Purpose: Neurodegenerative diseases are associated with neuroinflammation along with activation of microglia and oxidative stress, but currently lack effective treatments. Punicalagin is a natural ...bio-sourced product that exhibits anti-inflammatory effects on several chronic diseases; however, the anti-inflammatory and anti-oxidative effects on microglia have not been well examined. This study aimed to investigate the effects of punicalagin on LPS-induced inflammatory responses, NLRP3 inflammasome activation, and the production of ROS using murine microglia BV2 cells. Methods: BV2 cells were pre-treated with punicalagin following LPS treatment to induce inflammation. The secretion of NO and PGE2 was analyzed by Griess reagent and ELISA respectively, while the expressions of iNOS, COX-2, STAT3, ERK, JNK, and p38 were analyzed using Western blotting, the production of IL-6 was measured by ELISA, and the activity of NF-κB was detected using promoter reporter assay. To examine whether punicalagin affects NLRP3 inflammasome activation, BV2 cells were stimulated with LPS and then treated with ATP or nigericin. The secretion of IL-1β was measured by ELISA. The expressions of NLRP3 inflammasome-related proteins and phospho IκBα/IκBα were analyzed using Western blotting. The production of intracellular and mitochondrial ROS was analyzed by flow cytometry. Results: Our results showed that punicalagin attenuated inflammation with reduction of pro-inflammatory mediators and cytokines including iNOS, COX-2, IL-1β, and reduction of IL-6 led to inhibition of STAT3 phosphorylation by LPS-induced BV2 cells. Punicalagin also suppressed the ERK, JNK, and p38 phosphorylation, attenuated NF-κB activity, inhibited the activation of the NLRP3 inflammasome, and reduced the production of intracellular and mitochondrial ROS by LPS-induced BV2 cells. Conclusion: Our results demonstrated that punicalagin attenuated LPS-induced inflammation through suppressing the expression of iNOS and COX-2, inhibited the activation of MAPK/NF-κB signaling pathway and NLRP3 inflammasome, and reduced the production of ROS in microglia, suggesting that punicalagin might have the potential in treating neurodegenerative diseases. Graphical Abstract:
This study confirmed that the infection of nervous necrosis virus (NNV), belonging to the betanodavirus, can induce the expression of endogenous Mx in grouper fin-3 (GF-3), grouper brain (cGB), and ...barramundi brain (cBB) cells, but not in grouper fin-1 (GF-1) cells. In a co-sedimentation assay, RdRp appeared in the mitochondrial pellet of GF-1 cells without endogenous Mx expression. However, in GF-3, cGB, and cBB cells, RdRp was detected in the nuclear pellet accompanied by endogenous Mx. By immunostaining, RdRp was found to colocalize with not only endogenous Mx but also lysosomes and monodansylcadaverine (MDC)-labeled autophagic vacuoles. In GF-1 cells, the RdRp level continuously increased during 24–72 h post infection (hpi). When endogenous Mx expressed during 24–72 hpi in virus-infected GF-3, cGB, and cBB cells, the RdRp level peaked at 24 hpi but decreased at 48–72 hpi. The degradation of RdRp could be suppressed by treatment with 3-methyladenine (3MA), NH4Cl, and Mx-specific siRNA respectively. After poly I:C transfection, the endogenous Mx level peaked at 3 days post transfection (dpt) and then spontaneously decreased at 5–7 dpt. The poly I:C-indued Mx also colocalized with MDC-labeled autophagic vacuoles at 3 dpt, and its degradation could be inhibited by 3MA or NH4Cl treatments. Therefore, the anti-NNV mechanism of endogenous grouper and barramundi Mx is suggested to sequester RdRp for degradation through autophagy and lysosomes.
•Endogenous Mx colocalized with NNV RdRp.•NNV RdRp/Mx colocalized with MDC-labeled autophagic vacuoles and lysosomes.•NNV RdRp level peaked at 24 hpi and then decreased as endogenous Mx increased.•Poly I:C-induced endogenous Mx could be degraded through autophagy and lysosomes.•Endogenous Mx sequestered NNV RdRp to degrade through autophagy and lysosome.
Ubiquitin (Ub) regulates diverse functions in eukaryotes through its attachment to other proteins. The defining step in this protein modification pathway is the attack of a substrate lysine residue ...on Ub bound through its C‐terminus to the active site cysteine residue of a Ub‐conjugating enzyme (E2) or certain Ub ligases (E3s). So far, these E2 and E3 cysteine residues are the only enzyme groups known to participate in the catalysis of conjugation. Here we show that a strictly conserved E2 asparagine residue is critical for catalysis of E2‐ and E2/RING E3‐dependent isopeptide bond formation, but dispensable for upstream and downstream reactions of Ub thiol ester formation. In constrast, the strictly conserved histidine and proline residues immediately upstream of the asparagine are dispensable for catalysis of isopeptide bond formation. We propose that the conserved asparagine side chain stabilizes the oxyanion intermediate formed during lysine attack. The E2 asparagine is the first non‐covalent catalytic group to be proposed in any Ub conjugation factor.
•Green signal countdown timers have positive short-term effects on drivers’ behavior.•Driving speed and distance to intersection at yellow light affect drivers’ behavior.•Risk-taking and risk-averse ...drivers have different crossing speeds under the timer.•The timer encourages risk-taking drivers to cross intersections at higher speeds.•The timer causes risk-averse drivers to be indecisive while crossing intersections.
Introduction: Motor-vehicle crashes at signalized intersections are a significant traffic safety problem. To address this problem, many Asian cities have installed signal countdown displays at signalized intersections, aiming to assist drivers to make correct decisions in response to traffic signals. Method: In this study, we assessed the short-term and long-term effects of green signal countdown timers (GSCTs) on road safety, using a combination of driving simulator experiments and naturalistic observations. Results: In our driving simulator experiments, 80 participants drove at 50 km/h in scenarios in which a car either approached a signalized intersection alone or following another car. In naturalistic observations, short-term (1-week) and long-term (1-year) intersection safety in the presence and absence of GSCTs were compared. These observations revealed that GSCTs reduced the number of red-light-running violations over the short term, but not over the long term. In fact, GSCTs appeared to lead to an overall increase in rear-end crash risk at intersections, as their presence resulted in drivers exhibiting more sudden acceleration and braking, and altered intersection-crossing speeds and patterns. Conclusions: The results suggest that GSCTs worsen safety at signalized intersections, and thus their removal should be considered.
Lymph node (LN) metastasis is commonly associated with systemic distant organ metastasis in human breast cancer and is an important prognostic predictor for survival of breast cancer patients. ...However, whether tumor‐draining LNs (TDLNs) play a significant role in modulating the malignancy of cancer cells for distant metastasis remains controversial. Using a syngeneic mouse mammary tumor model, we found that breast tumor cells derived from TDLN have higher malignancy and removal of TDLNs significantly reduced distant metastasis. Up‐regulation of oncogenic Il‐17rb in cancer cells derived from TDLNs contributes to their malignancy. TGF‐β1 secreted from regulatory T cells (Tregs) in the TDLNs mediated the up‐regulation of Il‐17rb through downstream Smad2/3/4 signaling. These phenotypes can be abolished by TGF‐β1 neutralization or depletion of Tregs. Consistently, clinical data showed that the up‐regulation of IL‐17RB in cancer cells from LN metastases correlated with the increased prevalence of Tregs as well as the aggressive growth of tumors in mouse xenograft assay. Together, these results indicate that Tregs in TDLNs play an important role in modulating the malignancy of breast cancer cells for distant metastasis. Blocking IL‐17RB expression could therefore be a potential approach to curb the process.
Synopsis
Treg‐secreted TGF‐β1 in the tumor‐draining LN (TDLN) microenvironment up‐regulates IL‐17RB expression in breast cancer cells, thereby enhancing their metastatic potential.
Breast cancer cells isolated from TDLN displayed aggressive phenotypes; removal of TDLN reduced distant organ metastasis.
Tregs in TDLN was the major cell type contributing to the enhanced malignancy of breast cancer cells.
TGF‐β1 secreted from Tregs in TDLN up‐regulated IL‐17RB expression in breast cancer cells.
Up‐regulation of IL‐17RB by TGF‐β1 occurred through Smad2/3/4 signal pathway.
Treg‐secreted TGF‐β1 in the tumor‐draining LN (TDLN) microenvironment up‐regulates IL‐17RB expression in breast cancer cells, thereby enhancing their metastatic potential.
To describe the prevalence of daytime nap habit in participants of the China Kadoorie Biobank (CKB) study, across 10 study regions and explore its correlation with prevalence of major chronic ...diseases.
Participants with a self-reported pre-diagnosis of any cancer at baseline survey were excluded. Logistic regression models were used to analyze the differences in study regions and age distribution of the prevalence daytime nap habit, and its correlation with the prevalence of diabetes, hypertension, coronary heart disease (CHD), stroke, chronic obstructive pulmonary disease (COPD), and chronic liver diseases.
Among 510 145 participants, 39.9% had daytime nap habit in summer and 20.8% had daytime nap habit all the year round. Urban-rural differences were observed in the prevalence of summer nap habit and perennial nap habit. Daytime nap in summer was common in rural areas and Suzhou, with prevalence ranged from 32.9% to 73.3%. Haikou and Liuzhou had higher prevalence of perennial nap (60.4% and 63.3%). The prop
Infective endocarditis is a potentially fatal condition, and identifying the pathogen is crucial to optimizing antibiotic treatment. While a blood culture takes time and may yield negative results, ...it remains the gold standard for diagnosis, blood culture-negative endocarditis, which accounts for up to 20% of infective endocarditis cases, poses a clinical challenge with increasing mortality. To better understand the etiology of blood culture-negative infective endocarditis, we reviewed non-culture-based strategies and compared the results. Serology tests work best in limited pathogens, such as
and
infections. Most of the pathogens identified by broad-range PCR tests are
,
and
. adding specific real-time PCR assays to the systematic PCR testing of patients with blood culture-negative endocarditis will increase the efficiency of diagnosis. Recently, metagenomic next-generation sequencing has also shown promising results.
The transfer section between the cross passage and the main tunnel is the part that needs to be paid attention to during the underground excavation construction of the subway. Due to complex stress, ...the collapse of the horsehead gate and excessive surface settlement often occur. In order to determine the construction scheme of the transfer section between the cross passage and the main tunnel of Guanshui Road Station of Metro Line two in Guiyang, China, the numerical simulation method was used to analyze the “double-holes interval pillar method,” “sector expansion method,” and “gate climbing method,”, respectively. The mechanical response of the surrounding rock and supporting structure under each method was compared. The comparisons showed that the surface settlement, the displacement of the cave, and the plastic zone caused by the double-holes interval pillar method were the smallest, and the method can reduce the construction risk, shorten the construction period, and reduce the project cost. Therefore, it was recommended to use the double-holes interval pillar method to construct the transfer section. The comparison between the measured data and the numerical simulation results of the double-holes interval pillar method showed that the numerical simulation results were smaller than the measured data at each point, and the surface settlement and horizontal displacement in the tunnel both met the safety control standard.
Although highly heritable, environment also contributes to the etiology of autism spectrum disorder (ASD), with several specific environmental factors previously suggested. A registry-linked ...population-based twin cohort of 15,701 pairs (586 individuals with an ASD diagnosis), was established within the Child and Adolescent Twin Study in Sweden. Participants were evaluated for autistic symptoms at age 9 using the Autism-Tics, ADHD and other Comorbidities parental interview. A series of binary cut-offs indicated whether participants scored over various ASD symptom percentiles. Three early medical factors previously associated with ASD, beyond familial confounding (low birth weight, congenital malformations and perinatal hypoxia), were summed up creating an individual cumulative exposure load. A series of unconditional logistic regressions between all individuals and conditional regressions within twin pairs were performed for each outcome and exposure level. Between all individuals increasing cumulative early exposure loads were associated with increasing risk of ASD diagnosis (OR 3.33 (95%CI 1.79-6.20) for three exposures) and autistic symptoms (ranging from OR 2.12 (1.57-2.86) for three exposures at the 55th symptom percentile cut-off to OR 3.39 (2.2-5.24) at the 95th). Within twin pairs, the association between three exposures and an ASD diagnosis remained similar, but not statistically significant (OR 2.39 (0.62-9.24)). Having a higher load of early cumulative exposure was consistently associated with autistic symptoms after adjusting for familial confounding and sex (OR 3.45 (1.66-7.15) to OR 7.36 (1.99-27.18)). This study gives support to the cumulative stress hypothesis of ASD, and the dimensional model regarding environmental exposures, after adjustment for familial confounding.
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