In this study, we developed a facile gold nanozyme-based paper chip (AuNZ-PAD) for Hg
detection. This device has the advantages of being simple, rapid, cost effective, sensitive, selective, high ...throughput, and applicable to onsite detection. The colorimetric mercury assay on the AuNZ-PAD is established based on the enzyme-like catalytic activity of gold nanoparticles promoted by the formation of Au-Hg amalgam, which is correlated to the intensity of the colorimetric response resulting from the catalytic reaction of 3,3',5,5'-tetramethylbenzidine (TMB) and H
O
. Highly sensitive and selective detection of Hg
ions is achieved in both distilled and tap water samples, indicating the feasibility and applicability of our device for the determination of mercury pollution in real samples. Moreover, AuNZ-PAD analysis using a smartphone camera eliminates the need for expensive analytical equipment, thereby increasing the practicality of field monitoring of trace Hg
compared with other sensing methods.
Rationale
Various medium formulations contain essential fatty acids at concentrations ranging from 10 to 100 mg/L. Accurate and precise lipid measurement in media is crucial for monitoring media ...quality and conducting studies on lipids in the context of cell culture. This study employed two‐dimensional gas chromatography (GC × GC) analyses to offer enhanced resolution, sensitivity, and separation performance compared to GC.
Methods
Quantification of fatty acid methyl esters (FAMEs) in a medium was conducted using GC × GC combined with a high‐resolution mass spectrometer and flame ionization detector, considering potential interference from nonionic surfactant Tween 80, which was precipitated and removed by optimizing the concentration of cobalt thiocyanate (CTA) solution during pretreatment. This advanced analytical approach enabled identification of cis and trans isomers of identical molecular weights and determination of the location and number of double bonds in the same carbon number structure.
Results
Our analysis identified 36 FAMEs within the C6–C24 region, and a 5% CTA solution was optimal for efficient removal of Tween 80 during lipid extraction. Additionally, this advanced method minimized FAME contamination and loss during pretreatment, thereby significantly reducing the sample volume required to detect trace levels of FAMEs. This improvement led to a fatty acid recovery rate of 106% while maintaining the average relative standard deviation for the target FAMEs of about 3%.
Conclusions
Our research paves the way for future investigation into medium quality control and the role of fatty acids in cell culture. This offers the possibility for economical and effective trace quantification of fatty acids in complex media.
In this study, we developed a new type of paper-based analytical device (PAD), the three-dimensional (3D) slip-PAD, to detect infectious human norovirus for global healthcare. The 3D configuration of ...the papers combined with a slip design provides unique features and versatility that overcome the limitations of fluidic manipulation and sensitivity in point-of-care (POC) tests. The assay can be carried out in a single step based on a moveable slip design, making it suitable for unskilled users. The 3D fluidic network developed by layered construction of wax-patterned papers provides different fluidic paths for the sequential delivery of multiple fluids without the need for peripheral equipment. The release and mixing of enhancement reagents on the device improved the sensitivity and detection limit. The assay results could be visualized by naked eye within 10 min, with subsequent amplification of the signal over time (<60 min). The device showed a broad dynamic range of detection and high sensitivity, with a detection limit of 9.5 × 10(4) copies ml(-1) for human norovirus. These results demonstrate that the 3D slip-PAD is a sensitive diagnostic assay for detecting human norovirus infection that is particularly suitable for POC testing in regions where resources are scarce.
Tight regulation of Toll‐like receptor (TLR)‐mediated inflammatory responses is important for innate immunity. Here, we show that T‐cell death‐associated gene 51 (TDAG51/PHLDA1) is a novel regulator ...of the transcription factor FoxO1, regulating inflammatory mediator production in the lipopolysaccharide (LPS)‐induced inflammatory response. TDAG51 induction by LPS stimulation was mediated by the TLR2/4 signaling pathway in bone marrow‐derived macrophages (BMMs). LPS‐induced inflammatory mediator production was significantly decreased in TDAG51‐deficient BMMs. In TDAG51‐deficient mice, LPS‐ or pathogenic Escherichia coli infection‐induced lethal shock was reduced by decreasing serum proinflammatory cytokine levels. The recruitment of 14‐3‐3ζ to FoxO1 was competitively inhibited by the TDAG51‐FoxO1 interaction, leading to blockade of FoxO1 cytoplasmic translocation and thereby strengthening FoxO1 nuclear accumulation. TDAG51/FoxO1 double‐deficient BMMs showed significantly reduced inflammatory mediator production compared with TDAG51‐ or FoxO1‐deficient BMMs. TDAG51/FoxO1 double deficiency protected mice against LPS‐ or pathogenic E. coli infection‐induced lethal shock by weakening the systemic inflammatory response. Thus, these results indicate that TDAG51 acts as a regulator of the transcription factor FoxO1, leading to strengthened FoxO1 activity in the LPS‐induced inflammatory response.
Synopsis
Toll‐like receptor (TLR) signaling is tightly controlled during inflammatory responses. Here, TDAG51/PHLDA1 is identified as a new interactor and regulator of transcription factor FoxO1, stimulating inflammatory mediator production upon LPS challenge.
TDAG51 expression is induced during the LPS‐induced inflammatory response in macrophages.
TDAG51 interacts with the transcription factor FoxO1 and competitively inhibits 14‐3‐3𝝵 binding
TDAG51‐FoxO1 interaction and reduced 14‐3‐3𝝵 binding enhances FoxO1 nuclear accumulation, thereby promoting inflammatory mediator expression.
TDAG51 deficiency attenuates LPS‐induced lethal shock by weakening the inflammatory response in mice.
TLR‐mediated expression of inflammatory mediators is fine‐tuned by T‐cell death‐associated gene 51, which competes with inhibitory 14‐3‐3𝝵 binding to FoxO1.
In this study, a planar split-ring resonator (SRR)-based RF biosensor was developed for label-free detection of biomolecules such as the prostate cancer marker, prostate specific antigen (PSA), and ...cortisol stress hormone. The biosensor has a resonance-assisted transducer and is excited by a time-varying magnetic field component of a local high-impedance microstrip line. The resulting device exhibits an intrinsic S21 resonance with a quality-factor (or Q-factor) of 50. For the biomolecular interaction, anti-PSA and anti-cortisol were immobilized on the gold surface of the resonator by a protein-G mediated bioconjugation process and corresponding frequency shifts of Δf1p=30±2 MHz (for anti-PSA) and Δf1c=20±3 MHz (for anti-cortisol) were observed. The additional frequency shift of each PSA and cortisol antigen with a 100pg/ml concentration was about 5±1.5MHz and 3±1MHz, respectively. From the experimental results, we confirmed that our device is very effective RF biosensor with a limit of detection (LOD) of 100pg/ml and has sufficiently feasibility as a label-free biosensing scheme.
Postpartum depression is a severe emotional and mental disorder that involves maternal care defects and psychiatric illness. Postpartum depression is closely associated with a combination of physical ...changes and physiological stress during pregnancy or after parturition in stress-sensitive women. Although postpartum depression is relatively well known to have deleterious effects on the developing fetus, the influence of genetic risk factors on the development of postpartum depression remains unclear. In this study, we discovered a novel function of T cell death-associated gene 51 (TDAG51/PHLDA1) in the regulation of maternal and depressive-like behavior. After parturition, TDAG51-deficient dams showed impaired maternal behavior in pup retrieving, nursing and nest building tests. In contrast to the normal dams, the TDAG51-deficient dams also exhibited more sensitive depressive-like behaviors after parturition. Furthermore, changes in the expression levels of various maternal and depressive-like behavior-associated genes regulating neuroendocrine factor and monoamine neurotransmitter levels were observed in TDAG51-deficient postpartum brain tissues. These findings indicate that TDAG51 plays a protective role against maternal care defects and depressive-like behavior after parturition. Thus, TDAG51 is a maternal care-associated gene that functions as a crucial regulator of maternal and depressive-like behavior after parturition.
Extracellular signal-regulated kinase 3 (ERK3) is an atypical member of the mitogen-activated protein kinase (MAPK) family, members of which play essential roles in diverse cellular processes during ...carcinogenesis, including cell proliferation, differentiation, migration, and invasion. Unlike other MAPKs, ERK3 is an unstable protein with a short half-life. Although deubiquitination of ERK3 has been suggested to regulate the activity, its ubiquitination has not been described in the literature. Here, we report that FBXW7 (F-box and WD repeat domain-containing 7) acts as a ubiquitination E3 ligase for ERK3. Mammalian two-hybrid assay and immunoprecipitation results demonstrated that ERK3 is a novel binding partner of FBXW7. Furthermore, complex formation between ERK3 and the S-phase kinase-associated protein 1 (SKP1)-cullin 1-F-box protein (SCF) E3 ligase resulted in the destabilization of ERK3 via a ubiquitination-mediated proteasomal degradation pathway, and FBXW7 depletion restored ERK3 protein levels by inhibiting this ubiquitination. The interaction between ERK3 and FBXW7 was driven by binding between the C34D of ERK3, especially at Thr417 and Thr421, and the WD40 domain of FBXW7. A double mutant of ERK3 (Thr417 and Thr421 to alanine) abrogated FBXW7-mediated ubiquitination. Importantly, ERK3 knockdown inhibited the proliferation of lung cancer cells by regulating the G
/S-phase transition of the cell cycle. These results show that FBXW7-mediated ERK3 destabilization suppresses lung cancer cell proliferation in vitro.
Dasatinib is an inhibitor of Src that has anti‐tumour effects on many haematological and solid cancers. However, the anti‐tumour effects of dasatinib on human oral cancers remain unclear. In this ...study, we investigated the effects of dasatinib on different types of human oral cancer cells: the non‐tumorigenic YD‐8 and YD‐38 and the tumorigenic YD‐10B and HSC‐3 cells. Strikingly, dasatinib at 10 µM strongly suppressed the growth and induced apoptosis of YD‐38 cells and inhibited the phosphorylation of Src, EGFR, STAT‐3, STAT‐5, PKB and ERK‐1/2. In contrast, knockdown of Src blocked the phosphorylation of EGFR, STAT‐5, PKB and ERK‐1/2, but not STAT‐3, in YD‐38 cells. Dasatinib induced activation of the intrinsic caspase pathway, which was inhibited by z‐VAD‐fmk, a pan‐caspase inhibitor. Dasatinib also decreased Mcl‐1 expression and S6 phosphorylation while increased GRP78 expression and eIF‐2α phosphorylation in YD‐38 cells. In addition, to its direct effects on YD‐38 cells, dasatinib also exhibited anti‐angiogenic properties. Dasatinib‐treated YD‐38 or HUVEC showed reduced HIF‐1α expression and stability. Dasatinib alone or conditioned media from dasatinib‐treated YD‐38 cells inhibited HUVEC tube formation on Matrigel without affecting HUVEC viability. Importantly, dasatinib's anti‐growth, anti‐angiogenic and pro‐apoptotic effects were additionally seen in tumorigenic HSC‐3 cells. Together, these results demonstrate that dasatinib has strong anti‐growth, anti‐angiogenic and pro‐apoptotic effects on human oral cancer cells, which are mediated through the regulation of multiple targets, including Src, EGFR, STAT‐3, STAT‐5, PKB, ERK‐1/2, S6, eIF‐2α, GRP78, caspase‐9/3, Mcl‐1 and HIF‐1α.
Background: TNF receptor-associated factor 2 (TRAF2) is a key adaptor molecule in the TNF receptor (TNFR) signaling pathway.
Results: TRAF-interacting protein (TRIP) inhibits Lys63-linked TRAF2 ...ubiquitination by blocking the binding of the cofactor sphingosine 1-phosphate (S1P) to the TRAF2 RING domain.
Conclusion: TRIP negatively regulates the TRAF2 ubiquitin-dependent pathway by modulating the TRAF2-S1P interaction.
Significance: TRIP is an important cellular regulator of the TNFR-mediated inflammatory response.
The signaling pathway downstream of TNF receptor (TNFR) is involved in the induction of a wide range of cellular processes, including cell proliferation, activation, differentiation, and apoptosis. TNFR-associated factor 2 (TRAF2) is a key adaptor molecule in TNFR signaling complexes that promotes downstream signaling cascades, such as nuclear factor-κB (NF-κB) and mitogen-activated protein kinase activation. TRAF-interacting protein (TRIP) is a known cellular binding partner of TRAF2 and inhibits TNF-induced NF-κB activation. Recent findings that TRIP plays a multifunctional role in antiviral response, cell proliferation, apoptosis, and embryonic development have increased our interest in exploring how TRIP can affect the TNFR-signaling pathway on a molecular level. In our current study, we demonstrated that TRIP is negatively involved in the TNF-induced inflammatory response through the down-regulation of proinflammatory cytokine production. Here, we demonstrated that the TRAF2-TRIP interaction inhibits Lys63-linked TRAF2 ubiquitination by inhibiting TRAF2 E3 ubiquitin (Ub) ligase activity. The TRAF2-TRIP interaction inhibited the binding of sphingosine 1-phosphate, which is a cofactor of TRAF2 E3 Ub ligase, to the TRAF2 RING domain. Finally, we demonstrated that TRIP functions as a negative regulator of proinflammatory cytokine production by inhibiting TNF-induced NF-κB activation. These results indicate that TRIP is an important cellular regulator of the TNF-induced inflammatory response.
In the present study, to remove harmful cyanobacterial species Microcystis aeruginosa from aqueous phases, adsorption-based strategy was utilized. For this strategy, the surface of cotton fiber was ...modified using chitosan molecules to develop a highly efficient and ecofriendly adsorbent in removal of Microcystis aeruginosa from aqueous solution. The pristine cotton fiber could not remove M. aeruginosa, while the chitosan-modified cotton (CS-m-Cotton) showed the 95% of cell removal efficiency within 12 h. The surface characteristics of chitosan-modified cotton compared to the pristine cotton fiber was examined by various surface analysis methods. In addition, the pre-treatment of pristine cotton using sodium hydroxide solution was an important factor for enhancement of chitosan modification efficiency on the cotton fiber. The developed chitosan-modified cotton fiber could be reusable for M. aeruginosa cell removal after the simple desorption treatment using ultrasonication in alkaline solution. During the repeated adsorbent regeneration and reuse, the chitosan-modified cotton maintained its M. aeruginosa removal efficiencies (>90%). From the acute toxicity assessment using the chitosan-modified cotton and, the measurements of chemical oxygen demand and microcystin level changes in the M. aeruginosa treatment process using the adsorbent, the environmental safety of the adsorption strategy using the developed adsorbent could be confirmed. Based on our results, the chitosan-modified cotton fiber could be proposed as an efficient and ecofriendly solution for remediation of harmful cyanobacterial species occurring water resources.
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•Chemical free adsorbent can be fabricated by chitosan coating on the cotton fiber.•Fabricated CS-m-Cotton showed the high M. aeruginosa removal efficiency.•NaOH solution pretreatment of cotton improves chitosan modification efficiency.•CS-m-cotton could be reused five times for cell treatment even without crosslinking.•CS-m-cotton is non-toxic, and did not increase the COD and cyanotoxin levels.
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