HIV Reverse Transcriptase Fidelity And Inhibition Are Modulated By Divalent Cations In A Concentration-Dependent Manner In Vitro

Human immunodeficiency virus (HIV) rapidly evolves through generation and selection of mutants that can escape drug therapy. This process is fueled, in part, by the presumably highly error prone polymerase reverse transcriptase (RT). Fidelity of polymerases can be influenced by cation co-factors. Physiologically, magnesium (Mg2+) is used as a co-factor by RT to perform catalysis, however, alternative cations including manganese (Mn2+), cobalt (Co2+), and zinc (Zn2+) can also be used. I demonstrate here that fidelity and inhibition of HIV RT can be influenced differently, in vitro, by divalent cations depending on their concentration. The reported mutation frequency for purified HIV RT in vitro is typically in the 10-4 range (per nucleotide addition), making the enzyme several-fold less accurate than most polymerases. Paradoxically, results examining HIV replication in cells indicate an error frequency that is ~10 times lower than the error rate obtained in the test tube. Here, I reconcile, at least in part, these discrepancies by showing that HIV RT fidelity in vitro is in the same range as cellular results, in physiological concentrations of free Mg2+ (~0.25 mM). At low Mg2+, mutation rates were 5-10 times lower compared to high Mg2+ conditions (5-10 mM). Alternative divalent cations also have a concentration-dependent effect on RT fidelity. Presumed promutagenic cations Mn2+ and Co2+ decreases the fidelity of RT only at elevated concentrations, and Zn2+, when present in low concentration, increases the fidelity of HIV-1 RT by ~2.5 fold compared to Mg2+. HIV-1 and HIV-2 RT inhibition by nucleoside (NRTIs) and non-nucleoside RT inhibitors (NNRTIs) in vitro is also affected by the Mg2+ concentration. NRTIs lacking 3'-OH group inhibited both enzymes less efficiently in low Mg2+ than in high Mg2+; whereas inhibition by the “translocation defective RT inhibitor”, which retains the 3ʹ-OH, was unaffected by Mg2+ concentration, suggesting that NRTIs with a 3ʹ-OH group may be more potent than other NRTIs. In contrast, NNRTIs were more effective in low vs. high Mg2+ conditions. Overall, the studies presented reveal strategies for designing novel RT inhibitors and strongly emphasize the need for studying HIV RT and RT inhibitors in physiologically relevant low Mg2+ conditions.

Corrosion of stainless steel components in seawater reverse osmosis desalination plants-investigations on adapted internal cathodic protection

Stainless steel is widely used in seawater reverse osmosis units (SWRO) for both good mechanical and corrosion resistance properties. However, many corrosion failures of stainless steel in SWRO desalination units have been reported. These failures may often be attributed to un-adapted stainless steel grade selection and/or to the particular aggressive seawater conditions in "warm" regions (high ambient temperature, severe biofouling, etc.). Cathodic protection (CP) is a well-known efficient system to prevent corrosion of metallic materials in seawater. It is successfully used in the oil and gas industry to protect carbon steel structures exposed in open-sea. However, the specific service conditions of SWRO units may seriously affect the efficiency of such anti-corrosion system (high flow rates, large stainless steel surfaces affected by biofouling, confinement limiting protective cathodic current flow, etc.). Hence, CP in SWRO units should be considered with special care and modeling appears as useful tool to assess an appropriate CP design. However, there is a clear lack of CP data that could be transposed to SWRO service conditions (i.e. stainless steel, effect of biofouling, high flow rate, etc.). From this background a Join Industry Program was initiated including laboratory exposures, field measurements in a full scale SWRO desalination plant, and modeling work using PROCOR software. The present paper reviews the main parameters affecting corrosion of stainless steel alloys in seawater reverse osmosis units. CP on specific stainless steel devices was investigated in order to assess its actual efficiency for SWRO units. Severe environmental conditions were intentionally used to promote corrosion on the tested stainless steel products in order to evaluate the efficiency of CP. The study includes a modeling work aiming at predicting and designing adapted CP protection to modeled stainless steel units. An excellent correlation between modeling work and field measurements was found.

Integration of Social Concerns in Collaborative Logistics and Transportation Networks

Part 21: Mobility and Logistics

Forward - Green Virtual Enterprises and Their Breeding Environments: Sustainable Manufacturing, Logistics and Consumption

Part 9: Innovation Networks

A real-time reverse transcriptase polymerase chain reaction for detection and quantification of Vesiculovirus

Vesiculoviruses (VSV) are zoonotic viruses that cause vesicular stomatitis disease in cattle, horses and pigs, as well as sporadic human cases of acute febrile illness. Therefore, diagnosis of VSV infections by reliable laboratory techniques is important to allow a proper case management and implementation of strategies for the containment of virus spread. We show here a sensitive and reproducible real-time reverse transcriptase polymerase chain reaction (RT-PCR) for detection and quantification of VSV. The assay was evaluated with arthropods and serum samples obtained from horses, cattle and patients with acute febrile disease. The real-time RT-PCR amplified the Piry, Carajas, Alagoas and Indiana Vesiculovirus at a melting temperature 81.02 ± 0.8ºC, and the sensitivity of assay was estimated in 10 RNA copies/mL to the Piry Vesiculovirus. The viral genome has been detected in samples of horses and cattle, but not detected in human sera or arthropods. Thus, this assay allows a preliminary differential diagnosis of VSV infections.

Impaired reverse cholesterol transport and hepatic steatosis contribute to pathogenesis of high fat dietinduced hyperlipidemia in murine models

Purpose: To investigate the pathogenesis of high fat diet (HFD)-induced hyperlipidemia (HLP) in mice, rats and hamsters and to comparatively evaluate their sensitivity to HFD. Methods: Mice, rats and hamsters were fed with high-fat diet formulation (HFD, n = 8) or a control diet (control, n = 8) for 4 weeks. Changes in body weight, relative liver weight, serum lipid profile, expressions of hepatic marker gene of lipid metabolism and liver morphology were observed in three hyperlipidemic models. Results: Elevated total cholesterol (TC), triglyceride, low density lipoprotein-cholesterol (LDL-C) and high density lipoprotein-cholesterol (HDL-C) levels and body weight were observed in all hyperlipidemic animals (p < 0.05), while hepatic steatosis was manifested in rat and hamster HLP models, and increased hepatic TC level was only seen (p < 0.05) in hamster HLP model. Suppression of HMG-CoA reductase and up-regulation of lipoproteinlipase were observed in all HFD groups. Hepatic gene expression of LDLR, CYP7A1, LCAT, SR-B1, and ApoA I, which are a response to reverse cholesterol transport (RCT), were inhibited by HFD in the three models. Among these models, simultaneous suppression of HMG-CR, LCAT, LDLR and SR-BI and elevated LPL were features of the hamster model. Conclusion: As the results show, impaired RCT and excessive fat accumulation are major contributors to pathogenesis of HFD-induced murine HLP. Thus, the hamster model is more appropriate for hyperlipidemia research.

Culture shock and reverse culture shock: the moderating role of cultural intelligence in international students' adaptation

This study extends previous research by examining the role of cultural intelligence (CQ) in both culture shock and reverse culture shock. Specifically, this study asserts that CQ acts as a moderating mechanism that lessens the negative effects of both culture shock and reverse culture shock on psychological and sociocultural adaptation among international students. Two studies were conducted in Australia to test these assertions. Study 1 (n = 189) was participated in by new international students. An online survey was set up and disseminated. Results indicated that culture shock is significantly but negatively related to both psychological and sociocultural adaptation. In addition, results demonstrated that CQ moderates the relationship by lessening the impact of culture shock on students’ psychological and sociocultural adaptation. Study 2 (n = 123) was participated in by international students who had recently graduated and returned to their home countries. An online survey was also set up and disseminated. Results indicated that reverse culture shock is significantly but negatively related to both psychological and sociocultural adaptation. CQ also served as a moderator in lessening the impact of reverse culture shock on both forms of adaptation.

Hybrid threshold adaptable quantum secret sharing scheme with reverse Huffman-Fibonacci-tree coding

With prevalent attacks in communication, sharing a secret between communicating parties is an ongoing challenge. Moreover, it is important to integrate quantum solutions with classical secret sharing schemes with low computational cost for the real world use. This paper proposes a novel hybrid threshold adaptable quantum secret sharing scheme, using an m-bonacci orbital angular momentum (OAM) pump, Lagrange interpolation polynomials, and reverse Huffman-Fibonacci-tree coding. To be exact, we employ entangled states prepared by m-bonacci sequences to detect eavesdropping. Meanwhile, we encode m-bonacci sequences in Lagrange interpolation polynomials to generate the shares of a secret with reverse Huffman-Fibonacci-tree coding. The advantages of the proposed scheme is that it can detect eavesdropping without joint quantum operations, and permits secret sharing for an arbitrary but no less than threshold-value number of classical participants with much lower bandwidth. Also, in comparison with existing quantum secret sharing schemes, it still works when there are dynamic changes, such as the unavailability of some quantum channel, the arrival of new participants and the departure of participants. Finally, we provide security analysis of the new hybrid quantum secret sharing scheme and discuss its useful features for modern applications.

Silica fouling in high salinity waters in reverse osmosis desalination (sodium-silica system)

Silica fouling patterns in a sodium–silica system and the effect of pH on residual dissolved silica concentrations are reported. The unique chemical affinity between sodium and silica (SO4) prevented silica scale deposition on the membrane surface during reverse osmosis (RO) desalination. It was found that high concentrations of sodium in solutions depressed silica solubility to 81–84 mg L−1 for a maximum NaCl salinity of 60–65 g L−1. Using a range of membrane examination techniques, it was found that no silica scale formed on the RO membrane surfaces from NaCl solutions free from cations such as Ca, Al and Fe. This was considered to be the result of sodium ions acting as a barrier between polymeric silica and the membrane surface.

Reverse convertible bonds

I convertible bonds sono degli strumenti finanziari che conferiscono al suo possessore la facoltà di scegliere se, una volta scaduta l'obbligazione, essere rimborsato tramite una somma di denaro (valore nominale) oppure convertire l'obbligazione in un numero predefinito di azioni. Successivamente si è trattato l’argomento della tesi, i reverse convertible bonds. Queste obbligazioni sono simili ai convertible bonds con la differenza che in tal caso il diritto di scegliere se convertire o meno l'obbligazione in azioni è lasciato all'emittente e non al sottoscrittore. I reverse convertible si ottengono dalla combinazione di un coupon bond ordinario (senza l'opzione di conversione) a breve termine e di un'opzione put sulle azioni sottostanti. E' stata analizzata la formula di valutazione dei reverse convertible bonds, data dalla differenza tra il prezzo di un coupon-bond ordinario emesso dalla stessa società e il prezzo di un'opzione put (quest'ultimo moltiplicato per il rapporto di conversione, ossia per il numero di azioni che si ottengono dalla conversione di ciascuna obbligazione convertibile). E’ stata poi fatta un’analisi empirica dei prezzi dei reverse convertible bonds. Sono stati calcolati i prezzi di 7 reverse convertible utilizzando i dati forniti dal database finanziario e macroeconomico, Thomson Reuters Datastream.I prezzi calcolati sono stati poi confrontati con i prezzi di mercato di tali obbligazioni relativamente allo stesso istante temporale, il giorno 6/6/2016.

Tecniche geomatiche di reverse engineering nel campo dei Beni Culturali

La salvaguardia e conservazione del Patrimonio Artistico ed Architettonico rappresentano un aspetto imprescindibile di ogni cultura, e trovano le loro fondamenta nella coscienza e conoscenza dei Beni. Il rilievo è l’operazione basilare per acquisire una conoscenza rigorosa di un oggetto nella sua geometria e in altre sue caratteristiche. Le finalità delle operazioni di rilevamento sono molteplici, dall’archiviazione a scopo di documentazione fino all’indagine conservativa volta alla diagnostica e alla progettazione di interventi. I modelli digitali, introdotti dallo sviluppo tecnologico degli ultimi decenni, permettono una perfetta conoscenza del bene, non necessitano di contatto diretto durante la fase di rilevamento e possono essere elaborati secondo le esigenze del caso. Le tecniche adottate nel Reverse Engineering si differenziano per il tipo di sensore utilizzato: quelle fotogrammetriche utilizzano sensori di tipo “passivo” e trovano oggi largo impiego nel settore dei Beni Culturali grazie agli strumenti di Structure from Motion, mentre strumenti basati su sensori di tipo “attivo” utilizzano Laser o proiezione di luce strutturata e sono in grado di rilevare con grande precisione geometrie anche molto complesse. La costruzione del modello della fontana del Nettuno e della torre Garisenda di Bologna costituiscono un valido esempio di applicazione delle tecniche di rilievo digitale, e dimostrano la validità delle stesse su oggetti di diversa dimensione in due diversi ambiti applicativi: il restauro e il monitoraggio. Gli sviluppi futuri del Reverse Engineering in questo ambito sono molteplici, e la Geomatica rappresenta senza dubbio una disciplina fondamentale per poterli realizzare.

Distributed optimization in transportation and logistics networks

Many important problems in communication networks, transportation networks, and logistics networks are solved by the minimization of cost functions. In general, these can be complex optimization problems involving many variables. However, physicists noted that in a network, a node variable (such as the amount of resources of the nodes) is connected to a set of link variables (such as the flow connecting the node), and similarly each link variable is connected to a number of (usually two) node variables. This enables one to break the problem into local components, often arriving at distributive algorithms to solve the problems. Compared with centralized algorithms, distributed algorithms have the advantages of lower computational complexity, and lower communication overhead. Since they have a faster response to local changes of the environment, they are especially useful for networks with evolving conditions. This review will cover message-passing algorithms in applications such as resource allocation, transportation networks, facility location, traffic routing, and stability of power grids.

Conformational states of HIV-1 reverse transcriptase for nucleotide incorporation vs. pyrophosphorolysis – binding of foscarnet

HIV-1 reverse transcriptase (RT) catalytically incorporates individual nucleotides into a viral DNA strand complementing an RNA or DNA template strand; the polymerase active site of RT adopts multiple conformational and structural states while performing this task. The states associated are dNTP binding at the N site, catalytic incorporation of a nucleotide, release of a pyrophosphate, and translocation of the primer 3′-end to the P site. Structural characterization of each of these states may help in understanding the molecular mechanisms of drug activity and resistance and in developing new RT inhibitors. Using a 38-mer DNA template-primer aptamer as the substrate mimic, we crystallized an RT/dsDNA complex that is catalytically active, yet translocation-incompetent in crystals. The ability of RT to perform dNTP binding and incorporation in crystals permitted obtaining a series of structures: (I) RT/DNA (P-site), (II) RT/DNA/AZTTP ternary, (III) RT/AZT-terminated DNA (N-site), and (IV) RT/AZT-terminated DNA (N-site)/foscarnet complexes. The stable N-site complex permitted the binding of foscarnet as a pyrophosphate mimic. The Mg2+ ions dissociated after catalytic addition of AZTMP in the pretranslocated structure III, whereas ions A and B had re-entered the active site to bind foscarnet in structure IV. The binding of foscarnet involves chelation with the Mg2+ (B) ion and interactions with K65 and R72. The analysis of interactions of foscarnet and the recently discovered nucleotide-competing RT inhibitor (NcRTI) α-T-CNP in two different conformational states of the enzyme provides insights for developing new classes of polymerase active site RT inhibitors.

A dynamic pricing reverse auction-based resource allocation mechanism in cloud workflow systems

Market-oriented reverse auction is an efficient and cost-effective method for resource allocation in cloud workflow systems since it can dynamically allocate resources depending on the supply-demand relationship of the cloud market. However, during the auction the price of cloud resource is usually fixed, and the current resource allocation mechanisms cannot adapt to the changeable market properly which results in the low efficiency of resource utilization. To address such a problem, a dynamic pricing reverse auction-based resource allocation mechanism is proposed. During the auction, resource providers can change prices according to the trading situation so that our novel mechanism can increase the chances of making a deal and improve efficiency of resource utilization. In addition, resource providers can improve their competitiveness in the market by lowering prices, and thus users can obtain cheaper resources in shorter time which would decrease monetary cost and completion time for workflow execution. Experiments with different situations and problem sizes are conducted for dynamic pricing-based allocation mechanism (DPAM) on resource utilization and the measurement of Time∗Cost (TC). The results show that our DPAM can outperform its representative in resource utilization, monetary cost, and completion time and also obtain the optimal price reduction rates.

La influencia de la cultura nacional en el desempeño logístico de los 123 primeros países en el logistics performance index (LPI)

En esta tesis se presenta la influencia que tiene la Cultura Nacional en el desempeño Logístico de una Nación, en especial de los 123 primeros países evaluados por el Índice de Desempeño Logístico (LPI) realizado por el Banco Mundial. Este estudio se realiza a partir de los seis ítems evaluados en el LPI y de las seis Dimensiones Culturales de Hofstede, para relacionar estas calificaciones se empleará el programa estadístico SPSS Statistical Package for the Social Sciences obteniendo como resultado posibles correlaciones para probar la hipótesis del efecto de la cultura en el desempeño logístico y establecer patrones de comportamiento en los mejores países de cada correlación. De acuerdo a los resultados obtenidos de estas correlaciones, se encontró que si existe una relación entre la cultura nacional y el desempeño logístico de un país, al igual que las dimensiones culturales más influyentes para obtener dichos resultados y se identificaron los países que cuentan con condiciones sobresalientes que facilitan tener altas posiciones en el LPI.