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Prior constant-load exercise performed for 30-min at or above maximal lactate steady state (MLSSp) significantly impairs subsequent time-to-task failure (TTF) compared with TTF performed without prior exercise. We tested the hypothesis that TTF would decrease in relation to the intensity and the duration of prior exercise compared with a baseline TTF trial. Eleven individuals (6 males, 5 females, aged 28 ± 8 yrs) completed the following tests on a cycle ergometer (randomly assigned after MLSSp was determined): (i) a ramp-incremental test; (ii) a baseline TTF trial performed at 80% of peak power (TTFb); (iii) five 30-min constant-PO rides at 5% below lactate threshold (LT−5%), halfway between LT and MLSSp (Delta50), 5% below MLSSp (MLSS−5%), MLSSp, and 5% above MLSSp (MLSS+5%); and (iv) 15- and 45-min rides at MLSSp (MLSS15 and MLSS45, respectively). Each condition was immediately followed by a TTF trial at 80% of peak power. Compared with TTFb (330 ± 52 s), there was 8.0 ± 24.1, 23.6 ± 20.2, 41.0 ± 14.8, 52.2 ± 18.9, and 75.4 ± 7.4% reduction in TTF following LT−5%, Delta50, MLSS−5%, MLSSp, and MLSS+5%, respectively. Following MLSS15 and MLSS45 there were 29.0 ± 20.1 and 69.4 ± 19.6% reductions in TTF, respectively (P < 0.05). It is concluded that TTF is reduced following prior exercise of varying duration at MLSSp and at submaximal intensities below MLSS. Novelty: Prior constant-PO exercise, performed at intensities below MLSSp, reduces subsequent TTF performance. Subsequent TTF performance is reduced in a linear fashion following an increase in the duration of constant-PO exercise at MLSSp.

It has been stated that chronic cerebral hypoperfusion (CCH) markedly prompts neuronal damage and affects cognition. Dimethyl fumarate (DMF), a nuclear erythroid 2-related factor 2 (Nrf2) activator, represents a class of molecules exhibiting neuroprotection. We explored the effect of DMF on CCH using a model of permanent left common carotid occlusion. The left common carotid artery was occluded and then DMF (100mg.kg

Hyaluronan (HA) cross-linking is a conformational state of HA, a covalent complex between HA and heavy chains (HCs) from inter-α-trypsin inhibitor (I-α-I) mediated by tumor necrosis factor-induced protein 6 (TSG6). Cross-linked HA has been identified as a protective factor in physiological and inflammatory conditions. However, the state of HA cross-linking in tumor microenvironment has not been fully elucidated. As a major constituent of the extracellular matrix (ECM), HA is mainly synthesized by cancer-associated fibroblasts (CAFs). Our study aimed to clarify the role of HA cross-linking in breast cancer malignancy. Compared to normal mammary gland tissues, cross-linked HA levels were significantly decreased in breast cancer and associated with tumor malignancy. When NFbs were activated into CAFs, the levels of cross-linked HA and TSG6 were both suppressed. Through upregulating TSG6, CAFs restored the high level of cross-linked HA and significantly inhibited breast cancer malignancy, whereas NFbs promoted the malignancy when the cross-linked HA level was reduced. Furthermore, the inhibitory role of HA cross-linking in tumor malignancy was directly verified using the synthesized HA-HC complex. Collectively, our study found that the deficiency of cross-linked HA induced breast cancer malignancy in a CAF-dependent manner, suggesting that recovering HA cross-linking may be a potential therapeutic strategy.

Deubiquitinating enzymes (DUBs) play important roles in many physiological and pathological processes by modulating the ubiquitination of their substrates. DUBs undergo post-translational modifications including ubiquitination. However, whether DUBs can reverse their own ubiquitination and regulate their own protein stability requires further investigation. To answer this question, we screened an expression library of DUBs and their enzymatic activity mutants and found that some DUBs regulated their own protein stability in an enzymatic activity- and homomeric interaction-dependent manner. Taking Ubiquitin-specific-processing protease 29 (USP29) as an example, we found that USP29 deubiquitinates itself and protects itself from proteasomal degradation. We also revealed that the N-terminal region of USP29 is critical for its protein stability. Taken together, our work demonstrates that at least some DUBs regulate their own ubiquitination and protein stability. Our findings provide novel molecular insight into the diverse regulation of DUBs.

Regulation of hyaluronan (HA) is important for the maintenance of epidermal homeostasis. Here, we examined the mechanism by which 1-ethyl-β-N-acetylglucosaminide (β-NAG2), a newly developed N-acetylglucosamine (NAG) derivative, increases HA production in cultured human epidermal keratinocytes. When keratinocytes were treated with β-NAG2, mRNA expression of HA synthase 3, which is responsible for HA production in human keratinocytes, was not influenced, but the intracellular level of UDP-NAG, a substrate used for HA synthesis, was increased. By using a synthetic substrate for β-N-acetylglucosaminidase (β-NAGase), keratinocytes were found to possess β-NAGase activity, and treatment of o-(2-acetamido-2-deoxy-d-glucopyranosylidene) amino N-phenyl carbamate (PUGNAc), an inhibitor of β-NAGase, abolished the release of NAG from β-NAG2 in keratinocytes. Furthermore, PUGNAc attenuated the β-NAG2-induced intracellular UDP-NAG and HA production in keratinocytes. These results suggest that β-NAG2 is converted to NAG by endogenous β-NAGase in keratinocytes, and the resulting NAG is further metabolized to UDP-NAG and utilized for HA production.

Hydrogen sulfide (H2S) is a cytoprotective redox-active metabolite that signals through protein persulfidation (R-SSnH). Despite the known importance of persulfidation, tissue-specific persulfidome profiles and their associated functions are not well characterized, specifically under conditions and interventions known to modulate H2S production. We hypothesize that dietary restriction (DR), which increases lifespan and can boost H2S production, expands tissue-specific persulfidomes. Here, we find protein persulfidation enriched in liver, kidney, muscle, and brain but decreased in heart of young and aged male mice under two forms of DR, with DR promoting persulfidation in numerous metabolic and aging-related pathways. Mice lacking cystathionine γ-lyase (CGL) have overall decreased tissue protein persulfidation numbers and fail to functionally augment persulfidomes in response to DR, predominantly in kidney, muscle, and brain. Here, we define tissue- and CGL-dependent persulfidomes and how diet transforms their makeup, underscoring the breadth for DR and H2S to impact biological processes and organismal health., Dietary restriction (DR) can increase protein persulfidation but the tissue specificity of this process is not well understood. Here, the authors compare organ-specific protein persulfidomes in young and aged mice under DR, and show that DR-dependent persulfidome changes depend on cystathionine γ-lyase.

Many animals use the Earth’s geomagnetic field for orientation and navigation. Yet, the molecular and cellular underpinnings of the magnetic sense remain largely unknown. A biophysical model proposed that magnetoreception can be achieved through quantum effects of magnetically-sensitive radical pairs formed by the photoexcitation of cryptochrome (CRY) proteins. Studies in Drosophila are the only ones to date to have provided compelling evidence for the ultraviolet (UV)-A/blue light-sensitive type 1 CRY (CRY1) involvement in animal magnetoreception, and surprisingly extended this discovery to the light-insensitive mammalian-like type 2 CRYs (CRY2s) of both monarchs and humans. Here, we show that monarchs respond to a reversal of the inclination of the Earth’s magnetic field in an UV-A/blue light and CRY1, but not CRY2, dependent manner. We further demonstrate that both antennae and eyes, which express CRY1, are magnetosensory organs. Our work argues that only light-sensitive CRYs function in animal light-dependent inclination-based magnetic sensing., Exactly how some animals use magnetic fields to navigate is a longstanding puzzle. A study using a new behavioural assay and transgenic butterflies finds the cryptochrome gene necessary for inclination-based magnetic sensing, and shows that both antennae and eyes, which express this gene, are magnetosensory organs.

Chemical science 12(33), 11004-11012 (2021). doi:10.1039/D1SC03640H, Macrocyclic peptides are an important class of bioactive substances. When inserting an aromatic foldamer segment in a macrocyclic peptide, the strong folding propensity of the former may influence the conformation and alter the properties of the latter. Such an insertion is relevant because some foldamer���peptide hybrids have recently been shown to be tolerated by the ribosome, prior to forming macrocycles, and can thus be produced using an in vitro translation system. We have investigated the interplay of peptide and foldamer conformations in such hybrid macrocycles. We show that foldamer helical folding always prevails and stands as a viable means to stretch, i.e. unfold, peptides in a solvent dependent manner. Conversely, the peptide systematically has a reciprocal influence and gives rise to strong foldamer helix handedness bias as well as foldamer helix stabilisation. The hybrid macrocycles also show resistance towards proteolytic degradation., Published by RSC, Cambridge

Prunella vulgaris (PV) is a perennial plant which is widely grown around the world. It has been widely used as a medicinal treatment for generations. Previous studies showed extracts from this plant had a wide range of therapeutic efficacy, including anti-tumorous effect. However, the volatile organic compounds (VOCs) extracted from it were rarely explored. This paper reports on the characterization of a steam distillation process to extract VOCs in PV and also the anti-tumorous effects of the PV distillate using the tetrazolium-based Cell Counting Kit-8 (CCK-8) as the test agent, when the VOCs were used to treat oral squamous cancer cells, SSC154. It was found that most abundant VOCs came out steadily and continuously for as long as the duration of the steam extraction could extend. However, some compounds such as benzaldehyde did show depletion as the distillation process progressed, while some compounds such as caryophyllene oxide was only sparsely found at the beginning of distillation. The PV distillate was mildly effective in its cytotoxicity to cancer cells SCC154, in a dosage dependent manner.

Chitosan is a bio-based polysaccharide having promising biological and antitumor properties. The present study aims at the synthesis of chitosan using shrimp shell waste. Chitosan preparation consists of two consecutive steps such as demineralization and deproteinization. The structure, morphology and optical properties of the obtained chitosan were established using various instrumental techniques like XRD, FTIR, UV–Vis, SEM and TEM. Herein, we have shown that Significant bacterial activity was manifested by chitosan against both gram positive (S. aureus and S. mutans) and gram negative (E. coli and P. aeuriginosa) bacteria. The biosynthesized chitosan exerts an inhibitory effect on the proliferation of MCF-7 breast cancer cells in a dose –dependent manner while being non -toxic to fibroblast L929 normal cells.

Glioma is a pervasive malignancy and the main cause of cancer-related deaths worldwide. Circular RNA is an important subject of cancer research, and its role and function in glioma are poorly understood. This study demonstrated that hsa_circ_0091581 is upregulated in glioma tissues and cells. The results of the CCK-8, EdU, and transwell assays indicated that hsa_circ_0091581 promotes proliferation, migration, and invasion of glioma cells. The results of the luciferase reporter and RNA immunoprecipitation assays indicated that the mechanism of the effects of hsa_circ_0091581 on glioma cells involves sponging miR-1243-5p to regulate RMI1. The results of the rescue experiments indicated that hsa_circ_0091581 regulates proliferation, migration, and invasion of glioma cells by targeting RMI1 in a miR-1243-5p dependent manner. The results of the nude mice xenograft assays showed that knockdown of hsa_circ_0091581 inhibits glioma growth in vivo. Thus, our study determined the role of hsa_circ_0091581/miR-1243-5p/RMI1 in glioma and suggests that this axis may be a novel therapeutic target in glioma.

MicroRNAs (miRNAs) play critical roles in gene expression and numerous human diseases. The success of miRNA biogenesis is largely determined by the primary miRNA (pri-miRNA) processing by the DROSHA-DGCR8 complex, called Microprocessor. Here, we analysed the high-throughput pri-miRNA processing assays and secondary structures of pri-miRNAs to investigate the roles of bulges in the pri-miRNA processing. We found that bulges in multiple places control both the cleavage efficiency and accuracy of pri-miRNA processing. These bulges were shown to act on Microprocessor via its catalytic subunit, DROSHA, and function in a position and strand-dependent manner. Interestingly, we discovered that the enriched and conserved bulges, called midB, can correct DROSHA orientation on pri-miRNAs, thereby enhancing production of miRNAs. The revealed functions of the bulges help improve our understanding of pri-miRNA processing and suggest their potential roles in miRNA biogenesis regulation.

Studies have shown that exosomes can mediate the chemoresistance of drug-resistant cells by transmitting circular RNAs (circRNAs). However, the role of exosome-derived hsa_circ_103801 (exosomal hsa_circ_103801) in osteosarcoma (OS) remains unclear. The level of hsa_circ_103801 was upregulated in the serum exosomes from patients with OS, and OS patients with high hsa_circRNA_103801 expression had a shorter survival time relative to patients with low hsa_circ_103801 expression. The expression of hsa_circ_103801 was upregulated in cisplatin-resistant MG63 (MG63/CDDP) cells compared with that in MG63 cells. In addition, hsa_circ_103801 was highly enriched in exosomes derived from CDDP-resistant OS cells and could be delivered to MG63 and U2OS cells through exosomes. Exosomes derived from CDDP-resistant cells were shown to reduce the sensitivity of MG63 and U2OS cells to CDDP, inhibit apoptosis, and increase the expression of multidrug resistance-associated protein 1 and P-glycoprotein. Moreover, exosomal hsa_circ_103801 could strengthen the promotive effect of exosomes on the chemoresistance of MG63 and U2OS cells to CDDP. Hence, serum exosomal hsa_circ_103801 may serve as an effective prognostic biomarker for OS, and exosomal hsa_circ_103801 could be a potential target for overcoming OS chemoresistance.

The promising results of metal oxides nanoparticles in different areas including the biological system lead us to investigate the antioxidant and antimicrobial actions of chemically synthesized cobalt oxide (Co3O4) nanoparticles. The different concentrations of synthesized Co3O4 nanoparticles were prepared and evaluated for different parameters at different time intervals i.e. on day 1, 30 and 60 after preparations. Co3O4 nanoparticles synthesized in this study were of 52.2 nm average size with a polydispersity index of 0.465. We observed that Co3O4 nanoparticles scavenge different in vitro free radicals (DPPH, ABTS, superoxide anion and hydrogen peroxide radicals) in concentration dependent manner. The percentage of inhibitions of free radicals by Co3O4 nanoparticles was markedly more on day 1 as compared to day 30 and 60. The IC50 values of Co3O4 nanoparticles for these free radicals were also on day 1 as compared to day 30 and 60. The Co3O4 nanoparticles showed the antibacterial actions against both the bacterial strains i.e. S. aureus and E. coli. The MIC and MBC values revealed that action of Co3O4 nanoparticles was more against E. coli than S. aureus. The MIC and MBC values were lower on day 1 as compared to day 30 and 60 with respective to specific bacteria. In conclusions, the Co3O4 nanoparticles synthesized in this study showed potent antioxidant and antibacterial properties due to which it may serve as promising candidate for the combat the biological problems humans, animals and plants associated with reactive oxygen species and bacteria.

N6-methyladenosine (m6A) RNA methylation is the most prevalent modification of messenger RNAs (mRNAs) and catalyzed by a multicomponent methyltransferase complex (MTC), among which methyltransferase-like 3 (METTL3) and METTL14 are two core molecules. However, METTL3 and METTL14 play opposite regulatory roles in hepatocellular carcinoma (HCC). Based on The Cancer Genome Atlas (TCGA) database and Gene Expression Omnibus (GEO) database, we conducted a multi-omics analysis of METTL3 and METTL14 in HCC, including RNA-sequencing, m6ARIP-sequencing, and ribosome-sequencing profiles. We found that the expression and prognostic value of METTL3 and METTL14 are opposite in HCC. Besides, after METTL3 and METTL14 knockdown, most of the dysregulated mRNAs, signaling pathways and biological processes are distinct in HCC, which partly explains the contrary regulatory role of METTL3 and METTL14. Intriguingly, these mRNAs whose stability or translation efficiency are influenced by METTL3 or METTL14 in an m6A dependent manner, jointly regulate multiple signaling pathways and biological processes, which supports the cooperative role of METTL3 and METTL14 in catalyzing m6A modification. In conclusion, our study further clarified the contradictory role of METTL3 and METTL14 in HCC.

Ebola virus (EBOV) RNA has the potential to form hairpin structures at the transcription start sequence (TSS) and reinitiation sites of internal genes, both on the genomic and antigenomic/mRNA level. Hairpin formation involving the TSS and the spacer sequence between promotor elements (PE) 1 and 2 was suggested to regulate viral transcription. Here, we provide evidence that such RNA structures form during RNA synthesis by the viral polymerase and affect its activity. This was analysed using monocistronic minigenomes carrying hairpin structure variants in the TSS-spacer region that differ in length and stability. Transcription and replication were measured via reporter activity and by qRT-PCR quantification of the distinct viral RNA species. We demonstrate that viral RNA synthesis is remarkably tolerant to spacer extensions of up to ~54 nt, but declines beyond this length limit (~25% residual activity for a 66-nt extension). Minor incremental stabilizations of hairpin structures in the TSS-spacer region and on the mRNA/antigenomic level were found to rapidly abolish viral polymerase activity, which may be exploited for antisense strategies to inhibit viral RNA synthesis. Finally, balanced viral transcription and replication can still occur when any RNA structure formation potential at the TSS is eliminated, provided that hexamer phasing in the promoter region is maintained. Altogether, the findings deepen and refine our insight into structure and length constraints within the EBOV transcription and replication promoter and suggest a remarkable flexibility of the viral polymerase in recognition of PE1 and PE2.

International audience; Phosphorus (P) is an essential macronutrient for all living organisms. Importantly, plants require a large amount of P to grow, and P deficiency causes huge losses in plant production. Although this issue can be mitigated by the appropriate use of phosphate (Pi) rock-derived P fertilizers, phosphate rock is a finite natural resource. Moreover, the increased demand for food as a result of our growing global population is another factor contributing to a prospective P crisis. While creating crops that are resilient to Pi deficiency presents great scientific challenge, the current progress in our understanding of how plants regulate Pi homeostasis offers some opportunities for further study. In this review, we present the published research supporting these opportunities, which are based on the molecular mechanisms that plants have evolved to respond to P deficiency. First, we focus on recent advances in P sensing and signaling pathways in the regulation of root system architecture. Next, we describe the mechanisms that regulate Pi transport and accumulation, in a Pi- (or other nutrient) dependent manner. Integrating these data will help to design an innovative strategy for improving Pi nutrition in plants. In addition, this will help with Pi scarcity, one of the challenges facing agriculture in the twenty first century.

Objectives To investigate whether mechanical vibration at 30 or 60 Hz combined with compressive force alter IL-1β and TNF-α expression in human periodontal ligament (hPDL) cells. Methods hPDL cells isolated from the roots of first premolar teeth extracted from four independent donors were cultured and exposed to vibration (0.3 g, 20 min per cycle, every 24 h for 3 cycles) at 30 or 60 Hz (V30 or V60), 2.0 g/cm2 compressive force for 2 days (CF), or a combination of compressive force and vibration at 30 Hz or 60 Hz (V30CF or V60CF). Quantitative real-time polymerase chain reaction (qPCR) and enzyme-linked immunosorbent assays (ELISAs) were used to determine IL-1β and TNF-α mRNA and protein, respectively. Results The levels of IL-1β and TNF-α did not alter in groups V30 and V60. While, they were upregulated in groups CF, V30CF and V60CF. In addition, IL-1β mRNA and TNF-α mRNA and protein were expressed at significantly higher levels in group V30CF compared to CF group. However, IL-1β protein levels between V30CF and CF groups did not reach statistical significance. Conclusions 30 Hz vibration had the synergistic effects with compressive force on the upregulation of IL-1β mRNA and TNF-α mRNA and protein in PDL cells, while 60 Hz vibration did not have this synergistic effect.

The influence of mesenchymal stem cells (MSCs) on the phenotype and function of CD4+CD49b+FoxP3- regulatory cells has not been elucidated. We used Concanavalin A (ConA) - and α-galactosylceramide (α-GalCer)-induced acute liver injury to estimate the effects of MSCs on liver-infiltrating CD4+CD49b+FoxP3-regulatory cells. MSCs significantly reduced ConA- and α-GalCer-mediated liver injury in C57BL/6 mice, as demonstrated by biochemical tests, reduced influx of inflammatory CD4+ T cells, and increased presence of CD4+CD49b+FoxP3- regulatory cells in the injured livers. The number of CD4+CD49b+FoxP3-regulatory cells was also significantly increased in α-GalCer-treated mice that received MSC-derived conditioned medium (MSC-CM). The presence of 1-methyltryptophan, a specific inhibitor of indoleamine 2,3-dioxygenase (IDO), in MSC-CM completely abrogated the hepatoprotective eff ect of MSCs and significantly decreased the total number of liver-infiltrated CD4+CD49b+FoxP3- regulatory cells, indicating the crucial importance of MSC-derived IDO for the expansion of CD4+CD49b+FoxP3- regulatory cells and the consequent MSC-dependent attenuation of acute liver injury.

Previously, it was found that the hepatitis C virus NS5A interacted with ACBD3 in a genotype-dependent manner. However, the region in NS5A responsible for association with ACBD3 is not clear. Domain I of NS5A was identified as critical for ACBD3 binding. By comparing the differences of amino acids in domain I from different genotypes of NS5A, it was found that key amino acids potentially corresponded to the affinity of the NS5A-ACBD3 interaction. The findings not only revealed that domain I of NS5A associates with ACBD3 but they also shed mechanistic light on how NS5A is associated with ACBD3 in a genotype-dependent manner.

Natural antiangiogenic products interact with multiple molecular angiogenic pathways and possesses comparably less toxicities then chemically synthesized anti-angiogenic compounds. There is a need to screen varieties of medicinal plants to identify more sources of antiangiogenic compounds, as anticancer drugs with less side effects. This study was designed to explore the antiangiogenic property and cytotoxicity of Moricandia sinaica (Boiss.) in zebrafish embryos, as well as its anti-migratory potential in human umbilical vein endothelial cells (HUVECs). Gas chromatography coupled with mass spectrophotometry was performed in order to identify bioactive molecules present in the stem and leaves part of Moricandia sinaica.The methanol extract of the stem and leaves inhibited the formation of the inter-segmental, sub-intestinal vein, and dorsal longitudinal anastomotic veins formation in zebrafish embryos in a dose- dependent manner. It did not show cytotoxicity at lower concentration in HUVECs; however, it significantly (p

To investigate the role of YAP in cyclic mechanical stress induced up-regulation of HIF-1α in rat cartilage chondrocytes. Our in vitro and in vivo experiments demonstrated that cyclic mechanical stress promoted HIF-1α and YAP proteins expression in a magnitude dependent manner. Cyclic mechanical stress at 4000μ strain exhibited most significant effect in promoting HIF-1α and YAP up-regulation. Activation of YAP using LPA significantly promoted HIF-1α stabilization and expression, while YAP siRNA treatment suppressed the up-regulation of HIF-1α induced by cyclic mechanical stress. Our results indicated that cyclic mechanical stress promoted HIF-1α stabilization and YAP is involved in mechanical stress induced HIF-1α up-regulation.

Peroxisomes are metabolically active oxygen demanding organelles with a high abundance of oxidases making it vulnerable to low oxygen levels such as hypoxic conditions. However, the exact mechanism of peroxisome degradation in hypoxic condition remains elusive. In order to study the mechanism of peroxisome degradation in hypoxic condition, we use Dimethyloxaloylglycine (DMOG), a cell-permeable prolyl-4-hydroxylase inhibitor, which mimics hypoxic condition by stabilizing hypoxia-inducible factors. Here we report that DMOG degraded peroxisomes by selectively activating pexophagy in a HIF-2α dependent manner involving autophagy receptor p62. Furthermore, DMOG not only increased peroxisome turnover by pexophagy but also reduced HIF-2α dependent peroxisome proliferation at the transcriptional level. Taken together, our data suggest that hypoxic condition is a negative regulator for peroxisome abundance through increasing pexophagy and decreasing peroxisome proliferation in HIF-2α dependent manner.

Ionic Zn2+ has increasingly been recognized as an important neurotransmitter and signaling ion in glutamatergic neuron pathways. Intracellular Zn2+ transiently increases as a result of neuronal excitation, and this Zn2+ signal is essential for neuron plasticity, but the source and regulation of the signal is still unclear. In this study we rigorously quantified Zn2+, Ca2+ and pH dynamics in dissociated mouse hippocampal neurons stimulated with bath application of high KCl or glutamate. While both stimulation methods yielded Zn2+ signals, Ca2+ influx, and acidification, glutamate stimulation induced more sustained high intracellular Ca2+ and a larger increase in intracellular Zn2+. However, the stimulation-induced pH change was similar between conditions, indicating that a different cellular change is responsible for the stimulation-dependent difference in Zn2+ signal. This work provides the first robust quantification of Zn2+ dynamics in neurons using different methods of stimulation.

A practical synthetic route, consisting of 5 steps, has been developed and applied successfully for converting limonin/deoxylimonin into the corresponding amino derivatives I-5a–I-5e and II-5a–II-5e. Deoxylimonin analogs II-5a and II-5b bearing an open A ring structure underwent ring closure reaction by employing the Mitsunobu procedure to afford II-6a and II-6b. All of the 12 newly synthesized compounds were subjected to preliminary screening for evaluating their inflammation and nociception inhibition efficacy. The most promising compounds, I-5b and II-5d, were selected for further investigation by a carrageenan-induced mouse paw edema model, both of which displayed a dose–response dependent manner and demonstrated superior anti inflammation capacity to that of indomethacin in the first 2 hours.

Background Several findings suggest neuroinflammation as a contributing factor for the onset of psychiatric disorders such as Alzheimer’s disease, depression, and anxiety. There is increasing evidence pointing out that the Mediterranean diet influences brain and behavior. Mediterranean herbs and spices have been shown to be within those components of the Mediterranean diet involved in cognitive enhancement. Thus, we investigated the influence of Mediterranean natural extracts (MNE), Rosemary extract (RE) and Glycyrrhiza glabra root extract (GGRE), on cognitive behavior. Results Adult zebrafish were exposed to RE or GGRE (100 and 250 mg/L) treatments. Both MNE improved memory retention during the T-maze test, although no improvements were observed during the novel object preference. Similarly, chronic administration of RE (150 mg/Kg) and GGRE (150 mg/Kg) improved, respectively, spatial and retention memory, as assessed by the Morris Water Maze (MWM), and the Elevated Plus Maze (EPM) in healthy male rats. However, no improvements were observed during the novel object recognition. Finally, male, and female rats were chronically treated with lipopolysaccharide [(LPS) 300 ug/kg] and orally administered with RE. Interestingly, RE reversed LPS-induced cognitive deficit during the MWM and EPM in female rats. Conclusions We found that MNE improved cognition in both zebrafish and rats. Moreover, MNE rescued LPS-induced cognitive impairment in a gender-specific manner. Therefore, our study supports the view that zebrafish represent a valuable preclinical model for drug discovery in neuroscience. These findings contribute to an exciting and growing body of research suggesting that MNE may play an important role in the prevention of cognitive impairment.

Phosphoprotein enriched in astrocytes, 15 kDa (PEA-15) is a death-effector domain (DED) containing protein involved in regulating mitogen-activated protein kinase and apoptosis pathways. In this molecular dynamics study, we examined how phosphorylation of the PEA-15 C-terminal tail residues, Ser-104 and Ser-116, allosterically mediates conformational changes of the DED and alters the binding specificity from extracellular-regulated kinase (ERK) to Fas-associated death domain (FADD) protein. We delineated that the binding interfaces between the unphosphorylated PEA-15 and ERK2 and between the doubly phosphorylated PEA-15 and FADD are similarly composed of a scaffold that includes both the DED and the C-terminal tail residues of PEA-15. While the unphosphorylated serine residues do not directly interact with ERK2, the phosphorylated Ser-116 engages in strong electrostatic interactions with arginine residues on FADD DED. Upon PEA-15 binding, FADD repositions its death domain (DD) relative to the DED, an essential conformational change to allow the death-inducing signaling complex (DISC) assembly.

Objective To determine whether lumbar anatomy parameters are in dynamic change and related factors. Methods This is a retrospective study. Participants who did lumbar computed tomography (CT) scanning in Shandong University Qilu Hospital from October 2017 to March 2019 were selected. The 476 participants were randomly selected as male or female, with the age ranging from 17 to 87 years (mean, 55.19; standard deviation, 14.28 years). All the measurements were taken based on the CT scanning image and the measurement of lumbar morphology was conducted using picture archiving and communication systems (PACS). The angle between the horizontal alignment and pedicle center on median sagittal view, the angle between upper endplate and lower endplate on median sagittal view as well as transverse section angle (TSA) using Magerl point in the axial view was determined by reconstructive CT analysis. Results In the overall participants, the angle between the horizontal alignment and pedicle center on median sagittal view of lumbar one to three was significantly decreased with aging, from 3.90° ± 2.81° to −4.18° ± 6.86° (P = 0.002), 5.60° ± 2.89° to −4.14° ± 5.90° (P = 0.030), and 4.75° ± 2.95° to −2.87° ± 4.68° (P

The execution of functions on DNA relies on complex interactions between DNA and proteins in a sequence and structure dependent manner. Accurate quantification of the affinity and kinetics of these interactions is critical for understanding the molecular mechanisms underlying their corresponding biological functions. The development of single-molecule manipulation technologies in the last two decades has made it possible to apply a mechanical constraint to a single DNA molecule and measure the end-to-end extension changes with nanometer resolution in realtime. While it has been shown that such technologies can be used to investigate binding of ligands, which can be proteins or other molecules, to DNA in a fluorescence-label free manner, a systematic review on such applications has been lacking. Here, we provide a review on some of recently developed methods for fluorescence-label free single-molecule quantification of site-specific DNA binding by ligands and demonstrate their wide scope of applications using several examples of binding of ligands to dsDNA and ssDNA binding sites.

Heat activation at a sublethal temperature is widely applied to promote Bacillus species spore germination. This treatment also has the potential to be employed in food processing to eliminate undesired bacterial spores by enhancing their germination and then inactivating the less-heat-resistant germinated spores at a milder temperature. However, incorrect heat treatment could also generate heat damage in spores and lead to more heterogeneous spore germination. Here, the heat activation and heat damage profile of Bacillus subtilis spores was determined by testing spore germination and outgrowth at both population and single-spore levels. The heat treatments used were 40 to 80°C and for 0 to 300 min. The results were as follows. (i) Heat activation at 40 to 70°C promoted l-valine- and l-asparagine-glucose-fructose-potassium (AGFK)-induced germination in a time-dependent manner. (ii) The optimal heat activation temperatures for AGFK and l-valine germination via the GerB plus GerK or GerA germinant receptors were 65°C and 50 to 65°C, respectively. (iii) Heat inactivation of dormant spores appeared at 70°C, and the heat damage of molecules essential for germination and growth began at 70 and 65°C, respectively. (iv) Heat treatment at 75°C resulted in both activation of germination and damage to the germination apparatus, and 80°C treatment caused more pronounced heat damage. (v) For the spores that should withstand adverse environmental temperatures in nature, heat activation seemed functional for a subsequent optimal germination process, while heat damage affected both germination and outgrowth. IMPORTANCE Bacterial spores are thermal-stress-resistant structures that can thus survive food preservation strategies and revive through the process of spore germination. The more heat resistant spores are, the more heterogeneous their germination upon the addition of germinants. Upon germination, spores can cause food spoilage and food intoxication. Here, we provide new information on both heat activation and inactivation regimes and their effects on the (heterogeneity of) spore germination.

Vif is a lentiviral accessory protein that counteracts the antiviral activity of cellular APOBEC3 (A3) cytidine deaminases in infected cells. The exact contribution of each member of the A3 family for the restriction of HIV-2 is still unclear. Thus, the aim of this work was to identify the A3s with anti-HIV-2 activity and compare their restriction potential for HIV-2 and HIV-1. We found that A3G is a strong restriction factor of both types of viruses and A3C restricts neither HIV-1 nor HIV-2. Importantly, A3B exhibited potent antiviral activity against HIV-2, but its effect was negligible against HIV-1. Whereas A3B is packaged with similar efficiency into both viruses in the absence of Vif, HIV-2 and HIV-1 differ in their sensitivity to A3B. HIV-2 Vif targets A3B by reducing its cellular levels and inhibiting its packaging into virions, whereas HIV-1 Vif did not evolve to antagonize A3B. Our observations support the hypothesis that during wild-type HIV-1 and HIV-2 infections, both viruses are able to replicate in host cells expressing A3B but using different mechanisms, probably resulting from a Vif functional adaptation over evolutionary time. Our findings provide new insights into the differences between Vif protein and their cellular partners in the two human viruses. Of note, A3B is highly expressed in some cancer cells and may cause deamination-induced mutations in these cancers. Thus, A3B may represent an important therapeutic target. As such, the ability of HIV-2 Vif to induce A3B degradation could be an effective tool for cancer therapy. IMPORTANCE Primate lentiviruses encode a series of accessory genes that facilitate virus adaptation to its host. Among those, the vif-encoded protein functions primarily by targeting the APOBEC3 (A3) family of cytidine deaminases. All lentiviral Vif proteins have the ability to antagonize A3G; however, antagonizing other members of the A3 family is variable. Here, we report that HIV-2 Vif, unlike HIV-1 Vif, can induce degradation of A3B. Consequently, HIV-2 Vif but not HIV-1 Vif can inhibit the packaging of A3B. Interestingly, while A3B is packaged efficiently into the core of both HIV-1 and HIV-2 virions in the absence of Vif, it only affects the infectivity of HIV-2 particles. Thus, HIV-1 and HIV-2 have evolved two distinct mechanisms to antagonize the antiviral activity of A3B. Aside from its antiviral activity, A3B has been associated with mutations in some cancers. Degradation of A3B by HIV-2 Vif may be useful for cancer therapies.

SummaryDeveloping methods to modulate growth of the mucin-degrading gut bacterium Akkermansia muciniphila could benefit patients with different health needs, as A. muciniphila has been associated with both positive metabolic health outcomes and detrimental neurodegenerative outcomes. Growth of A. muciniphila is sensitive to plant-derived polyphenols, and particularly proanthocyanidins (PACs), when administered in isolated form at supraphysiological doses. However, it remains unclear whether doses sufficient for these effects are achievable via diet. Here, we explore the extent to which nutritionally relevant doses of common polyphenol-rich foods – berries, wine, and coffee – influence A. muciniphila abundance in C57BL/6J mice under varying dietary conditions. By administering polyphenol-rich whole foods, comparing polyphenol-depleted and PAC-rich versus PAC-poor food supplements, and through gradient PAC-dosing experiments, we show that PAC-rich foods uniquely induce A. muciniphila growth at doses that are feasibly achieved through routine diet. Notably, the effects of PAC supplementation were detected against a high-fat diet but not a low-fat control diet background, highlighting the importance of habitual diet strategies in either amplifying or mitigating the prebiotic effects of PAC-rich food consumption. Ultimately, our work suggests that both PACs and diet influence A. muciniphila abundance with downstream impacts for human health.

The aberrant expression of fat mass and obesity-associated protein (FTO) has been confirmed to be associated with a variety of cancers and participates in the regulation of multiple biological behaviours. FTO plays an oncogenic role in bladder cancer, but few studies have focused on how FTO promotes bladder cancer progression by regulating miRNA synthesis. Here, we confirmed that FTO expression was significantly increased in bladder cancer and was associated with a poor prognosis. FTO overexpression promoted bladder cancer cell proliferation, whereas FTO knockdown inhibited bladder cancer cell proliferation. We also demonstrated that FTO promoted bladder cancer cell proliferation via the FTO/miR-576/CDK6 pathways. Taken together, our work revealed that FTO plays a critical role in bladder cancer and could be a potential diagnostic or prognostic biomarker for this disease.

Background: Enterobacter cloacae (EC) is a commonly occurring opportunistic pathogen and is responsible for causing various infections in humans. Owing to its inducible chromosomal AmpC β-lactamase (AmpC), EC is inherently resistant to the 1st- and 2nd- generation cephalosporins. However, whether β-lactams antibiotics enhance EC resistance remains unclear.Results: In this study, we found that subinhibitory concentrations (SICs) of cefazolin (CFZ) and imipenem (IMP) are able to advance the expression of AmpC and improve its resistance towards β - lactams through NagZ in EC clinical isolate. Our work indicate that AmpC manifested a substantial upregulation in EC in response to SICs of CFZ and IMP. In nagZ knockout EC (ΔnagZ), we found that the resistance to β - lactam antibiotics was rather weakened and the effect of CFZ and IMP on induction of AmpC was completely abrogated. Ectopic expression of NagZ can rescue the induction effect of CFZ and IMP on AmpC and enhance resistance in ΔnagZ. More importantly, CFZ and IMP have the potential to bring about the target genes expressions of AmpR in a NagZ-dependent manner.Conclusions: Our findings show that NagZ is a critical determinant for CFZ and IMP to promote AmpC expression and improve resistance and that CFZ and IMP should be used with caution since they may aggravate EC resistance. At the same time, this study further improves our understanding of resistance mechanisms in EC.

Abstract. Background: Many studies have investigated the effect of ALA supplementation on lipid profile, and different results have been obtained from these studies. The current systematic review and dose-response meta-analysis was conducted to achive a strong conclusion about the effect of ALA supplementation on lipid profile including total cholesterol (TC), low- and high-density lipoprotein cholesterol (LDL, HDL) and triglyceride (TG). Methods: A systematic search was performed in PubMed, SCOPUS, ProQuest and Embase for randomized placebo-controlled human trials that examined the effect of ALA supplementation on lipid profile up to November 2020. The dose and duration of ALA supplementation for included studies were ranged between 300–1200 mg/d and 2–16 weeks respectively. Weighted mean differences (WMD) and 95% confidence intervals (CIs) were used to evaluate the effect size. Cochran’s Q and I2 tests were also used to assess between-study’s heterogeneity. In addition, subgroup analysis was performed to investigate potential sources of heterogeneity. Dose-response relationship was done using fractional polynomial modeling. Results: Among all eligible studies, 12 studies with a total number of 548 participants were selected. ALA caused a significant reduction on TC (WMD): −10.78 mg/dl, 95% CI: −20.81, −0.74, P=0.002), LDL (WMD: −10.88 mg/dl, 95% CI: −19.52, −2.24, P=0.014) and TG (WMD: −31.02 mg/dl, 95% CI: −49.63, −12.42, Pnon-linearity=0.026), TG (Pnon-linearity

The naïve epiblast transitions to a pluripotent primed state during embryo implantation. Despite the relevance of the FGF pathway during this period, little is known about the downstream effectors regulating this signaling. Here, we examined the molecular mechanisms coordinating the naïve to primed transition by using inducible ESC to genetically eliminate all RAS proteins. We show that differentiated RAS

Background: Previous studies have shown that TIFA (TNF receptor associated factor (TRAF)-interacting protein with a Forkhead-associated (FHA) domain) plays different roles in various tumor types. However, the function of TIFA in colorectal cancer (CRC) remains unclear. The goal of this study is to uncover the biological function and molecular mechanism of TIFA in CRC.Methods: Tissue microarrays were used to evaluate TIFA expression. Cancer cell proferation assays were performed in TIFA knockdown and overexpressing cells in vitro and in a xenograft model in vivo. Human phosphokinase array, immunoprecipitation assays were performed to explore the underlying mechanism.Results: We disclosed that the expression of TIFA was marked increased in CRC versus normal tissue, and positively correlated with CRC TNM stages. In agreement, we found that the CRC cell lines show increased TIFA expression levels versus normal control. The knockdown of TIFA inhibited cell proliferation but have no effects on cell apoptosis in vitro and in vivo. Moreover, the ectopic expression of TIFA enhanced cell proliferation ability in vitro and in vivo. In contrast, the expression of mutant TIFA (T9A, oligomerization site mutation; D6, TRAF6 binding site deletion) alternatively abolished TIFA mediated cell proliferation enhancement. Exploration of the underlying mechanism demonstrated that the protein synthesis associated kinase RSK and PRAS40 activation were all responsible for TIFA mediated CRC progression. Conclusions: The above results described a model of TIFA in mediating CRC progression. This may provide a promising target for CRC therapy.

The spiking activity of populations of cortical neurons is well described by the dynamics of a small number of population-wide covariance patterns, whose activation we refer to as ‘latent dynamics’. These latent dynamics are largely driven by the same correlated synaptic currents across the circuit that determine the generation of local field potentials (LFPs). Yet, the relationship between latent dynamics and LFPs remains largely unexplored. Here, we characterised this relationship for three different regions of primate sensorimotor cortex during reaching. The correlation between latent dynamics and LFPs was frequency-dependent and varied across regions. However, for any given region, this relationship remained stable throughout the behaviour: in each of primary motor and premotor cortices, the LFP-latent dynamics correlation profile was remarkably similar between movement planning and execution. These robust associations between LFPs and neural population latent dynamics help bridge the wealth of studies reporting neural correlates of behaviour using either type of recordings.

SUMMARYThe last stage of cell division involves two daughter cells remaining interconnected by a cytokinetic bridge that is cleaved in a process called abscission. During pre-abscission, we identified that the centrosome moves in a Rab11-dependent manner towards the cytokinetic bridge in human cells grown in culture and in an in vivo vertebrate model, Danio rerio (zebrafish). Rab11-endosomes are dynamically organized in a Rab11-GTP dependent manner at the centrosome during pre-abscission and this organization is required for the centrosome protein, pericentrin, to be enriched at the centrosome. Using zebrafish embryos, we found that reduction in pericentrin expression or optogenetically disrupting Rab11-endosome function inhibited centrosome movement towards the cytokinetic bridge and abscission resulting in daughter cells prone to being binucleated and/or having supernumerary centrosomes. These studies suggest that Rab11-endosomes contribute to centrosome function during pre-abscission by regulating pericentrin organization resulting in appropriate centrosome movement towards the cytokinetic bridge and subsequent abscission.

For the first time, the feasibility of creating morphed samples for attacking vascular biometrics is investigated, in particular finger vein recognition schemes are addressed. A conducted vulnerability analysis reveals that (i) the extent of vulnerability, (ii) the type of most vulnerable recognition scheme, and (iii) the preferred way to determine the best morph sample for a given target sample depends on the employed sensor. Digital morphs represent a significant threat as vulnerability in terms of IAPMR is often found to be > 0.8 or > 0.6 (in sensor dependent manner). Physical artefacts created from these morphs lead to clearly lower vulnerability (with IAPMR ≤ 0.25), however, this has to be attributed to the low quality of the artefacts (and is expected be increase for better artefact quality).

Reversible protein glycosylation and phosphorylation tightly modulate important cellular processes and are closely involved in pathological processes in a crosstalk dependent manner. Because of their significance and low abundances of glyco- and phosphopeptides, several strategies have been developed to simultaneously enrich and co-elute glyco- and phosphopeptides. However, phosphopeptides slowly hydrolyze during deglycosylation and the co-existence of deglycosylated peptides and phosphopeptides aggravates the mass spectrometry analysis. Herein we developed a novel strategy to analyze glyco- and phosphopeptides based on simultaneous enrichment with TiO2, on-line deglycosylation and collection of deglycosylated peptides, and subsequent elution of phosphopeptides. To optimize on-line deglycosylation conditions, the solution pH, buffer types and concentrations, and deglycosylation time were investigated. The application of this novel strategy to 100 μg mouse brain resulted in 1,405 glycopeptides and 2,022 phosphopeptides, which were 2.4 and 1.4 folds of those enriched with the reported method. This study will expand the application of TiO2 and may shed light on simultaneous monitoring of multiple protein post-translational modifications.

Pseudomonas aeruginosa, a major cause of nosocomial infection, can survive under diverse environmental conditions. Its great adaptive ability is dependent on its multiple signaling systems such as the two-component system (TCS). A TCS FleS/FleR has been previously identified to positively regulate a variety of virulence-related traits in P. aeruginosa PAO1 including motility and biofilm formation which are involved in the acute and chronic infections, respectively. However, the molecular mechanisms underlying these regulations are still unclear. In this study, we first analyzed the regulatory roles of each domains in FleS/FleR and characterized key residues in the FleS-HisKA, FleR-REC and FleR-AAA domains that are essential for the signaling. Next, we revealed that FleS/FleR regulates biofilm formation in a c-di-GMP and FleQ dependent manner. Lastly, we demonstrated that FleR can regulate flagellum biosynthesis independently without FleS, which explains the discrepant regulation of swimming motility by FleS and FleR.

Background: Human periodontal ligament stem cells (PDLSCs)-mediated regenerative periodontal therapy is a promising method for periodontitis. Besides, odontogenic stem cells are exposed to hypoxia in both physiological and pathological conditions. The behaviour of stem cells under hypoxia is worth exploring. So far, whether long noncoding RNAs (lncRNAs) can affect the proliferation and cementogenesis of PDLSCs, as well as their specific mechanism, remains to be elucidated. This study investigates lncRNA NEAT1 and its possible regulatory mechanisms of cementogenic differentiation of PDLSCs under hypoxia.Methods: CCK-8 assay, flow cytometry (FCM) and EdU assay was adopted to test the proliferation ability. Osteoblastic/cementogenic differentiation of PDLSCs under hypoxia was detected by western blot assay (WB), quantitative real-time polymerase chain reaction (RT-PCR), alkaline phosphatase (ALP) activity, alcian blue staining (ABS), alizarin red staining (ARS). The relationship between them was proved by dual-luciferase reporter assay and rescue experiments.Results: PDLSCs exhibited a more powerful cementogenic differentiation potential in hypoxic conditions. miR-214-5p may regulate SMAD4, while Lnc NEAT1 acts as a sponge, forming a ceRNA mechanism in which HIF-1α plays an important role. It may act as an enabler, jointly constitute the regulatory axis of Lnc NEAT1/ miR-214-5p/ SMAD4, and affect the cementogenic differentiation of PDLSCs under hypoxia.Conclusions: Our results showed that hypoxia could enhance the cementogenic differentiation potential of PDLSCs in vitro. In addition, we found that Lnc NEAT1 may be a potential target for improving the function of PDLSCs in regenerative periodontiology, and Lnc NEAT1/miR-214-5p/SMAD4 could regulate the cementogenic differentiation ability of PDLSCs under hypoxia.

Cripto is a small glycosylphosphatidylinositol-anchored signaling protein that can detach from the anchored membrane and stimulate proliferation, migration, differentiation, vascularization, and angiogenesis. In the present study, we demonstrated that Cripto positively affected proliferation and survival of mesenchymal stem cells (MSCs) without affecting multipotency. Cripto also increased expression of phosphorylated janus kinase 2 (p-JAK2), phosphorylated signal transducer and activator of transcription 3 (p-STAT3), 78 kDa glucose-regulated protein (GRP78), c-Myc, and cyclin D1. Notably, treatment with an anti-GRP78 antibody blocked these effects. In addition, pretreatment with STAT3 short interfering RNA (siRNA) inhibited the increase in p-JAK2, c-Myc, cyclin D1, and BCL3 levels caused by Cripto and attenuated the pro-survival action of Cripto on MSCs. We also found that incubation with Cripto protected MSCs from apoptosis caused by hypoxia or H2O2 exposure, and the level of caspase-3 decreased by the Cripto-induced expression of B-cell lymphoma 3-encoded protein (BCL3). These effects were sensitive to down-regulation of BCL3 expression by BCL3 siRNA. Finally, we showed that Cripto enhanced expression levels of vascular endothelial growth factor (VEGF), fibroblast growth factor (FGF), and hepatocyte growth factor (HGF). In summary, our results demonstrated that Cripto activated a novel biochemical cascade that potentiated MSC proliferation and survival. This cascade relied on phosphorylation of JAK2 and STAT3 and was regulated by GRP78. Our findings may facilitate clinical applications of MSCs, as these cells may benefit from positive effects of Cripto on their survival and biological properties.