Tyrosine Phosphorylation of the RUNX1 Transcription Factor by Src Kinase Activates C/EBP-Alpha in Granulopoiesis

The Runx1 transcription factor is a master regulator of hematopoiesis and myeloid transformation. Its function is tightly orchestrated by diverse and appropriate signaling events to regulate cell fate decision, including granulopoiesis. Src phosphorylated Runx1 at 5 tyrosine residues; Y260 located at the transactivation domain and the C-terminal tyrosine cluster Y375, Y378, Y379 and Y386 at the repression domain. Tyrosine phosphorylated Runx1 inhibits megakaryopoiesis and thymocyte development but its effect on granulopoiesis is yet to be reported. In luciferase reporter assay in HEK293T cells, Src tyrosine kinase synergized with wild-type (WT) Runx1 to increase its transactivation potency. Significant transactivation effect was also observed with Runx1 tyrosine (Y) to glutamate (E) or aspartate (D) phopho-mimetic mutant variants where the transactivity increased to about 2-fold compared to Runx1-WT. Conversely, tyrosine (Y) to phenylalanine (F) phospho-null mutant variants did not exhibit transactivity. Exogenous Runx1 and its C-terminal phospho-mimetic mutant variant induced the transcriptions of endogenous Cebpα and PU.1 in the 32Dcl3 myeloid progenitor cell line as well as their protein productions, but not with the phospho-null Runx1. At least two mechanisms contribute to the increased Runx1 activity upon tyrosine phosphorylation. Tyrosine phosphorylated Runx1 bound better to its consensus sequences as assessed in Gel Shift Assay, and to the endogenous Cebpα and PU.1 enhancers in Chromatin Immunoprecipitation Assay. In addition, after cycloheximide treatment, high levels of Runx1-5D and 4D protein expressions could sustain up to 8 hours while the Runx1-5F and 4F proteins were degraded between 2-4 hours. The stability of the Runx1-5D was associated with weaker interaction with ubiquitin and also its resistance to Cdh1-mediated ubiquitin proteosomal degradation. Combining these observations, we propose that Runx1 phosphorylation by Src at the five tyrosine sites increase its transactivity, binding affinity and stability, and therefore promoting the induction of mRNA and protein expressions of Cebpa and PU.1, which are the crucial early differentiation biomarkers in myelopoiesis and granulopoiesis. Further investigations on the regulation of tyrosine phosphorylation on Runx1 will shed more insights on the molecular mechanisms underlying granulopoiesis and its implications on myeloid transformation and diseases

Serial-integrated multi-criteria decision-making technique for resilient supplier selection in the manufacturing industry

A supply chain is an entire system of producing and delivering a product or service, from the very beginning stage of sourcing raw material to the final stage of delivering a product or service to end-users. Several global risks and disruptions brought massive and devastating impacts on the world economy including the Small and Medium Enterprises (SMEs). Since the supplier is one of the important elements in a supply chain, economic resilience can be implemented by selecting a resilient supplier. However, the literature shows that previous supplier selections only focused on traditional, green and sustainable suppliers’ criteria but resilience was rare to be discussed. Thus, the first objective of the study is to identify the generic criteria for selecting resilient suppliers. At the same time, there are problems in dealing with uncertainties and incomplete information while selecting suppliers. The second objective is to develop a new integrated Multi-Criteria Decision-Making (MCDM) model that considers incomplete data and uncertainties in selecting resilient suppliers. In this study, the proposed criteria were quality, lead time, cost, flexibility, visibility, responsiveness and financial stability. A serial-integrated MCDM technique was proposed by combining Grey Relational Analysis (GRA) from the grey theory and the Best Worst Method-Technique for Order Preference by Similarity to an Ideal Solution (BMW-TOPSIS) technique in serial to assess the suppliers and select the best alternative. The proposed criteria and technique were applied in the metal manufacturing company (Case 1) and the food manufacturing company (Case 2) which were facing economic problems to demonstrate its effectiveness. The result was generated using MATLAB. The result for Case 1 shows that Financial Stability has the largest weight and Supplier 1 is the best supplier for the company. For Case 2, Cost shows the largest weight, and the best supplier is Supplier 4. Then, the result was verified through manual calculation and validated with Analytic Hierarchy Process-VlseKriterijumska Optimizacija I Kompromisno Resenje (AHP-VIKOR). Through the identification of the generic resilience criteria and the suitable MCDM model, the managers can focus on resilience with the consideration of uncertainties and incomplete information to improve the supplier selection process. This can help to raise the supply chain performance of the companies

A new integrated multi-criteria decision-making model for resilient supplier selection

Unexpected worldwide disruptions brought various challenges to supply chain management thus manipulating the research direction towards resilience. Since the supplier is one of the important supply chain elements, the challenges can be overcome through resilient supplier selection. Supplier selection is a multi-criteria decision-making problem where several criteria are involved. In this study, GRA-BWM-TOPSIS was proposed to evaluate resilient suppliers. Seven resilience criteria which were Quality, Lead Time, Cost, Flexibility, Visibility, Responsiveness and Financial Stability have been proposed and five experts were selected to provide judgments for the selection process. By using the proposed method, the criteria importance levels were obtained using GRA and the criteria weights were computed using BWM, together with a consistency test. TOPSIS was applied to evaluate the suppliers’ performances. Through a case study in a food manufacturing company, 10 suppliers were evaluated and ranked. A validation process was carried out and the managerial implications were provided to ensure the effectiveness of the proposed model. GRA-BWM-TOPSIS is suitable for resilient supplier selection when there are uncertainties and incomplete data

The CRISPR/Cas9 System for Crop Improvement: Progress and Prospects

The global demand for high-quality crops is continuously growing with time. Crop improvement techniques have a long history and they had been applied since the beginning of domestication of the first agricultural plants. Since then, various new techniques have and are being developed to further increase the commercial value and yield of crops. The latest crop improvement technique known as genome editing is a technique that enables precise modification of the plant genome via knocking out undesirable genes or enabling genes to gain new function. The variants generated from the genome editing are indistinguishable from naturally occurring variation. It is also less time-consuming and more readily accepted in the market commercially. The usage of genome editing has proven to be advantages and plays a promising role in future crop improvement efforts. Therefore, in this chapter, we aim to highlight the progress and application of genome editing techniques, in particular, the CRISPR/Cas9 system as a powerful genome editing tool for crop improvement. In addition, the challenges and future prospects of this technology for crop improvement will also be discussed

Optimization and clinical validation of a pathogen detection microarray

New design and optimization of pathogen detection microarrays is shown to allow robust and accurate detection of a range of pathogens. The customized microarray platform includes a method for reducing PCR bias during DNA amplification

Diagnostic Accuracy of the Electrocardiogram for Heart Failure With Reduced or Preserved Ejection Fraction

Current heart failure (HF) guidelines recommend electrocardiography (ECG) as an essential initial investigation in a patient's workup. 1 However, these recommendations were based on studies primarily including patients with HF with reduced ejection fraction (HFrEF). 1 , 2 , 3 Guidelines do not distinguish HFrEF from HF with preserved and mid-range ejection fraction (HFpEF and HFmrEF) in their ECG recommendations. We hypothesized that a normal ECG does not exclude HFpEF and has a considerably lower sensitivity for diagnosing HFpEF than HFrEF

Plasmodium knowlesi malaria an emerging public health problem in Hulu Selangor, Selangor, Malaysia (2009–2013) : epidemiologic and entomologic analysis.

Background: While transmission of the human Plasmodium species has declined, a significant increase in Plasmodium knowlesi/Plasmodium malariae cases was reported in Hulu Selangor, Selangor, Malaysia. Thus, a study was undertaken to determine the epidemiology and the vectors involved in the transmission of knowlesi malaria. Methods: Cases of knowlesi/malariae malaria in the Hulu Selangor district were retrospectively reviewed and analyzed from 2009 to 2013. Mosquitoes were collected from areas where cases occurred in order to determine the vectors. Leucosphyrus group of mosquitoes were genetically characterized targeting the nuclear internal transcribed spacer 2 (ITS2) and mitochondrial cytochrome c oxidase subunit I (CO1). In addition, temporal and spatial analyses were carried out for human cases and vectors. Results: Of the 100 microscopy diagnosed P. knowlesi/P. malariae cases over the 5 year period in the Hulu Selangor district, there was predominance of P. knowlesi/P. malariae cases among the young adults (ages 20–39 years; 67 cases; 67%). The majority of the infected people were involved in occupations related to agriculture and forestry (51; 51%). No death was recorded in all these cases. Five hundred and thirty five mosquitoes belonging to 14 species were obtained during the study. Anopheles maculatus was the predominant species (49.5%) followed by Anopheles letifer (13.1%) and Anopheles introlatus (11.6%). Molecular and phylogenetic analysis confirmed the species of the Leucosphyrus group to be An. introlatus. In the present study, only An. introlatus was positive for oocysts. Kernel Density analysis showed that P. knowlesi hotspot areas overlapped with areas where the infected An. introlatus was discovered. This further strengthens the hypothesis that An. introlatusis is the vector for P. knowlesi in the Hulu Selangor district. Unless more information is obtained on the vectors as well as macaque involved in the transmission, it will be difficult to plan effective control strategies. The utilization of modern analytical tools such as GIS (Geographic Information System) is crucial in estimating hotspot areas for targeted control strategies. Conclusions: Anopheles introlatus has been incriminated as vector of P. knowlesi in Hulu Selangor. The cases of P. knowlesi are on the increase and further research using molecular techniques is needed

Identification of Giardia lamblia DHHC Proteins and the Role of Protein S-palmitoylation in the Encystation Process

Protein S-palmitoylation, a hydrophobic post-translational modification, is performed by protein acyltransferases that have a common DHHC Cys-rich domain (DHHC proteins), and provides a regulatory switch for protein membrane association. In this work, we analyzed the presence of DHHC proteins in the protozoa parasite Giardia lamblia and the function of the reversible S-palmitoylation of proteins during parasite differentiation into cyst. Two specific events were observed: encysting cells displayed a larger amount of palmitoylated proteins, and parasites treated with palmitoylation inhibitors produced a reduced number of mature cysts. With bioinformatics tools, we found nine DHHC proteins, potential protein acyltransferases, in the Giardia proteome. These proteins displayed a conserved structure when compared to different organisms and are distributed in different monophyletic clades. Although all Giardia DHHC proteins were found to be present in trophozoites and encysting cells, these proteins showed a different intracellular localization in trophozoites and seemed to be differently involved in the encystation process when they were overexpressed. dhhc transgenic parasites showed a different pattern of cyst wall protein expression and yielded different amounts of mature cysts when they were induced to encyst. Our findings disclosed some important issues regarding the role of DHHC proteins and palmitoylation during Giardia encystation.Fil: Merino, Maria Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; ArgentinaFil: Zamponi, Nahuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; ArgentinaFil: Vranych, Cecilia Verónica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; ArgentinaFil: Touz, Maria Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; ArgentinaFil: Ropolo, Andrea Silvana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; Argentin

Recurrent Fusion Genes in Gastric Cancer: CLDN18-ARHGAP26 Induces Loss of Epithelial Integrity.

Genome rearrangements, a hallmark of cancer, can result in gene fusions with oncogenic properties. Using DNA paired-end-tag (DNA-PET) whole-genome sequencing, we analyzed 15 gastric cancers (GCs) from Southeast Asians. Rearrangements were enriched in open chromatin and shaped by chromatin structure. We identified seven rearrangement hot spots and 136 gene fusions. In three out of 100 GC cases, we found recurrent fusions between CLDN18, a tight junction gene, and ARHGAP26, a gene encoding a RHOA inhibitor. Epithelial cell lines expressing CLDN18-ARHGAP26 displayed a dramatic loss of epithelial phenotype and long protrusions indicative of epithelial-mesenchymal transition (EMT). Fusion-positive cell lines showed impaired barrier properties, reduced cell-cell and cell-extracellular matrix adhesion, retarded wound healing, and inhibition of RHOA. Gain of invasion was seen in cancer cell lines expressing the fusion. Thus, CLDN18-ARHGAP26 mediates epithelial disintegration, possibly leading to stomach H(+) leakage, and the fusion might contribute to invasiveness once a cell is transformed. Cell Rep 2015 Jul 14; 12(2):272-285

Antimicrobial resistance among migrants in Europe: a systematic review and meta-analysis

BACKGROUND: Rates of antimicrobial resistance (AMR) are rising globally and there is concern that increased migration is contributing to the burden of antibiotic resistance in Europe. However, the effect of migration on the burden of AMR in Europe has not yet been comprehensively examined. Therefore, we did a systematic review and meta-analysis to identify and synthesise data for AMR carriage or infection in migrants to Europe to examine differences in patterns of AMR across migrant groups and in different settings. METHODS: For this systematic review and meta-analysis, we searched MEDLINE, Embase, PubMed, and Scopus with no language restrictions from Jan 1, 2000, to Jan 18, 2017, for primary data from observational studies reporting antibacterial resistance in common bacterial pathogens among migrants to 21 European Union-15 and European Economic Area countries. To be eligible for inclusion, studies had to report data on carriage or infection with laboratory-confirmed antibiotic-resistant organisms in migrant populations. We extracted data from eligible studies and assessed quality using piloted, standardised forms. We did not examine drug resistance in tuberculosis and excluded articles solely reporting on this parameter. We also excluded articles in which migrant status was determined by ethnicity, country of birth of participants' parents, or was not defined, and articles in which data were not disaggregated by migrant status. Outcomes were carriage of or infection with antibiotic-resistant organisms. We used random-effects models to calculate the pooled prevalence of each outcome. The study protocol is registered with PROSPERO, number CRD42016043681. FINDINGS: We identified 2274 articles, of which 23 observational studies reporting on antibiotic resistance in 2319 migrants were included. The pooled prevalence of any AMR carriage or AMR infection in migrants was 25·4% (95% CI 19·1-31·8; I2 =98%), including meticillin-resistant Staphylococcus aureus (7·8%, 4·8-10·7; I2 =92%) and antibiotic-resistant Gram-negative bacteria (27·2%, 17·6-36·8; I2 =94%). The pooled prevalence of any AMR carriage or infection was higher in refugees and asylum seekers (33·0%, 18·3-47·6; I2 =98%) than in other migrant groups (6·6%, 1·8-11·3; I2 =92%). The pooled prevalence of antibiotic-resistant organisms was slightly higher in high-migrant community settings (33·1%, 11·1-55·1; I2 =96%) than in migrants in hospitals (24·3%, 16·1-32·6; I2 =98%). We did not find evidence of high rates of transmission of AMR from migrant to host populations. INTERPRETATION: Migrants are exposed to conditions favouring the emergence of drug resistance during transit and in host countries in Europe. Increased antibiotic resistance among refugees and asylum seekers and in high-migrant community settings (such as refugee camps and detention facilities) highlights the need for improved living conditions, access to health care, and initiatives to facilitate detection of and appropriate high-quality treatment for antibiotic-resistant infections during transit and in host countries. Protocols for the prevention and control of infection and for antibiotic surveillance need to be integrated in all aspects of health care, which should be accessible for all migrant groups, and should target determinants of AMR before, during, and after migration. FUNDING: UK National Institute for Health Research Imperial Biomedical Research Centre, Imperial College Healthcare Charity, the Wellcome Trust, and UK National Institute for Health Research Health Protection Research Unit in Healthcare-associated Infections and Antimictobial Resistance at Imperial College London