Luminescent lanthanide-containing gelatin/polydextran/laponite nanocomposite double-network hydrogels for processing and sensing applications.

Lanthanide-containing nanomaterials have gained significant popularity for their utilization in polymeric networks, enabling the creation of luminescent nanocomposites for advanced applications. In this study, we developed a new type of lanthanide-containing nanocomposite hydrogels by incorporating terbium-containing laponite (Tb3+@Lap) into the networks of polyethyleneimine-modified gelatin/polydextran aldehyde (PG/PDA) through dynamic bonds. The structures and properties of the Tb3+@Lap-containing nanocomposite double-network (ncDN) hydrogels were comprehensively investigated in comparison with the DN hydrogels with a pure polymeric network and the Lap-containing ncDN hydrogels. The PG/PDA/Tb3+@Lap ncDN hydrogels with multiple dynamic bonds (i.e., imine bonds, coordination bonds, hydrogen bonds, and electrostatic interactions) exhibited remarkable characteristics of shear-thinning and self-healing, making them suitable for the construction of hydrogel scaffolds on a macroscale using fabrication techniques such as electrospinning and 3D printing. Moreover, the PG/PDA/Tb3+@Lap ncDN hydrogels have been demonstrated to act as sensitive and selective luminescent sensors for detecting copper ions. Taken together, a versatile lanthanide-containing ncDN hydrogel platform capable of dynamic features is developed for processing and sensing applications.

Machine Learning Classification for Assessing the Degree of Stenosis and Blood Flow Volume at Arteriovenous Fistulas of Hemodialysis Patients Using a New Photoplethysmography Sensor Device.

The classifier of support vector machine (SVM) learning for assessing the quality of arteriovenous fistulae (AVFs) in hemodialysis (HD) patients using a new photoplethysmography (PPG) sensor device is presented in this work. In clinical practice, there are two important indices for assessing the quality of AVF: the blood flow volume (BFV) and the degree of stenosis (DOS). In hospitals, the BFV and DOS of AVFs are nowadays assessed using an ultrasound Doppler machine, which is bulky, expensive, hard to use, and time consuming. In this study, a newly-developed PPG sensor device was utilized to provide patients and doctors with an inexpensive and small-sized solution for ubiquitous AVF assessment. The readout in this sensor was custom-designed to increase the signal-to-noise ratio (SNR) and reduce the environment interference via maximizing successfully the full dynamic range of measured PPG entering an analog-digital converter (ADC) and effective filtering techniques. With quality PPG measurements obtained, machine learning classifiers including SVM were adopted to assess AVF quality, where the input features are determined based on optical Beer-Lambert's law and hemodynamic model, to ensure all the necessary features are considered. Finally, the clinical experiment results showed that the proposed PPG sensor device successfully achieved an accuracy of 87.84% based on SVM analysis in assessing DOS at AVF, while an accuracy of 88.61% was achieved for assessing BFV at AVF.

A Portable, Wireless Photoplethysomography Sensor for Assessing Health of Arteriovenous Fistula Using Class-Weighted Support Vector Machine.

A portable, wireless photoplethysomography (PPG) sensor for assessing arteriovenous fistula (AVF) by using class-weighted support vector machines (SVM) was presented in this study. Nowadays, in hospital, AVF are assessed by ultrasound Doppler machines, which are bulky, expensive, complicated-to-operate, and time-consuming. In this study, new PPG sensors were proposed and developed successfully to provide portable and inexpensive solutions for AVF assessments. To develop the sensor, at first, by combining the dimensionless number analysis and the optical Beer Lambert's law, five input features were derived for the SVM classifier. In the next step, to increase the signal-noise ratio (SNR) of PPG signals, the front-end readout circuitries were designed to fully use the dynamic range of analog-digital converter (ADC) by controlling the circuitries gain and the light intensity of light emitted diode (LED). Digital signal processing algorithms were proposed next to check and fix signal anomalies. Finally, the class-weighted SVM classifiers employed five different kernel functions to assess AVF quality. The assessment results were provided to doctors for diagonosis and detemining ensuing proper treatments. The experimental results showed that the proposed PPG sensors successfully achieved an accuracy of 89.11% in assessing health of AVF and with a type II error of only 9.59%.

Functional regulation of Zfp36l1 and Zfp36l2 in response to lipopolysaccharide in mouse RAW264.7 macrophages.

BACKGROUND: The tristetraprolin (TTP) family of mRNA-binding proteins contains three major members, Ttp, Zfp36l1, and Zfp36l2. Ttp down-regulates the stability of AU-rich element-containing mRNAs and functions as an anti-inflammation regulator. METHODS: To examine whether other TTP family proteins also play roles in the inflammatory response, their expression profiles and the possible mRNA targets were determined in the knockdown cells. RESULTS: Ttp mRNA and protein were highly induced by lipopolysaccharide (LPS), whereas Zfp36l1 and Zfp36l2 mRNAs were down-regulated and their proteins were phosphorylated during early lipopolysaccharide stimulation. Biochemical and functional analyses exhibited that the decrease of Zfp36l2 mRNA was cross-regulated by Ttp. Knockdown of Zfp36l1 and Zfp36l2 increased the basal level of Mkp-1 mRNAs by prolonging its half-life. Increasing the expression of Mkp-1 inhibited the activation of p38 MAPK under lipopolysaccharide stimulation and down-regulated Tnfα, and Ttp mRNA. In addition, hyper-phosphorylation of Zfp36l1 might stabilize Mkp-1 expression by forming a complex with the adapter protein 14-3-3 and decreasing the interaction with deadenylase Caf1a. CONCLUSIONS: Our findings imply that the expression and phosphorylation of Zfp36l1 and Zfp36l2 may modulate the basal level of Mkp-1 mRNA to control p38 MAPK activity during lipopolysaccharide stimulation, which would affect the inflammatory mediators production. Zfp36l1 and Zfp36l2 are important regulators of the innate immune response.

Phosphorylation of mRNA decapping protein Dcp1a by the ERK signaling pathway during early differentiation of 3T3-L1 preadipocytes.

BACKGROUND: Turnover of mRNA is a critical step in the regulation of gene expression, and an important step in mRNA decay is removal of the 5' cap. We previously demonstrated that the expression of some immediate early gene mRNAs is controlled by RNA stability during early differentiation of 3T3-L1 preadipocytes. METHODOLOGY/PRINCIPAL FINDINGS: Here we show that the mouse decapping protein Dcp1a is phosphorylated via the ERK signaling pathway during early differentiation of preadipocytes. Mass spectrometry analysis and site-directed mutagenesis combined with a kinase assay identified ERK pathway-mediated dual phosphorylation at Ser 315 and Ser 319 of Dcp1a. To understand the functional effects of Dcp1a phosphorylation, we examined protein-protein interactions between Dcp1a and other decapping components with co-immunoprecipitation. Dcp1a interacted with Ddx6 and Edc3 through its proline-rich C-terminal extension, whereas the conserved EVH1 (enabled vasodilator-stimulated protein homology 1) domain in the N terminus of Dcp1a showed a stronger interaction with Dcp2. Once ERK signaling was activated, the interaction between Dcp1a and Ddx6, Edc3, or Edc4 was not affected by Dcp1a phosphorylation. Phosphorylated Dcp1a did, however, enhanced interaction with Dcp2. Protein complexes immunoprecipitated with the recombinant phosphomimetic Dcp1a(S315D/S319D) mutant contained more Dcp2 than did those immunoprecipitated with the nonphosphorylated Dcp1a(S315A/S319A) mutant. In addition, Dcp1a associated with AU-rich element (ARE)-containing mRNAs such as MAPK phosphatase-1 (MKP-1), whose mRNA stability was analyzed under the overexpression of Dcp1a constructs in the Dcp1a knockdown 3T3-L1 cells. CONCLUSIONS/SIGNIFICANCE: Our findings suggest that ERK-phosphorylated Dcp1a enhances its interaction with the decapping enzyme Dcp2 during early differentiation of 3T3-L1 cells.

Tristetraprolin inhibits poly(A)-tail synthesis in nuclear mRNA that contains AU-rich elements by interacting with poly(A)-binding protein nuclear 1.

BACKGROUND: Tristetraprolin binds mRNA AU-rich elements and thereby facilitates the destabilization of mature mRNA in the cytosol. METHODOLOGY/PRINCIPAL FINDINGS: To understand how tristetraprolin mechanistically functions, we biopanned with a phage-display library for proteins that interact with tristetraprolin and retrieved, among others, a fragment of poly(A)-binding protein nuclear 1, which assists in the 3'-polyadenylation of mRNA by binding to immature poly(A) tails and thereby increases the activity of poly(A) polymerase, which is directly responsible for polyadenylation. The tristetraprolin/poly(A)-binding protein nuclear 1 interaction was characterized using tristetraprolin and poly(A)-binding protein nuclear 1 deletion mutants in pull-down and co-immunoprecipitation assays. Tristetraprolin interacted with the carboxyl-terminal region of poly(A)-binding protein nuclear 1 via its tandem zinc finger domain and another region. Although tristetraprolin and poly(A)-binding protein nuclear 1 are located in both the cytoplasm and the nucleus, they interacted in vivo in only the nucleus. In vitro, tristetraprolin bound both poly(A)-binding protein nuclear 1 and poly(A) polymerase and thereby inhibited polyadenylation of AU-rich element-containing mRNAs encoding tumor necrosis factor α, GM-CSF, and interleukin-10. A tandem zinc finger domain-deleted tristetraprolin mutant was a less effective inhibitor. Expression of a tristetraprolin mutant restricted to the nucleus resulted in downregulation of an AU-rich element-containing tumor necrosis factor α/luciferase mRNA construct. CONCLUSION/SIGNIFICANCE: In addition to its known cytosolic mRNA-degrading function, tristetraprolin inhibits poly(A) tail synthesis by interacting with poly(A)-binding protein nuclear 1 in the nucleus to regulate expression of AU-rich element-containing mRNA.

Drosophila eyes absent is a novel mRNA target of the tristetraprolin (TTP) protein DTIS11.

The Tristetraprolin (TTP) protein family includes four mammalian members (TTP, TIS11b, TIS11d, and ZFP36L3), but only one in Drosophila melanogaster (DTIS11). These proteins bind target mRNAs with AU-rich elements (AREs) via two C3H zinc finger domains and destabilize the mRNAs. We found that overexpression of mouse TIS11b or DTIS11 in the Drosophila retina dramatically reduced eye size, similar to the phenotype of eyes absent (eya) mutants. The eya transcript is one of many ARE-containing mRNAs in Drosophila. We showed that TIS11b reduced levels of eya mRNA in vivo. In addition, overexpression of Eya rescued the TIS11b overexpression phenotype. RNA pull-down and luciferase reporter analyses demonstrated that the DTIS11 RNA-binding domain is required for DTIS11 to bind the eya 3' UTR and reduce levels of eya mRNA. Moreover, ectopic expression of DTIS11 in Drosophila S2 cells decreased levels of eya mRNA and reduced cell viability. Consistent with these results, TTP proteins overexpressed in MCF7 human breast cancer cells were associated with eya homologue 2 (EYA2) mRNA, and caused a decrease in EYA2 mRNA stability and cell viability. Our results suggest that eya mRNA is a target of TTP proteins, and that downregulation of EYA by TTP may lead to reduced cell viability in Drosophila and human cells.