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The RIIβ subunit of cAMP‐dependent protein kinase A (PKA) is expressed in the brain and adipose tissue. RIIβ‐knockout mice show leanness and increased UCP1 in brown adipose tissue. The authors have previously reported that RIIβ reexpression in hypothalamic GABAergic neurons rescues the leanness. However, whether white adipose tissue (WAT) browning contributes to the leanness and whether RIIβ‐PKA in these neurons governs WAT browning are unknown. Here, this work reports that RIIβ‐KO mice exhibit a robust WAT browning. RIIβ reexpression in dorsal median hypothalamic GABAergic neurons (DMH GABAergic neurons) abrogates WAT browning. Single‐cell sequencing, transcriptome sequencing, and electrophysiological studies show increased GABAergic activity in DMH GABAergic neurons of RIIβ‐KO mice. Activation of DMH GABAergic neurons or inhibition of PKA in these neurons elicits WAT browning and thus lowers body weight. These findings reveal that RIIβ‐PKA in DMH GABAergic neurons regulates WAT browning. Targeting RIIβ‐PKA in DMH GABAergic neurons may offer a clinically useful way to promote WAT browning for treating obesity and other metabolic disorders. [ABSTRACT FROM AUTHOR]

Thymosin β10 (TMSB10) has been found to be overexpressed and function as an oncogene in several types of cancer. However, there have been limited reports on the role of TMSB10 in bladder cancer (BCa). In the present study, reverse transcription-quantitative PCR was used to quantify the expression of TMSB10 in BCa cell lines, clinical specimens and their corresponding control samples. The protein expression of TMSB10 was also examined in archived tissues from 101 patients with pathologically confirmed BCa by immunohistochemistry. Univariate and multivariate Cox regression models were used to evaluate the prognostic significance of TMSB10 in patients with BCa. The data indicated that the mRNA levels of TMSB10 were significantly overexpressed in BCa cell lines. In addition, the protein levels of TMSB10 were overexpressed in BCa tissues compared with those in adjacent normal tissues. In 55/101 (54.5%) BCa specimens, high expression levels of TMSB10 were noted. Statistical analysis revealed that the high expression of TMSB10 was positively associated with muscular invasion (P<0.05). In addition, a high expression of TMSB10 was associated with shorter overall survival (OS) of patients (P<0.05; log-rank test). The univariate and multivariate analyses suggested that the protein overexpression of TMSB10 was an unfavorable prognostic factor for OS (P<0.05) in patients with BCa. Knockdown of the expression of TMSB10 significantly suppressed cell migration and invasion. In conclusion, TMSB10 can be considered an independent factor for the poor prognosis of patients with BCa. The targeting of TMSB10 can reduce the migration and invasion of BCa cells. [ABSTRACT FROM AUTHOR]

Summary Diet-induced obesity can induce low-level inflammation and insulin resistance. Interleukin-1β (IL-1β) is one of the key proinflammatory cytokines that contributes to the generation of insulin resistance and diabetes, but the mechanisms that regulate obesity-driven inflammation are ill defined. Here we found reduced expression of the E3 ubiquitin ligase Pellino3 in human abdominal adipose tissue from obese subjects and in adipose tissue of mice fed a high-fat diet and showing signs of insulin resistance. Pellino3-deficient mice demonstrated exacerbated high-fat-diet-induced inflammation, IL-1β expression, and insulin resistance. Mechanistically, Pellino3 negatively regulated TNF receptor associated 6 (TRAF6)-mediated ubiquitination and stabilization of hypoxia-inducible factor 1α (HIF1α), resulting in reduced HIF1α-induced expression of IL-1β. Our studies identify a regulatory mechanism controlling diet-induced insulin resistance by highlighting a critical role for Pellino3 in regulating IL-1β expression with implications for diseases like type 2 diabetes. [ABSTRACT FROM AUTHOR]

Mutations that result in loss of function of Nod2, an intracellular receptor for bacterial peptidoglycan, are associated with Crohn's disease. Here we found that the E3 ubiquitin ligase Pellino3 was an important mediator in the Nod2 signaling pathway. Pellino3-deficient mice had less induction of cytokines after engagement of Nod2 and had exacerbated disease in various experimental models of colitis. Furthermore, expression of Pellino3 was lower in the colons of patients with Crohn's disease. Pellino3 directly bound to the kinase RIP2 and catalyzed its ubiquitination. Loss of Pellino3 led to attenuation of Nod2-induced ubiquitination of RIP2 and less activation of the transcription factor NF-κB and mitogen-activated protein kinases (MAPKs). Our findings identify RIP2 as a substrate for Pellino3 and Pellino3 as an important mediator in the Nod2 pathway and regulator of intestinal inflammation. [ABSTRACT FROM AUTHOR]

Dopamine denervation in the striata of patients with Parkinson''s disease (PD) leads to changes in neural plasticity. However, the mechanisms leading to the changes are still poorly understood. In an effort to study the molecular events in the denervated striatum, we identified and cloned rat karyopherin α1 (KPNA1), a member of the importin/karyopherin α (KPNA) family. DNA sequence analysis revealed that the full-length cDNA, encoding rat KPNA1, was 4975 bp with a short 5′-untranslated region (UTR) of 70 bp, a putative coding sequence of 1617 bp, and an unusually long 3′-UTR of 3266 bp. The gene shared a high degree of similarity with its mouse and human homologs at both cDNA and protein levels. By computational analysis of its genomic sequence, the transcription unit was shown to span a 44-kb region and consist of 13 exons varying in size from 89 (6th exon) to 3454 bp (13th exon), and 12 introns varying in size from 0.3 to 8.9 kb. Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis demonstrated that KPNA1 transcript existed in various adult tissues. Both Northern blot and semi-quantitative RT-PCR analysis showed that the expression level of KPNA1 mRNA was altered in the denervated striatum post-lesion in a time-dependent manner, reaching the maximum at 2 weeks post-lesion. Our results suggest involvement of KPNA1 in the striatal responses to denervation following 6-hydroxydopamine (6-OHDA)-induced lesion. [Copyright &y& Elsevier]

Dopamine denervation in the striata of patients with Parkinson''s disease (PD) leads to changes in neural plasticity. However, the mechanisms leading to the changes are still poorly understood. In an effort to study the molecular events in the denervated striatum, we identified and cloned rat karyopherin α1 (KPNA1), a member of the importin/karyopherin α (KPNA) family. DNA sequence analysis revealed that the full-length cDNA, encoding rat KPNA1, was 4975 bp with a short 5′-untranslated region (UTR) of 70 bp, a putative coding sequence of 1617 bp, and an unusually long 3′-UTR of 3266 bp. The gene shared a high degree of similarity with its mouse and human homologs at both cDNA and protein levels. By computational analysis of its genomic sequence, the transcription unit was shown to span a 44-kb region and consist of 13 exons varying in size from 89 (6th exon) to 3454 bp (13th exon), and 12 introns varying in size from 0.3 to 8.9 kb. Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis demonstrated that KPNA1 transcript existed in various adult tissues. Both Northern blot and semi-quantitative RT-PCR analysis showed that the expression level of KPNA1 mRNA was altered in the denervated striatum post-lesion in a time-dependent manner, reaching the maximum at 2 weeks post-lesion. Our results suggest involvement of KPNA1 in the striatal responses to denervation following 6-hydroxydopamine (6-OHDA)-induced lesion. [Copyright &y& Elsevier]

Flexible pressure sensors are important components of electronic skin and flexible wearable devices. Most existing piezoresistive flexible pressure sensors have obtained high sensitivities, however, they have relatively small pressure detection ranges. Here, we report flexible pressure sensors with a wide detection range using polydimethylsiloxane (PDMS) as the substrate, carbon nanotube films as the electrode material, and self-assembled polystyrene microsphere film as the microstructure layer. The obtained pressure sensor had a sandwich structure, and had a wide pressure detection range (from 4 kPa to 270 kPa), a sensitivity of 2.49 kPa−1, and a response time of tens of milliseconds. Two hundred load–unload cycles indicated that the device had good stability. In addition, the sensor was obtained by large-area fabrication with a low power consumption. This pressure sensor is expected to be widely used in applications such as electronic skin and flexible wearable devices. [ABSTRACT FROM AUTHOR]

The increasing prevalence of obesity has resulted in demands for the development of new effective strategies for obesity treatment. Withaferin A (WA) shows a great potential for prevention of obesity by sensitizing leptin signaling in the hypothalamus. However, the mechanism underlying the weight- and adiposity-reducing effects of WA remains to be elucidated. In this study, we report that WA treatment induced white adipose tissue (WAT) browning, elevated energy expenditure, decreased respiratory exchange ratio, and prevented high-fat diet-induced obesity. The sympathetic chemical denervation dampened the WAT browning and also impeded the reduction of adiposity in WA-treated mice. WA markedly upregulated the levels of Prdm16 and FATP1 (Slc27a1) in the inguinal WAT (iWAT), and this was blocked by sympathetic denervation. Prdm16 or FATP1 knockdown in iWAT abrogated the WAT browning-inducing effects of WA and restored the weight gain and adiposity in WA-treated mice. Together, these findings suggest that WA induces WAT browning through the sympathetic nerve-adipose axis, and the adipocytic Prdm16-FATP1 pathway mediates the promotive effects of WA on white adipose browning. [ABSTRACT FROM AUTHOR]

Hexokinase (HK) catalyzes the first irreversible rate-limiting step in glycolysis that converts glucose to glucose-6-phosphate. HK1 is ubiquitously expressed in the brain, erythrocytes, and other tissues where glycolysis serves as the major source of ATP production. Spermatogenic cell-specific type 1 hexokinase (HK1S) is expressed in sperm but its physiological role in male mice is still unknown. In this study, we generate Hk1s knockout mice using the CRISPR/Cas9 system to study the gene function in vivo. Hk1s mRNA is exclusively expressed in testes starting from postnatal day 18 and continuing to adulthood. HK1S protein is specifically localized in the outer surface of the sperm fibrous sheath (FS). Depletion of Hk1s leads to infertility in male mice and reduces sperm glycolytic pathway activity, yet they have normal motile parameters and ATP levels. In addition, by using in vitro fertilization (IVF), Hk1s deficient sperms are unable to fertilize cumulus-intact or cumulus-free oocytes, but can normally fertilize zona pellucida-free oocytes. Moreover, Hk1s deficiency impairs sperm migration into the oviduct, reduces acrosome reaction, and prevents capacitation-associated increases in tyrosine phosphorylation, which are probable causes of infertility. Taken together, our results reveal that HK1S plays a critical role in sperm function and male fertility in mice. Author summary: About 8%-12% of couples worldwide are infertile, and male infertility is associated with about half of these cases. Common defects of male infertility include low sperm count, reduced sperm motility, abnormal sperm morphology, and unexplained male infertility with normal sperm parameters but infertility. The cause of over 50% of male infertility cases remains unidentified. Research on male reproduction has helped to unravel male infertility. In this study, we used a knockout mouse model to study the function of HK1S in the male mouse reproductive system. We showed that depletion of Hk1s leads to infertility in male mice but with normal sperm morphology and motility. These phenotypes are similar to patients with unexplained male infertility. Furthermore, HK1S is essential for sperm glycolysis. Hk1s deficiency in mice results in decreased glycolysis activity and leads to impaired sperm migration into the oviduct, reduced acrosome reaction, and defective capacitation-associated increase in tyrosine phosphorylation. These results suggested that HK1S is necessary for male fertility in mice. Our study suggests a new avenue for the development of new therapeutic strategies for male infertility. [ABSTRACT FROM AUTHOR]

Purpose: Prostate cancer is the second leading cause of cancer death in men worldwide. Olaparib is clinically approved for the treatment prostate cancer, but cytotoxicity and off-target effects including DNA damage limit its clinical applications. In the current study, new strategies to improve the therapeutic efficacy of olaparib for the treatment of prostate cancer were investigated. Methods: Two prostate cancer cell lines were exposed to the c-MET inhibitor PHA665752 and/or the PARP inhibitor olaparib. Cell counting kit-8, colony formation assays, and transwell assays were conducted to evaluate the cytotoxicity of olaparib alone or in combination with PHA665752 in prostate cancer cell lines. Western blotting, immunofluorescence staining, and the comet assay were used to assess the effects of PHA665752 on olaparib-induced DNA damage. Results: Combined inhibition of c-MET and PARP resulted in effective and synergistic blocking of the growth of prostate cancer cell lines. Invasion and migration were significantly suppressed when the agents were combined. Mechanistically, dual blocking of PARP and c-MET in prostate cancer cell lines was associated with an impaired DNA damage response. Interestingly, immunofluorescence staining analysis of RAD51 protein indicated that the c-MET inhibitor PHA665752 significantly impaired homologous repair via downregulated translocation of RAD51 into the nucleus in prostate cancer cells. Conclusion: The combination of the c-MET inhibitor PHA665752 and the PARP inhibitor olaparib may be a promising therapeutic strategy in patients with prostate cancer. [ABSTRACT FROM AUTHOR]

The intestinal epithelium is the fastest renewing tissue in mammals and its regenerative process must be tightly controlled to minimize the risk of dysfunction and tumorigenesis. The orderly expression and activation of Yes‐associated protein (YAP) are the key steps in driving intestinal regeneration and crucial for intestinal homeostasis. However, the regulatory mechanisms controlling this process remain largely unknown. Here, it is discovered that evolutionarily conserved signaling intermediate in Toll pathways (ECSIT), a multi‐functional protein, is enriched along the crypt–villus axis. Intestinal cell‐specific ablation of ECSIT results in the dysregulation of intestinal differentiation unexpectedly accompanied with enhanced YAP protein dependent on translation, thus transforming intestinal cells to early proliferative stem "‐like" cells and augmenting intestinal tumorigenesis. Loss of ECSIT leads to metabolic reprogramming in favor of amino acid–based metabolism, which results in demethylation of genes encoding the eukaryotic initiation factor 4F pathway and their increased expression that further promotes YAP translation initiation culminating in intestinal homeostasis imbalance and tumorigenesis. It is also shown that the expression of ECSIT is positively correlated with the survival of patients with colorectal cancer. Together, these results demonstrate the important role of ECSIT in regulating YAP protein translation to control intestinal homeostasis and tumorigenesis. [ABSTRACT FROM AUTHOR]

Background: Neuroinflammation has been widely accepted as a cause of the degenerative process. Increasing interest has been devoted to developing intervening therapeutics for preventing neuroinflammation in Parkinson's disease (PD). It is well known that virus infections, including DNA viruses, are associated with an increased risk of PD. In addition, damaged or dying dopaminergic neurons can release dsDNA during PD progression. However, the role of cGAS, a cytosolic dsDNA sensor, in PD progression remains unclear. Methods: Adult male wild‐type mice and age‐matched male cGAS knockout (cGas−/−) mice were treated with MPTP to induce neurotoxic PD model, and then behavioral tests, immunohistochemistry, and ELISA were conducted to compare disease phenotype. Chimeric mice were reconstituted to explore the effects of cGAS deficiency in peripheral immune cells or CNS resident cells on MPTP‐induced toxicity. RNA sequencing was used to dissect the mechanistic role of microglial cGAS in MPTP‐induced toxicity. cGAS inhibitor administration was conducted to study whether GAS may serve as a therapeutic target. Results: We observed that the cGAS‐STING pathway was activated during neuroinflammation in MPTP mouse models of PD. cGAS deficiency in microglia, but not peripheral immune cells, controlled neuroinflammation and neurotoxicity induced by MPTP. Mechanistically, microglial cGAS ablation alleviated the neuronal dysfunction and inflammatory response in astrocytes and microglia by inhibiting antiviral inflammatory signaling. Additionally, the administration of cGAS inhibitors conferred the mice neuroprotection during MPTP exposure. Conclusions: Collectively, these findings demonstrate microglial cGAS promote neuroinflammation and neurodegeneration during the progression of MPTP‐induced PD mouse models and suggest cGAS may serve as a therapeutic target for PD patients. Limitations of the Study: Although we demonstrated that cGAS promotes the progression of MPTP‐induced PD, this study has limitations. We identified that cGAS in microglia accelerate disease progression of PD by using bone marrow chimeric experiments and analyzing cGAS expression in CNS cells, but evidence would be more straightforward if conditional knockout mice were used. This study contributed to the knowledge of the role of the cGAS pathway in PD pathogenesis; nevertheless, trying more PD animal models in the future will help us to understand the disease progression deeper and explore possible treatments. [ABSTRACT FROM AUTHOR]

Apples quality monitoring and maturity measurement are important for yield evaluation, profit estimation, determination of harvest time and postharvest storage conditions. This study explored the monitoring ability of the micro-Unmanned Aerial Vehicle (UAV) to the orchard, the force sensor and gyroscope are used to observe the force and posture changes of apples during the spectral collection process. The UAV carries 18 channels of visible/near-infrared multispectral sensor to collect the internal information of growing apples at close range. A total of 200 apples were collected in the orchard, and then soluble solid content (SSC), titratable acid (TA), firmness, and starch pattern index (SPI) were destructively tested. Multiple scatter correction (MSC) and Standard normal variate (SNV) pretreatment combined with partial least squares regression (PLSR) were used to establish the internal quality prediction model. Streif index is used to evaluate the maturity of apples. The results showed that the detection of apples on trees by UAV would not affect the growth of apples and could collect the internal quality information. The correlation coefficient of the prediction model for apple maturity is 0.8593 and root mean square error is 0.0102. It shows that this study has potential in detecting the changes of quality and maturity of growing apples and is helpful to the overall perception of orchards. • Micro-flight detector is used to detect apple maturity indexes. • The detector will not damage the growth of apple. • PLSR was used to establish the prediction model of apple maturity indexes. [ABSTRACT FROM AUTHOR]

Landslides are one of the most prevalent environmental disasters in the Hengduan Mountain Region. Landslides lead to severe economic damage and property loss, as well as fatalities. Furthermore, they tend to increase in the context of climate change. The purpose of this study is to comprehensively assess landslide susceptibility across the Hengduan Mountain Region in southwest China. Specifically, the analysis is focused on the eastern boundary of the Tibetan Plateau within the context of future climate change scenarios, which are based on the latest Coupled Model Intercomparison Project Phase 6 (CMIP6) global climate model ensemble. The Generalized Additive Model (GAM), Random Forest (RF), and Light Gradient Boosting Machine (LightGBM) were selected in order to map landslide susceptibility within the context of 1.5–4.0 °C warming scenarios. This was achieved by considering the changes in extreme rainfall that exceeded the landslide triggering thresholds. The results show that the frequency over extreme rainfall thresholds (FOERT) tend to increase in conjunction with warming targets, thereby ranging from 2.3/a (at a 1.5 °C warming) to 9.0/a (at a 4.0 °C warming) on average. Such elevated extreme precipitation events contribute to an increase in projected future zones of high landslide susceptibility when compared to the historical baseline period ranging from −1.2% (at a 1.5 °C warming) to 4.0% (at a 4.0 °C warming) using different machine learning models. Moreover, the extent of high susceptibility zones increases more significantly in the context of 4.0 °C warming when compared to the historical baseline results. These results indicate the importance of limiting the global temperature rise to 1.5 as well as 2 °C. The high landslide susceptibility zones estimated by the CMIP6 multi-models ensemble are mainly located in the central and southeastern regions of the Hengduan Mountain Region. The possible changes in terms of introducing extreme precipitation in order to assess landslide susceptibility in the context of climate change that is proposed in this study may be further applied to additional study areas. These projections under different targets can provide scientific guidelines for the purposes of the development of climate change adaptation strategies. [ABSTRACT FROM AUTHOR]

In this work, the largest tributary of the South Branch in the upper reaches of the Xiong'an New Area is selected as the study area. The impact-based critical areal rainfall indices for early flood warnings are proposed from the perspective of the impacts of floods on socio-economic factors. Specifically, four steps, including the determination of the damage-causing discharges, the establishment of the rainfall–discharge relationship, the computation of the critical areal rainfall and the validation of the early warning indices, were used to determine the critical areal rainfall for early flood warnings in the watershed. The results showed that the 1-day critical areal rainfall amounts were 31 mm, 63 mm, 92 mm and 160 mm for early flood warning levels 4, 3, 2 and 1, respectively, when the 1-day antecedent areal rainfall was ≤10 mm. The critical areal rainfall amounts were 20 mm, 54 mm, 87 mm and 160 mm for early flood warning levels 4, 3, 2 and 1, respectively, when the 1-day antecedent areal rainfall was >10 mm. The early warning effectiveness of the proposed critical indices was validated with historical catastrophic flood events and precipitation data during recent flood seasons. The results demonstrated that the impact-based critical indices had a high accuracy and could release warnings 1–2 days in advance, which could effectively avoid the occurrence of missed and underestimated warnings. [ABSTRACT FROM AUTHOR]

The article discusses the regulatory approach of anticipatory governance concerning the issue of non-invasive prenatal testing (NIPT) for non-medical traits. It mentions the text emphasizes the importance of a regulatory approach based on lessons learned from the regulation of medically assisted reproduction. It highlights the governance recommendations for NIPT for non-medical traits and illustrate the necessity of such an approach for anticipatory governance.

Healthcare-associated infections (HAIs) have increasingly become a threat to global health. Rapid, accurate, and reliable detection of HAI-related bacteria is urgently needed. Here, we presented a novel antimicrobial peptide-based surface-enhanced Raman scattering lateral flow assay (SERS-LFA) for the detection of multiple HAI-related bacteria. Antimicrobial peptide cecropin 1(CP1) was immobilized on the surface of Fe 3 O 4 @Au by the streptavidin-biotin system to capture Pseudomonas aeruginosa (P. aeruginosa) and Escherichia coli O157:H7 (E. coli O157:H7) simultaneously. The synthesized Fe 3 O 4 @Au-CP1 exhibits excellent broad-spectrum capturing ability, rapid magnetic enrichment function, and superior antibacterial activity. LFA strips enable the specific detection of two target bacteria in a quick and simple manner, with bacterial type differentiation via corresponding monoclonal antibodies sprayed. The highly sensitive SERS signals and the amplification effect of streptavidin-biotin significantly improved the sensitivity of detection. The LODs of P. aeruginosa and E. coli O157:H7 were 12 and 16 cells/mL, respectively. This assay also exhibited good specificity and reproducibility when applied to urine samples. Therefore, Fe 3 O 4 @Au-CP1-based SERS-LFA has great potential for detecting multiple bacteria after infection as well as regularly monitoring therapeutic efficacy. • CP1 was first utilized as a broad-spectrum recognition element in SERS-LFA to detect multiple bacteria simultaneously. • The amplification effect of the streptavidin-biotin system enhances the binding efficiency of Fe 3 O 4 @Au-CP1 to bacteria. • Fe 3 O 4 @Au-CP1 MNPs enable quick capture and enrichment of bacteria. • CP1-based SERS-LFA can achieve highly sensitive and quantitative detection for P. aeruginosa and E. coli O157:H7. • The LODs for P. aeruginosa and E. coli O157:H7 were 12 cells/mL and 16 cells/mL, respectively. [ABSTRACT FROM AUTHOR]

The present study aims to investigate the influence of release rate, dose and co-administration on pharmacokinetics (PK) and pharmacodynamics (PD) of tanshinone IIA (TA) and tanshinol (TS), and reveal the changes in their PK-PD relationships. Sustained and immediate release pellets of TS and TA were prepared respectively, and oral administrated to angina model rabbits according to the experimental design. The administration dose of TS was 50, 35 or 20 mg/kg and that of TA was 30 mg/kg. Then, plasma concentrations of TS and TA were measured to evaluate the pharmacokinetics. Pharmacodynamic biomarkers including cardiac troponin (cTn-I), creatine kinase (CK-MB), superoxide dismutase (SOD) and nitric oxide (NO) were measured to evaluated the effects of cardioprotection, amelioration of oxidative stress and vasorelaxation of TS and TA. Parameters such as maximum plasma concentration (C max), maximum effect (E max), time to C max or E max (TC max or TE max), areas under the plasma concentration or effect curves (AUC 0-∞ or AUEC) and pharmacodynamic efficiency (EFF) were calculated based on non-compartmental analysis. Beside, PK-PD relationship/hysteresis was evaluated. The TE max was less sensitive than TC max to changes in release rate. The E max , AUEC and EFF showed increasing trend as the decrease of release rate even that the AUC 0-∞ showed no significant difference. In addition, slow drug release decreased the magnitude of hysteresis of TS and TA. The sensitivities of E max and AUEC of four biomarkers to changes in dose were varied and relatively lower than those of C max and AUC 0-∞. The EFF decreased and the magnitude of hysteresis increased for high dose. The C max and AUC 0-∞ of TS and TA showed little difference after co-administration. The E max and AUEC of four biomarkers increased for immediate release pellets (P < 0.05 or P < 0.01) and generally decreased for sustained release pellets (P < 0.05 or P < 0.01) after co-administration. In addition, the magnitudes of hysteresis of four biomarkers decreased for immediate release pellets and generally increased for sustained release pellets after co-administration. In summary, the dissociated and unstable PK-PD relationship should be considered during optimization of dosage forms and regimens to make sure the rationality, safety and efficacy. These findings could also provide some valuable information for the development and clinical therapy of other drugs. [Display omitted] [ABSTRACT FROM AUTHOR]

Point-of-care quantitative analysis of tracing microRNA disease-biomarkers remains a great challenge in the clinical diagnosis. In this paper, we developed a portable fluorescent lateral flow assay for ultrasensitive quantified detection of acute myocardial infarction related microRNAs in bio-samples. SiO 2 @DQD (bilayer quantum dots assembly with SiO 2 core) based fluorescent lateral flow strip was fabricated as the analysis tool. In order to quantify the tracing microRNA in biosamples, a catalytic hairpin assembly and CRISPR/Cas12a cascade amplification method was performed and combined with the fabricated SiO 2 @DQD lateral flow strip. Thus, our platform gathered double advantages of portability and ultrasensitive quantification. Based on our strips, target myocardial biomarker microRNA-133a can be detected with a detection limit of 0.32 fM, which was almost 1000-fold sensitive compared with previous reported microRNAs-lateral flow strips. Significantly, this portable fluorescent strip can directly detect microRNAs in serum without any pretreatment and PCR amplification steps. When spiked in serum samples, a recovery of 99.65 %-102.38 % can be obtained. Therefore, our method offers a potential tool for ultrasensitive quantification of diseases related microRNA in the point-of-care diseases diagnosis field. [Display omitted] • Ultrasensitive single-line based SERS lateral flow assay for multiple exosomal proteins analysis has been developed. • For the first time CHA and CRISPR/Cas12a strategy were combined with LFA to improve the sensitivity. • The LOD (0.3 fM) was almost 1000-fold sensitivity than the previous fluorescence strips for miRNAs. • MiRNAs can be directly detected in serum without pretreatment or PCR amplification. • This assay possesses universality for miRNAs related diseases diagnosis in clinical setting. [ABSTRACT FROM AUTHOR]

Temporal lobe epilepsy (TLE) is one of the most common and intractable neurological disorders in adults. Dysfunctional PKA signaling is causally linked to the TLE. However, the mechanism underlying PKA involves in epileptogenesis is still poorly understood. In the present study, we found the autophosphorylation level at serine 114 site (serine 112 site in mice) of PKA-RIIβ subunit was robustly decreased in the epileptic foci obtained from both surgical specimens of TLE patients and seizure model mice. The p-RIIβ level was negatively correlated with the activities of PKA. Notably, by using a P-site mutant that cannot be autophosphorylated and thus results in the released catalytic subunit to exert persistent phosphorylation, an increase in PKA activities through transduction with AAV-RIIβ-S112A in hippocampal DG granule cells decreased mIPSC frequency but not mEPSC, enhanced neuronal intrinsic excitability and seizure susceptibility. In contrast, a reduction of PKA activities by RIIβ knockout led to an increased mIPSC frequency, a reduction in neuronal excitability, and mice less prone to experimental seizure onset. Collectively, our data demonstrated that the autophosphorylation of RIIβ subunit plays a critical role in controlling neuronal and network excitabilities by regulating the activities of PKA, providing a potential therapeutic target for TLE. Jingliang Zhang et al. examine how dysfunctional PKA signaling impacts neuronal activity and seizure susceptibility in a mouse model of temporal lobe epilepsy. Their data suggest that autophosphorylation of the RIIβ subunit regulates neuronal excitability, and may act as a future therapeutic target. [ABSTRACT FROM AUTHOR]

The functional relevance and mechanistic basis of the effects of the neurotransmitter dopamine (DA) on inflammation remain unclear. Here we reveal that DA inhibited TLR2-induced NF-κB activation and inflammation via the DRD5 receptor in macrophages. We found that the DRD5 receptor, via the EFD and IYX(X)I/L motifs in its CT and IC3 loop, respectively, can directly recruit TRAF6 and its negative regulator ARRB2 to form a multi-protein complex also containing downstream signaling proteins, such as TAK1, IKKs, and PP2A, that impairs TRAF6-mediated activation of NF-κB and expression of pro-inflammatory genes. Furthermore, the DA-DRD5-ARRB2-PP2A signaling axis can prevent S. aureus -induced inflammation and protect mice against S. aureus -induced sepsis and meningitis after DA treatment. Collectively, these findings provide the first demonstration of DA-DRD5 signaling acting to control inflammation and a detailed delineation of the underlying mechanism and identify the DRD5-ARRB2-PP2A axis as a potential target for future therapy of inflammation-associated diseases such as meningitis and sepsis. • Dopamine inhibits TLR2-induced NF-κB activation and inflammation via the DRD5 receptor • DRD5 can directly recruit TRAF6 and its negative regulator ARRB2 • DA-DRD5-ARRB2 signaling axis impairs TRAF6-mediated NF-κB activation and inflammation • DA-DRD5 signaling can protect mice against S. aureus -induced sepsis and meningitis Wu et al. demonstrate that, upon DA-DRD5 signaling activation, the DRD5 receptor can directly recruit TRAF6 and its negative regulator ARRB2 to form a multi-protein complex also containing downstream signaling proteins, such as IKKs and PP2A, that impairs TRAF6-mediated activation of NF-κB and inflammation during acute S. aureus infection. [ABSTRACT FROM AUTHOR]

Activated macrophages undergo a metabolic switch to aerobic glycolysis, accumulating Krebs’ cycle intermediates that alter transcription of immune response genes. We extended these observations by defining fumarate as an inhibitor of pyroptotic cell death. We found that dimethyl fumarate (DMF) delivered to cells or endogenous fumarate reacts with gasdermin D (GSDMD) at critical cysteine residues to form S-(2-succinyl)cysteine. GSDMD succination prevents its interaction with caspases, limiting its processing, oligomerization, and capacity to induce cell death. In mice, the administration of DMF protects against lipopolysaccharide shock and alleviates familial Mediterranean fever and experimental autoimmune encephalitis by targeting GSDMD. Collectively, these findings identify GSDMD as a target of fumarate and reveal a mechanism of action for fumarate-based therapeutics that include DMF, for the treatment of multiple sclerosis. [ABSTRACT FROM AUTHOR]

Background and Objective. FBXW7, known as a general tumor suppressor, is commonly lowly expressed in metastatic malignancies. We aim to investigate the potential influence of FBXW7 overexpression on renal cell carcinoma (RCC) metastasis. Methods. We employed quantitative real-time PCR (qRT-PCR) and Western blotting (WB) to quantify the FBXW7 expression in RCC cell lines. Upregulation of FBXW7 was performed in vitro on RCC cells using the lentivirus covering coding region FBXW7 cDNA sequence, and functional tests were performed to verify FBXW7 overexpression on migration and invasion of RCC cells. Moreover, WB was employed to determine the expressions of MMP-2, MMP-9, and MMP-13, as well as EMT markers in the transfected RCC cells. Results. FBXW7 was significantly downregulated in RCC cell lines, dominated by 786-O and ACHN, when compared to normal renal cell line HK-2. Moreover, upregulation of FBXW7 in 786-O and ACHN cell lines significantly inhibited cell migration and invasion, as well as EMT. Present study also showed that FBXW7 was involved in the migration and invasion of RCC cells via regulating the expressions of MMP-2, MMP-9, and MMP-13. Conclusion. Our findings demonstrate that upregulation of FBXW7 inhibits RCC metastasis and EMT. FBXW7 is a potential therapeutic target for RCC patients. [ABSTRACT FROM AUTHOR]

The onset of variant angina (VA) shows circadian rhythmicity that its attacks occur most often from midnight to early morning. Thus, chronotherapeutic treatments should be tailored accordingly to its occurrence frequency. Tanshinol (TS), the bioactive component of Salvia miltiorrhiza was used as the model drug. The pharmacokinetics, pharmacodynamics and PK-PD relationship of TS was investigated in angina model rabbits. The therapeutic effect of TS was evaluated from different aspects including cardiac injury, oxidative stress and vascular endothelium by measuring the serum levels of cTn-I, CK-MB, SOD and NO. In addition, the change of cTn-I levels from baseline as the pharmacodynamic endpoint was used for establishing the pharmacodynamic model. To synchronize the therapeutic effect profile of TS to the occurrence frequency of VA, ideal time courses of therapeutic effect, plasma concentration and drug release were simulated and calculated based on pharmacodynamic/deconvolution integrated model method. Then, sustained release pellets of TS (TS-SRPs) were developed according to the above calculated results and evaluated in vitro-in vivo. The established pharmacodynamic model of TS could precisely quantify the relationship between its effect and concentration. Then, ideal time courses of therapeutic effect, plasma concentration and release of TS were simulated and calculated successfully. After formulation optimization, the prepared TS-SRPs exhibited similar in vitro and in vivo behaviors to the corresponding ideal ones. Meanwhile, the effect curves of TS were synchronous with the occurrence frequency of VA, implying that appropriate therapeutic effect could be provided according to the needs of patients. In conclusion, the tailor of therapeutic effect based on integrated model method is efficient, feasible and reliable. [Display omitted] [ABSTRACT FROM AUTHOR]

Toll-like receptors (TLRs) sense pathogen-associated molecules and respond by inducing cytokines and type I interferon. Here we show that genetic ablation of the E3 ubiquitin ligase Pellino3 augmented the expression of type I interferon but not of proinflammatory cytokines in response to TLR3 activation. Pellino3-deficient mice had greater resistance against the pathogenic and lethal effects of encephalomyocarditis virus (EMCV). TLR3 signaling induced Pellino3, which in turn interacted with and ubiquitinated TRAF6. This modification suppressed the ability of TRAF6 to interact with and activate IRF7, resulting in downregulation of type I interferon expression. Our findings highlight a new physiological role for Pellino3 and define a new autoregulatory network for controlling type I interferon expression. [ABSTRACT FROM AUTHOR]