Parasite Mitogen-Activated Protein Kinases as Drug Discovery Targets to Treat Human Protozoan Pathogens

Protozoan pathogens are a highly diverse group of unicellular organisms, several of which are significant human pathogens. One group of protozoan pathogens includes obligate intracellular parasites such as agents of malaria, leishmaniasis, babesiosis, and toxoplasmosis. The other group includes extracellular pathogens such as agents of giardiasis and amebiasis. An unfortunate unifying theme for most human protozoan pathogens is that highly effective treatments for them are generally lacking. We will review targeting protozoan mitogen-activated protein kinases (MAPKs) as a novel drug discovery approach towards developing better therapies, focusing on Plasmodia, Leishmania, and Toxoplasma, about which the most is known

The Spitzer c2d Survey of Nearby Dense Cores: VI. The Protostars of Lynds Dark Nebula 1221

Observations of Lynds Dark Nebula 1221 from the Spitzer Space Telescope are presented. These data show three candidate protostars towards L1221, only two of which were previously known. The infrared observations also show signatures of outflowing material, an interpretation which is also supported by radio observations with the Very Large Array. In addition, molecular line maps from the Five College Radio Astronomy Observatory are shown. One-dimensional dust continuum modelling of two of these protostars, IRS1 and IRS3, is described. These models show two distinctly different protostars forming in very similar environments. IRS1 shows a higher luminosity and larger inner radius of the envelope than IRS3. The disparity could be caused by a difference in age or mass, orientation of outflow cavities, or the impact of a binary in the IRS1 core.Comment: accepted for publication in Ap

Efficient Foreign Gene Expression in Epstein-Barr Virus-Transformed Human B-Cells

Epstein-Barr virus (EBV) is a herpesvirus that transforms B-cells (B-LCL) and has undergone intense scrutiny owing to its association with Burkitt's lymphoma, nasopharyngeal carcinoma, and immunoblastic lymphomas. B-LCL have also proven useful in the study of human immunology. We describe a novel system for inducing efficient foreign gene expression in B-LCL using biotinylated adenovirus as an endosome-disrupting agent. Plasmid DNA is coupled to the exterior of viral particles by streptavidin-polylysine chimeric proteins. Up to 67% of B-LCL may be induced to express foreign genes in vitro in transient expression systems, and gene expression lasts for at least 17 days. We have expressed firefly luciferase, β-galactosidase (β-gal), chloramphenicol acetyltransferase, HIV gag, and env genes, as well as infectious HIV, and the EBV-specific BZLF gene in B-LCL with this system. In vivo delivery of a β-gal reporter gene to B-LCL was documented in a SCID mouse model. Potential applications include study of genetic regulation of EBV infection and transformation events, study of potential gene therapies for EBV-related B-cell tumors, and production of antigen-presenting cells for use in immunologic assays. Because of the high percentage of cells transformed and the length of foreign gene expression, the possibility of examining foreign gene expression in transient assays, without selection for clonal populations, exists

The Detection and Characterization of cm Radio Continuum Emission from the Low-mass Protostar L1014-IRS

Observations by the Cores to Disk Legacy Team with the Spitzer Space Telescope have identified a low luminosity, mid-infrared source within the dense core, Lynds 1014, which was previously thought to harbor no internal source. Followup near-infrared and submillimeter interferometric observations have confirmed the protostellar nature of this source by detecting scattered light from an outflow cavity and a weak molecular outflow. In this paper, we report the detection of cm continuum emission with the VLA. The emission is characterized by a quiescent, unresolved 90 uJy 6 cm source within 0.2" of the Spitzer source. The spectral index of the quiescent component is $\alpha = 0.37\pm 0.34$ between 6 cm and 3.6 cm. A factor of two increase in 6 cm emission was detected during one epoch and circular polarization was marginally detected at the $5\sigma$ level with Stokes {V/I} $= 48 \pm 16$% . We have searched for 22 GHz H2O maser emission toward L1014-IRS, but no masers were detected during 7 epochs of observations between June 2004 and December 2006. L1014-IRS appears to be a low-mass, accreting protostar which exhibits cm emission from a thermal jet or a wind, with a variable non-thermal emission component. The quiescent cm radio emission is noticeably above the correlation of 3.6 cm and 6 cm luminosity versus bolometric luminosity, indicating more radio emission than expected. We characterize the cm continuum emission in terms of observations of other low-mass protostars, including updated correlations of centimeter continuum emission with bolometric luminosity and outflow force, and discuss the implications of recent larger distance estimates on the physical attributes of the protostar and dense molecular core.Comment: 14 pages. Accepted for publication in Ap

Attenuation of RNA polymerase II pausing mitigates BRCA1-associated R-loop accumulation and tumorigenesis.

Most BRCA1-associated breast tumours are basal-like yet originate from luminal progenitors. BRCA1 is best known for its functions in double-strand break repair and resolution of DNA replication stress. However, it is unclear whether loss of these ubiquitously important functions fully explains the cell lineage-specific tumorigenesis. In vitro studies implicate BRCA1 in elimination of R-loops, DNA-RNA hybrid structures involved in transcription and genetic instability. Here we show that R-loops accumulate preferentially in breast luminal epithelial cells, not in basal epithelial or stromal cells, of BRCA1 mutation carriers. Furthermore, R-loops are enriched at the 5' end of those genes with promoter-proximal RNA polymerase II (Pol II) pausing. Genetic ablation of Cobra1, which encodes a Pol II-pausing and BRCA1-binding protein, ameliorates R-loop accumulation and reduces tumorigenesis in Brca1-knockout mouse mammary epithelium. Our studies show that Pol II pausing is an important contributor to BRCA1-associated R-loop accumulation and breast cancer development

Extreme Active Molecular Jets in L1448C

The protostellar jet driven by L1448C was observed in the SiO J=8-7 and CO J=3-2 lines and 350 GHz dust continuum at ~1" resolution with the Submillimeter Array (SMA). A narrow jet from the northern source L1448C(N) was observed in the SiO and the high-velocity CO. The jet consists of a chain of emission knots with an inter-knot spacing of ~2" (500 AU) and a semi-periodic velocity variation. The innermost pair of knots, which are significant in the SiO map but barely seen in the CO, are located at ~1" (250 AU) from the central source, L1448C(N). Since the dynamical time scale for the innermost pair is only ~10 yr, SiO may have been formed in the protostellar wind through the gas-phase reaction, or been formed on the dust grain and directly released into the gas phase by means of shocks. It is found that the jet is extremely active with a mechanical luminosity of ~7 L_sun, which is comparable to the bolometric luminosity of the central source (7.5 L_sun). The mass accretion rate onto the protostar derived from the mass-loss rate is ~10^{-5} M_sun/yr. Such a high mass accretion rate suggests that the mass and the age of the central star are 0.03-0.09 M_sun and (4-12)x10^3 yr, respectively, implying that the central star is in the very early stage of protostellar evolution. The low-velocity CO emission delineates two V-shaped shells with a common apex at L1448C(N). The kinematics of these shells are reproduced by the model of a wide opening angle wind. The co-existence of the highly-collimated jets and the wide-opening angle shells can be explained by the unified X-wind model" in which highly-collimated jet components correspond to the on-axis density enhancement of the wide-opening angle wind. The CO $J$=3--2 map also revealed the second outflow driven by the southern source L1448C(S) located at ~8.3" (2000 AU) from L1448C(N).Comment: 45 pages, 13 figures, Accepted for the publication in the Astrophysical Journa

B7-H4 expression identifies a novel suppressive macrophage population in human ovarian carcinoma

Tumor-associated macrophages are a prominent component of ovarian cancer stroma and contribute to tumor progression. B7-H4 is a recently identified B7 family molecule. We show that primary ovarian tumor cells express intracellular B7-H4, whereas a fraction of tumor macrophages expresses surface B7-H4. B7-H4+ tumor macrophages, but not primary ovarian tumor cells, suppress tumor-associated antigen-specific T cell immunity. Blocking B7-H4-, but not arginase-, inducible nitric oxide synthase or B7-H1 restored the T cell stimulating capacity of the macrophages and contributes to tumor regression in vivo. Interleukin (IL)-6 and IL-10 are found in high concentrations in the tumor microenvironment. These cytokines stimulate macrophage B7-H4 expression. In contrast, granulocyte/macrophage colony-stimulating factor and IL-4, which are limited in the tumor microenvironment, inhibit B7-H4 expression. Ectopic expression of B7-H4 makes normal macrophages suppressive. Thus, B7-H4+ tumor macrophages constitute a novel suppressor cell population in ovarian cancer. B7-H4 expression represents a critical checkpoint in determining host responses to dysfunctional cytokines in ovarian cancer. Blocking B7-H4 or depleting B7-H4+ tumor macrophages may represent novel strategies to enhance T cell tumor immunity in cancer