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Objective: To construct a prognostic model based on MR features and clinical data to evaluate the progression free survival (PFS), overall survival (OS) and objective response rate (ORR) of pancreatic cancer patients with hepatic metastases who received chemoimmunotherapy. Methods: 105 pancreatic cancer patients with hepatic metastases who received chemoimmunotherapy were assigned to the training set (n = 52), validation set (n = 22), and testing set (n = 31). Multi-lesion volume of interest were delineated, multi-sequence radiomics features were extracted, and the radiomics models for predicting PFS, OS and ORR were constructed, respectively. Clinical variables were extracted, and the clinical models for predicting PFS, OS and ORR were constructed, respectively. The nomogram was jointly constructed by radiomics model and clinical model. Result: The ORR exhibits no significant correlation with either PFS or OS. The area under the curve (AUC) of nomogram for predicting 6-month PFS reached 0.847 (0.737–0.957), 0.786 (0.566–1.000) and 0.864 (0.735–0.994) in the training set, validation set and testing set, respectively. The AUC of nomogram for predicting 1-year OS reached 0.770 (0.635–0.906), 0.743 (0.479–1.000) and 0.818 (0.630–1.000), respectively. The AUC of nomogram for predicting ORR reached 0.914 (0.828–1.00), 0.938 (0.840–1.00) and 0.846 (0.689–1.00), respectively. Conclusion: The prognostic models based on MR imaging features and clinical data are effective in predicting the PFS, OS and ORR of chemoimmunotherapy in pancreatic cancer patients with hepatic metastasis, and can be used to evaluate the prognosis of patients. [ABSTRACT FROM AUTHOR]

The effect of low voltage electrostatic field (LVEF, 2500 V, 0.2 mA) on oxidative denaturation of myofibrillar proteins (MPs) in repeatedly freeze–thaw (F-T, both 24 h) lamb was investigated in this study. F-T cycles intensified MP oxidative denaturation and structural destruction, with the influences becoming more severe after 1, 3, and 5 F-T cycles. The samples of the LVEF-assisted F-T cycles (LFT) exhibited lower carbonyl/dityrosine content, "r" value, and surface hydrophobicity, higher sulfhydryl content, solubility, Ca2+-ATPase activity, α-helix content, and fluorescence intensity under the identical F-T cycles (P < 0.05) in comparison with the common F-T cycles (CFT). Although the suppression of LVEF on protein oxidation decreased with the boost in the amount of F-T cycles, the effectiveness was still remarkable compared to samples without LVEF treatment. These original study findings would assist supply theoretical proof for the usage of LVEF in the freezing, storing, and transporting of frozen meat. [ABSTRACT FROM AUTHOR]

A series of colloidal Ag3PO4 sphere loaded onto multi-walled carbon nanotube (CAPS-MWCNT) photocatalysts containing different mass ratio of MWCNT for the removal of RhB and phenol from waste water were synthesized. The photocatalysts were characterized by X-ray diffraction, UV-Vis diffuse reflectance spectroscopy, thermogravimetry, scanning electron microscopy (SEM), transmission electron microscopy (TEM), and X-ray photoelectron spectroscopy. By SEM and TEM test, the obtained Ag3PO4 form precipitated and colloidal method present different structure morphology and this showed an important effect on the photocatalytic activity. The CAPS-MWCNT photocatalysts were applied to the removal of Rhodamine B (RhB) and phenol from waste water; and the results indicated that CAPS-MWCNT sample can exhibit the high adsorption and photodegradation efficiency under visible light irradiation. For RhB, the removal efficiency of CAPS-MWCNT-4 reaches 99.8% within 50 min, of which adsorption efficiency is 76.5% within the former 30 min; for phenol, the removal efficiency of CAPS-MWCNT-4 reaches 90.6% within 110 min, of which adsorption efficiency is 14.2% within the former 30 min. The experiment results show that CAPS-MWCNT-4 has the highest removal efficiency for RhB while it presents an excellent reusability for removing phenol from waste water. The present work illustrates a new train for the design and synthesis of stable and high efficiency catalysts, applicable for the removal of various organic pollutants by adsorption and photodegradation. [ABSTRACT FROM AUTHOR]

Objectives: To assess the ability of renal blood oxygen level dependent (BOLD) MRI metrics to predict post-operative renal function.Methods: We studied 152 patients who underwent laparoscopic partial nephrectomy (LPN) and renal MRI examination including BOLD. Short-term and long-term renal function was evaluated using the glomerular filtration rate (GFR) derived from renal scintigraphy. Renal function decline was assessed as the absolute decline (AD), percentage decline (PD) and optimal renal function preservation (OP). T2* values were analysed in the renal cortex and medulla ipsilateral and contralateral to the tumour. Clinical characteristics and imaging metrics were evaluated using univariate and multivariate linear regression analyses. Risk factors obtained using BOLD metrics (determined by multivariate regression) were then combined and compared with RENAL scores to predict OP.Results: Increasing warm ischaemia time (WIT), resected and ischaemic volume (RAIV), larger tumour size, higher RENAL score and lower preoperative GFR were short-term risk factors for AD, while increasing WIT and lower preoperative GFR were significant for long-term outcomes. Increasing WIT, RAIV, lower T2* value in the cortex and higher T2* value in the medulla on the ipsilateral side were short-term risk factors for PD, while all of the above factors (except WIT and RAIV) were significant for long-term outcomes. The performance of the combination of T2* values in the cortex and medulla on the ipsilateral side to tumour in predicting OP was better than RENAL score (AUC 0.762 vs 0.634, p = 0.013).Conclusions: Renal BOLD-MRI metrics could provide useful information to the clinician in predicting post-operative renal function outcomes.Key Points: • Renal fMRI metrics may be useful for prediction of renal functional outcomes and merit further study. • Renal fMRI metrics may reflect degree of baseline disease and ability to tolerate warm ischaemia. • Combination of T2* values was better than RENAL score for predicting OP. [ABSTRACT FROM AUTHOR]

Purpose: To evaluate the utility of MR R2*-mapping and the optimal time-point for assessing the response of pulmonary metastatic renal cell carcinoma (mRCC) to anti-angiogenic targeted therapy (aATT).Materials and Methods: The exploration-sample group and the validation-sample group consisted of 22 and 16 patients. The parameters of MR R2*-mapping, including the R2* value at each time-point (R2*base, R2*1cyc and R2*2cyc) and change between different time-points (R2*(1cyc-base)/base, R2*(2cyc-base)/base and R2*(2cyc-1cyc)/1cyc), were evaluated with a receiver-operating-characteristic analysis, and a cut-off value derived from the clinical outcome was applied to the Kaplan-Meier method to assess the value of R2* mapping and Response-Evaluation-Criteria in Solid Tumours (RECIST) during treatment evaluation.Results: The inter-, intra-observer agreements and inter-scan consistency were excellent (p > 0.80). For the exploration-sample group, the areas under the curve for the parameters of MR R2* mapping were 0.55, 0.60, 0.83, 0.64, 0.88 and 0.83 for R2*base, R2*1cyc, R2*2cyc, R2*(1cyc-base)/base, R2*(2cyc-base)/base and R2*(2cyc-1cyc)/1cyc. For the validation-sample, R2*(2cyc-base)/base better predicted progression-free survival (p = 0.03) than RECIST and other R2* mapping parameters with a lower p value.Conclusion: Assessing aATT outcome based on changes in the R2* value between baseline and second treatment is more accurate than assessment at other time-points and assessment based on the RECIST.Key Points: • The inter-scan consistency of R2*-mapping in pulmonary mRCC are excellent. • The intra-/inter-observer agreement of R2* mapping in pulmonary mRCC are excellent. • Using changes in R2* value between baseline/after second-treatment is better than RECIST. • The choice of baseline/after second treatment is better than other time-points. [ABSTRACT FROM AUTHOR]

Purpose: To compare the renal function preservation between laparoscopic radio frequency ablation assisted tumor enucleation and laparoscopic partial nephrectomy. Methods: Data were analyzed from 246 patients who underwent laparoscopic radio frequency ablation assisted tumor enucleation and laparoscopic partial nephrectomy for solitary cT1a renal cell carcinoma from January 2013 to July 2015. To reduce the intergroup difference, we used a 1:1 propensity matching analysis. The functional renal parenchyma volume preservation were measured preoperative and 12 months after surgery. The total renal function recovery and spilt GFR was compared. Multivariable logistic analysis was used for predictive factors for renal function decline. Results: After 1:1 propensity matching, each group including 100 patients. Patients in the laparoscopic radio frequency ablation assisted tumor enucleation had a smaller decrease in estimate glomerular filtration rate at 1 day (−7.88 vs −20.01%, p < 0.001), 3 months (−2.31 vs −10.39%, p < 0.001), 6 months (−2.16 vs −7.99%, p = 0.015), 12 months (−3.26 vs −8.03%, p = 0.012) and latest test (−3.24 vs −8.02%, p = 0.040), also had better functional renal parenchyma volume preservation (89.19 vs 84.27%, p < 0.001), lower decrease of the spilt glomerular filtration rate (−9.41 vs −17.13%, p < 0.001) at 12 months. The functional renal parenchyma volume preservation, warm ischemia time and baseline renal function were the important independent factors in determining long-term functional recovery. Conclusions: The laparoscopic radio frequency ablation assisted tumor enucleation technology has unique advantage and potential in preserving renal parenchyma without ischemia damage compared to conventional laparoscopic partial nephrectomy, and had a better outcome, thus we recommend this technique in selected T1a patients. [ABSTRACT FROM AUTHOR]

A new probe based on oxadiazole derivative (OXD) containing Schiff base moiety was synthesised as fluorescence turn-on sensor S1 . Furthermore, its fluorescents spectra (FS) result demonstrate that S1 selectively senses bivalent lead ion (Pb) in HEPES buffer solution (10 mM, pH = 7.0,DMF-HO, 9:1, v/ v), so S1 was a viable candidate as fluorescent receptor for a new Pb sensor. And the results of fluorescent spectral titration showed this sensor formed 1:1complex with Pb. [ABSTRACT FROM AUTHOR]

Objectives: To develop an artificial intelligence (AI) model for prostate segmentation and prostate cancer (PCa) detection, and explore the added value of AI-based computer-aided diagnosis (CAD) compared to conventional PI-RADS assessment. Methods: A retrospective study was performed on multi-centers and included patients who underwent prostate biopsies and multiparametric MRI. A convolutional-neural-network-based AI model was trained and validated; the reliability of different CAD methods (concurrent read and AI-first read) were tested in an internal/external cohort. The diagnostic performance, consistency and efficiency of radiologists and AI-based CAD were compared. Results: The training/validation/internal test sets included 650 (400/100/150) cases from one center; the external test included 100 cases (25/25/50) from three centers. For diagnosis accuracy, AI-based CAD methods showed no significant differences and were equivalent to the radiologists in the internal test (127/150 vs. 130/150 vs. 125/150 for reader 1; 127/150 vs.132/150 vs. 131/150 for reader 2; all p > 0.05), whereas in the external test, concurrent-read methods were superior/equal to AI-first read (87/100 vs. 71/100, p < 0.001, for reader 2; 79/100 vs. 69/100, p = 0.076, for reader 1) and better than/equal to radiologists (79/100 vs. 72/100, p = 0.039, for reader 1; 87/100 vs. 86/100, p = 1.000, for reader 2). Moreover, AI-first read/concurrent read improved consistency in both internal test (κ = 1.000, 0.830) and external test (κ = 0.958, 0.713) compared to radiologists (κ = 0.747, 0.600); AI-first read method (8.54 s/7.66 s) was faster than readers (92.72 s/89.54 s) and concurrent-read method (29.15 s/28.92 s), respectively. Conclusion: AI-based CAD could improve the consistency and efficiency for accurate diagnosis; the concurrent-read method could enhance the diagnostic capabilities of an inexperienced radiologist in unfamiliar situations. Key Points: • For prostate cancer segmentation, the performance of multi-small Vnet displays optimal compared to small Vnet and Vnet (DSCmsvnet vs. DSCsvnet, p = 0.021; DSCmsvnet vs. DSCvnet, p < 0.001). • For prostate gland segmentation, the mean/median DSCs for fine and coarse segmentation were 0.91/0.91 and 0.88/0.89, respectively. Fine segmentation displays superior performance compared to coarse (DSCcoarse vs. DSCfine, p < 0.001). • For PCa diagnosis, AI-based CAD methods improve consistency in internal (κ = 1.000; 0.830) and external (κ = 0.958; 0.713) tests compared to radiologists (κ = 0.747; 0.600); the AI-first read (8.54 s/7.66 s) was faster than the readers (92.72 s/89.54 s) and the concurrent-read method (29.15 s/28.92 s). [ABSTRACT FROM AUTHOR]

A unique polymeric carbon nitride (CN) tubular sponge with an edge-functionalized phosphonic group (CNPC) has been obtained using a simple template-free method. Using phenylphosphonic acid as an additive promotes the formation of a cyanuric acid–melamine supramolecular precursor. Meanwhile, phosphonic groups are grafted to heptazine units. Experimental and characterization investigations reveal that the functional phosphonic group and tubular sponge morphology endow CNPCx with additional reaction active sites, negative conduction band levels, and highly efficient charge separation. The obtained CNPCx photocatalysts exhibit excellent catalytic performance with regard to H2 evolution and contaminant degradation. This study provides a new design based on the synergetic regulating effect of macro and microstructures of CN with high performance in energy conversion and environmental purification. [ABSTRACT FROM AUTHOR]

Unsatisfied visible light response and high recombination probability of photogenerated charge carriers greatly limit the advanced application of graphitic carbon nitride (CN). Here, the novel Cu and C co-doping CN (Cu/C-CN) photocatalyst is successfully fabricated by thermal polymerization of the melamine and chlorophyll sodium copper salt as precursors, which significantly improves the photocatalytic performance than pure CN. A series of characterizations confirm that the co-doped Cu and C in CN not only endow the material with improved light harvest abilities and narrowed band gap, but also facilitate the charge carriers separation. As a result, the Cu/C-CN-0.3 photocatalyst exhibits a highly efficient photocatalytic degradation of RhB, the reaction rate constant of Cu/C-CN-0.3 (kapp=0.036 min−1) is almost 13 times higher than CN (kapp = 0.0027 min−1). This study may open a new perspective for constructing other dual-elements co-doped CN materials with high performance and application in wastewater treatment. [ABSTRACT FROM AUTHOR]

A smart bioreactor (HPs-Ag), designed by protein-silver nanoclusters-PNIPAAm, could control the membrane permeability to determine the different molecular weight of materials and the speed of catalysis of enzyme reaction. Studies also showed that the molecular weight cutoff of the HPs-Ag membrane could be continuously turned up from 12 to 70 kDa. The catalytic reaction rate of the ALP loaded bared HPs-Ag by using pNPP as substrates was increased (18.14- and 9.23-fold) with different concentrations of H2O2. [ABSTRACT FROM AUTHOR]

G protein-coupled receptors (GPCRs) transmit signals by forming active-state complexes with heterotrimeric G proteins. It has been suggested that some GPCRs also assemble with G proteins before ligand-induced activation and that inactive-state preassembly facilitates rapid and specific G protein activation. However, no mechanism of preassembly has been described, and no functional consequences of preassembly have been demonstrated. Here we show that M3 muscarinic acetylcholine receptors (M3R) form inactive-state complexes with Gq heterotrimers in intact cells. The M3R C terminus is sufficient, and a six-amino-acid polybasic sequence distal to helix 8 (565KKKRRK570) is necessary for preassembly with Gq. Replacing this sequence with six alanine residues prevents preassembly, slows the rate of Gq activation and decreases steady-state agonist sensitivity. That other Gq-coupled receptors possess similar polybasic regions and also preassemble with Gq suggests that these GPCRs may use a common preassembly mechanism to facilitate activation of Gq heterotrimers. [ABSTRACT FROM AUTHOR]

Objectives: To develop a radiomics model using preoperative multiphasic CT for predicting distant metastasis after surgical resection in patients with localized clear cell renal cell carcinoma (ccRCC) and to identify key biological pathways underlying the predictive radiomics features using RNA sequencing data. Methods: In this multi-institutional retrospective study, a CT radiomics metastasis score (RMS) was developed from a radiomics analysis cohort (n = 184) for distant metastasis prediction. Using a gene expression analysis cohort (n = 326), radiomics-associated gene modules were identified. Based on a radiogenomics discovery cohort (n = 42), key biological pathways were enriched from the gene modules. Furthermore, a multigene signature associated with RMS was constructed and validated on an independent radiogenomics validation cohort (n = 37). Results: The 9-feature-based RMS predicted distant metastasis with an AUC of 0.861 in validation set and was independent with clinical factors (p < 0.001). A gene module comprising 114 genes was identified to be associated with all nine radiomics features (p < 0.05). Four enriched pathways were identified, including ECM-receptor interaction, focal adhesion, protein digestion and absorption, and PI3K-Akt pathways. Most of them play important roles in tumor progression and metastasis. A 19-gene signature was constructed from the radiomics-associated gene module and predicted metastasis with an AUC of 0.843 in the radiogenomics validation cohort. Conclusions: CT radiomics features can predict distant metastasis after surgical resection of localized ccRCC while the predictive radiomics phenotypes may be driven by key biological pathways related to cancer progression and metastasis. Key Points: • Radiomics features from primary tumor in preoperative CT predicted distant metastasis after surgical resection in patients with localized ccRCC. • CT radiomics features predictive of distant metastasis were associated with key signaling pathways related to tumor progression and metastasis. • Gene signature associated with radiomics metastasis score predicted distant metastasis in localized ccRCC. [ABSTRACT FROM AUTHOR]

The design and synthesis of economy and efficiency materials for oxygen evolution reaction (OER) have been a continuous hot spot in the field of scientific study. Herein, a high-valence-state two-dimension (2D) NiFe phosphonate-based (NiFeP) nanoribbons catalyst has been constructed through a one-step solvothermal process. The NiFeP nanoribbons exhibit highly active in both photocatalytic and electrocatalytic water oxidation due to the 2D nanoribbons and high-valence Ni3+ sites. The 2D nanoribbons not only provide more reactive sites for OER but also shorten bulk diffusion distance with better photoexcited carrier transport from the interior to the surface. Meanwhile, the existence of high-valence Ni3+ could be acted as an efficient redox site to reduce the overpotential and facilitate the catalytic reaction. In consequence, the NiFeP nanoribbons catalyst demonstrates a superior O2 yield of 65.7% and O2 production rate of 25.97 umol s−1 g−1, which are comparable or even much higher than those other reported transition metal oxide photocatalysts. At last, the possible proton-coupled electron transfer mechanism is also proposed. This study not only demonstrates the potential of a low-cost metal phosphonate OER catalyst but also provides a referential system for the fabrication of high activity and stability catalysts toward replacing noble metals for energy storage and conversion. High-valence-state 2D NiFe phosphonate-based nanoribbons catalyst has been constructed for the first time through a one-step solvothermal process without any organic additives or template agent. The as-prepared catalyst of NiFeP nanoribbons exhibits highly active in both photocatalytic and electrocatalytic water oxidation. [ABSTRACT FROM AUTHOR]

Purpose: To compare various models of diffusion-weighted imaging including mono-exponential, bi-exponential, diffusion kurtosis (DK) and fractional-order calculus (FROC) models in diagnosing prostate cancer (PCa) in transition zone (TZ) and distinguish the high-grade PCa [Gleason score (GS) ≥ 7] lesions from the total of low-grade PCa (GS ≤ 6) lesions and benign prostatic hyperplasia (BPH) in TZ. Methods: 80 Patients with 103 lesions were included in this study. Nine metrics [including apparent diffusion coefficient (ADC) derived from mono-exponential model, slow diffusion coefficient (Ds), fast diffusion coefficient (Df),, and f (the fraction of fast diffusion) from bi-exponential model; mean diffusivity (MD) and mean kurtosis (MK) from DK model; diffusion coefficient (D), fractional-order derivative in space (β), and spatial metric (μ) from FROC model] were calculated. Comparisons between BPH and PCa lesions as well as between clinically significant PCa (CsPCa) (GS ≥ 7, n = 31) and clinically insignificant lesions (Cins) (GS ≤ 6 and BPH, n = 72) of these metrics were conducted. Mann–Whitney U-test and receiver operating characteristic (ROC) analysis were used for statistical evaluations. Results: The areas under the ROC curve (AUC) values of β derived from FROC model were 0.778 and 0.853 in differentiating PCa from BPH and in differentiating CS (GS ≥ 7) from Cins (GS ≤ 6 and BPH), both were the highest compared to other metrics. The AUC value of β was significantly higher than that of ADC (P = 0.009) in differentiating CS from Cins, while the differentiation between BPH and PCa did not reach the statistical significance when comparing with ADC (P = 0.089). Conclusion: Although no significant difference was found in distinguishing PCa from BPH, the metric β derived from FROC model was superior to other diffusion metrics in differentiation between CS and Cins in TZ. [ABSTRACT FROM AUTHOR]

Purpose: To investigate the value of radiomics features from diffusion-weighted imaging (DWI) in differentiating muscle-invasive bladder cancer (MIBC) from non-muscle-invasive bladder cancer (NMIBC).Methods: This retrospective study included 218 pathologically confirmed bladder cancer patients (training set: 131 patients, 86 MIBC; validation set: 87 patients, 55 MIBC) who underwent DWI before biopsy through transurethral resection (TUR) between July 2014 and December 2018. Radiomics models based on DWI for discriminating state of muscle-invasive were built using random forest (RF) and all-relevant (AR) methods on the training set and were tested on validation set. Combination models based on TUR data were also built. Discrimination performances were evaluated with the area under the receiver operating characteristic (ROC) curve (AUC), accuracy, sensitivity, specificity, and F1 and F2 scores. Qualitative MRI evaluation based on morphology was performed for comparison.Results: No significant difference was found between RF and AR models. RF model was more sensitive than TUR (0.873 vs 0.655, p = 0.019) for discriminating muscle-invasive bladder cancer. When combining RF with TUR, the sensitivity increased to 0.964, significantly higher than TUR (0.655, p < 0.001), MRI evaluation (0.764, p = 0.006), and the combination of TUR and MRI (0.836, p = 0.046). Combining RF and TUR achieved the highest accuracy of 0.897 and F2 score of 0.946.Conclusion: Combining DWI radiomics features with TUR could improve the sensitivity and accuracy in discriminating the presence of muscle invasion in bladder cancer for clinical practice. Multicenter, prospective studies are needed to confirm our results.Key Points: • Twenty-seven to 51% of superficial bladder cancers diagnosed by transurethral resection are upstaged to muscle-invasive at radical cystectomy, suggesting its poor sensitivity for discriminating muscle-invasive bladder cancer. • A small subset of selected all-relevant radiomics features exhibited an equivalent performance compared to that of all the extracted features, confirming that radiomics data contained redundant or irrelevant features and that feature selection should be performed in building radiomics models. • Combining DWI radiomics features with transurethral resection could improve in clinical practice the sensitivity and accuracy for the detection of muscle invasion in bladder cancer. [ABSTRACT FROM AUTHOR]

The small GTPase Rab5 has been well defined to control the vesicle-mediated plasma membrane protein transport to the endosomal compartment. However, its function in the internalization of vascular endothelial (VE)-cadherin, an important component of adherens junctions, and as a result regulating the endothelial cell polarity and barrier function remain unknown. Here, we demonstrated that lipopolysaccharide (LPS) simulation markedly enhanced the activation and expression of Rab5 in human pulmonary microvascular endothelial cells (HPMECs), which is accompanied by VE-cadherin internalization. In parallel, LPS challenge also induced abnormal cell polarity and dysfunction of the endothelial barrier in HPMECs. LPS stimulation promoted the translocation of VE-cadherin from the plasma membrane to intracellular compartments, and intracellularly expressed VE-cadherin was extensively colocalized with Rab5. Small interfering RNA (siRNA)-mediated depletion of Rab5a expression attenuated the disruption of LPS-induced internalization of VE-cadherin and the disorder of cell polarity. Furthermore, knockdown of Rab5 inhibited the vascular endothelial hyperpermeability and protected endothelial barrier function from LPS injury, both in vitro and in vivo. These results suggest that Rab5 is a critical mediator of LPS-induced endothelial barrier dysfunction, which is likely mediated through regulating VE-cadherin internalization. These findings provide evidence, implicating that Rab5a is a potential therapeutic target for preventing endothelial barrier disruption and vascular inflammation. [ABSTRACT FROM AUTHOR]

Angiotensin II (Ang II), a potent precursor of hypertrophy and heart failure, upregulates neuronal nitric oxide synthase (nNOS or NOS1) in the myocardium. Here, we investigate the involvement of type 1 and 2 angiotensin receptors (AT1R and AT2R) and molecular mechanisms mediating Ang II-upregulation of nNOS. Our results showed that pre-treatment of left ventricular (LV) myocytes with antagonists of AT1R or AT2R (losartan, PD123319) and ROS scavengers (apocynin, tiron or PEG-catalase) blocked Ang II-upregulation of nNOS. Surface biotinylation or immunocytochemistry experiments demonstrated that AT1R expression in plasma membrane was progressively decreased (internalization), whereas AT2R was increased (membrane trafficking) by Ang II. Inhibition of AT1R or ROS scavengers prevented Ang II-induced translocation of AT2R to plasma membrane, suggesting an alignment of AT1R-ROS-AT2R. Furthermore, Ang II increased eNOS-Ser but decreased eNOS-Thr, indicating concomitant activation of eNOS. Intriguingly, ROS scavengers but not AT2R antagonist prevented Ang II-activation of eNOS. NOS inhibitor (L-NG-Nitroarginine Methyl Ester, L-NAME) or eNOS gene deletion (eNOS) abolished Ang II-induced membrane trafficking of AT2R, nNOS protein expression and activity. Mechanistically, S-nitrosation of AT2R was increased by sodium nitroprusside (SNP), a NO donor. Site-specific mutagenesis analysis reveals that C-terminal cysteine 349 in AT2R is essential in AT2R translocation to plasma membrane. Taken together, we demonstrate, for the first time, that Ang II upregulates nNOS protein expression and activity via AT1R/ROS/eNOS-dependent S-nitrosation and membrane translocation of AT2R. Our results suggest a novel crosstalk between AT1R and AT2R in regulating nNOS via eNOS in the myocardium under pathogenic stimuli. [ABSTRACT FROM AUTHOR]

Loss of cardiomyocytes through programmed cell death is a key event in the development of heart failure, but the inciting molecular mechanisms are largely unknown. We used microarray analysis to identify a genetic program for myocardial apoptosis in Gq-mediated and pressure-overload cardiac hypertrophy. A critical component of this apoptotic program was Nix/Bnip3L. Nix localized to mitochondria and caused release of cytochrome c, activation of caspase-3 and apoptotic cell death, when expressed in HEK293 fibroblasts. A previously undescribed truncated Nix isoform, termed sNix, was not targeted to mitochondria but heterodimerized with Nix and protected against Nix-mediated apoptosis. Forced in vivo myocardial expression of Nix resulted in apoptotic cardiomyopathy and rapid death. Conversely, sNix protected against apoptotic peripartum cardiomyopathy in Gαq-overexpressors. Thus, Nix/Bnip3L is upregulated in myocardial hypertrophy, and is both necessary and sufficient for Gq-mediated apoptosis of cardiomyocytes and resulting hypertrophy decompensation. [ABSTRACT FROM AUTHOR]

The regulatory mechanisms of cell surface targeting of nascent G protein-coupled receptors (GPCRs) en route from the endoplasmic reticulum through the Golgi remain poorly understood. We have recently demonstrated that three Golgi-localized, γ-adaptin ear domain homology, ADP ribosylation factor-binding proteins (GGAs) mediate the post-Golgi export of α2B-adrenergic receptor (α2B-AR), a prototypic GPCR, and directly interact with the receptor. In particular, GGA1 interaction with α2B-AR is mediated via its hinge domain. Here we determined the role of a naturally occurring truncated form of GGA1 (GGA1t) which lacks the N-terminal portion of the hinge domain in α2B-AR trafficking and elucidated the underlying mechanisms. We demonstrated that both GGA1 and GGA1t were colocalized and mainly expressed at the Golgi. In marked contrast to GGA1, the expression of GGA1t significantly attenuated the cell surface export of newly synthesized α2B-AR from the Golgi and in parallel receptor-mediated signaling. Furthermore, we found that GGA1t formed homodimers and heterodimers with GGA1. More interestingly, GGA1t was unable to bind the cargo α2B-AR and to recruit clathrin onto the trans-Golgi network. These data provide evidence implicating that the truncated form of GGA1 behaviors as a dominant-negative regulator for the cell surface export of α2B-AR and this function of GGA1t is attributed to its abilities to dimerize with its wide type counterpart and to inhibit cargo interaction and clathrin recruitment to form specialized transport vesicles. [ABSTRACT FROM AUTHOR]

Resistant starch (RS) has been reported to reduce body fat in obese mice. However, this effect has not been demonstrated in humans. In this study, we tested the effects of RS in 19 volunteers with normal body weights. A randomized, double-blinded and crossover design clinical trial was conducted. The study subjects were given either 40 g high amylose RS2 or energy-matched control starch with three identical diets per day throughout the study. The effect of RS was evaluated by monitoring body fat, glucose metabolism, gut hormones, gut microbiota, short-chain fatty acids (SCFAs) and metabolites. The visceral and subcutaneous fat areas were significantly reduced following RS intake. Acetate and early-phase insulin, C-peptide and glucagon-like peptide-1 (GLP-1) secretion were increased, and the low-density lipoprotein cholesterol (LDL-C) and blood urea nitrogen (BUN) levels were decreased after the RS intervention. Based on 16S rRNA sequencing, certain gut microbes were significantly decreased after RS supplementation, whereas the genus Ruminococcaceae_UCG-005 showed an increase in abundance. Other potential signatures of the RS intervention included Akkermansia, Ruminococcus_2, Victivallis, and Comamonas. Moreover, the baseline abundance of the genera Streptococcus, Ruminococcus_torques_group, Eubacterium_hallii_group, and Eubacterium_eligens_group was significantly associated with the hormonal and metabolic effects of RS. These observations suggest that a daily intake of 40 g of RS is effective in modulating body fat, SCFAs, early-phase insulin and GLP-1 secretion and the gut microbiota in normal-weight subjects. [ABSTRACT FROM AUTHOR]

The molecular mechanisms that control the targeting of newly synthesized G protein-coupled receptors (GPCRs) to the functional destinations remain poorly elucidated. Here, we have determined the role of Golgi-localized, γ-adaptin ear domain homology, ADP ribosylation factor-binding proteins 1 and 2 (GGA1 and GGA2) in the cell surface transport of α2B-adrenergic receptor (α2B-AR), a prototypic GPCR, and studied the underlying mechanisms. We demonstrated that knockdown of GGA1 and GGA2 by shRNA and siRNA significantly reduced the cell surface expression of inducibly expressed α2B-AR and arrested the receptor in the perinuclear region. Knockdown of each GGA markedly inhibited the dendritic expression of α2B-AR in primary cortical neurons. Consistently, depleting GGA1 and GGA2 attenuated receptor-mediated signal transduction measured as ERK1/2 activation and cAMP inhibition. Although full length α2B-AR associated with GGA2 but not GGA1, its third intracellular loop was found to directly interact with both GGA1 and GGA2. More interestingly, further mapping of interaction domains showed that the GGA1 hinge region and the GGA2 GAE domain bound to multiple subdomains of the loop. These studies have identified an important function and revealed novel mechanisms of the GGA family proteins in the forward trafficking of a cell surface GPCR. [ABSTRACT FROM AUTHOR]

Bioluminescence resonance energy transfer (BRET) is often used to study association of membrane proteins, and in particular oligomerization of G protein-coupled receptors (GPCRs). Oligomerization of class A GPCRs is controversial, in part because the methods used to study this question are not completely understood. Here we reconsider oligomerization of the class A β2 adrenergic receptor (β2AR), and reevaluate BRET titration as a method to study membrane protein association. Using inducible expression of the energy acceptor at multiple levels of donor expression we find that BRET between β2AR protomers is directly proportional to the density of the acceptor up to ~3,000 acceptors μm−2, and does not depend on the density of the donor or on the acceptor:donor (A:D) stoichiometry. In contrast, BRET between tightly-associating control proteins does not depend on the density of the acceptor, but does depend on the density of the donor and on the A:D ratio. We also find that the standard frameworks used to interpret BRET titration experiments rely on simplifying assumptions that are frequently invalid. These results suggest that β2ARs do not oligomerize in cells, and demonstrate a reliable method of assessing membrane protein association with BRET. [ABSTRACT FROM AUTHOR]