Infection prevention and control in conflict-affected areas in northeast Syria: A cross-sectional study.

Objectives: In northeastern Syria (NES), the adherence of health care facilities to infection prevention and control (IPC) standards remains underexplored. This study evaluates the IPC performance of various health facilities against World Health Organization (WHO) benchmarks using the IPC Assessment Framework (IPCAF) and the Hand Hygiene Self-Assessment Framework (HHSAF). Methods: We conducted a cross-sectional survey of 33 health care facilities, including primary (PHC), secondary (SHC), and tertiary health care centres (THC). Data were collected via on-site evaluations using the IPCAF and HHSAF tools. Results: A significant 91% of facilities did not meet half of the WHO IPC minimum requirements. Specifically, 57% of PHCs met 26-50% of the standards, while none exceeded 75%. Among SHCs, 71% met 26-50% of the standards, while 44% of THCs fell within this range. Notably, 81.8% of facilities were classified as 'inadequate' per the IPCAF, with none achieving 'intermediate' or 'advanced' levels. The HHSAF results were similarly concerning, with 34.4% deemed 'inadequate' and 65.6% at the 'basic' level. A weak positive correlation (0.137) was found between IPCAF and HHSAF scores. Conclusions: NES health care facilities demonstrate substantial deficiencies in IPC compliance, with critical gaps in IPC programmes, health care-associated infection surveillance, and training. Urgent interventions are required to enhance IPC practices, leveraging local strengths and fostering international collaborations to improve patient safety and health care quality in the region.

Expression of LOXL3, NES, and SNAI1 in Melanoma Genesis and Progression.

Melanoma is the most severe type of skin cancer and among the most malignant neoplasms in humans. With the growing incidence of melanoma, increased numbers of therapeutic options, and the potential to target specific proteins, understanding the basic mechanisms underlying the disease's progression and resistance to treatment has never been more important. LOXL3, SNAI1, and NES are key factors in melanoma genesis, regulating tumor growth, metastasis, and cellular differentiation. In our study, we explored the potential role of LOXL3, SNAI1, and NES in melanoma progression and metastasis among patients with dysplastic nevi, melanoma in situ, and BRAF+ and BRAF- metastatic melanoma, using immunofluorescence and qPCR analysis. Our results reveal a significant increase in LOXL3 expression and the highest NES expression in BRAF+ melanoma compared to BRAF-, dysplastic nevi, and melanoma in situ. As for SNAI1, the highest expression was observed in the metastatic melanoma group, without significant differences among groups. We found co-expression of LOXL3 and SNAI1 in the perinuclear area of all investigated subgroups and NES and SNAI1 co-expression in melanoma cells. These findings suggest a codependence or collaboration between these markers in melanoma EMT, suggesting new potential therapeutic interventions to block the EMT cascade that could significantly affect survival in many melanoma patients.

Engineering Cu-Ce-a nanozymes: Revolutionary alloy nanomaterials mimicking cytochrome c oxidase for ultra-sensitive cytochrome c detection.

The design of synthetic analogs of cytochrome c oxidase (CcO) is a formidable task due to its intricate structure encompassing multiple metal prosthetic sites and protein subunits. In recent years, artificial enzymes based on alloy nanomaterials have garnered significant attention due to the alloy design approach holds promise for the effective tuning of the properties of metal catalysts. In this study, we present copper-cerium alloy nanozymes (Cu-Ce-a NEs), where Cu mimics the active site of CcO, while Ce endows the alloy phase and enhances the capacity to catalyze the oxidation to cytochrome c (Cyt c). Cu-Ce-a NEs functionally mimics CcO, the terminal enzyme in the respiratory electron transport chain (ETC), by catalyzing the four-electron reduction of dioxygen to water. Utilizing the CcO-like properties of Cu-Ce-a NEs, we successfully implemented the electrochemical detection of Cyt c. The Cu-Ce-a NEs based electrochemical sensor revealed a favorable linear range spanning from 2 to 20 µM Cyt c, with a detection limit (LOD) of 2 µM. This method demonstrates high accuracy in Cyt c quantitation in pharmaceuticals, with results closely aligning with the actual concentrations. This finding not only offers new perspectives in the design of enzyme analogs, but also underscores the potential of this method for clinical Cyt c detection, highlighting its significance in biomedical research and diagnostics.

Exploring the manganese-dependent interaction between a transcription factor and its corresponding DNA: insights from gas-phase electrophoresis on a nES GEMMA instrument.

Manganese ion homeostasis is vital for bacteria and is achieved via manganese-dependent transcription factors. Manganese mediation of transcription factor attachment to the corresponding oligonucleotide sequences can be investigated, e.g. via electrophoretic mobility shift assays (EMSA). Formation of specific biocomplexes leads to differences in the migration pattern upon gel electrophoresis. Focusing on electrophoresis in the gas-phase, applying a nano electrospray gas-phase electrophoretic mobility molecular analyzer (nES GEMMA) also known as nES differential mobility analyzer (nES DMA), and on transcription factors (MntR proteins) from Bacillus subtilis and Mycobacterium tuberculosis, we took interest in the gas-phase electrophoresis of the corresponding biospecific complexes. We compared nES GEMMA, separating analytes in the nanometer regime (a few to several hundred nm in diameter) in the gas-phase in their native state according to particle size, to EMSA data. Indeed we were able to demonstrate manganese-mediated attachment of MntR to target genomic sequences with both analytical techniques. Despite some inherent pitfalls of the nES GEMMA method like analyte/instrument surface interactions, we were able to detect the target complexes. Moreover, we were able to calculate the molecular weight (MW) of the obtained species by application of a correlation function based on nES GEMMA obtained data. As gas-phase electrophoresis also offers the possibility of offline hyphenation to orthogonal analysis techniques, we are confident that nES GEMMA measurements are not just complementary to EMSA, but will offer the possibility of further in-depth characterization of biocomplexes in the future.

Pre-clinical evaluation of clinically relevant iPS cell derived neuroepithelial stem cells as an off-the-shelf cell therapy for spinal cord injury.

Preclinical transplantations using human neuroepithelial stem (NES) cells in spinal cord injury models have exhibited promising results and demonstrated cell integration and functional improvement in transplanted animals. Previous studies have relied on the generation of research grade cell lines in continuous culture. Using fresh cells presents logistic hurdles for clinical transition regarding time and resources for maintaining high quality standards. In this study, we generated a good manufacturing practice (GMP) compliant human iPS cell line in GMP clean rooms alongside a research grade iPS cell line which was produced using standardized protocols with GMP compliant chemicals. These two iPS cell lines were differentiated into human NES cells, from which six batches of cell therapy doses were produced. The doses were cryopreserved, thawed on demand and grafted in a rat spinal cord injury model. Our findings demonstrate that NES cells can be directly grafted post-thaw with high cell viability, maintaining their cell identity and differentiation capacity. This opens the possibility of manufacturing off-the-shelf cell therapy products. Moreover, our manufacturing process yields stable cell doses with minimal batch-to-batch variability, characterized by consistent expression of identity markers as well as similar viability of cells across the two iPS cell lines. These cryopreserved cell doses exhibit sustained viability, functionality, and quality for at least 2 years. Our results provide proof of concept that cryopreserved NES cells present a viable alternative to transplanting freshly cultured cells in future cell therapies and exemplify a platform from which cell formulation can be optimized and facilitate the transition to clinical trials.

The association between night eating syndrome and GERD symptoms among university students at An-Najah National University in Palestine: a cross-sectional study.

BACKGROUND: Night eating syndrome (NES) is a kind of eating disorder. NES association with gastroesophageal reflux disease (GERD) symptoms among university students is still not fully understood. We aimed to determine the relationship between NES and the presence of GERD symptoms among university students at An-Najah National University in Palestine. METHODS: This study involved undergraduate students from An-Najah National University. The data were collected through online surveys from November to December 2023. The sampling frame involved voluntary sampling, as the data were collected using a structured questionnaire to collect data on sociodemographic variables, medical history, lifestyle habits, nutritional status, GERD risk, and NES. The GERD questionnaire (GerdQ) was used to assess symptoms, while the Arabic version of the validated Night Eating Questionnaire (NEQ) was used to assess night eating. Physical activity was assessed using the short form of the International Physical Activity Questionnaire (SF-IPAQ), and adherence to a Mediterranean diet was assessed using the validated Arabic version of the MEDAS. Both univariate and multivariate analyses were also conducted to assess the study hypotheses. RESULTS: The study involved 554 participants, 59.9% female. A total of 33.4% reported GERD symptoms, with 10.3% having NES. A strong association was observed between GERD and NES and between GERD and physical activity. Night eating syndrome (AOR = 2.84, CI = 1.07-3.19), high physical activity (AOR = 0.473, CI = 1.05-3.19), and non-smoking (AOR = 0.586, CI = 1.27-7.89) were identified as independent predictors of GERD symptoms. CONCLUSION: This study revealed that 33.4% of undergraduate students were at risk of GERD, with night eaters having a greater risk. GERD risk was negatively associated with physical activity level and smoking status. No associations were found between GERD risk and weight status, Mediterranean diet adherence, sociodemographic factors, or sleep disturbances.

The Role of Cytokinins and Abscisic Acid in the Growth, Development and Virulence of the Pathogenic Fungus Stagonospora nodorum (Berk.).

Cytokinins (CKs) and abscisic acid (ABA) play an important role in the life of both plants and pathogenic fungi. However, the role of CKs and ABA in the regulation of fungal growth, development and virulence has not been sufficiently studied. We compared the ability of two virulent isolates (SnB and Sn9MN-3A) and one avirulent isolate (Sn4VD) of the pathogenic fungus Stagonospora nodorum Berk. to synthesize three groups of hormones (CKs, ABA and auxins) and studied the effect of exogenous ABA and zeatin on the growth, sporulation and gene expression of necrotrophic effectors (NEs) and transcription factors (TFs) in them. Various isolates of S. nodorum synthesized different amounts of CKs, ABA and indoleacetic acid. Using exogenous ABA and zeatin, we proved that the effect of these hormones on the growth and sporulation of S. nodorum isolates can be opposite, depends on both the genotype of the isolate and on the concentration of the hormone and is carried out through the regulation of carbohydrate metabolism. ABA and zeatin regulated the expression of fungal TF and NE genes, but correlation analysis of these parameters showed that this effect depended on the genotype of the isolate. This study will contribute to our understanding of the role of the hormones ABA and CKs in the biology of the fungal pathogen S. nodorum.

Modeling COVID-19 data with a novel neutrosophic Burr-III distribution.

In this study, we have presented a novel probabilistic model called the neutrosophic Burr-III distribution, designed for applications in neutrosophic surface analysis. Neutrosophic analysis allows for the incorporation of vague and imprecise information, reflecting the reality that many real-world problems involve ambiguous data. This ability to handle vagueness can lead to more robust and realistic models especially in situation where classical models fall short. We have also explored the neutrosophic Burr-III distribution in order to deal with the ambiguity and vagueness in the data where the classical Burr-III distribution falls short. This distribution offers valuable insights into various reliability properties, moment expressions, order statistics, and entropy measures, making it a versatile tool for analyzing complex data. To assess the practical relevance of our proposed distribution, we applied it to real-world data sets and compared its performance against the classical Burr-III distribution. The findings revealed that the neutrosophic Burr-III distribution outperformed than the classical Burr-III distribution in capturing the underlying data characteristics, highlighting its potential as a superior modeling toolin various fields.

Development of a NanoBRET assay for evaluation of 14-3-3σ molecular glues.

We report the development of a 384-well formatted NanoBRET assay to characterize molecular glues of 14-3-3/client interactions in living cells. The seven isoforms of 14-3-3 are dimeric hub proteins with diverse roles including transcription factor regulation and signal transduction. 14-3-3 interacts with hundreds of client proteins to regulate their function and is therefore an ideal therapeutic target when client selectivity can be achieved. We have developed the NanoBRET system for three 14-3-3σ client proteins CRAF, TAZ, and estrogen receptor α (ERα), which represent three specific binding modes. We have measured stabilization of 14-3-3σ/client complexes by molecular glues with EC50 values between 100 nM and 1 µM in cells, which align with the EC50 values calculated by fluorescence anisotropy in vitro. Developing this NanoBRET system for the hub protein 14-3-3σ allows for a streamlined approach, bypassing multiple optimization steps in the assay development process for other 14-3-3σ clients. The NanoBRET system allows for an assessment of PPI stabilization in a more physiologically relevant, cell-based environment using full-length proteins. The method is applicable to diverse protein-protein interactions (PPIs) and offers a robust platform to explore libraries of compounds for both PPI stabilizers and inhibitors.

Nucleocytoplasmic shuttling of BEFV M protein-modulated by lamin A/C and chromosome maintenance region 1 through a transcription-, carrier- and energy-dependent pathway.

This study demonstrates for the first time that the matrix (M) protein of BEFV is a nuclear targeting protein that shuttles between the nucleus and the cytoplasm in a transcription-, carrier-, and energy-dependent manner. Experiments performed in both intact cells and digitonin-permeabilized cells revealed that M protein targets the nucleolus and requires carrier, cytosolic factors or energy input. By employing sequence and mutagenesis analyses, we have determined both nuclear localization signal (NLS) 6KKGKSK11 and nuclear export signal (NES) 98LIITSYL TI106 of M protein that are important for the nucleocytoplasmic shuttling of M protein. Furthermore, we found that both lamin A/C and chromosome maintenance region 1 (CRM-1) proteins could be coimmunoprecipitated and colocalized with the BEFV M protein. Knockdown of lamin A/C by shRNA and inhibition of CRM-1 by leptomycin B significantly reduced virus yield. Collectively, this study provides novel insights into nucleocytoplasmic shuttling of the BEFV M protein modulated by lamin A/C and CRM-1 and by a transcription- and carrier- and energy-dependent pathway.

Noninvasive electrical stimulation as a neuroprotective strategy in retinal diseases: a systematic review of preclinical studies.

BACKGROUND: Electrical activity has a crucial impact on the development and survival of neurons. Numerous recent studies have shown that noninvasive electrical stimulation (NES) has neuroprotective action in various retinal disorders. OBJECTIVE: To systematically review the literature on in vivo studies and provide a comprehensive summary of the neuroprotective action and the mechanisms of NES on retinal disorders. METHODS: Based on the PRISMA guideline, a systematic review was conducted in PubMed, Web of Science, Embase, Scopus and Cochrane Library to collect all relevant in vivo studies on "the role of NES on retinal diseases" published up until September 2023. Possible biases were identified with the adopted SYRCLE's tool. RESULTS: Of the 791 initially gathered studies, 21 articles met inclusion/exclusion criteria for full-text review. The results revealed the neuroprotective effect of NES (involved whole-eye, transcorneal, transscleral, transpalpebral, transorbital electrical stimulation) on different retinal diseases, including retinitis pigmentosa, retinal degeneration, high-intraocular pressure injury, traumatic optic neuropathy, nonarteritic ischemic optic neuropathy. NES could effectively delay degeneration and apoptosis of retinal neurons, preserve retinal structure and visual function with high security, and its mechanism of action might be related to promoting the secretion of neurotrophins and growth factors, decreasing inflammation, inhibiting apoptosis. The quality scores of included studies ranged from 5 to 8 points (a total of 10 points), according to SYRCLE's risk of bias tool. CONCLUSION: This systematic review indicated that NES exerts neuroprotective effects on retinal disease models mainly through its neurotrophic, anti-inflammatory, and anti-apoptotic capabilities. To assess the efficacy of NES in a therapeutic setting, however, well-designed clinical trials are required in the future.

Risk of positive screening for anorexia nervosa, bulimia nervosa and night eating syndrome and associated risk factors in medical fellows in Northeastern Mexico: a multicenter study.

BACKGROUND: Eating disorders and food ingestion (EDs) are serious mental illnesses with a higher prevalence in young adults, with difficult diagnoses that cause serious morbidity and mortality problems. There is not much information about the risk of positive screening for EDs, specifically, anorexia nervosa (AN) and bulimia nervosa (BN) and night eating syndrome (NES) in undergraduate medical interns (UMI) and medical residents (MR) in Mexico. AIM: To determine the risk of AN, BN and NES and to determine the risk factors of such conditions such as age, body mass index (BMI) and gender of MR and UMI with AN/BN and NES at four private hospitals in northeastern Mexico. METHODS: A cross-sectional, descriptive, non-randomized survey in MR and UMI in four hospitals in Northeastern Mexico was conducted using an electronic questionnaire that included: informed consent signature, SCOFF questionnaire for AN and BN screening, NES questionnaire. Also, a survey on general sociodemographic data of each participant was included. Chi-square test and a logistic regression model were computed for analyses. RESULTS: The population included a total of 129 MR and UMI. It was observed that 48.8% were positive for AN or BN and 32.6% were positive for the NES. There was no difference between age, sex, BMI, or medical specialty (if they were MR); however, MR from the first year had a higher risk of AN or BN (OR 23.7, 95% CI 1.181-475.266). CONCLUSIONS: There was a higher risk of positive screening for AN or BN and NES in UMI and MR in our population. In the case of MR, those in first year have a higher risk of AN and BN. Timely diagnosis and treatment are mandatory in this population.

Eating disorders and food ingestion such as anorexia (AN) or bulimia (BN) nervosa and night eating syndrome (NES) are a group of mental illnesses that are frequently under diagnosed. Medical residents (MR) and undergraduate medical interns (UMI) are a high-risk population for such disorders due to their young age, stress environments, erratic eating patterns and long working hours. The aim of this study was to determine the risk of AN, BN and NES and to determine the risk factors of such conditions. One hundred twenty-nine UMI and MR were studied and showed that 48.8% were positive for AN or BN and 32.6% were positive for NES. MR in the first year of medical residency had a higher risk. Timely diagnosis and treatment are mandatory in this population.

Manufacture and Combustion Characteristics of Cellulose Flame-Retardant Plate through the Hot-Press Method.

This study focuses on the increased risk of high heat release and asphyxiation (toxic gas poisoning) in the event of a fire involving polyurethane (PU)- and MDF-based building materials, which are commonly used in buildings. Among them, polyurethane (PU) building materials are very commonly used in buildings, except in Europe and some other countries, due to their excellent thermal insulation performance. Still, problems of short-term heat release and the spread of toxic gases in the event of a fire continue to occur. To overcome these problems, researchers are actively working on introducing various flame retardants into building materials. Therefore, in this study, we produced a laboratory-sized (500 mm × 500 mm) plate-like flame-retardant board that can be utilized as a building material with a lower heat release rate and a lower toxicity index. The material was made by mixing expanded graphite and ceramic binder as flame retardants in a material that is formulated based on the cellulose of waste paper, replacing the existing building materials with a hot-press method. According to the ISO-5660-1 test on the heat release rate of the plate-like flame-retardant board, the Total Heat Release (THR) value was 2.9 (MJ/m2) for 10 min, showing an effect of reducing the THR value by 36.3 (MJ/m2) compared to the THR value of 39.2 (MJ/m2) of the specimen made using only paper. In addition, the toxicity index of the flame-retardant board was checked through the NES (Naval Engineering Standards)-713 test. As a result, the test specimen showed a toxicity index of 0.7, which is 2.4 lower than the toxicity index of 3.1 of MDF, which is utilized as a conventional building material. Based on the results of this study, the cellulose fire-retardant board showed the effect of reducing the heat release rate and toxicity index of building materials in a building fire, which reduces the risk of rapid heat spread and smoke toxicity. This has the potential to improve the evacuation time (A-SET) of evacuees in fires. It is also important to show that recycling waste paper and utilizing it as the main material for building materials can be an alternative in terms of sustainable development.

In vitro Quality Assessments of Perfluorocarbon Nanoemulsions for Near-infrared Fluorescence Imaging of Inflammation in Preclinical Models.

Tracking macrophages by non-invasive molecular imaging can provide useful insights into the immunobiology of inflammatory disorders in preclinical disease models. Perfluorocarbon nanoemulsions (PFC-NEs) have been well documented in their ability to be taken up by macrophages through phagocytosis and serve as 19F magnetic resonance imaging (MRI) tracers of inflammation in vivo and ex vivo. Incorporation of near-infrared fluorescent (NIRF) dyes in PFC-NEs can help monitor the spatiotemporal distribution of macrophages in vivo during inflammatory processes, using NIRF imaging as a complementary methodology to MRI. Here, we discuss in depth how both colloidal and fluorescence stabilities of the PFC-NEs are essential for successful and reliable macrophage tracking in vivo and for their detection in excised tissues ex vivo by NIRF imaging. Furthermore, PFC-NE quality assures NIRF imaging reproducibility and reliability across preclinical studies, providing insights into inflammation progression and therapeutic response. Previous studies focused on assessments of colloidal property changes in response to stress and during storage as a means of quality control. We recently focused on the joint evaluation of both colloidal and fluorescence properties and their relationship to NIRF imaging outcomes. In this protocol, we summarize the key assessments of the fluorescent dye-labeled nanoemulsions, which include long-term particle size distribution monitoring as the measure of colloidal stability and monitoring of the fluorescence signal. Due to its simplicity and reproducibility, our protocols are easy to adopt for researchers to assess the quality of PFC-NEs for in vivo NIRF imaging applications.

La réduction du risque d'infection périnatale chez les nouveau-nés de mères dont les soins prénatals étaient inappropriés.

Le risque que des infections maternelles ne soient ni décelées ni traitées augmente lorsque les soins prénatals sont inappropriés, ce qui met la santé de la mère et de son nouveau-né à risque. Lorsqu'une femme enceinte se présente tardivement pour recevoir des soins, les tests systématiques qui influent sur la prise en charge du nouveau-né devraient inclure l'antigène de surface de l'hépatite B (AgHBs), la sérologie du virus de l'hépatite C (VHC), du virus de l'immunodéficience humaine (VIH) et de la syphilis, de même que le dépistage de la Chlamydia trachomatis et de la Neisseria gonorrhoeae. Si la mère ne s'est pas soumise aux dépistages avant ou après l'accouchement et qu'elle n'est pas disponible pour s'y soumettre, il faudrait procéder au dépistage du VIH, du virus de l'hépatite B (VHB), du VHC et de la syphilis chez le nouveau-né. Le dépistage de la C. trachomatis et de la N. gonorrhoeae est toutefois réservé aux cas où le nouveau-né démontre des manifestations cliniques compatibles avec ces infections. Il est optimal d'obtenir rapidement les résultats du dépistage du VIH, du VHB et de la syphilis, car l'utilisation des traitements préventifs est circonscrite dans le temps. Il existe des interventions préventives précoces et efficaces pour les nouveau-nés à risque de VIH, de VHB, de syphilis ou de gonorrhée. Un suivi clinique étroit et des tests de suivi s'imposent auprès des nouveau-nés de mères dont les soins prénatals étaient inappropriés, car il est impossible d'exclure pleinement toutes les infections pendant la période périnatale.

Adeno-Associated Virus-like Particles' Response to pH Changes as Revealed by nES-DMA.

Gas-phase electrophoresis on a nano-Electrospray Gas-phase Electrophoretic Mobility Molecular Analyzer (nES GEMMA) separates single-charged, native analytes according to the surface-dry particle size. A volatile electrolyte, often ammonium acetate, is a prerequisite for electrospraying. Over the years, nES GEMMA has demonstrated its unique capability to investigate (bio-)nanoparticle containing samples in respect to composition, analyte size, size distribution, and particle numbers. Virus-like particles (VLPs), being non-infectious vectors, are often employed for gene therapy applications. Focusing on adeno-associated virus 8 (AAV8) based VLPs, we investigated the response of these bionanoparticles to pH changes via nES GEMMA as ammonium acetate is known to exhibit these changes upon electrospraying. Indeed, slight yet significant differences in VLP diameters in relation to pH changes are found between empty and DNA-cargo-filled assemblies. Additionally, filled VLPs exhibit aggregation in dependence on the applied electrolyte's pH, as corroborated by atomic force microscopy. In contrast, cryogenic transmission electron microscopy did not relate to changes in the overall particle size but in the substantial particle's shape based on cargo conditions. Overall, we conclude that for VLP characterization, the pH of the applied electrolyte solution has to be closely monitored, as variations in pH might account for drastic changes in particles and VLP behavior. Likewise, extrapolation of VLP behavior from empty to filled particles has to be carried out with caution.

APEX3 - An Optimized Tool for Rapid and Unbiased Proximity Labeling.

Macromolecular interactions regulate all aspects of biology. The identification of interacting partners and complexes is important for understanding cellular processes, host-pathogen conflicts, and organismal development. Multiple methods exist to label and enrich interacting proteins in living cells. Notably, the soybean ascorbate peroxidase, APEX2, rapidly biotinylates adjacent biomolecules in the presence of biotin-phenol and hydrogen peroxide. However, during initial experiments with this system, we found that APEX2 exhibits a cytoplasmic-biased localization and is sensitive to the nuclear export inhibitor leptomycin B (LMB). This led us to identify a putative nuclear export signal (NES) at the carboxy-terminus of APEX2 (NESAPEX2), structurally adjacent to the conserved heme binding site. This putative NES is functional as evidenced by cytoplasmic localization and LMB sensitivity of a mCherry-NESAPEX2 chimeric construct. Single amino acid substitutions of multiple hydrophobic residues within NESAPEX2 eliminate cytoplasm-biased localization of both mCherry-NESAPEX2 as well as full-length APEX2. However, all but one of these NES substitutions also compromises peroxide-dependent labeling. This unique separation-of-function mutant, APEX2-L242A, is termed APEX3. Localization and functionality of APEX3 are confirmed by fusion to the nucleocytoplasmic shuttling transcriptional factor, RELA. APEX3 is therefore an optimized tool for unbiased proximity labeling of cellular proteins and interacting factors..

Nuclear export signal mutation of epidermal growth factor receptor enhances malignant phenotypes of cancer cells.

Nuclear epidermal growth factor receptor (EGFR) has been shown to be correlated with drug resistance and a poor prognosis in patients with cancer. Previously, we have identified a tripartite nuclear localization signal (NLS) within EGFR. To comprehensively determine the functions and underlying mechanism of nuclear EGFR and its clinical implications, we aimed to explore the nuclear export signal (NES) sequence of EGFR that is responsible for interacting with the exportins. We combined in silico prediction with site-directed mutagenesis approaches and identified a putative NES motif of EGFR, which is located in amino acid residues 736-749. Mutation at leucine 747 (L747) in the EGFR NES led to increased nuclear accumulation of the protein via a less efficient release of the exportin CRM1. Interestingly, L747 with serine (L747S) and with proline (L747P) mutations were found in both tyrosine kinase inhibitor (TKI)-treated and -naïve patients with lung cancer who had acquired or de novo TKI resistance and a poor outcome. Reconstituted expression of the single NES mutant EGFRL747P or EGFRL747S, but not the dual mutant along with the internalization-defective or NLS mutation, in lung cancer cells promoted malignant phenotypes, including cell migration, invasiveness, TKI resistance, and tumor initiation, supporting an oncogenic role of nuclear EGFR. Intriguingly, cells with germline expression of the NES L747 mutant developed into B cell lymphoma. Mechanistically, nuclear EGFR signaling is required for sustaining nuclear activated STAT3, but not for Erk. These findings suggest that EGFR functions are compartmentalized and that nuclear EGFR signaling plays a crucial role in tumor malignant phenotypes, leading to tumorigenesis in human cancer.

Psychological Well-Being, Self-Esteem, Quality of Life and Gender Differences as Determinants of Post-Traumatic Growth in Long-Term Knee Rotationplasty Survivors: A Cohort Study.

Rotationplasty (RP) is a special surgical technique for bone tumors of the lower limb and is the chosen procedure for children under 6 with bone sarcoma in the distal femur. Leg reconstruction results in an unusual aspect of the limb potentially giving life-long emotional outcomes, especially considering the young age of most RP patients. Although the high level of the quality of life of these patients has been previously reported, aspects related to long-term psychological well-being, self-esteem and life satisfaction, particularly regarding the gender, procreation and parenting, have never been explored. The aim of this study was to assess the general degree of psychological well-being of RP patients, with specific reference to gender, procreation and parenting. Twenty long-term RP survivors of high-grade bone sarcoma participated in the study. They were administered the following validated questionnaires: HADS for psychological well-being (degree of anxiety and depression), Temperament and Character Inventory (TCI), RSES for self-esteem, SF-36 for quality of life, SWLS extended to life satisfaction, and ABIS for body image integration. Data on education, marriage, employment and parenthood were gathered. All the scores obtained were very close to normal references. The only gender difference was found for the TCI Cooperativeness scale, which was higher in women than in men. A satisfactory psychological well-being in terms of both self-esteem and integration of the prosthetic joint limb into one's body image, with relatively limited amount of anxiety/depression, good quality of life, and good temperament and character traits, was found. No major gender differences were reported.

Night eating syndrome subtypes: differences in binge eating and food addiction symptoms.

PURPOSE: The purpose of the current study was to examine differences in binge eating and food addiction symptoms between Night Eating Syndrome (NES) latent subtypes: evening hyperphagia with nocturnal ingestions (EHNI), evening hyperphagia-only (EHO), and nocturnal ingestions-only (NIO). It was hypothesized that the EHNI group would report more binge eating behaviors and more food addiction symptoms than both the EHO and NIO groups. Further, it was hypothesized that the EHO and NIO groups would differ with the EHO group reporting more binge eating behaviors and the NIO group reporting more food addiction symptoms. METHODS: Participants completed measures online relating to night eating, binge eating, and food addiction. Average age of the final sample was 34.3 (SD = 10.5) and 62.0% were men. Responses to the Night Eating Questionnaire (NEQ; Allison et al., 2008) were used to create an EHNI group (n = 65), an EHO group (n = 32), and a NIO group (n = 69). ANOVAs were conducted to examine between-group differences on disordered eating symptoms. RESULTS: Participants in the EHNI group reported more severe binge eating and food addiction symptoms than those in the EHO and NIO groups. However, there were no significant differences in binge eating or food addiction between the EHO and NIO groups. CONCLUSION: Individuals who meet both NES core criteria (evening hyperphagia and nocturnal ingestions) are likely at a higher risk for experiencing other, more severe disordered eating pathologies. Implications concerning assessment and future research on NES typology are discussed. LEVEL OF EVIDENCE: Level V, cross-sectional descriptive study.