Phase 1 trial of malaria transmission blocking vaccine candidates Pfs25 and Pvs25 formulated with montanide ISA 51

Background

Pfs25 and Pvs25, surface proteins of mosquito stage of the malaria parasites P. falciparum and P. vivax, respectively, are leading candidates for vaccines preventing malaria transmission by mosquitoes. This single blinded, dose escalating, controlled Phase 1 study assessed the safety and immunogenicity of recombinant Pfs25 and Pvs25 formulated with Montanide ISA 51, a water-in-oil emulsion.

Methodology/Principal Findings

The trial was conducted at The Johns Hopkins Center for Immunization Research, Washington DC, USA, between May 16, 2005–April 30, 2007. The trial was designed to enroll 72 healthy male and non-pregnant female volunteers into 1 group to receive adjuvant control and 6 groups to receive escalating doses of the vaccines. Due to unexpected reactogenicity, the vaccination was halted and only 36 volunteers were enrolled into 4 groups: 3 groups of 10 volunteers each were immunized with 5 µg of Pfs25/ISA 51, 5 µg of Pvs25/ISA 51, or 20 µg of Pvs25/ISA 51, respectively. A fourth group of 6 volunteers received adjuvant control (PBS/ISA 51). Frequent local reactogenicity was observed. Systemic adverse events included two cases of erythema nodosum considered to be probably related to the combination of the antigen and the adjuvant. Significant antibody responses were detected in volunteers who completed the lowest scheduled doses of Pfs25/ISA 51. Serum anti-Pfs25 levels correlated with transmission blocking activity.

Conclusion/Significance

It is feasible to induce transmission blocking immunity in humans using the Pfs25/ISA 51 vaccine, but these vaccines are unexpectedly reactogenic for further development. This is the first report that the formulation is associated with systemic adverse events including erythema nodosum.

Trial Registration

ClinicalTrials.gov {"type":"clinical-trial","attrs":{"text":"NCT00295581","term_id":"NCT00295581"}}NCT00295581     

Resistance exercise training reduces hypertriglyceridemia in HIV-infected men treated with antiviral therapy

Hypertriglyceridemia, peripheral insulin resistance,and trunk adiposity are metabolic complications recently recognized in people infected with human immunodeficiency virus (HIV) and treated with highly active antiretroviral therapy (HAART). These complications may respond favorably to exercise training. Using a paired design, wedetermined whether 16 wk of weight-lifting exercise increased musclemass and strength and decreased fasting serum triglycerides and adiposetissue mass in 18 HIV-infected men. The resistance exercise regimenconsisted of three upper and four lower body exercises done for1-1.5 h/day, 4 days/wk for 64 sessions. Dual-energy X-rayabsorptiometry indicated that exercise training increased whole bodylean mass 1.4 kg (P = 0.005) but did not reduce adipose tissue mass (P = NS). Axial proton-magnetic resonanceimaging indicated that thigh muscle cross-sectional area increased5-7 cm² (P < 0.005). Muscle strengthincreased 23-38% (P < 0.0001) on all exercises.Fasting serum triglycerides were decreased at the end of training(281-204 mg/dl; P = 0.02). These findings imply that resistance exercise training-induced muscle hypertrophy may promote triglyceride clearance from the circulation ofhypertriglyceridemic HIV-infected men treated with antiviral therapy.

     

SVachra: a tool to identify genomic structural variation in mate pair sequencing data containing inward and outward facing reads

Background

Characterization of genomic structural variation (SV) is essential to expanding the research and clinical applications of genome sequencing. Reliance upon short DNA fragment paired end sequencing has yielded a wealth of single nucleotide variants and internal sequencing read insertions-deletions, at the cost of limited SV detection. Multi-kilobase DNA fragment mate pair sequencing has supplemented the void in SV detection, but introduced new analytic challenges requiring SV detection tools specifically designed for mate pair sequencing data. Here, we introduce SVachra – Structural Variation Assessment of CHRomosomal Aberrations, a breakpoint calling program that identifies large insertions-deletions, inversions, inter- and intra-chromosomal translocations utilizing both inward and outward facing read types generated by mate pair sequencing.

Results

We demonstrate SVachra’s utility by executing the program on large-insert (Illumina Nextera) mate pair sequencing data from the personal genome of a single subject (HS1011). An additional data set of long-read (Pacific BioSciences RSII) was also generated to validate SV calls from SVachra and other comparison SV calling programs. SVachra exhibited the highest validation rate and reported the widest distribution of SV types and size ranges when compared to other SV callers.

Conclusions

SVachra is a highly specific breakpoint calling program that exhibits a more unbiased SV detection methodology than other callers.

     

Quantitative-Trait Loci Influencing Body-Mass Index Reside on Chromosomes 7 and 13: The National Heart, Lung, and Blood Institute Family Heart Study

Obesity is a risk factor for many chronic diseases, including glucose intolerance, lipid disorders, hypertension, and coronary heart disease. Even though the body-mass index (BMI) is a heterogeneous phenotype reflecting the amount of fat, lean mass, and body build, several studies have provided evidence of one or two major loci contributing to the variation in this complex trait. We sought to identify loci with potential influence on BMI in the data obtained from National Heart, Lung, and Blood Institute Family Heart Study. Two complementary samples were studied: (a) 1,184 subjects in 317 sibships, with 243 markers typed by the Utah Molecular Genetics Laboratory (UMGL) and (b) 3,027 subjects distributed among 401 three-generation families, with 404 markers typed by the Mammalian Genotyping Service (MGS). A genome scan using a variance-components-based linkage approach was performed for each sample, as well as for the combined sample, in which the markers from each analysis were placed on a common genetic map. There was strong evidence for linkage on chromosome 7q32.3 in each sample: the maximum multipoint LOD scores were 4.7 (P<10-5) at marker GATA43C11 and 3.2 (P=.00007) at marker D7S1804, for the MGS and UMGL samples, respectively. The linkage result is replicated by the consistent evidence from these two complementary subsets. Furthermore, the evidence for linkage was maintained in the combined sample, with a LOD score of 4.9 (P<10-5) for both markers, which map to the same location. This signal is very near the published location for the leptin gene, which is the most prominent candidate gene in this region. For the combined-sample analysis, evidence of linkage was also found on chromosome 13q14, with D13S257 (LOD score 3.2, P=.00006), and other, weaker signals (LOD scores 1.5-1.9) were found on chromosomes 1, 2, 3, 5, 6, 14, and 15.     

1alpha,25-dihydroxyvitamin D3 stimulates phosphorylation of IkappaBalpha and synergizes with TPA to induce nuclear translocation of NFkappaB during monocytic differentiation of NB4 leukemia cells.

Treatment of NB4 acute promyelocytic leukemia cells with 1,25-dihydroxyvitamin D3 (1,25D3) or analogs 20-epi-22-oxa-24a,26a,27a-trihomo-1alpha,25-dihydroxyvitamin D3, 1,24-dihydroxy-22-ene-24-cyclopropylvitamin D3, 1alpha,25-dihydroxylumisterol3, or 1alpha,25(OH)2-d5-previtamin D3 in combination with TPA induces monocytic differentiation. The role of 1,25D3 in the induction of maturation has been shown to be a priming effect. Differentiation in response to these agents requires VDR-independent signaling of 1,25D3, PKC signaling, intracellular calcium, and calpain activity. In this study we identify the NFkappaB/IkappaB signaling pathway as a target of 1,25D3 and TPA action. One of the priming effects of 1,25D3 appears to be the rapid phosphorylation of serine residues on IkappaBalpha. On their own, 1,25D3, its analogs, and TPA do not alter IkappaBalpha expression; however, combinations of analogs with TPA result in a synergistic decrease in IkappaBalpha expression. Decreased expression of IkappaBalpha likely results from enhanced degradation, which allows the observed subsequent nuclear translocation of NFkappaB subunit p65. Since nuclear-localized NFkappaB was observed only in combination-treated cells, it is proposed that nuclear targets of NFkappaB are required for monocytic differentiation. Intracellular calcium and proteolytic activity are both necessary for the induction of IkappaB regulation and translocation of NFkappaB and are critical components of the nongenomic signaling cascades of the 1,25D3-induced differentiation pathway.     

Mercury in Fish-eating Communities of the Andean Amazon,Napo River Valley,Ecuador

This exploratory study aimed to examine the relationship between fish eating habits, human mercury levels, and mercury levels in fish in three communities of the Napo River Valley, Ecuadorian Andean Amazon, a region without gold mining but with significant deforestation and volcanic soils with naturally high mercury levels. By recognizing the politicoeconomic factors which cause deforestation, the cultural factors which influence diet, and the biogeochemical factors which contribute to mercury levels, this study employs an ecosystem approach. Interviews on diet were conducted, hair samples from 99 individuals were collected, and samples of commonly eaten fish were taken. Samples were analyzed using cold vapor atomic fluorescence spectrometry (CVAFS). Two rural communities were found to have higher fish consumption and hair mercury levels (8.71 μg/g and 5.32 μg/g) as compared to an urban community (1.87 μg/g). A sequential analysis of hair established mercury levels by month. No seasonal tendencies were noted. Piscivorous fish (0.36 μg/g) were found to be more contaminated in mercury than herbivorous fish (0.05 μg/g). The study shows that sociocultural factors are important in determining mercury exposure. The two village communities consume different species of fish with different frequencies, leading to differential exposure and mercury concentrations in hair samples. The levels of mercury in these two villages were similar to those found in Brazil where neurobehavioral tests showed a correlation between these relatively low levels of mercury and decreased psychomotor capacities. These findings are concerning and should be followed by further studies on the multiple factors that affect the health status of these exposed communities.     

Toxicity Caused by Hydroxycinnamoyl-Coenzyme A Thioester Accumulation in Mutants of Acinetobacter sp. Strain ADP1

Hydroxycinnamates, aromatic compounds that play diverse roles in plants, are dissimilated by enzymes encoded by the hca genes in the nutritionally versatile, naturally transformable bacterium Acinetobacter sp. strain ADP1. A key step in the hca-encoded pathway is activation of the natural substrates caffeate, p-coumarate, and ferulate by an acyl:coenzyme A (acyl:CoA) ligase encoded by hcaC. As described in this paper, Acinetobacter cells with a knockout of the next enzyme in the pathway, hydroxycinnamoyl-CoA hydratase/lyase (HcaA), are extremely sensitive to the presence of the three natural hydroxycinnamate substrates; Escherichia coli cells carrying a subclone with the hcaC gene are hydroxycinnamate sensitive as well. When the hcaA mutation was combined with a mutation in the repressor HcaR, exposure of the doubly mutated Acinetobacter cells to caffeate, p-coumarate, or ferulate at 10⁻⁶ M totally inhibited the growth of cells. The toxicity of p-coumarate and ferulate to a ΔhcaA strain was found to be a bacteriostatic effect. Although not toxic to wild-type cells initially, the diphenolic caffeate was itself converted to a toxin over time in the absence of cells; the converted toxin was bactericidal. In an Acinetobacter strain blocked in hcaA, a secondary mutation in the ligase (HcaC) suppresses the toxic effect. Analysis of suppression due to the mutation of hcaC led to the development of a positive-selection strategy that targets mutations blocking HcaC. An hcaC mutation from one isolate was characterized and was found to result in the substitution of an amino acid that is conserved in a functionally characterized homolog of HcaC.     

Mediators of Weight Loss in a Family‐Based Intervention Presented over the Internet

Objective: To assess the process variables involved in a weight loss program for African‐American adolescent girls. Several process variables have been identified as affecting success in in vivo weight loss programs for adults and children, including program adherence, self‐efficacy, and social support. The current study sought to broaden the understanding of these process variables as they pertain to an intervention program that is presented using the Internet. It was hypothesized that variables such as program adherence, dietary self‐efficacy, psychological factors, and family environment factors would mediate the effect of the experimental condition on weight loss. Research Methods and Procedures: Participants were 57 adolescent African‐American girls who joined the program with one obese parent; family pairs were randomized to either a behavioral or control condition in an Internet‐based weight loss program. Outcome data (weight loss) are reported for the first 6 months of the intervention. Results: Results partially supported the hypotheses. For weight loss among adolescents, parent variables pertaining to life and family satisfaction were the strongest mediating variables. For parental weight loss, changes in dietary practices over the course of 6 months were the strongest mediators. Discussion: The identification of factors that enhance or impede weight loss for adolescents is an important step in improving weight loss programs for this group. The current findings suggest that family/parental variables exert a strong influence on weight loss efforts for adolescents and should be considered in developing future programs.