Cloning, expression and purification of pwo polymerase from pyrococcus woesei

Background and objectives: Pyrococcus woesei is a hyperthermophilic archaea and produces a heat stable polymerase (Pwo polymerase) that has proofreading activity. Materials and Methods: In this study, this microorganism was cultured, its DNA was extracted and the pwo gene polymerase was cloned, expressed and purified. The DNA sequence of the cloned gene was verified by sequencing. The pwo polymerase gene consists of 2,328 bps (775 amino acids with about 90 kD molecular weight). Cloning was done by GATEWAY TM Cloning System and for purification of recombinant protein; His6x-Tag was added to the C-terminus of the recombinant protein. Results and Conclusion: We could purify Pwo polymerase enzyme by Ni-NTA resin. PCR assay showed that Pwo polymerase activity is comparable to a commercial Pfu polymerase activity

Comparing the Mini-BESTest with the Berg Balance Scale to Evaluate Balance Disorders in Parkinson's Disease

Objective. The purpose of this study was to explore the usefulness of the Mini-BESTest compared to the Berg Balance Scale in evaluating balance in people with PD of varying severity. We evaluated (1) the distribution of patients scores to look for ceiling effects, (2) concurrent validity with severity of disease, and (3) the sensitivity/specificity of separating people with or without postural response deficits. Subjects. Ninety-seven people with PD were tested for balance deficits using the Berg, Mini-BESTest, Unified Parkinson's Disease Rating Scale (UPDRS) III and the Hoehn & Yahr (H&Y) disease severity classification. Setting. Clinical research facility at Oregon Health & Science University. Results. The Mini-BESTest is highly correlated with the Berg (r = 0.79, P < 0.001), but avoids the ceiling compression effect of the Berg for mild PD (skewness −2.30 Berg, −0.93 Mini-BESTest). Consequently, the Mini-BESTest is more effective than the Berg for predicting UPDRS Motor score (P < 0.001 Mini-BESTest versus P = 0.86 Berg), and for discriminating between those with and without postural response deficits as measured by the H&Y (ROC differential P = 0.06). Conclusion. The Mini-BESTest is a promising tool for discerning balance deficits in patients with PD, most importantly those with more subtle deficits

A case of rhinolithiasis

Rhinolith is an uncommon nasal mass in children and adolescents. We report a 8 years old boy with a history of long standing foul smelling right sided nasal discharge, progressive nasal obstruction and recurrent epistaxis, which was clinically suspected as being due to the presence of a long standing foreign body. Rhinolith in the right nasal cavity was diagnosed on rigid endoscopic examination. Rhinolith was removed by using a 00 rigid nasal endoscope.Key Words: Rhinolith, Endoscop

Red cell distribution width elevation and sepsis in pediatric critically ill patients

Background: Recently, a relationship has been demonstrated between red blood cell distribution width (RDW) and mortality risk in critically ill patients although the exact mechanism of this association is still vague. However, the impact of changes in RDW on sepsis and its outcome in critically ill patients has not been widely studied. Therefore, we studied the prognostic impact of changes in RDW in critically ill pediatric patients with sepsis. Methods: A total of 304 patients who were admitted to pediatric intensive care unit were selected to participate in this study. The changes in RDW on the day of admission and 4 and 8 days after admission in PICU were documented and their relationship with SIRS positivity, sepsis, and mortality were analyzed. Results: The mortality rate in our patients was 10.5. In total, 39.8 of patients were SIRS positive and 50.4 fulfilled the criteria of sepsis. The mean of RDW at the time of admission, on Day 4 and on Day 8 of admission was 14.8, 16.1, and 16.6, respectively. At the time of admission, RDW had a significant correlation with mortality and SIRS positivity, but RDW measured on Days 4 and 8 of admission did not correlate with neither of them. Neither of RDW0, RDW4, nor RDW8 did correlate with sepsis criteria fulfillment. �RDW day 4-adm &gt; 0.2, �RDW day 8-adm &gt; 0.2, �RDW day 8-day 4 &gt; 0.2 exhibited no correlation with SIRS positivity, sepsis, and mortality. Conclusions:We found that an increase in RDW from baseline during the first 4 and 8 days after admission of critically ill pediatric patients did not correlate with their mortality, SIRS positivity, and sepsis. However, elevated baseline RDW is a valuable prognostic marker in patients with sepsis. © 2018, Archives of Pediatric Infectious Diseases

Red cell distribution width elevation and sepsis in pediatric critically ill patients

Background: Recently, a relationship has been demonstrated between red blood cell distribution width (RDW) and mortality risk in critically ill patients although the exact mechanism of this association is still vague. However, the impact of changes in RDW on sepsis and its outcome in critically ill patients has not been widely studied. Therefore, we studied the prognostic impact of changes in RDW in critically ill pediatric patients with sepsis. Methods: A total of 304 patients who were admitted to pediatric intensive care unit were selected to participate in this study. The changes in RDW on the day of admission and 4 and 8 days after admission in PICU were documented and their relationship with SIRS positivity, sepsis, and mortality were analyzed. Results: The mortality rate in our patients was 10.5. In total, 39.8 of patients were SIRS positive and 50.4 fulfilled the criteria of sepsis. The mean of RDW at the time of admission, on Day 4 and on Day 8 of admission was 14.8, 16.1, and 16.6, respectively. At the time of admission, RDW had a significant correlation with mortality and SIRS positivity, but RDW measured on Days 4 and 8 of admission did not correlate with neither of them. Neither of RDW0, RDW4, nor RDW8 did correlate with sepsis criteria fulfillment. �RDW day 4-adm &gt; 0.2, �RDW day 8-adm &gt; 0.2, �RDW day 8-day 4 &gt; 0.2 exhibited no correlation with SIRS positivity, sepsis, and mortality. Conclusions:We found that an increase in RDW from baseline during the first 4 and 8 days after admission of critically ill pediatric patients did not correlate with their mortality, SIRS positivity, and sepsis. However, elevated baseline RDW is a valuable prognostic marker in patients with sepsis. © 2018, Archives of Pediatric Infectious Diseases

GRIDDS - A Gait Recognition Image and Depth Dataset

Several approaches based on human gait have been proposed in the literature, either for medical research reasons, smart surveillance, human-machine interaction, or other purposes, whose validation highly depends on the access to common input data through available datasets, enabling a coherent performance comparison. The advent of depth sensors leveraged the emergence of novel approaches and, consequently, the usage of new datasets. In this work we present the GRIDDS - A Gait Recognition Image and Depth Dataset, a new and publicly available gait depth-based dataset that can be used mostly for person and gender recognition purposes. (c) Springer Nature Switzerland AG 2019

Integrated analysis of gene expression profiles reveals deregulation of the immune response genes during different phases of chronic hepatitis B infection

© 2017, Kowsar Corp.Background: The natural history of chronic hepatitis B (CHB) infection is divided into different phases including immune tolerance (IT), immune clearance (or immune active [IA]), inactive carrier (IC), and reactivation. Despite utilizing high-throughput data, the distinct immunological mechanisms of these phases have been insufficiently investigated. Objectives: The aim of the present study was to determine candidate disease-associated genes and significantly altered biological processes for each phase of CHB infection. Methods: The gene expression profiles of 83 CHB patients (22 IT, 50 IA, and 11 IC phases) were obtained from gene expression omnibus (GEO dataset: GSE65359) and analyzed by bioinformatics tools. Several plugins of Cytoscape software were used to construct protein-protein interaction (PPI) networks and measure their topological properties. Subsequently, functional annotation and signaling pathway enrichment were carried out using the database for annotation, visualization and integrated discovery (DAVID) and Kyoto encyclopedia of genes and genomes (KEGG). Results: 449 and 452 deregulated genes were identified in IT-IA and IA-IC patients, respectively. Gene ontology and KEGG pathway analyses showed that several immune response-associated genes and signaling pathways (i.e. cytokine-cytokine receptor interaction, chemokine signaling pathway and T cell receptor signalling pathway) were upregulated in the IA phase, but downregulated in the IC phase. The LCK (encoding a tyrosine kinase) was determined as the most important hub gene of both constructed PPI networks. Furthermore, other immune response-associated genes such as CXCR3, VCAN, MYC, and STAT1 were found to be the important hub genes in clinical phases of CHB. Conclusions: The immune response-related pathways were found to be up and downregulated in the immune clearance phase and inactive carrier phase of CHB, respectively. The LCK hub gene might help the pathogenesis of different phases of CHB and serve as a therapeutic target for the treatment of hepatitis B virus

Regulatory network analysis of Epstein-Barr virus identifies functional modules and hub genes involved in infectious mononucleosis

© 2017, Springer-Verlag Wien.Epstein-Barr virus (EBV) is the most common cause of infectious mononucleosis (IM) and establishes lifetime infection associated with a variety of cancers and autoimmune diseases. The aim of this study was to develop an integrative gene regulatory network (GRN) approach and overlying gene expression data to identify the representative subnetworks for IM and EBV latent infection (LI). After identifying differentially expressed genes (DEGs) in both IM and LI gene expression profiles, functional annotations were applied using gene ontology (GO) and BiNGO tools, and construction of GRNs, topological analysis and identification of modules were carried out using several plugins of Cytoscape. In parallel, a human-EBV GRN was generated using the Hu-Vir database for further analyses. Our analysis revealed that the majority of DEGs in both IM and LI were involved in cell-cycle and DNA repair processes. However, these genes showed a significant negative correlation in the IM and LI states. Furthermore, cyclin-dependent kinase 2 (CDK2) – a hub gene with the highest centrality score – appeared to be the key player in cell cycle regulation in IM disease. The most significant functional modules in the IM and LI states were involved in the regulation of the cell cycle and apoptosis, respectively. Human-EBV network analysis revealed several direct targets of EBV proteins during IM disease. Our study provides an important first report on the response to IM/LI EBV infection in humans. An important aspect of our data was the upregulation of genes associated with cell cycle progression and proliferation

Discovering the typing behaviour of Parkinson's patients using topic models

Sensing health-related behaviours in an unobtrusive, ubiquitous and cost-effective manner carries significant benefits to healthcare and patient management. In this paper, we focus on detecting typing behaviour that is characteristic of patients suffering from Parkinson’s disease. We consider typing data obtained from subjects with and without Parkinson’s, and we present a framework based on topic models that determines the differing behaviours between these two groups based on the key hold time. By learning a topic model on each group separately and measuring the dissimilarity between topic distributions, we are able to identify particular topics that emerge in Parkinson’s patients and have low probability for the control group, demonstrating a clear shift in terms of key stroke duration. Our results further support the utilisation of key stroke logs for the early onset detection of Parkinson’s disease, while the method presented is straightforwardly generalisable to similar applications