Deep Sequencing in Pre- and Clinical Vaccine Research

Vaccine research has experienced a quantum leap after the beginning of the genomics era. High-throughput sequencing techniques, unlimited computing resources, as well as new bioinformatic algorithms are now changing the way we perform genomic studies. Whole genome sequencing will soon become the gold standard for phylogenetic and epidemiology studies and is already shedding new light on the dynamics of bacterial evolution. We believe that deep sequencing projects, together with structural studies on vaccine candidates, will allow targeting constant epitopes and avoid vaccine failure due to antigenic variability. Systems biology, which is expected to revolutionize vaccine research and clinical studies, greatly relies on high-throughput technologies such as RNA-seq. Furthermore, genomics is a key element to develop safer vaccines, and the accuracy of deep sequencing will allow monitoring vaccine coverage after their introduction on the market

Syndecan 4 Is Involved in Mediating HCV Entry through Interaction with Lipoviral Particle-Associated Apolipoprotein E

Hepatitis C virus (HCV) is a major cause of liver disease worldwide and HCV infection represents a major health problem. HCV associates with host lipoproteins forming host/viral hybrid complexes termed lipoviral particles. Apolipoprotein E (apoE) is a lipoprotein component that interacts with heparan sulfate proteoglycans (HSPG) to mediate hepatic lipoprotein uptake, and may likewise mediate HCV entry. We sought to define the functional regions of apoE with an aim to identify critical apoE binding partners involved in HCV infection. Using adenoviral vectors and siRNA to modulate apoE expression we show a direct correlation of apoE expression and HCV infectivity, whereas no correlation exists with viral protein expression. Mutating the HSPG binding domain (HSPG-BD) of apoE revealed key residues that are critical for mediating HCV infection. Furthermore, a novel synthetic peptide that mimics apoE's HSPG-BD directly and competitively inhibits HCV infection. Genetic knockdown of the HSPG proteins syndecan (SDC) 1 and 4 revealed that SDC4 principally mediates HCV entry. Our data demonstrate that HCV uses apoE-SDC4 interactions to enter hepatoma cells and establish infection. Targeting apoE-SDC interactions could be an alternative strategy for blocking HCV entry, a critical step in maintaining chronic HCV infection

Soluble Serum CD81 Is Elevated in Patients with Chronic Hepatitis C and Correlates with Alanine Aminotransferase Serum Activity

Aim: Cellular CD81 is a well characterized hepatitis C virus (HCV) entry factor, while the relevance of soluble exosomal CD81 in HCV pathogenesis is poorly defined. We performed a case-control study to investigate whether soluble CD81 in the exosomal serum fraction is associated with HCV replication and inflammatory activity. Patients and Methods: Four cohorts were investigated, patients with chronic hepatitis C (n = 37), patients with chronic HCV infection and persistently normal ALT levels (n = 24), patients with long term sustained virologic response (SVR, n = 7), and healthy volunteers (n = 23). Concentration of soluble CD81 was assessed semi-quantitatively after differential centrifugation ranging from 200 g to 100,000 g in the fifth centrifugation fraction by immunoblotting and densitometry. Results: Soluble CD81 was increased in patients with chronic hepatitis C compared to healthy subjects (p = 0.03) and cured patients (p = 0.017). Patients with chronic HCV infection and persistently normal ALT levels and patients with long term SVR had similar soluble CD81 levels as healthy controls (p>0.2). Overall, soluble CD81 levels were associated with ALT levels (r = 0.334, p = 0.016) and severe liver fibrosis (p = 0.027). Conclusion: CD81 is increased in the exosomal serum fraction in patients with chronic hepatitis C and appears to be associated with inflammatory activity and severity of fibrosis

Hepatocyte Permissiveness to Plasmodium Infection Is Conveyed by a Short and Structurally Conserved Region of the CD81 Large Extracellular Domain

Invasion of hepatocytes by Plasmodium sporozoites is a prerequisite for establishment of a malaria infection, and thus represents an attractive target for anti-malarial interventions. Still, the molecular mechanisms underlying sporozoite invasion are largely unknown. We have previously reported that the tetraspanin CD81, a known receptor for the hepatitis C virus (HCV), is required on hepatocytes for infection by sporozoites of several Plasmodium species. Here we have characterized CD81 molecular determinants required for infection of hepatocytic cells by P. yoelii sporozoites. Using CD9/CD81 chimeras, we have identified in CD81 a 21 amino acid stretch located in a domain structurally conserved in the large extracellular loop of tetraspanins, which is sufficient in an otherwise CD9 background to confer susceptibility to P. yoelii infection. By site-directed mutagenesis, we have demonstrated the key role of a solvent-exposed region around residue D137 within this domain. A mAb that requires this region for optimal binding did not block infection, in contrast to other CD81 mAbs. This study has uncovered a new functionally important region of CD81, independent of HCV E2 envelope protein binding domain, and further suggests that CD81 may not interact directly with a parasite ligand during Plasmodium infection, but instead may regulate the function of a yet unknown partner protein

El nacionalismo musical en Manuel de Falla

Colofó

Anuario nº1. Panorama histórico del litoral argentino, 1516-1820

• F. Adolfo Masciopinto: Historia del Litoral Argentino • Luis Alberto Candioti: Significación y proyección histórica del Litoral Argentino • Pierina Pasotti: Paisajes del Litoral • Antonio Serrano: Panorama étnico del Litoral en el siglo XVI • F. Adolfo Masciopinto: La epopeya en la conquista del Litoral • Leoncio Gianello: La acción pobladora en nuestro Litoral • Agustín Zapata Gollán: Las ruinas de la primitiva ciudad de Santa Fe. Las tres Iglesias. • Ricardo Orta Nadal: Un aspecto de la historiografía y etnología jesuítica del Litoral • Héctor A. Di Bitetti: Los arquitectos jesuitas Primoli y Bianchi en el Litoral • Owen Usinger: La economía del Litoral • Francisco Villamil: La sociedad del Litoral • José Carmelo Busaniche: Los pueblos libres del Litoral y la estructura virreinal • Actividades del Instituto de Investigaciones Históricas durante 1952Fil: Masciopinto, Adolfo F. Instituto de Investigaciones Históricas. Facultad de Filosofía, Letras y Ciencias de la Educación. Universidad del Litoral. Rosario, Argentin

A Decision Support System for Artificial Recharge Plant

The need for artificial recharge plants is the result of the qualitative and quantitative worsening of groundwater resources due to increased pumping and wastewater discharge. This paper described a system that uses artificial intelligence techniques for designing an artificial recharge plant. The system can be used as a training tool for new engineers, as well as an aid in the choices for expert engineers. The system is an application of an expert system shell running on common p.c. machine. The model is made up of two knowledge bases, respectively denoted as Quantity artificial recharge and Quality artificial recharge. The former is related to the quantitative aspects, such as geology, climate and land availability, the latter to qualitative aspects, such as water use and treatment plant. Two case studies have been implemented in order to confirm the validity of this kind of systemic approach. The need for artificial recharge plants is the result of the qualitative and quantitative worsening of groundwater resources due to increased pumping and wastewater discharge. This paper described a system that uses artificial intelligence techniques for designing an artificial recharge plant. The system can be used as a training tool for new engineers, as well as an aid in the choices for expert engineers. The system is an application of an expert system shell running on a common p.c. machine. The model is made up of two knowledge bases, respectively denoted as Quantity artificial recharge and Quality artificial recharge. The former is related to the quantitative aspects, such as geology, climate and land availability, the latter to qualitative aspects, such as water use and treatment plant. Two case studies have been implemented in order to confirm the validity of this kind of systemic approach

Shooting the Numerical Solution of Moisture Flow Equation with Root Water Uptake Models: A Python Tool

Modeling the water uptake by plant roots is a key issue in studying soil processes, which are governed by water dynamics: a comprehensive understanding and forecast of such dynamics is a relevant issue in managing water resources. Typically, movement of water in soils and uptake by roots are described by the Richards’ equation with a sink term, and numerical treatment of this problem is still a challenge, together with its practical implementations in user-friendly softwares. In order to tackle this problem, in the present paper we propose a simple and computationally fast algorithm developed as a Python code, implementing a numerical approach based on the shooting method, a classical tool for handling boundary value problems (BVPs) arising here from a discretization recently introduced for Richards’ equation: such a method is applied to the linearized Richards’ equation with Gardner’s hydraulic functions. This method is implemented also in MATLAB, in order to accomplish comparisons with built-in MATLAB solver for parabolic partial differential equations. The Python code is made available to readers, and is intended to be an easy tool for handling this problem in the framework of Gardner’s constitutive relations, filling the gap of other commercial codes, which do not provide choice of Gardner functions. Many numerical simulations are performed: the results are promising, since the proposed method behaves efficiently and in some cases it is able to converge even when the MATLAB solver fails; mass balance properties and order of accuracy issues are also investigated

Current Surgical Therapy of Locally Advanced cSCC: From Patient Selection to Microsurgical Tissue Transplant. Review

Among the non-melanoma skin cancers (NMSC) the squamous cell carcinoma (SCC) is one of the most challenging for the surgeon. Local aggressiveness and a tendency to metastasize to regional lymph nodes characterize the biologic behavior. The variants locally advanced and metastatic require wide excision and node dissection. Such procedures can be extremely detrimental for patients. The limit of the surgery can be safely pushed forward with a multidisciplinary approach. The concept of skin oncoplastic surgery, the ablative procedures and the reconstructive options (skin graft, pedicled flap, microsurgical free flap) are discussed together with a literature review

Neonatal anesthesia for major thoracic surgery

Neonatal anesthesia for major thoracic surger