In vitro synergy and enhanced murine brain penetration of saquinavir coadministered with mefloquine.

Highly active antiretroviral therapy has substantially improved prognosis in human immunodeficiency virus (HIV). However, the integration of proviral DNA, development of viral resistance, and lack of permeability of drugs into sanctuary sites (e.g., brain and lymphocyte) are major limitations to current regimens. Previous studies have indicated that the antimalarial drug chloroquine (CQ) has antiviral efficacy and a synergism with HIV protease inhibitors. We have screened a panel of antimalarial compounds for activity against HIV-1 in vitro. A limited efficacy was observed for CQ, mefloquine (MQ), and mepacrine (MC). However, marked synergy was observed between MQ and saquinavir (SQV), but not CQ in U937 cells. Furthermore, enhancement of the antiviral activity of SQV and four other protease inhibitors (PIs) by MQ was observed in MT4 cells, indicating a class specific rather than a drug-specific phenomenon. We demonstrate that these observations are a result of inhibition of multiple drug efflux proteins by MQ and that MQ also displaces SQV from orosomucoid in vitro. Finally, coadministration of MQ and SQV in CD-1 mice dramatically altered the tissue distribution of SQV, resulting in a >3-fold and >2-fold increase in the tissue/blood ratio for brain and testis, respectively. This pharmacological enhancement of in vitro antiviral activity of PIs by MQ now warrants further examination in vivo

An Iterative and Toolchain-Based Approach to Automate Scanning and Mapping Computer Networks

As today's organizational computer networks are ever evolving and becoming more and more complex, finding potential vulnerabilities and conducting security audits has become a crucial element in securing these networks. The first step in auditing a network is reconnaissance by mapping it to get a comprehensive overview over its structure. The growing complexity, however, makes this task increasingly effortful, even more as mapping (instead of plain scanning), presently, still involves a lot of manual work. Therefore, the concept proposed in this paper automates the scanning and mapping of unknown and non-cooperative computer networks in order to find security weaknesses or verify access controls. It further helps to conduct audits by allowing comparing documented with actual networks and finding unauthorized network devices, as well as evaluating access control methods by conducting delta scans. It uses a novel approach of augmenting data from iteratively chained existing scanning tools with context, using genuine analytics modules to allow assessing a network's topology instead of just generating a list of scanned devices. It further contains a visualization model that provides a clear, lucid topology map and a special graph for comparative analysis. The goal is to provide maximum insight with a minimum of a priori knowledge.Comment: 7 pages, 6 figure

Sequence and gene expression of chloroquine resistance transporter (pfcrt) in the association of in vitro drugs resistance of Plasmodium falciparum

<p>Abstract</p> <p>Background</p> <p><it>Plasmodium falciparum </it>chloroquine resistance (CQR) transporter protein (PfCRT) is known to be the important key of CQR. Recent studies have definitively demonstrated a link between mutations in the gene <it>pfcrt </it>and resistance to chloroquine in <it>P. falciparum</it>. Although these mutations are predictive of chloroquine resistance, they are not quantitatively predictive of the degree of resistance.</p> <p>Methods</p> <p>In this study, a total of 95 recently adapted <it>P. falciparum </it>isolates from Thailand were included in the analysis. Parasites were characterized for their drug susceptibility phenotypes and genotypes with respect to <it>pfcrt</it>. From the original 95 isolates, 20 were selected for complete <it>pfcrt </it>sequence analysis.</p> <p>Results</p> <p>Almost all of the parasites characterized carried the previously reported mutations K76T, A220S, Q271E, N326S, I356T and R371I. On complete sequencing, isolates were identified with novel mutations at K76A and E198K. There was a suggestion that parasites carrying E198K were less resistant than those that did not. In addition, <it>pfcrt </it>and <it>pfmdr1 </it>gene expression were investigated by real-time PCR. No relationship between the expression level of either of these genes and response to drug was observed.</p> <p>Conclusion</p> <p>Data from the present study suggest that other genes must contribute to the degree of resistance once the resistance phenotype is established through mutations in <it>pfcrt</it>.</p

Recognising the small Ree groups in their natural representations

We present Las Vegas algorithms for constructive recognition and constructive membership testing of the Ree groups 2G_2(q) = Ree(q), where q = 3^{2m + 1} for some m > 0, in their natural representations of degree 7. The input is a generating set X. The constructive recognition algorithm is polynomial time given a discrete logarithm oracle. The constructive membership testing consists of a pre-processing step, that only needs to be executed once for a given X, and a main step. The latter is polynomial time, and the former is polynomial time given a discrete logarithm oracle. Implementations of the algorithms are available for the computer algebra system MAGMA

Glycerol: An unexpected major metabolite of energy metabolism by the human malaria parasite

<p>Abstract</p> <p>Background</p> <p>Malaria is a global health emergency, and yet our understanding of the energy metabolism of the principle causative agent of this devastating disease, <it>Plasmodium falciparum</it>, remains rather basic. Glucose was shown to be an essential nutritional requirement nearly 100 years ago and since this original observation, much of the current knowledge of <it>Plasmodium </it>energy metabolism is based on early biochemical work, performed using basic analytical techniques (e.g. paper chromatography), carried out almost exclusively on avian and rodent malaria. Data derived from malaria parasite genome and transcriptome studies suggest that the energy metabolism of the parasite may be more complex than hitherto anticipated. This study was undertaken in order to further characterize the fate of glucose catabolism in the human malaria parasite, <it>P. falciparum</it>.</p> <p>Methods</p> <p>Products of glucose catabolism were determined by incubating erythrocyte-freed parasites with D-[1-¹³C] glucose under controlled conditions and metabolites were identified using ¹³C-NMR spectroscopy.</p> <p>Results</p> <p>Following a 2 h incubation of freed-<it>P. falciparum </it>parasites with 25 mM D-[1-¹³C] glucose (<it>n </it>= 4), the major metabolites identified included; [3-¹³C] lactate, [1,3-¹³C] glycerol, [3-¹³C] pyruvate, [3-¹³C] alanine and [3-¹³C] glycerol-3-phosphate. Control experiments performed with uninfected erythrocytes incubated under identical conditions did not show any metabolism of D-[1-¹³C] glucose to glycerol or glycerol-3-phosphate.</p> <p>Discussion</p> <p>The identification of glycerol as a major glucose metabolite confirms the view that energy metabolism in this parasite is more complex than previously proposed. It is hypothesized here that glycerol production by the malaria parasite is the result of a metabolic adaptation to growth in O₂-limited (and CO₂elevated) conditions by the operation of a glycerol-3-phosphate shuttle for the re-oxidation of assimilatory NADH. Similar metabolic adaptations have been reported previously for other microaerobic/anaerobic organisms, such as yeast, rumen protozoa and human parasitic protozoa.</p> <p>Conclusion</p> <p>These data highlight the need to re-evaluate the carbon and redox balance of this important human pathogen, ultimately leading to a better understanding of how the parasite is able to adapt to the variable environments encountered during parasite development and disease progression.</p

Common Genetic Variants Found in HLA and KIR Immune Genes in Autism Spectrum Disorder

The “common variant—common disease” hypothesis was proposed to explain diseases with strong inheritance. This model suggests that a genetic disease is the result of the combination of several common genetic variants. Common genetic variants are described as a 5% frequency differential between diseased vs. matched control populations. This theory was recently supported by an epidemiology paper stating that about 50% of genetic risk for autism resides in common variants. However, rare variants, rather than common variants, have been found in numerous genome wide genetic studies and many have concluded that the “common variant—common disease” hypothesis is incorrect. One interpretation is that rare variants are major contributors to genetic diseases and autism involves the interaction of many rare variants, especially in the brain. It is obvious there is much yet to be learned about autism genetics. Evidence has been mounting over the years indicating immune involvement in autism, particularly the HLA genes on chromosome 6 and KIR genes on chromosome 19. These two large multigene complexes have important immune functions and have been shown to interact to eliminate unwanted virally infected and malignant cells. HLA proteins have important functions in antigen presentation in adaptive immunity and specific epitopes on HLA class I proteins act as cognate ligands for KIR receptors in innate immunity. Data suggests that HLA alleles and KIR activating genes/haplotypes are common variants in different autism populations. For example, class I allele (HLA-A2 and HLA-G 14 bp-indel) frequencies are significantly increased by more than 5% over control populations (Table 2). The HLA-DR4 Class II and shared epitope frequencies are significantly above the control populations (Table 2). Three activating KIR genes: 3DS1, 2DS1, and 2DS2 have increased frequencies of 15, 22, and 14% in autism populations, respectively. There is a 6% increase in total activating KIR genes in autism over control subjects. And, more importantly there is a 12% increase in activating KIR genes and their cognate HLA alleles over control populations (Torres et al., 2012a). These data suggest the interaction of HLA ligand/KIR receptor pairs encoded on two different chromosomes is more significant as a ligand/receptor complex than separately in autism

A Thermodynamic Model for Receptor Clustering

Intracellular signaling often arises from ligand-induced oligomerization of cell surface receptors. This oligomerization or clustering process is fundamentally a cooperative behavior between near-neighbor receptor molecules; the properties of this cooperative process clearly affects the signal transduction. Recent investigations have revealed the molecular basis of receptor-receptor interactions, but a simple theoretical framework for using this data to predict cluster formation has been lacking. Here, we propose a simple, coarse-grained, phenomenological model for ligand-modulated receptor interactions and discuss its equilibrium properties via mean-field theory. The existence of a first-order transition for this model has immediate implications regarding the robustness of the cellular signaling response.Comment: Biophysical Journal, to appea