Cell entosis is a novel cell death process starting from cell-in-cell invasion. In general, cancer cells own higher incidence rate of cell entosis comparing to non-cancerous cells. Studies arguing ...whether cell entosis is a tumor suppressing process or a tumor accelerating process can deepen our understanding of tumor development. Cell elasticity is recognized as one of tumor malignant biomarkers. There have been some researchers studying cell elasticity in cell entosis. However, existing cell elasticity sensing technique (i.e. micropipette aspiration) can hardly be reliable neither high-throughput. In this work, we introduce an elasticity sensing method for quantifying both cell elasticity in cell-in-cell structures and single floating cells using a microfluidic cytometer. We not only argue our cell elasticity sensing method is reliable for already occurred entosis but also apply such method on predicting the “outer” cells in entosis of different cell types. The elasticity sensing method proposed in this manuscript is able to provide an effective and reliable way to further study deeper mechanism in cell entosis.
Age-related hearing loss is a complex disease caused by a combination of genetic and environmental factors, and a study have conducted animal experiments to explore the association between BCL11B ...heterozygosity and age-related hearing loss. The present study used established genetic models to examine the association between BCL11B gene polymorphisms and age-related hearing loss. A total of 410 older adults from two communities in Qingdao, China, participated in this study. The case group comprised individuals aged greater than or equal to 60 years with age-related hearing loss, and the control group comprised individuals without age-related hearing loss from the same communities. The groups were matched 1:1 for age and sex. The individual characteristics of the participants were analyzed descriptively using the Mann-Whitney U test and the chi-square test. To explore the association between BCL11B gene polymorphisms and age-related hearing loss, conditional logistic regression was performed to construct genetic models for two single-nucleotide-polymorphisms (SNPs) of BCL11B, and haplotype analysis was conducted to construct their haplotype domains. Two SNP sites of the BCL11B gene, four genetic models of rs1152781 (additive, dominant, recessive, and codominant), and five genetic models of rs1152783 (additive, dominant, recessive, codominant, and over dominant) were significantly associated with age-related hearing loss in the models both unadjusted and adjusted for all covariates (P < 0.05). Additionally, a linkage disequilibrium between rs1152781 and rs1152783 was revealed through haplotype analysis. Our study revealed that BCL11B gene polymorphisms were significantly associated with age-related hearing loss.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Abstract
Human host-associated microbial communities in body sites can reflect health status based on the population distribution and specific microbial properties in the heterogeneous community. ...Bacteria identification at the single-cell level provides a reliable biomarker and pathological information for clinical diagnosis. Nevertheless, biosamples obtained from some body sites cannot offer sufficient sample volume and number of target cells as required by most of the existing single-cell isolation methods such as flow cytometry. Herein we report a novel integrated microfluidic system, which consists of a microemulsion module for single-bacteria encapsulation and a sequential microdroplet capture and release module for selectively extracting only the single-bacteria encapsulated in microdroplets. We optimize the system for a success rate of the single-cell extraction to be > 38%. We further verify applicability of the system with prepared cell mixtures (
Methylorubrum extorquens
AM1 and
Methylomicrobium album
BG8) and biosamples collected from human skin, to quantify the population distribution of multiple key species in a heterogeneous microbial community. Results indicate perfect viability of the single-cell extracts and compatibility with downstream analyses such as PCR. Together, this research demonstrates that the reported single-bacteria extraction system can be applied in microbiome and pathology research and clinical diagnosis as a clinical or point-of-care device.
Investigating floating cells along a narrow microchannel (e.g., a blood vessel) for their transiting speeds and the corresponding roles of cell physical properties can deepen our understanding of ...circulating tumor cells (CTCs) metastasis via blood vessels. Many existing studies focus on the cell transiting process in blood vessel-like microchannels; further analytical studies are desired to summarize behaviors of the floating cell movement under different conditions. In this work, we perform a theoretical analysis to establish a relation between the transiting speed and key cell physical properties. We also conduct computational fluid dynamics simulation and microfluidic experiments to verify the theoretical model. This work reveals key cell physical properties and the channel configurations determining the transiting speed. The reported model can be applied to other works with various dimensions of microchannels as a more general way to evaluate the cancer cell metastasis ability with microfluidics.
Single-cell level coculture facilitates the study of cellular interactions for uncovering unknown physiological mechanisms, which are crucial for the development of new therapies for diseases. ...However, efficient approaches for high-throughput deterministic pairing of single cells and traceable coculture remain lacking. In this study, we report a new microfluidic device, which combines hydrodynamic and recirculation flow captures, to achieve high-throughput and deterministic pairing of single cells in a microwell array for traceable coculture. Compared with the existing techniques, the developed device exhibits advantages with regard to pairing efficiency, throughput, determinacy, and traceability. Through repeating a two-step method, which sequentially captures single cells in a meandering channel and a microwell array, cell number and type can be easily controlled. Double and triple single-cell pairings have been demonstrated with an efficiency of 72.2% and 38.0%, respectively. Cellular engulfment using two breast cell lines is investigated on a developed microfluidic chip as a biological case study, in which the morphological characteristics and the incidence rate are analyzed. This research provides an efficient and reliable alternative for the coculture of single cells on the microfluidic platform for various biomedical applications, such as studying cellular engulfment and tumor sphere formation under single-cell pairing condition.
The tumor microenvironment (TME) is considered to be one of the vital mediators of tumor progression. Extracellular matrix (ECM), infiltrating immune cells, and stromal cells collectively constitute ...the complex ecosystem with varied biochemical and biophysical properties. The development of liver cancer is strongly tied with fibrosis and cirrhosis that alters the microenvironmental landscape, especially ECM composition. Enhanced deposition and cross‐linking of type I collagen are frequently detected in patients with liver cancer and have been shown to facilitate tumor growth and metastasis by epithelial‐to‐mesenchymal transition. However, information on the effect of collagen enrichment on drug resistance is lacking. Thus, the present study has comprehensively illustrated phenotypical and mechanistic changes in an in vitro mimicry of collagen‐enriched TME and revealed that collagen enrichment could induce 5‐fluorouracil (5FU) and sorafenib resistance in liver cancer cells through hypoxia‐induced up‐regulation of lysyl oxidase‐like 2 (LOXL2). LOXL2, an enzyme that facilitates collagen cross‐linking, enhances cell adhesion‐mediated drug resistance by activating the integrin alpha 5 (ITGA5)/focal adhesion kinase (FAK)/phosphoinositide 3‐kinase (PI3K)/rho‐associated kinase 1 (ROCK1) signaling axis. Conclusion: We demonstrated that inhibition of LOXL2 in a collagen‐enriched microenvironment synergistically promotes the efficacy of sorafenib and 5FU through deterioration of focal adhesion signaling. These findings have clinical implications for developing LOXL2‐targeted strategies in patients with chemoresistant liver cancer and especially for those patients with advanced fibrosis and cirrhosis.
In this study, we revealed the collagen‐mediated chemoresistance via activation of HIF1A/LOXL2/ITGA5/FAK/PI3K/ROCK1 signaling and showed that LOXL2 is a promising therapeutic target to enhance the efficacy of 5FU and sorafenib in resistant liver cancer patients, especially those with advanced fibrosis and cirrhosis.
OBJECTIVETo explore the differences in DNA methylation associated with age-related hearing loss in a study of 57 twin pairs from China. DESIGNMonozygotic twins were identified through the Qingdao ...Twin Registration system. The median age of participants was > 50 years. Their hearing thresholds were measured using a multilevel pure-tone audiometry assessment. The pure-tone audiometry was calculated at low frequencies (0.5, 1.0, and 2.0 kHz), speech frequencies (0.5, 1.0, 2.0, and 4.0 kHz), and high frequencies (4.0 and 8 kHz). The CpG sites were tested using a linear mixed-effects model, and the function of the cis-regulatory regions and ontological enrichments were predicted using the online Genomic Regions Enrichment of Annotations Tool. The differentially methylated regions were identified using a comb-p python library approach. RESULTSIn each of the PTA categories (low-, speech-, high-frequency), age-related hearing loss was detected in 25.9%, 19.3%, and 52.8% of participants. In the low-, speech- and high-frequency categories we identified 18, 42, and 12 individual CpG sites and 6, 11, and 6 differentially methylated regions. The CpG site located near DUSP4 had the strongest association with low- and speech-frequency, while the strongest association with high-frequency was near C21orf58. We identified associations of ALG10 with high-frequency hearing, C3 and LCK with low- and speech-frequency hearing, and GBX2 with low-frequency hearing. Top pathways that may be related to hearing, such as the Notch signaling pathway, were also identified. CONCLUSIONOur study is the first of its kind to identify these genes and their associated with DNA methylation may play essential roles in the hearing process. The results of our epigenome-wide association study on twins clarify the complex mechanisms underlying age-related hearing loss.
Phagocytic cells recognize and phagocytose invading microbes for destruction. However, bacterial pathogens can remain hidden at low levels from conventional detection or replicate intracellularly ...after being phagocytosed by immune cells. Current phagocytosis-detection approaches involve flow cytometry or microscopic search for rare bacteria-internalized phagocytes among large populations of uninfected cells, which poses significant challenges in research and clinical settings. Hence it is imperative to develop a rapid, non-disruptive, and label-free phagocytosis detection approach. Using deformability assays and microscopic imaging, we have demonstrated for the first time that the presence of intracellular bacteria in phagocytic blood cells led to aberrant physical properties. Specifically, human monocytes with internalized bacteria of various species were stiffer and larger compared with uninfected monocytes. Taking advantage of these physical differences, a novel microfluidics-based biosensor platform was developed to passively sort, concentrate and quantify rare monocytes with internalized pathogens (MIP) from uninfected monocyte populations for phagocytosis detection. The clinical utility of the MIP platform was demonstrated by enriching and detecting bacteria-internalized monocytes from spiked human blood samples within 1.5 h. Patient-derived clinical isolates were used to validate the utility of the MIP platform further. This proof-of-concept presents a phagocytosis detection platform that could be used to rapidly diagnose microbial infections, especially in bloodstream infections (BSIs), thereby improving the clinical outcomes for point-of-care management.
•Development of the first label-free biosensor to detect phagocytosis, based on physical differences in ininfected cells.•Circumvent search for rare infected cells. Identification of internalized bacteria-positive samples could be completed within 1.5 h.•Broad applications to infection caused by different bacteria (gram-positive and gram-negative, extracellular and intracellular bacteria).•Demonstration of clinical relevance with P. aeruginosa-based pneumonia isolates from cystic fibrosis patients.•Application for clinically relevant infected blood samples at low sample volume (~1 ml).
Stretchable electronic circuits can seamlessly conform to irregular and dynamic surfaces with high integration. However, current stretchable configurations typically have limited stretchability due ...to the lack of robust connections between soft interconnects and rigid electronics. Here, we printed highly stretchable metal–polymer conductors on thermoplastic elastomers as interconnects. We developed electronic vests with porous surfaces for rigid electronics and introduced polyester hot-melt adhesives to strengthen connections between soft interconnects and rigid electronics. After thermal bonding, the adhesive penetrates the porous surface of electronic vests, creating a mechanical interlock and providing an adhesion force of 8.34 N/cm for the connection (3× higher than conductive adhesives). Thus, rigid electronics of different sizes and different pin counts can form strong connections to soft interconnects, achieving a maximum strain tolerance of ~700% (10× higher than conductive adhesives). We achieved highly integrated ultra-stretchable displays that can withstand stretching up to 220% without dead pixels.
This work describes a two-chip acoustofluidic platform for two-dimensional (2D) manipulation of microparticles in a closed microchamber on a reusable surface acoustic wave (SAW) device. This platform ...comprises two microfabricated chips: (1) a detachable silicon superstrate enclosed by a PDMS microfluidic chamber and (2) a reusable SAW device for generating standing SAW (SSAW), which is typically an expensive component. Critical to such a two-chip acoustofluidic platform is the selection of a suitable coupling agent at the interface of the SAW device and superstrate. To this end, we applied a polymer thin film as a coupling agent that balances between acoustic coupling efficiency, stability over time, and reusability. Recycling of the SAW device lowers the cost-barrier for acoustofluidic particle manipulation. The SSAW is transmitted into the silicon superstrate
via
the coupling agent to form a standing Lamb wave (SLW) to trap and move microparticles. The reported two-chip strategy enables the single-use microfluidic superstrates to avoid chemical and biological contaminations, while maintaining the merits of acoustofluidic manipulation of being noncontact and label-free and applicable to a wide range of microparticles with different shapes, density, polarity, and electrical properties.
A two-chip acoustofluidic particle manipulation platform with a detachable and reusable surface acoustic wave device enables manipulation of microparticles in 2D on a replaceable silicon superstrate.
PDF
