Cysteinyl Leukotrienes and Experimental Glomerulonephritis in the Rat

Note:The involvement of cysteinylleukotrienes (LT) in the development of renal dysfunction was investigated in a model of experimental anti-glomerular basement membrane (GBM) glomerulonephritis (GN). Rats, injected iv with rabbit sera containing GBM antibodies, were monitored with respect to renal function, urinary cysteinyl LT excretion and morphological change over 7 days. Renal LTC4 synthase, the enzyme which catalyzes the formation of cysteinyl LTs, was partially purified and characterized with respect to optimal assay conditions. Biochemical and molecular studies demonstrated LTC4 synthase's distinction from known cellular glutathione S-transferase enzymes. […]Au moyen d'un modèle expérimental de glomérulonéphrite (GN) par auto-anticorps anti-membrane basale (AMB), nous avons étudié le rôle des cysteinyl-leucotriènes (LT) dans l'apparition et l'évolution de la dysfonction rénale. Des rats auxquels on avait administré, par injection intraveineuse, un sérum de lapin contenant des anticorps d'AMB ont fait l'objet pendant 7 jours d'une observation portant sur les fonctions rénales, l'excrétion de cysteinyl-leucotriènes et les changements morphologiques. Nous avons procédé à une purification partielle et a une caractérisation de la synthase rénale du LTC4, l'enzyme qui catalyse la formation des peptidoleucotriènes, et ce, dans les conditions optimales de dosage. Les études biochimiques et moléculaires ont permis de distinguer la synthase du LTC4 des enzymes cellulaires S-transférase glutathion déjà connues. […

Therapeutics for dengue: recommendations for design and conduct of early-phase clinical trials.

10.1371/journal.pntd.0001752PLoS Neglected Tropical Diseases69e175

LoFreq: a sequence-quality aware, ultra-sensitive variant caller for uncovering cell-population heterogeneity from high-throughput sequencing datasets.

The study of cell-population heterogeneity in a range of biological systems, from viruses to bacterial isolates to tumor samples, has been transformed by recent advances in sequencing throughput. While the high-coverage afforded can be used, in principle, to identify very rare variants in a population, existing ad hoc approaches frequently fail to distinguish true variants from sequencing errors. We report a method (LoFreq) that models sequencing run-specific error rates to accurately call variants occurring in <0.05% of a population. Using simulated and real datasets (viral, bacterial and human), we show that LoFreq has near-perfect specificity, with significantly improved sensitivity compared with existing methods and can efficiently analyze deep Illumina sequencing datasets without resorting to approximations or heuristics. We also present experimental validation for LoFreq on two different platforms (Fluidigm and Sequenom) and its application to call rare somatic variants from exome sequencing datasets for gastric cancer. Source code and executables for LoFreq are freely available at http://sourceforge.net/projects/lofreq/

A randomized, double-blind placebo controlled trial of balapiravir, a polymerase inhibitor, in adult dengue patients.

BACKGROUND: Dengue is the most common arboviral infection of humans. There are currently no specific treatments for dengue. Balapiravir is a prodrug of a nucleoside analogue (called R1479) and an inhibitor of hepatitis C virus replication in vivo. METHODS: We conducted in vitro experiments to determine the potency of balapiravir against dengue viruses and then an exploratory, dose-escalating, randomized placebo-controlled trial in adult male patients with dengue with <48 hours of fever. RESULTS: The clinical and laboratory adverse event profile in patients receiving balapiravir at doses of 1500 mg (n = 10) or 3000 mg (n = 22) orally for 5 days was similar to that of patients receiving placebo (n = 32), indicating balapiravir was well tolerated. However, twice daily assessment of viremia and daily assessment of NS1 antigenemia indicated balapiravir did not measurably alter the kinetics of these virological markers, nor did it reduce the fever clearance time. The kinetics of plasma cytokine concentrations and the whole blood transcriptional profile were also not attenuated by balapiravir treatment. CONCLUSIONS: Although this trial, the first of its kind in dengue, does not support balapiravir as a candidate drug, it does establish a framework for antiviral treatment trials in dengue and provides the field with a clinically evaluated benchmark molecule. CLINICAL TRIALS REGISTRATION: NCT01096576

Suggested minimum schedule of assessments for a trial of a candidate therapeutic in a patient population with <48 h of fever at the time of commencing treatment.

<p>Suggested minimum schedule of assessments for a trial of a candidate therapeutic in a patient population with <48 h of fever at the time of commencing treatment.</p

Standard analyses and reporting of clinical and virological endpoints in early-phase trials of dengue therapeutics.

*<p>Comparisons of laboratory and virological features between study groups should be adjusted for the baseline value, the day of illness on admission, and serotype/serology (for virological features) to maximize power.</p>a<p>AUC calculated based on the trapezoidal rule with values below the limit of detection replaced by half of the detection limit.</p

Reduction of HBV replication prolongs the early immunological response to IFNα therapy.

BACKGROUND & AIMS: The interaction between HBV replication and immune modulatory effects mediated by IFNα therapy is not well understood. We characterized the impact of HBV DNA replication on the early IFNα-induced immunomodulatory mechanisms. METHODS: We interrogated the transcriptional, serum cytokine/chemokine and cellular immune profiles of 28 patients with HBeAg+ chronic HBV infection (CHB) randomly assigned to one of 4 treatment cohorts (untreated n=5, weekly dosing of 360 μg Pegasys [PegIFNα] n=11, daily dose of 300 mg Viread [tenofovir disoproxil fumarate, TDF] n=6, or a combination of both n=6). Samples were characterized at multiple early time points through day 14 of therapy, after which all patients were given standard of care (180 μg Pegasys injected subcutaneously, weekly). RESULTS: PegIFNα induced a distinct and rapid up-regulation of IFN signaling pathway that coincided with increase detection of distinct serum cytokines/chemokines (IL-15, IL-6, and CXCL-10) and the up-regulation of the frequency of proliferating NK and activated total CD8+ T cells. IFNα treatment alone did not result in rapid decay of HBV replication and was not able to restore the defective HBV-specific T cell response present in CHB patients. In addition, the IFNα immune-stimulatory effects diminished after the first dose, but this refractory effect was reduced in patients where HBV replication was simultaneously inhibited with TDF. CONCLUSIONS: We present here the first comprehensive description of the early effects of IFNα treatment on immune and viral biomarkers in HBeAg+ CHB patients. Our results show that PegIFNα-induced innate immune activation directly benefits from the suppression of HBV replication