The new paradigm of hepatitis C therapy: integration of oral therapies into best practices.

Emerging data indicate that all-oral antiviral treatments for chronic hepatitis C virus (HCV) will become a reality in the near future. In replacing interferon-based therapies, all-oral regimens are expected to be more tolerable, more effective, shorter in duration and simpler to administer. Coinciding with new treatment options are novel methodologies for disease screening and staging, which create the possibility of more timely care and treatment. Assessments of histologic damage typically are performed using liver biopsy, yet noninvasive assessments of histologic damage have become the norm in some European countries and are becoming more widespread in the United States. Also in place are new Centers for Disease Control and Prevention (CDC) initiatives to simplify testing, improve provider and patient awareness and expand recommendations for HCV screening beyond risk-based strategies. Issued in 2012, the CDC recommendations aim to increase HCV testing among those with the greatest HCV burden in the United States by recommending one-time testing for all persons born during 1945-1965. In 2013, the United States Preventive Services Task Force adopted similar recommendations for risk-based and birth-cohort-based testing. Taken together, the developments in screening, diagnosis and treatment will likely increase demand for therapy and stimulate a shift in delivery of care related to chronic HCV, with increased involvement of primary care and infectious disease specialists. Yet even in this new era of therapy, barriers to curing patients of HCV will exist. Overcoming such barriers will require novel, integrative strategies and investment of resources at local, regional and national levels.

Beta-lapachone-mediated apoptosis in human promyelocytic leukemia (HL-60) and human prostate cancer cells: a p53-independent response.

beta-Lapachone and certain of its derivatives directly bind and inhibit topoisomerase I (Topo I) DNA unwinding activity and form DNA-Topo I complexes, which are not resolvable by SDS-K+ assays. We show that beta-lapachone can induce apoptosis in certain cells, such as in human promyelocytic leukemia (HL-60) and human prostate cancer (DU-145, PC-3, and LNCaP) cells, as also described by Li et al. (Cancer Res., 55: 0000-0000, 1995). Characteristic 180-200-bp oligonucleosome DNA laddering and fragmented DNA-containing apoptotic cells via flow cytometry and morphological examinations were observed in 4 h in HL-60 cells after a 4-h, > or = 0.5 microM beta-lapachone exposure. HL-60 cells treated with camptothecin or topotecan resulted in greater apoptotic DNA laddering and apoptotic cell populations than comparable equitoxic concentrations of beta-lapachone, although beta-lapachone was a more effective Topo I inhibitor. beta-Lapachone treatment (4 h, 1-5 microM) resulted in a block at G0/G1, with decreases in S and G2/M phases and increases in apoptotic cell populations over time in HL-60 and three separate human prostate cancer (DU-145, PC-3, and LNCaP) cells. Similar treatments with topotecan or camptothecin (4 h, 1-5 microM) resulted in blockage of cells in S and apoptosis. Thus, beta-lapachone causes a block in G0/G1 of the cell cycle and induces apoptosis in cells before, or at early times during, DNA synthesis. These events are p53 independent, since PC-3 and HL-60 cells are null cells, LNCaP are wild-type, and DU-145 contain mutant p53, yet all undergo apoptosis after beta-lapachone treatment. Interestingly, beta-lapachone treatment of p53 wild type-containing prostate cancer cells (i.e., LNCaP) did not result in the induction of nuclear levels of p53 protein, as did camptothecin-treated cells. Like other Topo I inhibitors, beta-lapachone may induce apoptosis by locking Topo I onto DNA, blocking replication fork movement, and inducing apoptosis in a p53-independent fashion. beta-Lapachone and its derivatives, as well as other Topo I inhibitors, have potential clinical utility alone against human leukemia and prostate cancers.

The Python pit organ: imaging and immunocytochemical analysis of an extremely sensitive natural infrared detector.

The Python infrared-sensitive pit organ is a natural infrared imager that combines high sensitivity, ambient temperature function, microscopic dimensions, and self-repair. We are investigating the spectral sensitivity and signal transduction process in snake infrared-sensitive neurons, neither of which is understood. For example, it is unknown whether infrared receptor neurons function on a thermal or a photic mechanism. We imaged pit organs in living Python molurus and Python regius using infrared-sensitive digital video cameras. Pit organs were significantly more absorptive and/or emissive than surrounding tissues in both 3-5 microns and 8-12 microns wavelength ranges. Pit organs exhibited greater absorption/emissivity in the 8-12 microns range than in the 3-5 microns range. To directly test the relationship between photoreceptors and pit organ infrared-sensitive neurons, we performed immunocytochemistry using antisera directed against retinal photoreceptor opsins. Retinal photoreceptors were labeled with antisera specific for retinal opsins, but these antisera failed to label terminals of infrared-sensitive neurons in the pit organ. Infrared-receptive neurons were also distinguished from retinal photoreceptors on the basis of their calcium-binding protein content. These results indicate that the pit organ absorbs infrared radiation in two major atmospheric transmission windows, one of which (8-12 microns) matches emission of targeted prey, and that infrared receptors are biochemically distinct from retinal photoreceptors. These results also provide the first identification of prospective biochemical components of infrared signal transduction in pit organ receptor neurons.

Prey targeting by the infrared-imaging snake Python molurus: effects of experimental and congenital visual deprivation.

Boid and crotaline snakes possess two distinct types of organ evolved to image radiant electromagnetic energy: the lateral eye, which responds to visible light, and the pit organ, which responds to infrared radiation. While infrared imaging may allow accurate predatory targeting in complete absence of visual information, both infrared and visual information are probably normally involved in prey targeting. We examined the roles of vision and infrared imaging in Python molurus predatory performance under conditions of (1) high visual contrast; (2) very low visual contrast; (3) complete blinding; (4) experimental monocular occlusion; and (5) congenital monocularity. Normally sighted pythons were equally successful at targeting white (BALB/c) and black (C57BL6/J) mice (Mus domesticus) against a black background. Binocularly occluded snakes exhibited strike angles and distances similar to non-occluded snakes, but exhibited lower strike success, suggesting that high visible contrast is not required for accurate targeting, but that precise targeting depends to some degree upon visual information. Strike angles, distances and latencies were indistinguishable between snakes subjected to experimental monocular occlusion and normally sighted snakes. However, snakes congenitally lacking one eye preferentially targeted on the sighted side. Thus, accurate targeting of highly mobile homeothermic prey by Python can be accomplished with little or no visual information, but performance can be affected by complete visual deprivation or by alteration of visual input during development. The developmental effects of early visual deprivation in this system provide a novel opportunity to investigate the neural integration of two electromagnetic radiation-imaging systems in a single animal.

Interleukin-1 beta reduces daily activity level in male lizards, Sceloporus occidentalis.

In mammals, interleukin-1 (IL-1) mediates many of the behavioral consequences of pathogen infection. Other vertebrates show behavioral changes when infected, but the neuroendocrine bases of these changes are seldom known. Here we report that IL-1 beta alters the daily activity cycle of lizards (Sceloporus occidentalis) similar to that seen in lizards infected with malaria. To our knowledge, this is the first report of behavioral effects of interleukin in lower vertebrates. Male lizards were injected with human IL-1 beta (10 ng/g animal), saline, or nothing (control) and the activity level (proportion of lizards above ground) was monitored for 48 h. IL-1 beta-injected lizards showed a decrease in activity compared to saline-injected and control lizards within 2 h after treatment. Activity levels were equivalent among treatment groups during the middle of the day (1200-1600 h), but IL-1-treated animals were significantly less active in the morning (0800-1200 h) and evening (1600-2200 h). This decrease in activity is similar that seen in free-ranging lizards infected with malaria, supporting the hypothesis that IL-1 mediates the pathogen-induced alterations to activity behavior under natural conditions.