutes the standard practices of organizations serving feral and community cats in the United States.
PubMed: 34970620
DOI: 10.3389/fvets.2021.791134 -
Animals : An Open Access Journal From... May 2022The frequent monitoring of a horse’s body temperature post strenuous exercise is critical to prevent or alleviate exertional heat illness (EHI) from occurring....
The frequent monitoring of a horse’s body temperature post strenuous exercise is critical to prevent or alleviate exertional heat illness (EHI) from occurring. Percutaneous thermal sensing microchip (PTSM) technology has the potential to be used as a means of monitoring a horse’s body temperature during and post-exercise. However, the accuracy of the temperature readings obtained, and their relationship to core body temperature are dependent on where they are implanted. This study aimed to document the relationship between core body temperature, and temperature readings obtained using PTSM implanted in different muscles, during exercise and post application of different cool-down methods. PTSMs were implanted into the right pectoral, right gluteal, right splenius muscles, and nuchal ligament. The temperatures were monitored during treadmill exercise, and post application of three different cool-down methods: no water application (Wno), water application only (Wonly), and water application following scraping (Wscraping). Central venous temperature (TCV) and PTSM temperatures from each region were obtained to investigate the optimal body site for microchip implantation. In this study, PTSM technology provided a practical, safe, and quick means of measuring body temperature in horses. However, its temperature readings varied depending on the implantation site. All muscle temperature readings exhibited strong relationships with TCV (r = 0.85~0.92, p < 0.05) after treadmill exercise without human intervention (water application), while the nuchal ligament temperature showed poor relationship with TCV. The relationships between TCV and PTSM temperatures became weaker with water application. Overall, however the pectoral muscle temperature measured by PTSM technology had the most constant relationships with TCV and showed the best potential to act as an alternate means of monitoring body temperature in horses for 50 min post-exercise, when there was no human intervention with cold water application.
PubMed: 35625113
DOI: 10.3390/ani12101267 -
Frontiers in Bioengineering and... 2022In oral and maxillofacial bone reconstruction, autografts from the iliac crest represent the gold standard due to their superior clinical performance, compared to...
In oral and maxillofacial bone reconstruction, autografts from the iliac crest represent the gold standard due to their superior clinical performance, compared to autografts derived from other extraoral regions. Thus, the aim of our study was to identify putative differences between osteoblasts derived from alveolar (hOB-A) and iliac crest (hOB-IC) bone of the same donor (nine donors) by means of their molecular properties in 2D and 3D culture. We thereby focused on the gene expression of biomarkers involved in osteogenic differentiation, matrix formation and osteoclast modulation. Furthermore, we examined the transcriptional response to Vit.D3 in hOB-A and hOB-IC. Our results revealed different modulation modes of the biomarker expression in osteoblasts, namely cell origin/bone entity-dependent, and culture configuration- and/or time-dependent modulations. SEMA3A, SPP1, BGLAP and PHEX demonstrated the strongest dependence on cell origin. With respect to Vit.D3-effects, BGLAP, SPP1 and ALPL displayed the highest Vit.D3-responsiveness. In this context we demonstrated that the transcriptional Vit.D3-response concerning SPP1 and ALPL in human osteoblasts depended on the cell origin. The results indicate a higher bone remodeling activity of iliac crest than alveolar osteoblasts and support the growing evidence that a high osteoclast activity at the host-/donor bone interface may support graft integration.
PubMed: 36246375
DOI: 10.3389/fbioe.2022.918866 -
Biochip Journal Apr 2023A novel integrated detection system that introduces a paper-chip-based molecular detection strategy into a polydimethylsiloxane (PDMS) microchip and temperature control...
UNLABELLED
A novel integrated detection system that introduces a paper-chip-based molecular detection strategy into a polydimethylsiloxane (PDMS) microchip and temperature control system was developed for on-site colorimetric detection of DNA. For the paper chip-based detection strategy, a padlock probe DNA (PLP)-mediated rolling circle amplification (RCA) reaction for signal amplification and a radial flow assay according to the Au-probe labeling strategy for visualization were optimized and applied for DNA detection. In the PDMS chip, the reactions for ligation of target-dependent PLP, RCA, and labeling were performed one-step under isothermal temperature in a single chamber, and one drop of the final reaction solution was loaded onto the paper chip to form a radial colorimetric signal. To create an optimal analysis environment, not only the optimization of molecular reactions for DNA detection but also the chamber shape of the PDMS chip and temperature control system were successfully verified. Our results indicate that a detection limit of 14.7 nM of DNA was achieved, and non-specific DNAs with a single-base mismatch at the target DNA were selectively discriminated. This integrated detection system can be applied not only for single nucleotide polymorphism identification, but also for pathogen gene detection. The adoption of inexpensive paper and PDMS chips allows the fabrication of cost-effective detection systems. Moreover, it is very suitable for operation in various resource-limited locations by adopting a highly portable and user-friendly detection method that minimizes the use of large and expensive equipment.
SUPPLEMENTARY INFORMATION
The online version contains supplementary material available at 10.1007/s13206-023-00101-7.
PubMed: 37363267
DOI: 10.1007/s13206-023-00101-7 -
Journal of Diabetes and Metabolic... Dec 2019In this paper, the effect of the position of the inlet and outlet microchannels on the flow profile and the geometry of the recognition chamber for sample pre-treatment...
OBJECTIVES
In this paper, the effect of the position of the inlet and outlet microchannels on the flow profile and the geometry of the recognition chamber for sample pre-treatment in an electrochemical biosensor to be used in osteoporosis management were investigated.
METHODS
All numerical computation presented in this work were performed using COMSOL Multiphysics and Fluent. Simulation was performed for a three-dimensional, incompressible Navier-Stokes flow and so explicit biphasic volume of fluid (VOF) equations were used.
RESULTS
In the designed microfluidic system, a pressure-driven laminar flow with no-slip boundary condition was responsible for fluid actuation through microchannels in a reproducible approach. Based on the simulation results, the number of outlets was increased and the angel through which the inlets and outlets were attached to the microchamber was changed so that the dead volume would be eliminated and the fluid flow trajectory, the velocity field and pressure were evenly distributed across the chamber. The Re number in the inlets was equal to 4.41, suggesting a laminar flow at this site.
CONCLUSION
The simulation results along with the fact that the design change was tested using laser ablated tape and a color dye at different steps provided the researchers with the opportunity to study the changes in a fast and accurate but cheap method. The absence of backflow helps with the cross-talk concern in the channels and the lack of bubbles and complete coverage of the chamber helps with a better surface modification and thus better sensing performance.
PubMed: 31890659
DOI: 10.1007/s40200-019-00418-x