Small Peptides Able to Suppress Prostaglandin E₂ Generation in Renal Mesangial Cells.

Peptide drug discovery may play a key role in the identification of novel medicinal agents. Here, we present the development of novel small peptides able to suppress the production of PGE₂ in mesangial cells. The new compounds were generated by structural alterations applied on GK115, a novel inhibitor of secreted phospholipase A₂, which has been previously shown to reduce PGE₂ synthesis in rat renal mesangial cells. Among the synthesized compounds, the tripeptide derivative 11 exhibited a nice dose-dependent suppression of PGE₂ production, similar to that observed for GK115

The orphan works problem and the future of intellectual property

The paper describes the orphan works problem and advocates putting into practice in the EU the ideas of Prof. Laurence Lessig of Stanford Law School who proposes to address the orphan works problem, along with the problem of the excessive duration of copyright, by going back to the idea of the original copyright and the subsequent copyright renewal embedded in the Statute of Anne. After the initial 14-year term of automatic protection, the rightholder would be required to register the work for a marginal fee to be grated another 14-year protection term. The system would result in the creation of a publicly searchable database, similar to those that exist under patent law.peer-reviewe

A Prokaryotic S1P Lyase Degrades Extracellular S1P In Vitro and In Vivo: Implication for Treating Hyperproliferative Disorders

Sphingosine-1-phosphate (S1P) regulates a broad spectrum of fundamental cellular processes like proliferation, death, migration and cytokine production. Therefore, elevated levels of S1P may be causal to various pathologic conditions including cancer, fibrosis, inflammation, autoimmune diseases and aberrant angiogenesis. Here we report that S1P lyase from the prokaryote Symbiobacterium thermophilum (StSPL) degrades extracellular S1P in vitro and in blood. Moreover, we investigated its effect on cellular responses typical of fibrosis, cancer and aberrant angiogenesis using renal mesangial cells, endothelial cells, breast (MCF-7) and colon (HCT 116) carcinoma cells as disease models. In all cell types, wild-type StSPL, but not an inactive mutant, disrupted MAPK phosphorylation stimulated by exogenous S1P. Functionally, disruption of S1P receptor signaling by S1P depletion inhibited proliferation and expression of connective tissue growth factor in mesangial cells, proliferation, migration and VEGF expression in carcinoma cells, and proliferation and migration of endothelial cells. Upon intravenous injection of StSPL in mice, plasma S1P levels rapidly declined by 70% within 1 h and then recovered to normal 6 h after injection. Using the chicken chorioallantoic membrane model we further demonstrate that also under in vivo conditions StSPL, but not the inactive mutant, inhibited tumor cell-induced angiogenesis as an S1P-dependent process. Our data demonstrate that recombinant StSPL is active under extracellular conditions and holds promise as a new enzyme therapeutic for diseases associated with increased levels of S1P and S1P receptor signaling

Ukraine: Electronic filing of tax returns

Oleksandr Pastukhov sets out the position in relation to the submission of electronic tax returns in the Ukraine, finally made possible once the state public key infrastructure was put into place and became effective in June 2009

International Taxation of Electronic Commerce

INTRODUCTION 6 PART I THE CURRENT INTERNATIONAL TAX PARADIGM 16 1. KEY DEFINITIONS 16 1.1. The notion of tax 16 1.2. Taxpayers 17 1.3. Types of taxes 18 1.3.1. Direct taxes 19 1.3.2. Indirect taxes 25 2. AN OVERVIEW OF THE CURRENT SYSTEM OF INTERNATIONAL TAXATION 27 2.1. Is there a system of international taxation? 27 2.2. Double taxation 30 2.3. International tax sharing: The 1920s compromise 33 2.4. Tax treaties 44 2.5. Principles of international taxation 51 3. THE (HARMFUL) TAX COMPETITION PHENOMENON 53 3.1. The disappearing taxpayer 53 3.2. In search of the definition 57 3.3. Tax havens 64 3.4. How harmful is it? 69 4. CONCLUSIONS TO THE PART 70 PART II THE TECHNOLOGICAL AND BUSINESS ASPECTS OF ELECTRONIC COMMERCE THAT CAUSE TAX PROBLEMS 76 1. THE NATURE OF ELECTRONIC COMMERCE AND THE WAYS IT GENERATES INCOME 76 1.1. The phenomenon of electronic commerce 76 1.1.1. Background 76 1.1.2. Definition 78 1.2. What is new about electronic commerce? 80 1.2.1. Speed 80 1.2.2. Interactivity 82 1.2.3. Electronic payment 83 1.2.4. Peculiarities of the Internet economy 84 1.3. Types of electronic commerce 86 1.4. How it works: A typical online business model 89 2. THE WAYS THE INTERNET UNDERMINES TRADITIONAL TAX CONCEPTS 91 2.1. Absence of central operation control and knowledge of the information transmitted 91 2.2. Trans-border character 93 2.3. Automation and remote control 94 2.4. Lack of physical presence 96 2.5. Digital products 97 2.6. No need in intermediaries 99 2.7. Unclear identity 101 2.8. Tax compliance 104 2.9. Tax administration 106 3. MARRIED IN HAVEN: ELECTRONIC COMMERCE AND OFFSHORE BUSINESS 109 3.1. Introduction to offshore electronic commerce 110 3.2. What can be sent offshore? 111 3.2.1. Digital sales 112 3.2.2. Physical distribution 113 3.2.3. Banking and other financial services 114 3.2.4. Corporate functions 116 4. FACTORS INFLUENCING THE DECISION TO (RE)LOCATE 120 4.1. Fiscal considerations 120 4.2. Non-tax aspects 125 4.2.1. Legal factors 126 4.2.2. Technical conditions 127 4.2.3. Skilled labor 129 4.2.4. Accessibility 130 4.2.5. Investment climate 130 4.3. Conclusions on (re)location 138 5. CONCLUSIONS TO THE PART 140 PART III DISTORTIONS TO THE SYSTEM OF INTERNATIONAL TAXATION CAUSED BY THE ADVENT OF ELECTRONIC COMMERCE 144 1. JURISDICTION TO TAX: THE CONCEPT OF NEXUS 144 2. DIRECT TAXATION 145 2.2. Permanent establishment in electronic commerce 145 2.2.1. Web server as a permanent establishment 145 2.2.2. Permanent establishment by agency 157 2.2.3. Conclusions on permanent establishment in the electronic commerce context 162 2.3. Attribution of profits to a Web server 163 2.4. Capital mobility 166 2.5. Income characterization 167 2.6. Disintermediation 171 2.7. Profit allocation 172 3. INDIRECT TAXATION 178 3.1. Generally 178 3.2. Customs duties 181 4. HOW ELECTRONIC COMMERCE AGGRAVATES THE PROBLEM OF HARMFUL TAX COMPETITION 187 4.1. Corporate tax aspects of offshore electronic commerce 187 4.2. Offshore electronic commerce and competition in turnover taxes 189 5. MEASURES AIMED AT CURBING HARMFUL TAX COMPETITION 195 5.1. National Measures 195 5.2. Bilateral Measures 198 5.3. Regional measures (exemplified by the EU) 199 5.3.1. Tax competition in the Common Market 199 5.3.2. The EU tax harmonization initiatives 204 5.3.3. The Code of Conduct 206 5.3.4. Other measures designed to counteract harmful tax competition 210 5.3.5. The role of the European Court of Justice 213 5.3.6. The future of the EU efforts aimed at restraining harmful tax competition 221 5.4. International measures 224 6. WHY A CRACKDOWN ON HARMFUL TAX PRACTICES MIGHT NOT WORK AND WHAT SHOULD BE DONE INSTEAD 238 7. CONCLUSIONS TO THE PART 245 PART IV PROPOSALS FOR CHANGES TO THE SYSTEM OF INTERNATIONAL TAXATION IN RESPONSE TO THE RISE OF ELECTRONIC COMMERCE AND PROBLEMS OF THEIR IMPLEMENTATION 250 1. LEGAL SOLUTIONS PROPOSED AND PROBLEMS ASSOCIATED WITH IMPLEMENTATION THEREOF 250 1.1. Conservative approach: Preserving the current mix of residence- and source-based taxation 251 1.2. Evolutionary approach: Switching to residence-based taxation 253 1.3. Revolutionary ideas: Switching the emphasis to consumption-based taxation, changing transfer pricing rules, sui generis taxes and no taxes at all 256 1.3.1. Shifting to consumption-based taxation 257 1.3.2. Capturing Internet tax revenues through transfer pricing rules 259 1.3.3. “Bit tax” 260 1.3.4. Other alternative taxes 263 1.3.5. Ban on Internet taxes 265 2. THE EFFECT OF TAXPAYERS’ EDUCATION AND ENCOURAGEMENT. 273 3. POSSIBLE TECHNOLOGY-ASSISTED SOLUTIONS 276 3.1. Technology as a regulation tool 276 3.2. Identifying the location of taxpayers and the place of consumption 279 3.3. International tax extranet 281 3.4. International online tax clearinghouse 288 4. THE LAISSEZ-FAIRE APPROACH AND SELF-REGULATION 292 4.1. Free market solutions 292 4.2. Self-regulation 294 4.2.1. User self-regulation 296 4.2.2. Industry self-regulation 299 4.3. The new lex mercatoria 301 4.4. Government still has a role to play 306 5. THE NEED FOR GREATER INTERNATIONAL COOPERATION IN TAX MATTERS 312 6. STAKEHOLDERS’ ATTITUDES TOWARDS POSSIBLE CHANGES IN THE STATUS QUO 314 6.1. Governments 314 6.2. Business generally 316 6.3. Small and medium-sized enterprises 321 6.4. Developing countries 325 7. OPPORTUNITIES ELECTRONIC COMMERCE CREATES FOR TAX AUTHORITIES 327 7.1. Streamlining tax administration 328 7.2. The OECD guidelines 329 7.3. The Streamlined Sales Tax Project 332 7.4. The E-Invoicing Directive 340 8. CONCLUSIONS TO THE PART 342 OVERALL CONCLUSIONS AND RECOMMENDATIONS 346 BIBLIOGRAPHY 360status: publishe

CRISPR-Directed Therapeutic Correction at the NCF1 Locus Is Challenged by Frequent Incidence of Chromosomal Deletions

Resurrection of non-processed pseudogenes may increase the efficacy of therapeutic gene editing, upon simultaneous targeting of a mutated gene and its highly homologous pseudogenes. To investigate the potency of this approach for clinical gene therapy of human diseases, we corrected a pseudogene-associated disorder, the immunodeficiency p47 phox -deficient chronic granulomatous disease (p47 phox CGD), using clustered regularly interspaced short palindromic repeats-associated nuclease Cas9 (CRISPR-Cas9) to target mutated neutrophil cytosolic factor 1 (NCF1). Being separated by less than two million base pairs, NCF1 and two pseudogenes are closely co-localized on chromosome 7. In healthy people, a two-nucleotide GT deletion (ΔGT) is present in the NCF1B and NCF1C pseudogenes only. In the majority of patients with p47 phox CGD, the NCF1 gene is inactivated due to a ΔGT transfer from one of the two non-processed pseudogenes. Here we demonstrate that concurrent targeting and correction of mutated NCF1 and its pseudogenes results in therapeutic CGD phenotype correction, but also causes potentially harmful chromosomal deletions between the targeted loci in a p47 phox -deficient CGD cell line model. Therefore, development of genome-editing-based treatment of pseudogene-related disorders mandates thorough safety examination, as well as technological advances, limiting concurrent induction of multiple double-strand breaks on a single chromosome

Small Peptides Able to Suppress Prostaglandin E2 Generation in Renal Mesangial Cells

Peptide drug discovery may play a key role in the identification of novel medicinal agents. Here, we present the development of novel small peptides able to suppress the production of PGE2 in mesangial cells. The new compounds were generated by structural alterations applied on GK115, a novel inhibitor of secreted phospholipase A2, which has been previously shown to reduce PGE2 synthesis in rat renal mesangial cells. Among the synthesized compounds, the tripeptide derivative 11 exhibited a nice dose-dependent suppression of PGE2 production, similar to that observed for GK115

Ceramide Kinase Contributes to Proliferation but not to Prostaglandin E2 Formation in Renal Mesangial Cells and Fibroblasts

Background/Aims: Ceramide kinase (CerK) catalyzes the generation of the sphingolipid ceramide-1-phosphate (C1P) which regulates various cellular functions including cell growth and death, and inflammation. Here, we used a novel catalytic inhibitor of CerK, NVP-231, and CerK knockout cells to investigate the contribution of CerK to proliferation and inflammation in renal mesangial cells and fibroblasts. Methods: Cells were treated with NVP-231 and [3H]-thymidine incorporation into DNA, [3H]-arachidonic acid release, prostaglandin E2 (PGE2) synthesis, cell cycle distribution, and apoptosis were determined. Results: Treatment of rat mesangial cells and mouse renal fibroblasts with NVP-231 decreased DNA synthesis, but not of agonist-stimulated arachidonic acid release or PGE2 synthesis. Similarly, proliferation but not arachidonic acid release or PGE2 synthesis was reduced in CERK knockout renal fibroblasts. The anti-proliferative effect of NVP-231 on mesangial cells was due to M phase arrest as determined using the mitosis markers phospho-histone H3, cdc2 and polo-like kinase-1, and induction of apoptosis. Moreover, loss of CerK sensitized cells towards stress-induced apoptosis. Conclusions: Our data demonstrate that CerK induces proliferation but not PGE2 formation of renal mesangial cells and fibroblasts, and suggest that targeted CerK inhibition has potential for treating mesangioproliferative kidney diseases

Ceramide Kinase Contributes to Proliferation but not to Prostaglandin E₂ Formation in Renal Mesangial Cells and Fibroblasts

Background/Aims: Ceramide kinase (CerK) catalyzes the generation of the sphingolipid ceramide-1-phosphate (C1P) which regulates various cellular functions including cell growth and death, and inflammation. Here, we used a novel catalytic inhibitor of CerK, NVP-231, and CerK knockout cells to investigate the contribution of CerK to proliferation and inflammation in renal mesangial cells and fibroblasts. Methods: Cells were treated with NVP-231 and [3H]-thymidine incorporation into DNA, [3H]-arachidonic acid release, prostaglandin E2 (PGE2) synthesis, cell cycle distribution, and apoptosis were determined. Results: Treatment of rat mesangial cells and mouse renal fibroblasts with NVP-231 decreased DNA synthesis, but not of agonist-stimulated arachidonic acid release or PGE2 synthesis. Similarly, proliferation but not arachidonic acid release or PGE2 synthesis was reduced in CERK knockout renal fibroblasts. The anti-proliferative effect of NVP-231 on mesangial cells was due to M phase arrest as determined using the mitosis markers phospho-histone H3, cdc2 and polo-like kinase-1, and induction of apoptosis. Moreover, loss of CerK sensitized cells towards stress-induced apoptosis. Conclusions: Our data demonstrate that CerK induces proliferation but not PGE2 formation of renal mesangial cells and fibroblasts, and suggest that targeted CerK inhibition has potential for treating mesangioproliferative kidney diseases