cAMP Signaling Enhances HIV-1 Long Terminal Repeat (LTR)-directed Transcription and Viral Replication in Bone Marrow Progenitor Cells.

CD34+ hematopoietic progenitor cells have been shown to be susceptible to HIV-1 infection, possibly due to a low-level expression of CXCR4, a coreceptor for HIV-1 entry. Given these observations, we have explored the impact of forskolin on cell surface expression of CXCR4 in a cell line model (TF-1). The elevation of intracellular cyclic adenosine monophosphate (cAMP) by forskolin through adenylyl cyclase (AC) resulted in transcriptional upregulation of CXCR4 with a concomitant increase in replication of the CXCR4-utilizing HIV-1 strain IIIB. Transient expression analyses also demonstrated an increase in CXCR4-, CCR5-, and CXCR4-/CCR5-utilizing HIV-1 (LAI, YU2, and 89.6, respectively) promoter activity. Studies also implicated the protein kinase A (PKA) pathway and the downstream transcription factor CREB-1 in interfacing with cAMP response elements located in the CXCR4 and viral promoter. These observations suggest that the cAMP signaling pathway may serve as a regulator of CXCR4 levels and concomitantly of HIV-1 replication in bone marrow (BM) progenitor cells. © 2017, © The Author(s) 2017

Evolution of Thermotolerance and Variation in the Heat Shock Protein, Hsp70

Low to moderate levels of stress induce a class of molecular chaperones called heat shock proteins (Hsps), which protect cells, tissues and whole organisms from more severe stress. In higher Eukaryotes, Hsp70 is one of the principle heat-induced chaperones. This response is general, and how much Hsp70 an animal produces correlates with the level of stress to which it is exposed. Nonetheless, definitively linking high Hsp70 expression as an adaptation to stress tolerance is problematic, because organisms and cells respond to stress in many ways. By molecular manipulation of Hsp70 in one animal group, Drosophila, differences in hsp70 copy number are shown to directly influence heat-induced expression of Hsp70 and tolerance of heat. However, too high an expression level of Hsp70 can harm individuals during periods of rapid growth. This strong physiological relationship between Hsp70 concentration and thermotolerance, along with Hsp70\u27s remarkable degree of interspecific coding sequence conservation, suggest that hsp70 regulatory elements may evolve as an adaptation in diverse species to their thermal environments. To examine this possibility, correlative studies within species and research on phylogenetic covariation between these traits is reviewed with a focus on Drosophila species. However, the techniques and results discussed should broadly apply to other animal groups where evolutionary approaches can be used to test whether genetic variation in both thermotolerance and Hsp expression within and among species select locally on either hsp70 sequence and/or expression

THE SYNTHSEQ APPROACH TO PERSONAL GENOTYPING

Inspired by the Archon X Prize for Genomics, our research project involves implementing a novel strategy for sequencing the human genome. This prize worth $10 million will be awarded to the first company to sequence 100 human genomes with 99.999$10,000 each. However, the possibility of winning the X Prize is secondary to the prospect of revolutionizing medical diagnostics. Currently, the genomic state of‐the‐art involves identifying SNPs (single nucleotide polymorphisms) that are correlated to certain diseases. Compared to such existing diagnostics, the genome‐wide, sequence‐based biomarkers that will be made possible by fast and affordable human genome sequencing are staggering. After six months of thorough investigation and development, we are pleased to present SynthSeq, a cutting‐edge, whole‐genome sequencing venture based on the novel sequencing‐bysynthesis technology. In contrast to high‐priced competitors, our inexpensive and comparatively error‐free whole genome sequencing solution will prove to be an invaluable diagnostic resource, and it will only become more valuable as advances are made in the field of molecular diagnostics. At our intended retail price of $5,000, series A investors can expect a worst‐case MIRR of 22$700 thousand. We are confident that our innovative SynthSeq technology will deliver high‐fidelity, low‐cost whole genome sequencing to as many as 3,000 customers per year as currently envisioned, with the potential for scale‐up to millions

Changes in the Freshwater Mussel (Bivalvia: Unionidae) Fauna of the Cuyahoga River, Ohio, Since Late Prehistory

We provide new information from archaeological samples on the historical freshwater mussel fauna (Mollusca: Bivalvia: Unionoidea) of the Cuyahoga River (South Park site: occupied between ca. A.D. 950 and 1650) and Black River (White Fort site, occupation centered at ca. A.D. 1350), northeast Ohio. Data from these prehi storic sites are compared with information on extant mussel populations of the Cuyahoga River published between 1890 and 2000. The high representation at both archaeological sites of the species Actinonaias ligamentina, Elliptio dilatata, and Ptychobranchus fasciolaris suggests that these were· among the important clean water species in northeast Ohio prior to European settlement. By comparison, the modem mussel fauna of the lower Cuyahoga River (between Cleveland and Akron) contains none of these relatively abundant species, or any of the species represented in the archaeological material. The modern fauna of the lower river was established during the 20\u27 century. This fauna is a low diversity assemblage of pollution tolerant species represented by rare live individuals. The modem mussel fauna of the upper Cuyahoga River (between Akron and the source) suggests that the upper and lower reaches are effectively isolated from each other. Published records indicate little change in the fauna during the last three quarters of the 20\u27 century. Nevertheless, overall diversity, although substantially higher than that of the lower river, is considerably lower than that of the Grand River, which is located to the east of the Cuyahoga. Overall, the mussel fauna of the Cuyahoga River has changed greatly over time, most notably in terms of losses in diversity of clean water species and overall abundance

A Styrene-alt-Maleic Acid Copolymer Is an Effective Inhibitor of R5 and X4 Human Immunodeficiency Virus Type 1 Infection

An alternating copolymer of styrene and maleic acid (alt-PSMA) differs from other polyanionic antiviral agents in that the negative charges of alt-PSMA are provided by carboxylic acid groups instead of sulfate or sulfonate moieties. We hypothesized that alt-PSMA would have activity against human immunodeficiency virus type 1 (HIV-1) comparable to other polyanions, such as the related compound, poly(sodium 4-styrene sulfonate) (PSS). In assays using cell lines and primary immune cells, alt-PSMA was characterized by low cytotoxicity and effective inhibition of infection by HIV-1 BaL and IIIB as well as clinical isolates of subtypes A, B, and C. In mechanism of action assays, in which each compound was added to cells and subsequently removed prior to HIV-1 infection (“washout” assay), alt-PSMA caused no enhancement of infection, while PSS washout increased infection 70% above control levels. These studies demonstrate that alt-PSMA is an effective HIV-1 inhibitor with properties that warrant further investigation

Slc15a4, a gene required for pDC sensing of TLR ligands, is required to control persistent viral infection

Plasmacytoid dendritic cells (pDCs) are the major producers of type I IFN in response to viral infection and have been shown to direct both innate and adaptive immune responses in vitro. However, in vivo evidence for their role in viral infection is lacking. We evaluated the contribution of pDCs to acute and chronic virus infection using the feeble mouse model of pDC functional deficiency. We have previously demonstrated that feeble mice have a defect in TLR ligand sensing. Although pDCs were found to influence early cytokine secretion, they were not required for control of viremia in the acute phase of the infection. However, T cell priming was deficient in the absence of functional pDCs and the virus-specific immune response was hampered. Ultimately, infection persisted in feeble mice. We conclude that pDCs are likely required for efficient T cell priming and subsequent viral clearance. Our data suggest that reduced pDC functionality may lead to chronic infection

Cervicovaginal Safety of the Formulated, Biguanide-Based Human Immunodeficiency Virus Type 1 (HIV-1) Inhibitor NB325 in a Murine Model

Vaginal microbicides that reduce or eliminate the risk of HIV-1 sexual transmission must do so safely without adversely affecting the integrity of the cervicovaginal epithelium. The present studies were performed to assess the safety of the biguanide-based antiviral compound NB325 in a formulation suitable for topical application. Experiments were performed using a mouse model of cervicovaginal microbicide application, which was previously shown to be predictive of topical agent toxicity revealed in microbicide clinical trials. Mice were exposed vaginally to unformulated NB325 or NB325 formulated in the hydroxyethyl cellulose “universal placebo.” Following exposures to formulated 1% NB325 for 10 min to 24 h, the vaginal and cervical epithelia were generally intact, although some areas of minimal vaginal epithelial damage were noted. Although formulated NB325 appeared generally safe for application in these studies, the low but observable level of toxicity suggests the need for improvements in the compound and/or formulation