Evaluation of Viremia Frequencies of a Novel Human Pegivirus by Using Bioinformatic Screening and PCR

Next-generation sequencing has critical applications in virus discovery, diagnostics, and environmental surveillance. We used metagenomic sequence libraries for retrospective screening of plasma samples for the recently discovered human hepegivirus 1 (HHpgV-1). From a cohort of 150 hepatitis C virus (HCV)-positive case-patients, we identified 2 persons with HHpgV-1 viremia and a high frequency of human pegivirus (HPgV) viremia (14%). Detection of HHpgV-1 and HPgV was concordant with parallel PCR-based screening using conserved primers matching groups 1 (HPgV) and 2 (HHPgV-1) nonstructural 3 region sequences. PCR identified 1 HHPgV-1-positive person with viremia from a group of 195 persons with hemophilia who had been exposed to nonvirally inactivated factor VII/IX; 18 (9%) were HPgV-positive. Relative to HCV and HPgV, active infections with HHpgV-1 were infrequently detected in blood, even in groups that had substantial parenteral exposure. Our findings are consistent with lower transmissibility or higher rates of virus clearance for HHpgV-1 than for other bloodborne human flaviviruses

Genetic Variants in Nuclear-Encoded Mitochondrial Genes Influence AIDS Progression

Background: The human mitochondrial genome includes only 13 coding genes while nuclear-encoded genes account for 99% of proteins responsible for mitochondrial morphology, redox regulation, and energetics. Mitochondrial pathogenesis occurs in HIV patients and genetically, mitochondrial DNA haplogroups with presumed functional differences have been associated with differential AIDS progression. Methodology/Principal Findings: Here we explore whether single nucleotide polymorphisms (SNPs) within 904 of the estimated 1,500 genes that specify nuclear-encoded mitochondrial proteins (NEMPs) influence AIDS progression among HIV-1 infected patients. We examined NEMPs for association with the rate of AIDS progression using genotypes generated by an Affymetrix 6.0 genotyping array of 1,455 European American patients from five US AIDS cohorts. Successfully genotyped SNPs gave 50% or better haplotype coverage for 679 of known NEMP genes. With a Bonferroni adjustment for the number of genes and tests examined, multiple SNPs within two NEMP genes showed significant association with AIDS progression: acyl-CoA synthetase medium-chain family member 4 (ACSM4) on chromosome 12 and peroxisomal D3,D2-enoyl- CoA isomerase (PECI) on chromosome 6. Conclusions: Our previous studies on mitochondrial DNA showed that European haplogroups with presumed functional differences were associated with AIDS progression and HAART mediated adverse events. The modest influences of nuclearencoded mitochondrial genes found in the current study add support to the idea that mitochondrial function plays a role in AIDS pathogenesis

The Role of Fibrocytes in Sickle Cell Lung Disease

<div><h3>Background</h3><p>Interstitial lung disease is a frequent complication in sickle cell disease and is characterized by vascular remodeling and interstitial fibrosis. Bone marrow-derived fibrocytes have been shown to contribute to the pathogenesis of other interstitial lung diseases. The goal of this study was to define the contribution of fibrocytes to the pathogenesis of sickle cell lung disease.</p> <h3>Methodology/Principal Findings</h3><p>Fibrocytes were quantified and characterized in subjects with sickle cell disease or healthy controls, and in a model of sickle cell disease, the NY1DD mouse. The role of the chemokine ligand CXCL12 in trafficking of fibrocytes and phenotype of lung disease was examined in the animal model. We found elevated concentration of activated fibrocytes in the peripheral blood of subjects with sickle cell disease, which increased further during vaso-occlusive crises. There was a similar elevations in the numbers and activation phenotype of fibrocytes in the bone marrow, blood, and lungs of the NY1DD mouse, both at baseline and under conditions of hypoxia/re-oxygenation. In both subjects with sickle cell disease and the mouse model, fibrocytes expressed a hierarchy of chemokine receptors, with CXCR4 expressed on most fibrocytes, and CCR2 and CCR7 expressed on a smaller subset of cells. Depletion of the CXCR4 ligand, CXCL12, in the mouse model resulted in a marked reduction of fibrocyte trafficking into the lungs, reduced lung collagen content and improved lung compliance and histology.</p> <h3>Conclusions</h3><p>These data support the notion that activated fibrocytes play a significant role in the pathogenesis of sickle cell lung disease.</p> </div

From theory to practice: Applying current clinical knowledge and treatment strategies to the care of hemophilia A patients with inhibitors

Two bypassing agents are currently available to circumvent the need for factor FVIII in hemophilia A patients with inhibitors: the activated prothrombin complex FEIBA VH and recombinant activated factor VII (NovoSeven (R)). Both products are highly effective in controlling bleeding in the presence of inhibitory alloantibodies, yet their hemostatic efficacy can be unpredictable. As the results of the FEIBA NovoSeven (R) Comparative (FENOC) study illustrate, patients may respond better to one bypassing agent than the other. Furthermore, guidelines from an expert panel reflect that responsiveness to bypassing therapy may change from one bleed to the next in the same patient and even from hour to hour during the course of a single bleeding event. These findings underscore the need to have both bypassing products available to treat bleeding episodes in inhibitor patients, to frequently evaluate the efficacy of hemostasis during the course of a bleeding event, and to switch products early if the response to treatment is unsatisfactory. (C) 2008 Elsevier Ltd. All rights reserved

The Pharmacokinetic Evaluation Of Oral Administered Carbon Monoxide Instilled In a Liquid Formulation (CO-LF) To Rats To Determine Carbon Monoxide Hemoglobin Levels With Potential Efficacy In Patients With Sickle Cell Disease (SCD)

Abstract There is a large published literature presenting information regarding the toxicology of CO in humans. There is information showing that CO-Hb levels below 15% are not associated with clinical adverse signs or symptoms in healthy individuals and smokers have CO-Hb levels around 6%, but may be above 10% on occasion. It has also been found that CO at low levels may have positive effects in SCD. It has been reported that non-toxic levels of CO extend the red cell life span in SCD patients (Beutler E, 1975.). Further, CO might play a role in preventing sickle cell formation by preventing the polymerization of hemoglobin (Sirs JA, 1963), and it has been reported that CO might have a preventative effect on the occurrence of clinical symptoms of SCD (Yallop D et al, 2007). Inhaled CO administered in low concentrations to transgenic sickle mice for one hour per day, 3 days per week for 10 weeks, was non-toxic and substantially decreased white cell counts and cellular inflammatory signals (Beckman J et al, 2009). This research program hypothesizes that CO, when administered orally at non-toxic levels, results in CO-Hb levels that will result in benefit to patients with SCD through two mechanisms: reducing the sickling phenomenon of red cells and by reducing the inflammatory response in these patients. It also hypothesizes that the safest and most efficient mode of administration of CO is by the oral route via a solution with a known dose of CO. Methods The carbon monoxide (CO) is obtained from a commercial source. The liquid formulation (LF) is composed of a mixture of the following components: water, proteins, lipids and carbohydrates ( PLC Mixture). The CO is instilled into this LF and tested for CO concentration. The principle of the method used for CO measurement is to allow the CO and added Hb to interact and to measure the CO-Hb formed by using the dithionite reduction method employing a spectrophotometer. (A.H. Chalmers 1991).The container is weighed before and after CO instillation to determine the occurrence of CO gas leakage. The container for the CO-LF is of low gas permeability. Once CO instillation in the LF is complete, the containers of CO-LF are sealed and labeled. The prepared containers are stored at 2-8°C for no longer than 72 hours. The CO content in blood is expressed as ‘%’ of total hemoglobin (CO-Hb) and mg/L in the CO-LF. CO-LF was administered via esophageal-placed gavage using Sprague-Dawley rats with jugular venous catheters. Results In the first study, there were 5 groups of 4 non-anemic rats studied, with each group comprising of a different LF made with the following percent of PLC Mixture combined with water: 50% PLC Mixture, 20% PLC Mixture, 10% PLC Mixture, 5% PLC Mixture, and 0% PLC Mixture (water only). Two 3 mL doses of CO-LF were administered to each rat one hour apart. Data generated showed levels of CO-Hb at baseline varying between 0 and 1.1 %. After the administration of the test article, the highest CO-Hb levels were 6.1% in one animal at 1 hour after the first dose, and 9.6% one hour after the second dose in one animal. The levels of CO-Hb fell at different rates depending upon the dose and formulation. The PK data for the first study is shown in Table 1. Differences among the test articles were not statistically different. The PK parameters evaluated over 24 hours were not impacted by the administered PLC Mixture, but animals receiving CO-50% PLC Mixture showed an initial fall to baseline levels, then a second rise in CO-Hb levels peaking in one animal at 3.1% at 48 hours. Such late increments were not seen with other CO-LF’s. The second PK study was comprised of 2 groups of 4 anemic rates, and employed CO-0% and CO-10% PLC Mixture. This study was carried out on rats made anemic by blood draw to an estimated 50% blood volume. The PK characteristics show data similar to that seen in non-anemic rats. Conclusion This research has documented the feasibility in a proof-of-concept rat model that orally administered CO via a CO liquid formulation can produce CO-Hb levels that would be expected to impact the pathophysiology of the Sickle Cell Disease-associated micro-vasculature. The terminal half-life of orally administered CO in the rat is similar to that reported via inhaled CO. Disclosures: Gomperts: Hillhurst Biopharmaceuticals Inc: Equity Ownership, Patents &amp; Royalties. Forman:Hillhurst Biopharmaceuticals Inc: Equity Ownership, Patents &amp; Royalties

Delays in maturation among adolescents with hemophilia and a history of inhibitors

Inhibitory antibodies to factors VIII or IX have the potential to affect a broad range of outcomes among people with hemophilia; however, their possible effect on growth and maturation has not been explored. We evaluated skeletal maturation (bone age), pubertal progression, serum testosterone levels, height velocity, and stature in the multicenter Hemophilia Growth and Development Study. A total of 333 children and adolescents (mean age, 12.4 years) were enrolled from 1989 to 1990 and followed for 7 years. Of these, 18% (n = 60) had a history of inhibitors. Bone age among HIV− adolescents with a history of inhibitors lagged 9 or more months behind those without inhibitors at every age from 12 to 15 years. Those with a history of inhibitors were older at every Tanner stage transition, attained a lower maximum growth velocity, and their serum testosterone levels were significantly lower compared with those without inhibitors. Delays were greater among HIV+ patients with a history of inhibitors compared with those without inhibitors; however, the differences were generally small and not statistically significant. The results of this investigation underscore the importance of monitoring the growth and maturation of children and adolescents with hemophilia, particularly those with inhibitors