Miracle-dz/UniFAR-FTUs huggingface dataset:
Control of rectification and permeation by two distinct sites after the second transmembrane region in Kir2.1 K+ channel

1 The rectification property of the inward rectifier K+ channel is chiefly due to the block of outward current by cytoplasmic Mg2+ and polyamines. In the cloned inward rectifier K+ channel Kir2.1 (IRK1), Asp172 in the second transmembrane region (M2) and Glu224 in the putative cytoplasmic region after M2 are reported to be critical for the sensitivity to these blockers. However, the difference in the inward rectification properties between Kir2.1 and a very weak inward rectifier sWIRK could not be explained by differences at these two sites. 2 Following sequence comparison of Kir2.1 and sWIRK, we focused this study on Glu299 located in the centre of the putative cytoplasmic region after M2. Single‐point mutants of Kir2.1 (Glu224Gly and Glu299Ser) and a double‐point mutant (Glu224Gly‐Glu299Ser) were made and expressed in Xenopus oocytes or in HEK293T cells. 3 Their electrophysiological properties were compared with those of wild‐type (WT) Kir2.1 and the following observations were made. (a) Glu299Ser showed a weaker inward rectification, a slower activation upon hyperpolarization, a slower decay of the outward current upon depolarization, a lower sensitivity to block by cytoplasmic spermine and a smaller single‐channel conductance than WT. (b) The features of Glu224Gly were similar to those of Glu299Ser. (c) In the double mutant (Glu224Gly‐Glu299Ser), the differences from WT described above were more prominent. 4 These results demonstrate that Glu299 as well as Glu224 control rectification and permeation, and suggest the possibility that the two sites contribute to the inner vestibule of the channel pore. The slowing down of the on‐ and off‐blocking processes by mutation of these sites implies that Glu224 and Glu299 function to facilitate the entry (and exit) of spermine to (and from) the blocking site.
Reconciling Entrepreneurial Orientation and Dynamic Capabilities: A Strategic Entrepreneurship Perspective

Strategic entrepreneurship (SE) emphasises the complementary roles played by entrepreneurship and strategic management in promoting firm growth. This article adopts two dominant concepts from each field—entrepreneurial orientation (EO) and dynamic capabilities (DCs)—to investigate their interaction effects on firm performance (FP). It further examines three contingencies—firm, market and product innovation—that significantly affect the levels of EO and DCs that firms pursue. This study analyses the influence of EO and DCs on performance using hierarchical regression models. Interaction effects of EO and DCs on FP demonstrate a positive relationship. This study found that DCs are more critical for incumbent firms than for small firms. Both EO and DCs enhance performance in dynamic markets. The EO increases performance under radical product innovation, while DCs show no effects. This study provides important and unique implications for the complementary roles of entrepreneurship and strategic management.
Macroeconomic Conditions at Entry and Injury Risk in the Workplace

Using a unique dataset from Italy, we show that the local unemployment rate at entry has a persistent positive effect on severe and non‐severe workplace injuries of young workers. Entrants during recessions, despite receiving marginally higher entry wages, also experience slower wage growth. The observed pattern in the differences between severe and non‐severe injuries indicates that entrants during recessions might under‐report non‐severe workplace injuries. Our findings suggest that workers entering during recessions are persistently locked into low‐quality jobs and that the mix of hazardous tasks endogenously adjusts to the business cycle.
Coping with functional collective action dilemma: functional fragmentation and administrative integration

ABSTRACT Functional fragmentation among city departments has been a critical public administration problem in practice and theory. This paper investigates the political and administrative motivations for the reform from functionally fragmented water governance model towards an integrative agency design. Drawing on the Institutional Collective Action (ICA) framework and the literature on bureaucratic structures, we hypothesize that cities’ decisions for administrative reform are influenced by different types of transactions costs incurred in the negotiation process among local agencies. Empirical results of this study provide strong support for the transaction cost hypotheses on the roles of bargaining cost, enforcement cost, and information cost.
Embryonic Stem Cells Exhibit mRNA Isoform Specific Translational Regulation

The presence of multiple variants for many mRNAs is a major contributor to protein diversity. The processing of these variants is tightly controlled in a cell-type specific manner and has a significant impact on gene expression control. Here we investigate the differential translation rates of individual mRNA variants in embryonic stem cells (ESCs) and in ESC derived Neural Precursor Cells (NPCs) using polysome profiling coupled to RNA sequencing. We show that there are a significant number of detectable mRNA variants in ESCs and NPCs and that many of them show variant specific translation rates. This is correlated with differences in the UTRs of the variants with the 5’UTR playing a predominant role. We suggest that mRNA variants that contain alternate UTRs are under different post-transcriptional controls. This is likely due to the presence or absence of miRNA and protein binding sites that regulate translation rate. This highlights the importance of addressing translation rate when using mRNA levels as a read out of protein abundance. Additional analysis shows that many annotated non-coding mRNAs are present on the polysome fractions in ESCs and NPCs. We believe that the use of polysome fractionation coupled to RNA sequencing is a useful method for analysis of the translation state of many different RNAs in the cell.
Muscle-specific Knock-out of NUAK Family SNF1-like Kinase 1 (NUAK1) Prevents High Fat Diet-induced Glucose Intolerance*

Background: The physiological roles of NUAK1 are poorly understood because of embryonic lethality in NUAK1 null mice. Results: Negative feedback regulation of insulin signaling was abrogated in skeletal muscle of muscle-specific NUAK1 knock-out mice. Conclusion: NUAK1 controls glucose metabolism through negative regulation of insulin signal transduction in skeletal muscle. Significance: This is the first report of a physiological role of NUAK1 in adult tissues. NUAK1 is a member of the AMP-activated protein kinase-related kinase family. Recent studies have shown that NUAK1 is involved in cellular senescence and motility in epithelial cells and fibroblasts. However, the physiological roles of NUAK1 are poorly understood because of embryonic lethality in NUAK1 null mice. The purpose of this study was to elucidate the roles of NUAK1 in adult tissues. We determined the tissue distribution of NUAK1 and generated muscle-specific NUAK1 knock-out (MNUAK1KO) mice. For phenotypic analysis, whole body glucose homeostasis and muscle glucose metabolism were examined. Quantitative phosphoproteome analysis of soleus muscle was performed to understand the molecular mechanisms underlying the knock-out phenotype. Nuak1 mRNA was preferentially expressed in highly oxidative tissues such as brain, heart, and soleus muscle. On a high fat diet, MNUAK1KO mice had a lower fasting blood glucose level, greater glucose tolerance, higher insulin sensitivity, and higher concentration of muscle glycogen than control mice. Phosphoproteome analysis revealed that phosphorylation of IRS1 Ser-1097 was markedly decreased in NUAK1-deficient muscle. Consistent with this, insulin signaling was enhanced in the soleus muscle of MNUAK1KO mice, as evidenced by increased phosphorylation of IRS1 Tyr-608, AKT Thr-308, and TBC1D4 Thr-649. These observations suggest that a physiological role of NUAK1 is to suppress glucose uptake through negative regulation of insulin signaling in oxidative muscle.
Identification of a Novel Element Involved in Regulation of the Lytic Switch BZLF1 Gene Promoter of Epstein-Barr Virus

ABSTRACT Epstein-Barr virus (EBV) is a human herpesvirus capable of establishing a latent state in B lymphocytes. EBV's BZLF1 gene product plays a central role in regulating the switch from latency to productive infection. Here, we identify a sequence element, 5′-CAGGTA-3′, called ZV, located at nucleotides −17 to −12 relative to the transcription initiation site of the BZLF1 promoter. ZV sequence-specifically binds a cellular nuclear factor(s), ZVR. ZVR DNA-binding activity was present in the EBV-negative B-lymphocytic cell line DG75, the EBV-positive B-lymphocytic cell lines GG68 and 721, the cervical cell line C33A, and the kidney cell line CV-1 but not in the breast carcinoma cell line MCF-7. Mutations in ZV that relieve binding of ZVR lead to a two- to fourfold increase in basal expression of the BZLF1 promoter in DG75, C33A, and CV-1 cells. The same mutants exhibited a 40- to 180-fold increase in tetradecanoyl phorbol acetate-ionomycin-induced expression in DG75 cells and a 22-fold increase in C33A cells. Thus, ZVR functions as a regulator of the BZLF1 promoter, repressing transcription when bound to the ZV site in the absence of inducers. No differences in basal or induced transcription between wild-type and ZV mutant BZLF1 promoters were observed in ZVR-negative MCF-7 cells. ZVR failed to bind any of the previously identified negative regulatory elements within the BZLF1 promoter. We conclude that ZV functions as an important regulatory element of the BZLF1 promoter, with ZVR likely playing important roles in the maintenance of latency and reactivation of EBV.
Risk factors for adverse events related to polypectomy in the English Bowel Cancer Screening Programme

Background and study aims: The English National Health Service Bowel Cancer Screening Programme (NHSBCSP) is one of the world’s largest organized screening programs. Minimizing adverse events is essential for any screening program. Study aims were to determine rates and to examine risk factors for adverse events. Patients and methods: Bleeding and perforations in NHSBCSP colonoscopies between August 2006 and January 2012 were examined. Logistic regression was used to examine risk factors for adverse events, including age, gender, polyp size, morphology, and location. For accurate attribution of adverse events, procedures with resection of only one polyp (“single-polypectomy”) were analyzed in detail. Results: 130 831 colonoscopies (167 208 polypectomies) were analyzed, including 30 881 single-polypectomies. Overall bleeding rate was 0.65 %, rate of bleeding requiring transfusion was 0.04 % and perforation rate was 0.06 %. Polypectomy increased bleeding risk 11.14-fold and perforation risk 2.97-fold. Cecal location (but not elsewhere in the proximal colon) and increasing polyp size were the two most important risk factors for bleeding and perforation. After adjustment for polyp size, the odds ratio (OR) relative to the distal colon for bleeding requiring transfusion after cecal snare polypectomy was 13.5 (95 %CI 3.9 – 46.4) and for perforation after cecal nonpedunculated polypectomy it was 12.2 (95 %CI 1.2 – 119.5). Conclusion: This is the largest study focusing on polyp-specific risk factors. We have confirmed that the greatest risk factor for both post-polypectomy bleeding and perforation is polyp size. This is the first demonstration of substantial and significantly increased risk for both noteworthy bleeding (requiring transfusion) and perforation from cecal polypectomy for a given polyp size, compared with elsewhere in the colon.
The Influence of Estradiol and Progesterone and Melatonin Supplementation on TNF-α Levels in Ovariectomized and Pinealectomized Rats

The aim of this study was to investigate the effect of estradiol and progesterone and melatonin supplementation on TNF-α levels in ovariectomized and pinealectomized rats. The study was carried out on 42 adult, Spraque-Dawley strain female rats aged 6 months and weighing 200–250 grams. The rats were divided into 6 groups, each group contained 7 rats. Group 1: Sham-ovariectomized (Sham-Ovx), Group 2: Ovariectomized (Ovx), Group 3: Ovx and estradiol (E) and progesterone (P) supplemented (Ovx+E-P) group, Group 4: Ovx+E-P+Melatonin (M) supplemented group, Group 5: Ovx — Pinealectomized (Pnx) group, Group 6: Ovx - Pnx+E-P supplemented group. Serum TNF-α levels were determined after 4 weeks application period. Group 6 (Ovx-Pnx + E - P) has the highest serum TNF-α compared with other groups while group 2 (ovariectomized), has the lowest levels (P<0.001). Group 5 was higher than groups 1, 2, 3 and 4 (P<0.001). The results of the study show that ovariectomy reduces the serum level of TNF-α, but estradiol and progesterone application prevents this reduction in ovariectomized rats. However, pinealectomy intensifies the increases in TNF-α levels in ovariectomized and estradiol and progesterone supplemented rats, whereas melatonin reduces TNF-α levels in ovariectomized rats.
Liver Transplantation in Patients with Cryptogenic Cirrhosis Provides Excellent Long-Term Outcome.

BACKGROUND Cryptogenic cirrhosis (CC) is an indication for liver transplantation in 5-9% of recipients. Diagnosis is made when other diagnostic possibilities have been ruled out. The aim of this study was to present long-term outcomes of liver transplantation for CC. MATERIAL AND METHODS There have been 1367 liver transplantations performed during the years 1994-2013 in the Department of General, Transplant and Liver Surgery at the Medical University of Warsaw. This retrospective study comprised 55 patients after liver transplantation for CC (4.0%). Perioperative mortality (30 days) and patient and graft 1-, 5-, and 10-year survival rates were set as outcome measures. RESULTS Peri-operative mortality reached 10.9% (6 of 55). The 1-, 5- and 10-year patient and graft survival rates were 85.2%, 78.8%, and 73.9%, respectively, and 83.3%, 74.5%, and 74.5%, respectively. In univariate analyses, the following parameters significantly influenced patient survival: pre-operative aspartate (AST; p=0.013) and alanine (ALT; p=0.043) aminotransferases activity, INR (p=0.040), bilirubin concentration (p=0.045), and donor age (p=0.033). Similarly, graft survival was significantly associated with AST (p=0.013), ALT (p=0.043), bilirubin concentration (p=0.044), INR (p=0.038), and recipient sex (p=0.049). In multivariable analyses, a tendency towards worse patient and graft survival was observed in patients with higher pre-operative AST (p=0.078 for patient survival and p=0.063 for graft survival). Analyses of the pathological reports indicated that underlying immunological processes were the most probable cause of liver damage in 16 of 51 patients (31.4%). CONCLUSIONS Long-term outcomes of liver transplantation in patients with cryptogenic liver cirrhosis are encouraging. Analysis of the clinical course, biochemical parameters, and factors influencing outcomes suggest an underlying autoimmunological etiology of cirrhosis in this population of patients.
In Situ Flow Activates Endothelial Nitric Oxide Synthase in Luminal Caveolae of Endothelium with Rapid Caveolin Dissociation and Calmodulin Association*

Acute changes in pressure or shear stress induce the rapid release of nitric oxide (NO) from the vascular endothelium resulting in vasodilation. Endothelial nitric oxide synthase (eNOS) regulates this flow-induced NO secretion. The subcellular location of flow-induced eNOS activity in the endothelium in vivo as well as the mechanisms by which hemodynamic forces regulate eNOS activity are unknown. The luminal cell surface of the endothelium, which is directly exposed to circulating blood stressors, has been examined for eNOS expression and functional activity. Immunoelectron microscopy of rat lung tissue shows eNOS labeling on the endothelial cell surface primarily within caveolae. Subcellular fractionation to purify luminal endothelial cell plasma membranes and their caveolae directly from rat lungs reveals that eNOS is not only concentrated but also enzymatically active in caveolae. Increasing vascular flow and pressure in situ rapidly activates caveolar eNOS with apparent eNOS dissociation from caveolin and association with calmodulin. Hemodynamic forces resulting from increased flow appear to transmit through caveolae to release eNOS from its inhibitory association with caveolin, apparently to allow more complete activation by calmodulin and other possible effectors. These data demonstrate a physiological relevant mechanotransduction event directly in caveolae at the luminal endothelial cell surface. Caveolae may serve as flow-sensing organelles with the necessary molecular machinery to transduce rapidly, mechanical stimuli and thereby regulate eNOS activity.
Tumour multifocality and grade predict intravesical recurrence after nephroureterectomy in patients with upper urinary tract urothelial carcinoma without a history of bladder cancer.

OBJECTIVE Patients with upper urinary tract urothelial carcinoma (UUT-UC) without a history of bladder cancer have a different natural history of intravesical recurrence after nephroureterectomy compared with those with a history of bladder cancer. The aim of this study was to identify predictive factors for post-operative intravesical recurrence in patients with non-metastatic upper urinary tract-localized urothelial carcinoma without a history of bladder cancer and who were not taking medication during the perioperative period. METHODS This retrospective study included 133 patients who were treated between 1995 and 2012. Univariate and multivariate analyses were used to evaluate the clinical and pathological factors associated with the cumulative incidence of bladder cancer. RESULTS Of the 133 patients, 51 (38.3%) developed intravesical recurrence during a median follow-up of 71 months (range, 0.8-210.8). In the multivariate analysis, multifocality (P = 0.03) and high tumour grade (P = 0.007) were significantly associated with the cumulative incidence of bladder cancer. We constructed a prediction classification model on the basis of the total number of risk factors. The 2-year cumulative incidence rates were 5.6, 34.8 and 50.0% in individuals with no, one and two risk factors, respectively. There was a significant difference between patients with no risk factors and those with two risk factors (P = 0.01). CONCLUSIONS Although this retrospective study had several limitations, tumour multifocality and tumour grade were found to be potential risk factors for intravesical recurrence in our cases.
Comparative study of the plasma pharmacokinetics and tissue concentrations of danofloxacin and enrofloxacin in broiler chickens.

The plasma pharmacokinetics of danofloxacin and enrofloxacin in broiler chickens was investigated following single intravenous (i.v.) or oral administration (p.o.) and the steady-state plasma and tissue concentrations of both drugs were investigated after continuous administration via the drinking water. The following dosages approved for the treatment of chickens were used: danofloxacin 5 mg/kg and enrofloxacin 10 mg/kg of body weight. Concentrations of danofloxacin and enrofloxacin including its metabolite ciprofloxacin were determined in plasma and eight tissues by specific and sensitive high performance liquid chromatography methods. Pharmacokinetic parameter values for both application routes calculated by noncompartmental methods were similar for danofloxacin compared to enrofloxacin with respect to elimination half-life (t1/2: approximately 6-7 h), mean residence time (MRT; 6-9 h) and mean absorption time (MAT; 1.44 vs. 1.20 h). However, values were twofold higher for body clearance (ClB; 24 vs. 10 mL/min. kg) and volume of distribution at steady state (VdSS; 10 vs. 4 L/kg). Maximum plasma concentration (Cmax) after oral administration was 0.5 and 1.9 micrograms/mL for danofloxacin and enrofloxacin, respectively, occurring at 1.5 h for both drugs. Bioavailability (F) was high: 99% for danofloxacin and 89% for enrofloxacin. Steady-state plasma concentrations (mean +/- SD) following administration via the drinking water were fourfold higher for enrofloxacin (0.52 +/- 0.16 microgram/mL) compared to danofloxacin (0.12 +/- 0.01 microgram/mL). The steady-state AUC0-24 h values of 12.48 and 2.88 micrograms.h/mL, respectively, derived from these plasma concentrations are comparable with corresponding area under the plasma concentration-time curve (AUC) values after single oral administration. For both drugs, tissue concentrations markedly exceeded plasma concentrations, e.g. in the target lung, tissue concentrations of 0.31 +/- 0.07 microgram/g for danofloxacin and 0.88 +/- 0.24 microgram/g for enrofloxacin were detected. Taking into account the similar in vitro activity of danofloxacin and enrofloxacin against important pathogens in chickens, a higher therapeutic efficacy of water medication for enrofloxacin compared to danofloxacin can be expected when given at the approved dosages.
Role of IL-2 in cancer immunotherapy

ABSTRACT Interleukin-2 (IL-2) is one of the key cytokines with pleiotropic effects on immune system. It has been approved for the treatment of metastatic renal cell carcinoma and metastatic melanoma. Recent progress has been made in our understanding of IL-2 in regulating lymphocytes that has led to exciting new directions for cancer immunotherapy. While improved IL-2 formulations might be used as monotherapies, their combination with other anticancer immunotherapies, such as adoptive cell transfer regimens, antigen-specific vaccination, and blockade of immune checkpoint inhibitory molecules, for example cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) and programmed death 1 (PD-1) mono-antibodies, would held the promise of treating metastatic cancer. Despite the comprehensive studies of IL-2 on immune system have established the application of IL-2 for cancer immunotherapy, a number of poignant obstacles remain for future research. In the present review, we will focus on the key biological features of IL-2, current applications, limitations, and future directions of IL-2 in cancer immunotherapy.
On-Chip Antenna Integration for Millimeter-Wave Single-Chip FMCW Radar, Providing High Efficiency and Isolation

A complete functional 60-GHz BiCMOS single-chip millimeter-wave frequency-modulated continuous wave (FMCW) radar with on-chip integrated antennas is designed, realized, and experimentally validated. The chip is configured with one transmitting and two receiving antennas. A high antenna efficiency and isolation between the transmitting and receiving antennas is achieved by using a cavity-backed on-chip monopole and by optimizing the package environment. Surface-wave losses and back radiation are reduced, and a high antenna gain is obtained with small ripples in the antenna pattern by implementing special structures on the PCB of the integrated circuit package. The experimental results from the single-chip radar show that the on-chip antennas provide a gain of 2 dBi with very low back radiation. The realized antennas achieve an impedance bandwidth of more than 50% with a corresponding simulated efficiency of 45%. In addition, the proposed approach suppresses surface waves and meets the specific FMCW radar requirements, such as a more than 25-dB isolation between the transmitting and receiving antennas.