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Background: Thoracic empyema is a serious infectious disease worldwide. Video-assisted thoracoscopic surgery is strongly recommended as a treatment, and pleural fluid and tissue cultures can be obtained intraoperatively. The combination of a pleural peels tissue culture and a pleural fluid culture improves the positive culture rate. We aimed to investigate the role of sputum cultures to determine the optimal management for improving surgical outcome. Methods: This retrospective study identified 1197 patients with phase II or III thoracic empyema from our institution. Patients who underwent decortication of the pleura from April 2011 to May 2022 with a positive pleural culture were included. Results: There were 225 empyema patients with either a positive pleural fluid culture or a positive pleural peel tissue culture. Of these, 76 patients had positive sputum culture findings during hospitalization. The most common species of pathogens were Pseudomonas aeruginosa (44%) and Klebsiella pneumoniae (16%) in the sputum cultures and Streptococcus spp. (20%) and Staphylococcus aureus(11%) in the pleural cultures. There were 30 patients who had a common pathogen in a sputum culture and in the pleural fluid/tissue culture. Poor outcome measures were found in these patients, including the longer use of antibiotics preoperatively (19.03±37.66 days versus 9.59±27.09 days, p=0.006) and a higher mortality rate during hospitalization (40.0% versus 17.4%, p=0.002). Conclusions: The sputum culture plays an essential role in diagnosis of bacterial empyema and in prediction of surgical outcome. Obtaining the sputum specimen promptly through proper methods helps to improve the survival of empyema patients.

We characterize a new chaos lidar system configuration and demonstrate its capability for high-speed 3D imaging. Compared with a homodyned scheme employing single-element avalanche photodetectors (APDs), the proposed scheme utilizes a fiber Bragg grating and quadrant APDs to substantially increase the system throughput, frame rate, and field-of-view. By quantitatively analyzing the signal-to-noise ratio, peak-to-standard deviation of the sidelobe level, precision, and detection probability, we show that the proposed scheme has better detection performance suitable for practical applications. To show the feasibility of the chaos lidar system, while under the constrain of eye-safe regulation, we demonstrate high-speed 3D imaging with indoor and outdoor scenes at a throughput of 100 kHz, a frame rate of 10 Hz, and a FOV of 24.5$$^\circ $$ ∘ $$\times $$ × 11.5$$^\circ $$ ∘ for the first time.

Purpose Speech-language pathologists (SLPs) are faced with the challenge of quickly and accurately identifying children who present with speech sound disorders (SSD) compared to typically developing (TD) children. The goal of this study was to compare the clinical relevance of two speech sampling methods (single-word vs. connected speech samples) in how sensitive they are in detecting atypical speech sound development in children, and to know whether the information obtained from single-word samples is representative enough of children's overall speech sound performance. Method We compared the speech sound performance of 37 preschool children with SSD ( M age = 4;11 years) and 37 age-sex-matched typically developing children ( M age = 5;0 years) by eliciting their speech in two ways: (a) a picture-naming task to elicit single words, and (b) a story-retelling task to elicit connected speech. Four speech measures were compared across sample type (single words vs. connected speech) and across groups (SSD vs. TD): intelligibility, speech accuracy, phonemic inventory, and phonological patterns. Results Interaction effects were found between sample type and group on several speech sound performance measures. Single-word speech samples were found to differentiate the SSD group from the TD group, and were more sensitive than connected speech samples across various measures. The effect size of single-word samples was consistently higher than connected speech samples for three measures: intelligibility, speech accuracy, and phonemic inventory. The gap in sample type informativeness may be attributed to salience and avoidance effects, given that children tend to avoid producing unfamiliar phonemes in connected speech. The number of phonological patterns produced was the only measure that revealed no gap between two sampling types for both groups. Conclusions On measures of intelligibility, speech accuracy, and phonemic inventory, obtaining a single-word sample proved to be a more informative method of differentiating children with SSD from TD children than connected speech samples. This finding may guide SLPs in their choice of sampling type when they are under time pressure. We discuss how children's performance on the connected speech sample may be biased by salience and avoidance effects and/or task design, and may, therefore, not necessarily reveal a poorer performance than single-word samples, particularly in intelligibility, speech accuracy, and the number of phonological patterns, if these task limitations are circumvented. Our findings show that the performance gap, typically observed between the two sampling types, largely depends on which performance measures are evaluated with the speech sample. Our study is the first to address sampling type differences in SSD versus TD children and has significant clinical implications for SLPs looking for sampling types and measures that reliably identify SSD in preschool-aged children.

Efficacy of therapies that target the downstream nitric oxide (NO) pathway in pulmonary arterial hypertension (PAH) depends on the bioavailability of NO. Reduced NO level in PAH is secondary to "uncoupling" of endothelial nitric oxide synthase (eNOS). Stimulation of β3 adrenergic receptors (β3 ARs) may lead to the recoupling of NOS and therefore be beneficial in PAH. We aimed to examine the efficacy of β3 AR agonism as a novel pathway in experimental PAH. In hypoxia (5 weeks) and Sugen hypoxia (hypoxia for 5 weeks + SU5416 injection) models of PAH, we examined the effects of the selective β3 AR agonist CL316243. We measured echocardiographic indices and invasive right ventricular (RV)-pulmonary arterial (PA) hemodynamics and compared CL316243 with riociguat and sildenafil. We assessed treatment effects on RV-PA remodeling, oxidative stress, and eNOS glutathionylation, an oxidative modification that uncouples eNOS. Compared with normoxic mice, RV systolic pressure was increased in the control hypoxic mice (p 0.0001) and Sugen hypoxic mice (p 0.0001). CL316243 reduced RV systolic pressure, to a similar degree to riociguat and sildenafil, in both hypoxia (p 0.0001) and Sugen hypoxia models (p 0.03). CL316243 reversed pulmonary vascular remodeling, decreased RV afterload, improved RV-PA coupling efficiency and reduced RV stiffness, hypertrophy, and fibrosis. Although all treatments decreased oxidative stress, CL316243 significantly reduced eNOS glutathionylation. β3 AR stimulation improved RV hemodynamics and led to beneficial RV-PA remodeling in experimental models of PAH. β3 AR agonists may be effective therapies in PAH.

Background: Volume doubling time (VDT) has been proven to be a powerful predictor of lung cancer progression. In non-small cell lung cancer (NSCLC) patients receiving sublobar resection, the discussion of correlation between VDT and surgery was absent. We propose to investigate the surgical outcomes according to VDT, to see if it could serve as a prognostic predictor preoperatively. Methods: We retrospectively studied 96 NSCLC patients post sublobar resection from 2012 to 2018, collecting two chest CT scans preoperatively of each case and calculating VDT. The receiver operating characteristic curve was constructed to identify the optimal cut-off point of VDTs as 133 days. We divided patients into two groups: VDT < 133 days (n=22) and VDT ≥ 133 days (n=74). Univariable and multivariable analyses were performed for comparative purposes. Results: Clinical characteristics including gender, smoking history, chronic obstructive pulmonary disease, epidermal growth factor receptor gene mutation, tumor stage, and histological type were significantly different between the two groups. Univariable and multivariable analyses revealed that the consolidation and tumor diameter ratio was the factor of overall survival (OS), and VDT was the only factor of disease-free survival (DFS). The 5-year OS rates of patients with VDTs ≥ 133 days and VDTs < 133 days, respectively, were 89.9% and 71.9% (p = 0.003), and the 5-year DFS rates were 95.9% and 61.5% (p = 0.002). Conclusions: As the VDT serves as a powerful prognostic predictor and provides an essential role in planning surgical procedures, the evaluation of VDT preoperatively is highly suggested.

Background: The objective of our study was to assess if 3D reconstructed images could be extrapolated to reflect pathologies, as evaluated by early-stage lung adenocarcinoma tumor size and simulated segmentectomy resection margin. Methods: Retrospectively selected patients (n = 18) who underwent segmentectomy at Changhua Christian Hospital between 2012 and 2018 and then had pulmonary 3D reconstruction using Ziostation2 were included in our study. Tumor size and simulated segmentectomy resection distance on a 3D model were measure and compared to pathology. Results: Both tumor size and segmentectomy resection margin showed positive correlations between 3D image measurements and pathological measurements. The resection margin showed a stronger correlation and was beneficial in pre-operative planning. Conclusions: A 3D reconstructed model aided understanding of pulmonary anatomy, prompting confidence in surgical approaches and ensured segmentectomy outcome success. Regardless of age and pulmonary function, 3D simulation can accurately mimic segmentectomy, making it a simple, effective and feasible pre-operative planning tool.

Nitric oxide (NO) production by eNOS (endothelial NO synthase) is critical for vascular health. Oxidative stress-induced uncoupling of eNOS leads to decreased NO bioavailability, compounded by increased superoxide generation. FXYD1 (FXYD domain containing ion transport regulator 1), a caveolar protein, protects against oxidative inhibition of the Na + -K + -ATPase. We hypothesized that FXYD1 may afford a similar inhibition of oxidative dysregulation of eNOS, providing a broader protection within caveolae. FXYD1-eNOS colocalization was demonstrated by co-immunoprecipitation in heart protein and by proximity ligation assay in human umbilical vein endothelial cells. The functional nature of this partnership was shown by silencing FXYD1 in human umbilical vein endothelial cells, where 50% decreased NO and 2-fold augmented superoxide was shown. Three-dimensional cocultured cardiac spheroids generated from FXYD1 knockout mice were incapable of acetylcholine-induced NO production. Overexpression of FXYD1 in HEK293 cells revealed a possible mechanism, where FXYD1 protected against redox modification of eNOS cysteines. In vivo, vasodilation in response to increasing doses of bradykinin was impaired in knockout mice, and this was rescued in mice by delivery of FXYD1 protein packaged in exosomes. Bloods vessels extracted from knockout mice exhibited increased oxidative and nitrosative stress with evidence of reduce eNOS phosphorylation. Impaired vascular function and augmented superoxide generation were also evident in diabetic knockout mice. Despite this, blood pressure was similar in wildtype and knockout mice, but after chronic angiotensin II infusion, knockout of FXYD1 was associated with a heightened blood pressure response. FXYD1 protects eNOS from dysregulated redox signaling and is protective against both hypertension and diabetic vascular oxidative stress.

Context Reduced Na+-K+ pump activity is widely reported in preeclampsia and may be caused by a reversible oxidative modification that is a novel pathological feature of preeclampsia. Objective This work aims to determine whether β 1 subunit (GSS-β 1) protein glutathionylation of the Na+-K + pump occurs in preeclampsia. Methods The GSS-β1 of the Na+-K+ pump and its subunit expression in human placentas were compared between women with healthy pregnancies and women with preeclampsia. Human placental samples of pregnant women with preeclampsia (n = 11, mean gestational age 36.5 weeks) were used to examine the GSS-β 1 of the Na+-K+ pump, compared to healthy pregnancies (n = 11, mean gestational age 39 weeks). The potential pathogenetic role of GSS-β 1-mediated Na+-K+ pump dysfunction in preeclampsia was investigated. Results Protein expression of the β 1 subunit was unchanged in placentas from women with preeclampsia vs those with normotensive pregnancies. Preeclamptic placentas had a significantly increased GSS-β 1 of the Na+-K+ pump compared to those from healthy pregnancies, and this was linked to a decrease in α 1/β 1 subunit coimmunoprecipitation. The cytosolic p47phox nicotinamide adenine dinucleotide phosphate hydrogen (NADPH) oxidase subunit and its coimmunoprecipitation with the α 1 Na+-K+ pump subunit was increased in preeclamptic placentas, thus implicating NADPH oxidase–dependent pump inhibition. Conclusions The high level of β 1 pump subunit glutathionylation provides new insights into the mechanism of Na+-K+ pump dysfunction in preeclampsia.

The seven mammalian FXYD proteins associate closely with α/β heterodimers of Na+/K+-ATPase. Most of them protect the β1 subunit against glutathionylation, an oxidative modification that destabilizes the heterodimer and inhibits Na+/K+-ATPase activity. A specific cysteine (Cys) residue of FXYD proteins is critical for such protection. One of the FXYD proteins, FXYD3, confers treatment resistance when overexpressed in cancer cells. We developed two FXYD3 peptide derivatives. FXYD3-pep CKCK retained the Cys residue that can undergo glutathionylation and that is critical for protecting the β1 subunit against glutathionylation. FXYD3-pep SKSK had all Cys residues mutated to Serine (Ser). The chemotherapeutic doxorubicin induces oxidative stress, and suppression of FXYD3 with siRNA in pancreatic- and breast cancer cells that strongly express FXYD3 increased doxorubicin-induced cytotoxicity. Exposing cells to FXYD3-pep SKSK decreased co-immunoprecipitation of FXYD3 with the α1 Na+/K+-ATPase subunit. FXYD3-pep SKSK reproduced the increase in doxorubicin-induced cytotoxicity seen after FXYD3 siRNA transfection in pancreatic- and breast cancer cells that overexpressed FXYD3, while FXYD3-pep CKCK boosted the native protein’s protection against doxorubicin. Neither peptide affected doxorubicin’s cytotoxicity on cells with no or low FXYD3 expression. Fluorescently labeled FXYD3-pep SKSK was detected in a perinuclear distribution in the cells overexpressing FXYD3, and plasmalemmal Na+/K+-ATPase turnover could not be implicated in the increased sensitivity to doxorubicin that FXYD3-pep SKSK caused. FXYD peptide derivatives allow rapid elimination or amplification of native FXYD protein function. Here, their effects implicate the Cys residue that is critical for countering β1 subunit glutathionylation in the augmentation of cytotoxicity with siRNA-induced downregulation of FXYD3.

Objectives Integrins are cell adhesion receptors that participated in endovascular invasion by cytotrophoblasts in preeclampsia. This study aimed to investigate the effect of calcium on cellular pathways influencing the trophoblast integration into endothelial cellular networks in vitro. Study design Red fluorescent-labelled human uterine myometrial microvascular endothelial cells (UtMVECs) were seeded on Matrigel. Green fluorescent-labelled HTR-8/SVneo trophoblast cells were then co-cultured with endothelial cells in different concentrations of calcium for 24 h. Main outcome measures The calcium effects on HTR-8/SVneo cell integration were quantified by Image J. Quantitative PCR was performed to measure mRNA expression of integrins α1, α5, α6, β1 and β4. The concentrations of interleukin IL-6, matrix metalloproteinase-2 (MMP-2), MMP-9, PlGF and sFlt-1 in the conditioned medium were measured by ELISA while levels of cytokines IL-1β, IL-8, IL-10, TNF-α and INF-γ were assessed by magnetic Luminex assays™. Results Both calcium depletion (0.4 mM) and low calcium (1.8 mM) groups demonstrated inhibited integration of trophoblast cells into endothelial cellular networks, compared with the normal calcium group (2.4 mM). The IL-6 production was reduced from conditioned media in both calcium depletion and low calcium groups. In calcium depletion group, mRNA expression of integrin α5 and β4 in trophoblasts was increased while integrin α1 was decreased. Conclusions The in vitro trophoblast cell integration into endothelial cellular networks could be modified by altering media calcium through integrin switch away from integrins α5 and β4 and towards integrin α1 which may be required for healthy early trophoblast integration.

Background/aim Maslinic acid (MA), a pentacyclic triterpene extracted from wax-like coatings of olives, has been shown to reduce cancer cell number through induction of autophagy and apoptosis in many human cancer cells including human leukemia HL-60 cells. In the present study, we investigated whether or not MA affects immune responses in a leukemia mouse model. Materials and methods WEHI-3 cells were intraperitonealIy (i.p.) injected into normal BALB/c mice to develop leukemia. Mice were then treated by i.p. injection with MA at different doses (0, 8, 16 and 32 mg/kg) for 2 weeks. After treatment, all animals were weighed and blood, liver and spleen tissues were weighed. Blood or spleen both were used for determination of cell markers or phagocytosis, natural killer (NK) cell activities and T- and B-cell proliferation, respectively, by using a flow cytometric assay. Results MA did not significantly affect body, liver, and spleen weights. However, MA increased markers of T-cells (at 16 mg/kg treatment) and monocytes (at 32 mg/kg treatment), but reduced B-cell markers (at 8 mg/kg treatment); MA did not significantly affect cell marker of macrophages. Furthermore, MA increased phagocytosis by macrophages from peripheral blood mononuclear cells and peritoneal cavity at 32 mg/kg treatment and increased NK cell activity at target cell:splenocyte ratio of 25:1 but did not affect B- and T-cell proliferation. Conclusion MA increased immune responses by enhancing macrophage phagocytosis and NK cell activities in leukemic mice.

Patients with critical illnesses face an elevated risk of medical adhesive relation skin injuries (MARSI), which have negative, subsequent impacts on recovery and quality of healthcare.The aim of this project was to decrease the incidence of MARSI in the surgical intensive care unit and to improve the accuracy of MARSI preventive care implementation.The intervention included the implementation of product-use cards, high-risk warning slogans, education programs, experience workshops, and a standard prevention-care protocol for MARSI.After project implementation, the incidence rate of MARSI decreased from 18.2% to 0%-9.3%, and the accuracy rate of preventive care increased from 38.6% to 95.5%.This project effectively reduced the incidence of skin injury and improved the quality of critical care. The skills related to the care and prevention of MARSI have been implemented throughout the hospital.外科加護病房醫療黏性相關皮膚損傷發生率之改善專案.重症病人常因使用醫療黏性產品黏貼及撕除時不注意，而造成醫療黏性相關皮膚損傷（medical adhesive-related skin injury, MARSI），其後續影響甚鉅。期望透過專案改善降低皮膚損傷發生率，增進重症照護品質。.旨在降低外科加護病房醫療黏性相關皮膚損傷發生率及提高預防照護正確率。.介入措施涵蓋設計產品圖示卡、高風險警示標語、舉辦在職教育、拍攝MARSI預防教育影片、體驗工作坊及建構預防MARSI照護標準。.2017年11月至2018年4月醫療黏性相關皮膚損傷發生率由前測18.2%下降至後測0%−9.3%，預防照護平均正確率由38.6%提升至95.5%。.專案推行後降低重症病人皮膚損傷，提升重症照護品質，MARSI照護及預防技巧也納入傷口換藥護理標準內容，並平行推展至全院，讓皮膚照護更加完善。.

Background: Abnormally high cytosolic Na + concentrations in advanced heart failure impair myocardial contractility. Stimulation of the membrane Na + -K + pump should lower Na + concentrations, and the β3 adrenoceptor (β3 AR) mediates pump stimulation in myocytes. We examined if β3 AR-selective agonists given in vivo increase myocyte Na + -K + pump activity and reverse organ congestion in severe heart failure (HF). Methods: Indices for HF were lung-, heart-, and liver: body weight ratios and ascites after circumflex coronary artery ligation in rabbits. Na + -K + pump current, I p , was measured in voltage-clamped myocytes from noninfarct myocardium. Rabbits were treated with the β3 AR agonists CL316,243 or ASP9531, starting 2 weeks after coronary ligation. Results: Coronary ligation caused ascites in most rabbits, significantly increased lung-, heart-, and liver: body weight ratios, and decreased I p relative to that for 10 sham-operated rabbits. Treatment with CL316,243 for 3 days significantly reduced lung-, heart-, and liver: body weight ratios and prevalence of ascites in 8 rabbits with HF relative to indices for 13 untreated rabbits with HF. It also increased I p significantly to levels of myocytes from sham-operated rabbits. Treatment with ASP9531 for 14 days significantly reduced indices of organ congestion in 6 rabbits with HF relative to indices of 6 untreated rabbits, and it eliminated ascites. β3 AR agonists did not significantly change heart rates or blood pressures. Conclusions: Parallel β3 AR agonists-induced reversal of Na + -K + pump inhibition and indices of congestion suggest pump inhibition is a useful target for treatment with β3 AR agonists in congestive HF.

Abnormally high cytosolic NaIndices for HF were lung-, heart-, and liver: body weight ratios and ascites after circumflex coronary artery ligation in rabbits. NaCoronary ligation caused ascites in most rabbits, significantly increased lung-, heart-, and liver: body weight ratios, and decreased IParallel β3 AR agonists-induced reversal of Na

BackgroundFXYD proteins associate closely with- and protect plasmalemmal Na+/K+-ATPase against oxidative inhibition. One of them, FXYD3, is often overexpressed in cancers, including those of breast and pancreas. Down-regulation of overexpression in MCF-7 breast cancer cells with siRNA augments doxorubicin-induced cytotoxicity. Because down-regulation with siRNA is not readily translated therapeutically, we developed a peptide as an alternative for suppression of FXYD3.MethodsA shortened peptide derivative of the wild-type (WT) FXYD3 protein, FXYD3-pep has the four cysteine residues in the WT protein replaced by serine, which eliminates the WT protein’s protection against oxidative Na+/K+-ATPase inhibition. We exposed human cancer cells to FXYD3-pep and measured cytotoxicity and caspase 3/7 activity with co-exposure to doxorubicin. We also measured effects of the peptide on expression glutathione-S-transferase π (GST-π), implicated in treatment resistance, and on expression of tumor suppressor p53. Selected experiments were performed with parallel FXYD3 suppression with siRNA or FXYD3-pep.ResultsExposure of cells to FXYD3-pep displaced WT FXYD3 from Na+/K+-ATPase. Exposure of MCF-7 breast or pancreatic BxPC-3 cancer cells that highly express FXYD3 to the peptide had little effect alone but combined with doxorubicin it significantly (P < 0.05) increased cytotoxicity. A peptide not mutated to eliminate FXYD3’s protective effect of Na+/K+-ATPase had no effect. FXYD3-pep did not augment doxorubicin’s cytotoxicity against MDA-MB-468 breast and Panc-1 pancreatic cancer cells that have low- or no FXYD3 expression. Cellular FXYD3 expressions was reflected by expression of the α1 Na+/K+-ATPase subunits but not by plasmalemmal Na+/K+-ATPase function. Signals from fluorescently labeled FXYD3-pep were detected in a perinuclear distribution in BxPC-3 cells as reported for overexpressed FXYD3, α- and β Na+/K+-ATPase subunits in cancer. Exposure to FXYD3-pep or to FXYD3 siRNA almost eliminated expression of GST-π. FXYD3-pep alone had no effect on p53 levels but significantly augmented a doxorubicin-induced increase, and, while the peptide alone had no effect on caspase 3/7 activity, it significantly augmented a doxorubicin-induced increase. ConclusionsOverexpressed FXYD3 has intracellular roles beyond its accepted modulation of plasmalemmal Na+/K+-ATPase. These roles can be countered with a membrane-permeable peptide derivative of FXYD3 that suppresses GST-π and enhances chemosensitivity of cancer cells overexpressing FXYD3.

In this study, biomimetic Mg-N

Conventional gas sensors work upon changes in mechanical or conductive properties of sensing materials during a chemical process, which may limit availabilities of size miniaturization and design simplification. However, fabrication of miniaturized sensors with superior sensitivities in real-time and label-free probing of chemical reactions or catalytic processes remains highly challenging, in particular with regard to integration of materials into a desired smaller volume without losing the recyclability of sensing properties. Here, we demonstrate a unique bimetallic nanostructure, the Au–Pd–Au core–shell–frame nanobrick, as a promising archetype for fabrication of miniaturized sensors at nanoscale. Upon analysis of the aqueous synthesis, both ex situ and in situ, the formation of Au frames is consistent with selective deposition and aggregation of NaBH4-reduced Au nanoparticles at the corners and edges of cubic Pd shells, where the {100} surfaces, capped by iodide ions, are growth-limited. By virtue of ...

We apply charged-particle simulation to determine the constraints imposed additionally on the photoemission electron microscope (PEEM) when x rays are applied to image buried objects. PEEM initiates its image acquisition by reading the electron-emission profile on the specimen surface, but the profile is not invariant if the electrons originate from a buried object. In this work, we show that, other than the anticipated intensity reduction, the electron-emission profile projected from a buried object displays a blurrier edge for a deeper burial, whereas the distribution of an emission angle becomes narrower for electrons carrying greater kinetic energy. The burial not only redefines the objects seen by PEEM but also introduces additional depth and energy dependence to the resulted images. We demonstrate that inserting a contrast aperture in the PEEM lens can improve the image quality and ease the burial dependence.

Herein, we report the synthesis of a non-precious metal dual-doped catalyst. Melamine and lipoic acid were used as precursors for the nitrogen and sulfur dopant atoms, respectively, which were mixed with iron precursors and carbon black and pyrolyzed at 700 °C to yield the catalyst Fe-M-LA/C. Fe-M-LA/C shows high oxygen reduction reaction (ORR) capability, with an almost-ideal electron transfer number of 3.99. It shows almost no degradation after potential cycling for 30 000 cycles, and the structural analysis indicates that a graphene-like structure of Fe-M-LA/C can improve its ORR activity, electrical conductivity, and corrosion resistance. X-ray photoelectron spectroscopy (XPS) results show that the catalyst has high contents of pyridinic- and quaternary N atoms and thiophene S atoms that can significantly enhance the ORR performance of Fe-M-LA/C. X-ray absorption spectroscopy (XAS) data show that the oxidation state of iron and the interatomic distance of the heteroatoms in Fe-M-LA/C play an important role in determining the ORR ability. These data confirm that the presence of a nitrogen and sulfur dual-doped –Fe–N–S– structure can enhance the ORR kinetics.

Background Raised intracellular Na+ impairs cardiac contractility in severe heart failure (HF). β3 adrenoceptor (β3 AR) agonists activate cardiomyocyte Na+-K+ pump-mediated Na+ export. Treatment with a selective β3 AR agonist in patients with severely reduced left ventricular ejection fraction (LVEF) and stable HF improves LVEF. We examined effects of β3 AR agonists on Na+-K+ pump function in experimental severe HF characterized by organ congestion and, in parallel, the clinical response in human treatment-refractory worsening chronic HF (WCHF). Methods The circumflex coronary artery was ligated in rabbits to induce HF. Treatment with β3 AR agonists was started 2 weeks later and effects on congestive signs were assessed, including ascites and the primary outcome, lung: body weight ratio. Na+-K+ pump current was measured in voltage-clamped myocytes from non-infarcted myocardium. The β3 AR agonist mirabegron was prescribed off-label to patients hospitalized with advanced HF (stage D) that was refractory to maximally tolerated guideline-directed treatment and intravenous furosemide, with or without inotropes, as assessed by ≥2 cardiologists. Results Rabbits developed severe HF with markedly increased lung:body weight ratio and ascites. Na+-K+ pump current was decreased in myocytes from rabbits with HF. β3 AR agonists eliminated pump inhibition, significantly reduced lung:body weight ratio and prevalence of ascites. Mirabegron was given to 9 patients aged 74 ± 7 years (mean ± SD) with advanced HF. LVEF was 26 ± 5%. Signs and symptoms improved within 1-2 days of treatment. Mirabegron was well-tolerated and improvement of ≥1 NYHA Class maintained by all early after discharge. One patient died from HF at 16 months, 4 died from other causes at 2 – 30 months and 4 remain alive at 30 ± 5 months with NYHA Class II symptoms. Conclusion Treatment with β3 AR agonist in a pre-clinical model of severe HF can reverse Na+-K+ pump inhibition and improve organ congestion implicating pump inhibition as a reversible myocardial dysfunction that is a useful treatment target. The changed in-hospital clinical trajectory and a more favourable post-discharge course than expected in advanced HF, suggests efficacy of β3 AR agonist shown in the animal model might translate into efficacy in advanced human HF.

Background There are limited effective drugs that can reach the brain to target brain tumors, in particular glioblastoma, which is one of the most difficult cancers to be cured from. Because the overexpression of the sigma-2 receptor is frequently reported in glioma clinical samples and associated with poor prognosis and malignancy, we herein studied the anti-tumor effect of the sigma-2 receptor agonist PB221 (4-cyclohexyl-1-[3-(5-methoxy-1,2,3,4-tetrahydronaphthalen-1-yl)propyl]piperidine) on an anaplastic astrocytoma tumor model based on previous encouraging results in pancreatic cancer and neuroblastoma SK-N-SH cells. Methods The expression of the sigma-2 receptor, transmembrane protein 97 (TMEM97), in ALTS1C1 and UN-KC6141 cell lines was measured by RT-PCR and quantitative RT-PCR. The binding of sigma-2 receptor fluorescent ligands PB385 (6-[5-[3-(4-cyclohexylpiperazin-1-yl)propyl]-5,6,7,8-tetrahydronaphthalen-5-yloxy]-N-(7-nitro-2,1,3-benzoxadiazol-4-yl)hexanamine) and NO1 (2-{6-[2-(3-(6,7-dimethoxy-3,4-dihydroisoquinolin-2(1H)-yl)propyl)-3,4-dihydroisoquinolin-1(2H)-one-5-yloxy]hexyl}-5-(dimethylamino)isoindoline-1,3-dione) was examined by flow cytometry and the fluorescent plate reader. The antitumor activity of PB221 was initially examined in the murine brain tumor cell line ALTS1C1 and then in the murine pancreatic cell line UN-KC6141. The potential therapeutic efficacy of PB221 for murine brain tumors was examined by in vitro migration and invasion assays and in vivo ectopic and orthotopic ALTS1C1 tumor models. Results: The IC50 of PB221 for ALTS1C1 and UN-KC6141 cell lines was 10.61 ± 0.96 and 13.13 ± 1.15 μM, respectively. A low dose of PB221 (1 μM) significantly repressed the migration and invasion of ALTS1C1 cells, and a high dose of PB221 (20 μM) resulted in the apoptotic cell death of ALTS1C1 cells. These effects were reduced by the lipid antioxidant α-tocopherol, but not by the hydrophilic N-acetylcysteine, suggesting mitochondrial oxidative stress is involved. The in vivo study revealed that PB221 effectively retarded tumor growth to 36% of the control tumor volume in the ectopic intramuscular tumor model and increased the overall survival time by 20% (from 26 to 31 days) in the orthotopic intracerebral tumor model. Conclusions This study demonstrates that the sigma-2 receptor agonist PB221 has the potential to be an alternative chemotherapeutic drug for brain tumors with comparable side effects as the current standard-of-care drug, temozolomide. Electronic supplementary material The online version of this article (10.1186/s12885-019-5700-7) contains supplementary material, which is available to authorized users.

To understand the effect of an iron precursor on the structure and oxygen reduction activity of an Iron–Nitrogen–Carbon (Fe–N–C) catalyst, several iron precursors, FeCl3, FeCl2, Fe(C2H3O2)2, and FeSO4, were mixed with polyaniline (PANI) and carbon black and then pyrolyzed at 700 °C under a nitrogen atmosphere; the obtained nonprecious metal catalysts were denoted as FeCl3–PANI/C, FeCl2–PANI/C, Fe(C2H3O2)2–PANI/C, and Fe2SO4–PANI/C, which were compared for the activity of oxygen reduction reaction (ORR) in acidic media and characterized through X-ray diffraction, X-ray photoelectron spectroscopy, X-ray absorption near-edge spectroscopy, and extended X-ray absorption fine structure. The ORR activity of the catalysts was ranked in the following order: FeCl3–PANI/C, FeCl2–PANI/C, Fe(C2H3O2)2–PANI/C, and Fe2SO4–PANI/C, where the electron-transfer numbers were 3.99, 3.98, 3.94, and 3.83, respectively. FeCl3–PANI/C and FeCl2–PANI/C exhibited Fe–N4 structures and Fe3+ chelates, in which a large amount of quaternary-type nitrogen and pyrrolic-type nitrogen but a small amount of cyanide were formed in the catalysts. Fe(C2H3O2)2–PANI/C showed that Fe–N4 structures and Fe2+ chelates were formed mostly in the catalyst, but showed the lowest nitrogen content among all catalysts. Fe2SO4–PANI/C was composed of FeS and Fe2O3 compounds. These results indicated that iron precursors affected the carbon–nitrogen composition, iron oxidation state of iron chelate, and chelate structure of Fe–N–C catalysts, resulting in the change of the ORR activity.

FXYD3, also known as mammary tumor protein 8, is overexpressed in several common cancers, including in many breast cancers. We examined if such overexpression might protect Na(+)/K(+)-ATPase and cancer cells against the high levels of oxidative stress characteristic of many tumors and often induced by cancer treatments. We measured FXYD3 expression, Na(+)/K(+)-ATPase activity and glutathionylation of the β1 subunit of Na(+)/K(+)-ATPase, a reversible oxidative modification that inhibits the ATPase, in MCF-7 and MDA-MB-468 cells. Expression of FXYD3 was suppressed by transfection with FXYD3 siRNA. A colorimetric end-point assay was used to estimate cell viability. Apoptosis was estimated by caspase 3/7 (DEVDase) activation using a Caspase fluorogenic substrate kit. Expression of FXYD3 in MCF-7 breast cancer cells was ~eightfold and ~twofold higher than in non-cancer MCF-10A cells and MDA-MB-468 cancer cells, respectively. A ~50 % reduction in FXYD3 expression increased glutathionylation of the β1 Na(+)/K(+)-ATPase subunit and reduced Na(+)/K(+)-ATPase activity by ~50 %, consistent with the role of FXYD3 to facilitate reversal of glutathionylation of the β1 subunit of Na(+)/K(+)-ATPase and glutathionylation-induced inhibition of Na(+)/K(+)-ATPase. Treatment of MCF-7 and MDA-MB- 468 cells with doxorubicin or γ-radiation decreased cell viability and induced apoptosis. The treatments upregulated FXYD3 expression in MCF-7 but not in MDA-MB-468 cells and suppression of FXYD3 in MCF-7 but not in MDA-MB-468 cells amplified effects of treatments on Na(+)/K(+)-ATPase activity and treatment-induced cell death and apoptosis. Overexpression of FXYD3 may be a marker of resistance to cancer treatments and a potentially important therapeutic target.

Laws that mandate reporting of child abuse have stirred global controversy. Mandatory reporting to authorities for different types of child abuse includes physical abuse, sexual abuse, emotional abuse, physical neglect, educational neglect, and emotional neglect. While the definition of child abuse differs from jurisdiction-to-jurisdiction, which covers a wide spectrum of abuse and maltreatment, employees who work in professions required to report child abuse include health care workers, educational employees, social workers, religious leaders, employees of divisions of child protective services, criminal justice and other governmental personnel, and child care professionals. Lack of a uniform definition of what constitutes child abuse causes problems for mandatory reporters, ranging from false arrest and malicious prosecution to failure to investigate ongoing child abuse. This article reviews international norms associated with mandatory reporting of child abuse and assesses court cases within the United States that involve mandatory reporting, identifying important issues and underlying problems with the current mandatory reporting system; it concludes with policy recommendations and suggestions for future research.

Protein kinase C can activate NADPH oxidase and induce glutathionylation of the β1-Na+-K+pump subunit, inhibiting activity of the catalytic α-subunit. To examine if signaling of nitric oxide-induced soluble guanylyl cyclase (sGC)/cGMP/protein kinase G can cause Na+-K+pump stimulation by counteracting PKC/NADPH oxidase-dependent inhibition, cardiac myocytes were exposed to ANG II to activate NADPH oxidase and inhibit Na+-K+pump current ( Ip). Coexposure to 3-(5′-hydroxymethyl-2′-furyl)-1-benzylindazole (YC-1) to stimulate sGC prevented the decrease of Ip. Prevention of the decrease was abolished by inhibition of protein phosphatases (PP) 2A but not by inhibition of PP1, and it was reproduced by an activator of PP2A. Consistent with a reciprocal relationship between β1-Na+-K+pump subunit glutathionylation and pump activity, YC-1 decreased ANG II-induced β1-subunit glutathionylation. The decrease induced by YC-1 was abolished by a PP2A inhibitor. YC-1 decreased phosphorylation of the cytosolic p47phoxNADPH oxidase subunit and its coimmunoprecipitation with the membranous p22phoxsubunit, and it decreased O2·−-sensitive dihydroethidium fluorescence of myocytes. Addition of recombinant PP2A to myocyte lysate decreased phosphorylation of p47phoxindicating the subunit could be a substrate for PP2A. The effects of YC-1 to decrease coimmunoprecipitation of p22phoxand p47phoxNADPH oxidase subunits and decrease β1-Na+-K+pump subunit glutathionylation were reproduced by activation of nitric oxide-dependent receptor signaling. We conclude that sGC activation in cardiac myocytes causes a PP2A-dependent decrease in NADPH oxidase activity and a decrease in β1pump subunit glutathionylation. This could account for pump stimulation with neurohormonal oxidative stress expected in vivo.

This paper presents an adaptive equalizer used for 1.5-Gb/s phase-locked loop (PLL)-based point-to-point (P2P) high-speed interface with periodically embedded clock encoding (PECE). The adaptive equalizer uses a spectrum-balancing technique to detect the loss of energy without using the regulating comparator. The power detector combines current steering circuits and a preamplifier circuit to enhance the voltage swing. Implemented by 0.18-um CMOS process, this equalizer consumes 14.9 mW (excluding the output buffers) at 1.5 Gb/s with an output swing of 400 mV (p-p). The core area occupied is 0.05 mm2 (including output buffers), and the measured output peak-to-peak jitter is below 0.2 UI. The equalizer achieves high-frequency compensation, small area, and low power consumption.

Rationale: Cardiotonic steroids (CTS), such as digoxin, have been used to treat heart failure (HF) for over 200 years. They inhibit the sodium-potassium pump (NKP), and increase cardiac contractility by inhibiting efflux of sodium through the pump (“digitalis hypothesis”). CTS possess three structural components: a saturated/unsaturated lactone ring, steroid core, and sugar moiety, each of which may be involved in NKP inhibition/stimulation. It is now known that inhibition of the NKP in patients with HF increases mortality, and all major beneficial treatments increase its activity. Endogenous circulating CTS such as ouabain are generally thought to inhibit the NKP, despite studies sporadically reporting ouabain-induced pump stimulation. This study aims to identify whether ouabain-induced pump stimulation occurs, and if so, which structural components are involved in causing pump stimulation. Methods & Results: Cardiac myocytes were isolated from male New Zealand White rabbits, placed in a Tyrode’s solution, and whole-cell patch clamped. They were exposed to 0-30nM ouabain, 0-50nM dihydroouabain (ouabain with a saturated lactone ring) or 0-500nM ouabagenin (ouabain lacking a sugar moiety) for 1 min, followed by a potassium-free solution, with the difference in current yielding the NKP current. Compared to the 0.47±0.05 pA/pF Tyrode’s solution control (n=11), 5nM ouabain significantly increased NKP current to 0.69±0.09 pA/pF ( P P Conclusion: Low-dose ouabain uniquely stimulates NKP activity. Low-dose dihydroouabain and ouabagenin do not, suggesting that a sugar moiety and unsaturated lactone ring are required for pump stimulation. Ouabain in its unaltered form may be a potential treatment for HF.

By the time it was appreciated that the positive inotropic effect of cardiac glycosides is due to inhibition of the membrane Na+-K+ pump, glycosides had been used for treatment of heart failure on an empiric basis for ~200 years. The subsequent documentation of their lack of clinical efficacy and possible harmful effect largely coincided with the discovery that a raised Na+ concentration in cardiac myocytes plays an important role in the electromechanical phenotype of heart failure syndromes. Consistent with this, efficacious pharmacological treatments for heart failure have been found to stumulate the Na+-K+ pump, effectively the only export route for intracellular Na+ in heart failure. A paradigm has emerged that implicates pump inhibition in the raised Na+ levels in heart failure. It invokes protein kinase-dependent activation of nicotinamide adenine dinucleotide phosphate oxidase (NADPH oxidase) and glutathionylation, a reversible oxidative modification, of the Na+-K+ pump molecular complex that inhibits its activity. Since treatments of proven efficacy reverse the oxidative Na+-K+ pump inhibition, the pump retains its status as a key pharmacological target in heart failure. Its role as a target is well integrated with the paradigms of neurohormonal abnormalities, raised myocardial oxidative stress and energy deficiency implicated in the pathophysiology of the failing heart. We propose that targeting oxidative inhibition of the pump is useful for the exploration of future treatment strategies. This article is part of a Special Issue entitled "Na+ Regulation in Cardiac Myocytes". Australian Research Council, National Health and Medical Research Council (Australia), Heart Research Australia, National Heart Foundation of Australia, Medical Foundation (University of Sydney), Viertel Charitable Foundation

Key points • The widely believed effects of β1 adrenergic receptors and protein kinase A (PKA) to stimulate the membrane Na+–K+ pump in cardiac myocytes are not easily reconciled with the effects of activation of the receptor and PKA on contractility in normal heart or with the benefit of β1 adrenergic blockade in heart failure. • We show that reduction in PKA activity by β1 adrenergic blockade in vivo stimulates the Na+–K+ pump by reducing glutathionylation of one of its subunits, a reversible oxidative modification that inhibits pump activity. • Na+–K+ pump stimulation induced by β1 adrenergic blockade in vivo is reversed by activation of PKA-dependent signalling in isolated cardiac myocytes studied ex vivo. • Inhibition of the myocyte Na+–K+ pump mediated by PKA- and redox-dependent signalling pathways and downstream glutathionylation of a subunit of the Na+–K+ pump is readily reconciled with effects of PKA on contractility in normal heart and efficacy of β1 adrenergic blockade in heart failure. Abstract The widely reported stimulation of the cardiac Na+–K+ pump by protein kinase A (PKA) should oppose other effects of PKA to increase contractility of the normal heart. It should also reduce harmful raised myocyte Na+ levels in heart failure, yet blockade of the β1 adrenergic receptor (AR), coupled to PKA signalling, is beneficial. We treated rabbits with the β1 AR antagonist metoprolol to modulate PKA activity and studied cardiac myocytes ex vivo. Metoprolol increased electrogenic pump current (Ip) in voltage clamped myocytes and reduced glutathionylation of the β1 pump subunit, an oxidative modification causally related to pump inhibition. Activation of adenylyl cyclase with forskolin to enhance cAMP synthesis or inclusion of the catalytic subunit of PKA in patch pipette solutions abolished the increase in Ip in voltage clamped myocytes induced by treatment with metoprolol, supporting cAMP/PKA-mediated pump inhibition. Metoprolol reduced myocardial PKA and protein kinase C (PKC) activities, reduced coimmunoprecipitation of cytosolic p47phox and membranous p22phox NADPH oxidase subunits and reduced myocardial O2•−-sensitive dihydroethidium fluorescence. Treatment also enhanced coimmunoprecipitation of the β1 pump subunit with glutaredoxin 1 that catalyses de-glutathionylation. Since angiotensin II induces PKC-dependent activation of NADPH oxidase, we examined the effects of angiotensin-converting enzyme inhibition with captopril. This treatment had no effect on PKA activity but reduced the activity of PKC, reduced β1 subunit glutathionylation and increased Ip. The PKA-induced Na+–K+ pump inhibition we report should act with other mechanisms that enhance contractility of the normal heart but accentuate the harmful effects of raised cytosolic Na+ in the failing heart. This scheme is consistent with the efficacy of β1 AR blockade in the treatment of heart failure.