Miracle-dz/UniFAR-FTUs huggingface dataset:
Calnuc Function in Endosomal Sorting of Lysosomal Receptors

Calnuc is a ubiquitous Ca2+‐binding protein present on the trans‐Golgi network (TGN) and endosomes. However, the precise role of Calnuc in these organelles is poorly characterized. We previously highlighted the role of Calnuc in the transport of LRP9, a new member of a low‐density lipoprotein (LDL) receptor subfamily that cycles between the TGN and endosomes. The objective of this study was to explore the role of Calnuc in the endocytic sorting of mannose‐6‐phosphate receptor (MPR) and Sortilin, two well‐characterized lysosomal receptors that transit between the TGN and endosomes. Using biochemical and microscopy assays, we showed that Calnuc depletion [by small interfering RNA (siRNA)] causes the misdelivery to and degradation in lysosomes of cationic‐independent mannose‐6‐phosphate receptor (CI‐MPR) and Sortilin due to a defect in the endosomal recruitment of retromers, which are key components of the endosome‐to‐Golgi retrieval machinery. Indeed, we demonstrated that Calnuc depletion impairs the activation and membrane association of Rab7, a small G protein required for the endosomal recruitment of retromers. Overall, our data indicate a novel role for Calnuc in the endosome‐to‐TGN retrograde transport of lysosomal receptors through the regulation of Rab7 activity and the recruitment of retromers to endosomes.
Characterization of sPARP-1

Poly(ADP-ribose) polymerase-1 (PARP-1) is an abundant nuclear enzyme that catalyzes the synthesis of poly(ADP-ribose) (pADPr) from its substrate NAD+ upon binding to DNA strand breaks. Poly(ADP-ribosyl)ation has been implicated in many cellular processes including replication, transcription, and the maintenance of genomic stability. However, studies with mice and cells lacking PARP-1 reveal a critical role for the enzyme in the maintenance of genomic integrity only. Recently, a significant level of poly(ADP-ribose) polymerase activity has been detected in fibroblasts derived from mice lacking PARP-1 following treatment with genotoxic agents (Shieh, W. M., Amé, J-C., Wilson, M. V., Wang, Z-Q., Koh, D. W., Jacobson, M. K., and Jacobson, E. L. (1998) J. Biol. Chem. 273, 30069–30072). We have isolated a cDNA that originates from PARP-1 (−/−) fibroblasts and encodes a polypeptide of 493 amino acid residues bearing poly(ADP-ribose) polymerase activity. This protein, that we named sPARP-1 for shortpoly(ADP-ribose)polymerase-1, has a calculated mass of 55.3 kDa and is identical in deduced amino acid sequence to the catalytic domain of PARP-1. Radiation hybrid analysis assigned the sPARP-1 gene to the chromosome 1H5-H6 in an immediate proximity to the known location of PARP-1 gene, indicating that sPARP-1 and PARP-1 are most probably products of the same gene. Active sPARP-1 is present in both PARP-1 (+/+) and PARP-1 (−/−) cells as demonstrated by activity-Western blotting and immunostaining using a specific antibody developed against sPARP-1. Like PARP-1, sPARP-1 is localized in the cell nucleus, uses NAD+ as a substrate and is inhibited by nicotinamide analogues. sPARP-1 produces pADPr of similar length and structure to that of PARP-1. However, contrary to PARP-1, sPARP-1 does not require DNA strand breaks for its activation, although it is stimulated following genotoxic treatments.
Lightning-Related Fatalities in Romania from 1999 to 2015

AbstractLightning-related fatalities in Romania are analyzed and presented for the first time using data from the Romanian National Institute of Statistics. The database contains 724 lightning fatalities that occurred between 1999 and 2015 in Romania, corresponding to an average of 42.6 fatalities per year. The annual number of lightning fatalities decreased from 65 fatalities per year between 1999 and 2003 to 23.2 fatalities per year between 2011 and 2015. The majority of fatalities occurred in May–August (42% of all fatalities) with a peak in June (31%) and July (28%). The highest fatality rates (>2.6 fatalities per million inhabitants per year) are observed over southwestern Romania, a region characterized by high values of cloud-to-ground lightning density (>2 flashes per square kilometer per year) and by a relatively high percentage (>40%) of the population living in rural areas. The majority of fatalities (78%) were reported in rural areas. Approximately 78% of the victims were male. The most vulner.
Advanced heart failure and nocturnal hypoxaemia due to central sleep apnoea are associated with increased serum erythropoietin

Central sleep apnoea (CSA) and increased serum erythropoietin (EPO) concentration have each been associated with adverse prognosis in heart failure (HF) patients. The aim of this study was to examine the relationship between nocturnal hypoxaemia due to CSA and the serum EPO concentration in patients with HF.
Neural stem cell transplantation and melatonin treatment in a 6‐hydroxydopamine model of Parkinson’s disease

Abstract: Melatonin has multiple roles including neuroprotection. Melatonin signaling involves diverse targets including two G‐protein‐coupled receptors, MT1 and MT2, which have both been localized to the nigrostriatal pathway. Previous studies in our laboratory demonstrated preservation of tyrosine hydroxylase immunoreactivity, following chronic treatment with a physiological dose of melatonin, in the 6‐hydroxydopamine rat model of Parkinson’s disease. Additionally, we reported the presence of the melatonin MT1 receptor subtype in cultured C17.2 neural stem cells (NSCs). In the present study, we examined the effects of C17.2 NSC transplantation on dopaminergic denervation following 6‐hydroxydopamine lesioning in the rat striatum. Moreover, based on our detection of the MT1 in these cells, we examined the effects of combined C17.2 NSC transplantation and melatonin treatment, following striatal lesioning. Behavioral studies indicated a marked inhibition of apomorphine‐induced rotations in lesioned animals that received C17.2 NSC transplantation, melatonin, or the combined regimen. In addition, these treatments resulted in a significant protection of tyrosine hydroxylase immunoreactivity in the striatum and substantia nigra of lesioned animals, when compared with untreated controls. Lesioned animals treated with C17.2 NSCs, melatonin or a combination of both agents exhibited no significant differences in the number of tyrosine hydroxylase‐positive cells in the substantia nigra or ventral tegmental area ipsilateral or contralateral to the lesioned striatum. These findings suggest that stem cell therapy and concomitant use of neuroprotective agents such as melatonin could be a viable approach in Parkinson’s disease.
Ellagic Acid and Embelin Affect Key Cellular Components of Pancreatic Adenocarcinoma, Cancer, and Stellate Cells

Ellagic acid is a polyphenolic phytochemical present in many fruits and nuts with anticancer properties demonstrated in experimental tumor studies. Embelin is a benzoquinone phytochemical isolated from the Japanese herb Ardisiae Japonicae and has been shown to induce apoptosis in cancer cells. We found that ellagic acid and embelin each dose-dependently increased apoptosis and inhibited proliferation in human pancreatic cancer cells, MIA PaCa-2 and HPAF-II cells, and in pancreatic stellate cells, which are progenitors of pancreatic cancer desmoplasia. In each of these cell types, combinations of ellagic acid and embelin at low micromolar concentrations (0.5–3 μM) induced synergistic increases in apoptosis and decreases in proliferation. Ellagic acid decreased NF-κB transcriptional activity, whereas embelin decreased STAT-3 phosphorylation and protein expression of its downstream target survivin in cancer cells. In vivo dietary ellagic acid alone or in combination with embelin decreased tumor size and tumor cellularity in a subcutaneous xenograft mouse model of pancreatic cancer. These results show that ellagic acid and embelin interact with divergent intracellular signaling pathways resulting in augmentation of apoptosis and inhibition of proliferation at low micromolar concentrations for the key cellular components of pancreatic adenocarcinoma.
Altering the Regioselectivity of Cytochrome P450 CYP102A3 of Bacillus subtilis by Using a New Versatile Assay System

A novel monooxygenase (CYP102A3) has been discovered within the Bacillus subtilis genome that reveals a similarity of 76 % to the well‐known cytochrome P450 BM‐3 of B. megaterium (CYP102A1). Both enzymes are natural fusion proteins consisting of a heme domain and a FAD/FMN‐reductase domain. Because of their high turnover rates, these biocatalysts are of special interest for industrial applications, but show only limited regioselectivity. In this work, the regioselectivity of CYP102A3 was changed by directed evolution and protein design to hydroxylate substrates not only in different subterminal, but also to a high extent, in terminal carbon chain positions. To enable a high‐throughput screening procedure, a very versatile assay was developed that is capable of discriminating between terminal and subterminal hydroxylation of carbon chains. A double mutant of CYP102A3 was obtained that produces 48 % octan‐1‐ol as the main product of the reaction.
Functional Interplay Between Nuclear Factor‐κB and c‐Jun Integrated by Coactivator p300 Determines the Survival of Nerve Growth Factor‐Dependent PC12 Cells

Abstract: Nerve growth factor (NGF) activates the transcription factors nuclear factor κB (NF‐κB) and activator protein‐1 (AP‐1) in sympathetic neurons. Whereas NGF‐inducible NF‐κB is required for the survival of neurons, c‐Jun has the ability to promote neuronal death. In this report, we have examined the effect of NGF withdrawal on c‐Jun and NF‐κB transcription factors in PC12 cells differentiated to a neuronal phenotype. We show that the withdrawal of NGF from these cultures results in de novo synthesis of c‐Jun, increase in AP‐1 activity, and down‐regulation of NF‐κB activity. To investigate how the signal transduction pathways activating c‐Jun and NF‐κB are differentially regulated by NGF, we performed transcriptional analyses. Expression of RelA (NF‐κB) suppressed the c‐Jun‐dependent transcription of c‐jun, and this effect was reversed by overexpression of the coactivator p300. RelA’s effects on c‐Jun transcription were mediated by competitive binding of the carboxy‐terminal region of RelA to the CH1 domain of p300, which also binds to c‐Jun; deletion of this region abrogated the ability of RelA to inhibit c‐Jun activity. Furthermore, the inhibition of endogenous NF‐κB in NGF‐maintained neuronal PC12 cells led to the induction of c‐Jun synthesis and a marked increase in cell death. Together, these studies demonstrate a functional interaction between NF‐κB and c‐Jun and suggest a novel mechanism of NF‐κB‐mediated neuroprotection.
Technical Advance: The rat aorta contains resident mononuclear phagocytes with proliferative capacity and proangiogenic properties

Angiogenesis in the aortic ring model is preceded by activation of the immune system and impaired by ablation of adventitial macrophages. Treatment of aortic cultures with M‐CSF induced extensive periaortic outgrowth of CD45+ CD68+ mononuclear cells with ultrastructural features of macrophages and DCs. Periaortic lysis of collagen caused many CD45+ CD68+ cells to attach to the bottom of the culture dish. Lifting the collagen gels left behind patches of CD45+ CD68+ cells, which focally organized into branching cords. These cells also expressed CD14, CD169, F4/80, and α‐SMA but not CD31, vWF, desmin, or CD163. DNA synthesis studies showed that M‐CSF‐stimulated cells were actively proliferating. Aortic patch cells showed phagocytic properties and responded to IL‐4 and GM‐CSF by expressing MHC II, differentiating into DCs, and forming multinucleated giant cells. They also stimulated angiogenesis and VEGF production in aortic ring cultures. This study demonstrates that the rat aorta contains a distinct subset of immature immunocytes capable of proliferating, differentiating into macrophages and DCs, and stimulating angiogenesis. Isolation of these cells in patches from M‐CSF‐stimulated aortic rings provides a reproducible system to study the biology and angiogenic role of the resident immune system of the aortic wall.
Stress Processes: An Essential Ingredient in the Entrepreneurial Process

The entrepreneurial process is associated with high uncertainty, and uncertainty is a major source of stress. Therefore, a core aim of entrepreneurs is to reduce uncertainty to allow the entrepreneurial process to unfold. However, entrepreneurship scholars have insufficiently addressed stress processes that may be associated with this uncertainty. We argue that uncertainty is the concept connecting both the entrepreneurial and stress processes. We discuss the links between these two processes in terms of (1) opportunity recognition, (2) opportunity exploitation, and (3) associated outcomes. We then illustrate how future research should incorporate the interaction between the two processes using a morphological box and discuss how such research would change the way we specify entrepreneurial process models and study entrepreneurial behavior.
A shared promoter region suggests a common ancestor for the human VCX/Y, SPANX, and CSAG gene families and the murine CYPT family

Many testis‐specific genes from the sex chromosomes are subject to rapid evolution, which can make it difficult to identify murine genes in the human genome. The murine CYPT gene family includes 15 members, but orthologs were undetectable in the human genome. However, using refined homology search, sequences corresponding to the shared promoter region of the CYPT family were identified at 39 loci. Most loci were located immediately upstream of genes belonging to the VCX/Y, SPANX, or CSAG gene families. Sequence comparison of the loci revealed a conserved CYPT promoter‐like (CPL) element featuring TATA and CCAAT boxes. The expression of members of the three families harboring the CPL resembled the murine expression of the CYPT family, with weak expression in late pachytene spermatocytes and predominant expression in spermatids, but some genes were also weakly expressed in somatic cells and in other germ cell types. The genomic regions harboring the gene families were rich in direct and inverted segmental duplications (SD), which may facilitate gene conversion and rapid evolution. The conserved CPL and the common expression profiles suggest that the human VCX/Y, SPANX, and CSAG2 gene families together with the murine SPANX gene and the CYPT family may share a common ancestor. Finally, we present evidence that VCX/Y and SPANX may be paralogs with a similar protein structure consisting of C terminal acidic repeats of variable lengths. Mol. Reprod. Dev. 75: 219–229, 2008. © 2007 Wiley‐Liss, Inc.
Unidimensional Modulation Technique for Cascaded Multilevel Converters

This paper presents a simple and low-computational-cost modulation technique for multilevel cascaded H-bridge converters. The technique is based on geometrical considerations considering a unidimensional control region to determine the switching sequence and the corresponding switching times. In addition, a simple strategy to control the dc-voltage ratio between the H-bridges of the multilevel cascaded converter is presented. Examples for the two-cell topology are shown, but the proposed technique can be applied to develop modulation methods for a higher number of H-bridges. Experimental results are presented to validate the proposed technique.
SHAPE-DEFINING SCAFFOLDS FOR MINIMALLY INVASIVE TISSUE ENGINEERING

Background. Minimally invasive surgical procedures are increasingly important in medicine, but biomaterials consistent with this delivery approach that allow one to control the structure of the material after implantation are lacking. Biomaterials with shape-memorizing properties could permit minimally invasive delivery of cell transplantation constructs and enable the formation of new tissues or structures in vivo in desired shapes and sizes. Methods. Macroporous alginate hydrogel scaffolds were prepared in a number of predefined geometries, compressed into significantly smaller, different “temporary” forms, and introduced into immunocompromised mice by means of minimally invasive surgical delivery through a small catheter. Scaffolds were rehydrated in situ with a suspension of cells (primary bovine articular chondrocytes) or cell-free medium and delivered through the same catheter. Specimens were harvested at 1 hr to evaluate the efficacy of cell delivery and the recovery of scaffold geometry, and at 8 and 24 weeks to evaluate neotissue formation. Results. A high percentage (88%) of scaffolds that were introduced with a catheter and rehydrated with cells had recovered their original shape and size within 1 hr. This delivery procedure resulted in cartilage structures with the geometry of the original scaffold by 2 months and histologically mature appearing tissue at 6 months. Conclusions. Shaped hydrogels, formed by covalently cross-linking, can be structurally collapsed into smaller, temporary shapes that permit their minimally invasive delivery in vivo. The rapid recovery of scaffold properties facilitates efficient cell seeding in vivo and permits neotissue formation in desired geometries.
Open-label escitalopram treatment for pathological skin picking

Pathological skin picking is characterized by dysfunctional, repetitive and excessive manipulation of the skin resulting in noticeable tissue damage. This study sought to assess the effectiveness of escitalopram in treating pathological skin picking. Twenty-nine individuals with pathological skin picking were enrolled in an 18-week, open-label trial of escitalopram. Study measures assessing skin picking severity and impact, anxiety, depression, and quality of life were given at baseline and weeks 2, 4, 6, 10, 14, and 18. The mean maximally tolerated dose was 25.0 mg (standard deviation=8.4). For the 19 study completers, pre-post-treatment analyses revealed significant improvements (P<0.05) on measures of skin picking severity and impact, quality of life, and self-rated anxiety and depression. Completer as well as intent-to-treat analyses indicated that approximately half of the sample satisfied full medication response criteria and one-quarter were partial medication responders. Correlational analyses indicated that changes in depression, anxiety, and quality of life co-occurred with reductions in skin picking severity but not impact. A high percentage of variance in severity, however, remained unexplained. These results suggest that escitalopram can be an effective agent in reducing pathological skin picking. The lack of medication response in a subset of our sample suggests the possibility of pathological skin picking subtypes.
Where’s the demand? Explaining the EU’s surprisingly constructive response to adverse WTO rulings

ABSTRACT The political significance of international institutions depends on whether states comply when doing so is costly. The IR literature suggests several reasons a state might do so, but does a poor job of explaining the actual choice. This article addresses that weakness by analyzing the causal pathways though which adverse World Trade Organization rulings prompt the European Union to change its policies. Using Qualitative Comparative Analysis, it analyses all EU policies successfully challenged before the WTO through 2017. It also presents two case studies of policies that the QCA suggests should be least likely cases for my argument. The article demonstrates that, contrary to many depictions, the EU almost always changes its policies in response to an adverse WTO ruling, although those changes frequently fall short of compliance. Further, it argues that the preferences of policymakers, rather than the demands of societal actors, provide the primary impetus for policy change.
The NGC 3341 minor merger: a panchromatic view of the active galactic nucleus in a dwarf companion

We present X-ray (Chandra), radio (EVLA and EVN), and archival optical data of the triple-merging system in NGC3341. Our panchromatic analysis confirms the presence of a Seyfert 2 AGN in NGC3341B, one of the secondary dwarf companions. On the other hand, the nucleus of the primary galaxy, consistent with a star-forming region of a few solar masses per year, and NGC3341C are very unlikely to host an AGN. We therefore suggest that NGC3341 is an exceptional example of an AGN triggered in the satellite galaxy of a minor-merging system. The existence of such a system can have important implications in the models of hierarchical growth of structures. Further observational and theoretical efforts on NGC3341 and potentially similar sources are needed in order to understand the role of minor mergers on the onset of AGN activity, and in the evolution of massive galaxies.
An update on PPAR activation by cannabinoids

Some cannabinoids activate the different isoforms of PPARs (α, β and γ), as shown through the use of reporter gene assays, binding studies, selective antagonists and knockout studies. Activation of all isoforms, but primarily PPARα and γ, mediates some (but not all) of the analgesic, neuroprotective, neuronal function modulation, anti‐inflammatory, metabolic, anti‐tumour, gastrointestinal and cardiovascular effects of some cannabinoids, often in conjunction with activation of the more traditional target sites of action such as the cannabinoid CB1 and CB2 receptors and the TRPV1 ion channel. PPARs also mediate some of the effects of inhibitors of endocannabinoid degradation or transport. Cannabinoids may be chaperoned to the PPARs by fatty acid binding proteins. The aims of this review are to update the evidence supporting PPAR activation by cannabinoids and to review the physiological responses to cannabinoids that are mediated, and not mediated, by PPAR activation.
Mycobacterium‐containing phagosomes are accessible to early endosomes and reflect a transitional state in normal phagosome biogenesis.

The success of Mycobacterium as a pathogen hinges on its ability to modulate its intracellular environment. Mycobacterium avium reside in vacuoles with limited proteolytic activity, maintain cathepsin D in an immature form and remain accessible to internalized transferrin. Artificial acidification of isolated phagosomes facilitated processing of cathepsin D, demonstrating that pH alone limits proteolysis in these vacuoles. Moreover, analysis of IgG‐bead phagosomes at early time points during their formation indicates that these phagosomes also acquire LAMP 1 and cathepsin D prior to the accumulation of proton‐ATPases, and are transiently accessible to sorting endosomes. This suggests that the anomolous distribution of endosomal proteins in M. avium‐containing vacuoles results from their arrested differentiation in an early transitional stage through which all phagosomes pass.