A Generalizable, Tunable Microfluidic Platform for Delivering Fast Temporally Varying Chemical Signals to Probe Single-Cell Response Dynamics

Understanding how biological systems transduce dynamic, soluble chemical cues into physiological processes requires robust experimental tools for generating diverse temporal chemical patterns. The advent of microfluidics has seen the development of platforms for rapid fluid exchange allowing ease of changes in the cellular microenvironment and precise cell handling. Rapid exchange is important for exposing systems to temporally varying signals. However, direct coupling of macroscale fluid flow with microstructures is potentially problematic due to the high shear stresses that inevitably add confounding mechanical perturbation effects to the biological system of interest. Here, we have devised a method of translating fast and precise macroscale flows to microscale flows using a monolithically integrated perforated membrane. We integrated a high-density cell trap array for nonadherent cells that are challenging to handle under flow conditions with a soluble chemical signal generator module. The platform enables fast and repeatable switching of stimulus and buffer at low shear stresses for quantitative live, single-cell fluorescent studies. This modular design allows facile integration of any cell-handling chip design with any chemical delivery module. We demonstrate the utility of this device by characterizing heterogeneity of oscillatory response for cells exposed to alternating Ca²⁺ waveforms at various periodicities. This platform enables the analysis of cell responses to chemical perturbations at a single-cell resolution that is necessary in understanding signal transduction pathways

Identification of miRNAs and Their Target Genes Associated with Sweet Corn Seed Vigor by Combined Small RNA and Degradome Sequencing

High seed vigor is significant for agriculture. Low seed vigor of sweet corn hindered the popularization of sweet corn (Zea mays L. saccharata Sturt). To better understand the involvement and regulatory mechanism of miRNAs with seed vigor, small RNA libraries from seeds non-artificially aged and artificially aged for 2 days were generated by small RNA sequencing. A total of 27 differentially expressed miRNAs were discovered, of which 10 were further confirmed by real-time quantitative polymerase chain reaction. Furthermore, targets of miRNAs were identified by degradome sequencing. A total of 1142 targets that were potentially cleaved by 131 miRNAs were identified. Gene ontology (GO) annotations of target transcripts indicated that 26 target genes cleaved by 9 differentially expressed miRNAs might play roles in the regulation of seed vigor, such as peroxidase superfamily protein targeted by PC-5p-213179_17 playing a role in the oxidation–reduction process and response to oxidative stress. These findings provide valuable information to understand the involvement of miRNAs with seed vigor

pone.0289248.t001 - Agomelatine prevented depression in the chronic restraint stress model through enhanced catalase activity and halted oxidative stress

pone.0289248.t001 - Agomelatine prevented depression in the chronic restraint stress model through enhanced catalase activity and halted oxidative stress</p

Effects of the CRS, AGO, and DFX intervention on immunohistochemistry of CAT protein in the hippocampus of mice.

Immunohistochemistry shows the changes in CAT protein expression in the hippocampus of mice after CRS, AGO, and DFX intervention. (a) Staining of CAT protein. (B) Positive rate of CAT protein expression in the mouse hippocampus. *P<0.05; **P<0.01; ***P<0.001; CAT, catalase; CRS, chronic restraint stress; AGO, agomelatine; DFX, deferasirox; ns, no significant difference. Data are expressed as the mean ± standard deviation and were analyzed by one-way ANOVA, followed by post hoc multiple comparisons (LSD method). n = 3.</p

Effects of the CRS, AGO, and DFX intervention on body weight and behavioral tests.

Depression-like behavior in mice induced by CRS. (a) Initial body weight of the mice. (b) Body weight of the mice after different treatments. (c) Weight gain of the mice. (d) Sucrose preference rate in mice. (e) Forced swimming rest time in mice. (f) The total distance of mice in the central region of the open field test. (g) Total time of mice in the central region in the open field test. (h) The total distance of mice in the open field test. *P<0.05; ** P<0.01; *** P<0.001; CRS, chronic restraint stress; AGO, agomelatine; DFX, deferasirox; ns, no significant difference. Data are expressed as the mean ± standard deviation and were analyzed by one-way ANOVA, followed by post hoc multiple comparisons (LSD method). n = 10.</p

Reliable Facility Location Problem with Facility Protection

<div><p>This paper studies a reliable facility location problem with facility protection that aims to hedge against random facility disruptions by both strategically protecting some facilities and using backup facilities for the demands. An Integer Programming model is proposed for this problem, in which the failure probabilities of facilities are site-specific. A solution approach combining Lagrangian Relaxation and local search is proposed and is demonstrated to be both effective and efficient based on computational experiments on random numerical examples with 49, 88, 150 and 263 nodes in the network. A real case study for a 100-city network in Hunan province, China, is presented, based on which the properties of the model are discussed and some managerial insights are analyzed.</p></div

ELISA, Western blot, and PCR results.

Effect of agomelatine on oxidative stress and the NF-кB pathway in CRS mice. (a) CAT activity in the serum of mice under CRS, AGO, and DFX intervention. (b-c) SOD and MDA concentrations in the serum of mice under CRS, AGO, and DFX intervention. (d, g) Representative protein blot results of CAT, NF-кB p65, and IкBα in the mouse hippocampus under CRS and AGO intervention, showing the quantitative density analysis of CAT in the hippocampus (e, f, h). (i-k) mRNA expression levels of CAT, NF-кB, and IкBα in the mouse hippocampus. *P<0.05; **P<0.01; ***P<0.001; CAT, catalase; CRS, chronic restraint stress; AGO, agomelatine; DFX, deferasirox; SOD, superoxide dismutase; MDA, malondialdehyde; ns, no significant difference. Data are expressed as the mean ± standard deviation and were analyzed by one-way ANOVA, followed by post hoc multiple comparisons (LSD method). n = 3–5.</p

S1 File -

BackgroundAgomelatine (AGO) is an antidepressant with unique pharmacological effects; however, its underlying mechanisms remain unknown. In this study, we examined agomelatine’s effects on catalase activity, oxidative stress, and inflammation.MethodsChronic restraint stress (CRS) model mice were established over 4 weeks, and AGO 50 mg/kg was administered to different groups alongside a deferasirox (DFX) 10 mg/kg gavage treatment. Behavioral tests were performed to assess the effect of AGO on the remission of depression-like behaviors. Meanwhile, the expression of CAT, the oxidative stress signaling pathway and inflammatory protein markers were assessed using ELISA, qRT-PCR, Western blot, and immunohistochemistry.ResultsFour weeks of AGO treatment significantly improved depression-like behavior in mice through the activation of catalase in the hippocampus and serum of the model mice, increased superoxide dismutase expression, reduced malondialdehyde expression, and reduced oxidative stress damage. Deferasirox was found to offset this therapeutic effect partially. In addition, the inflammatory pathway (including nuclear factor-κB and nuclear factor of kappa light polypeptide gene enhancer in B cells inhibitor, alpha) was not significantly altered.ConclusionsAGO can exert antidepressant effects by altering oxidative stress by modulating catalase activity.</div

Number of sites with facilities opened corresponding to different W.

<p>Number of sites with facilities opened corresponding to different W.</p

Number of opened unreliable and reliable facilities corresponding to different q.

<p>Number of opened unreliable and reliable facilities corresponding to different q.</p