Cas9+ conditionally-immortalized macrophages as a tool for bacterial pathogenesis and beyond.

Macrophages play critical roles in immunity, development, tissue repair, and cancer, but studies of their function have been hampered by poorly-differentiated tumor cell lines and genetically-intractable primary cells. Here we report a facile system for genome editing in non-transformed macrophages by differentiating ER-Hoxb8 myeloid progenitors from Cas9-expressing transgenic mice. These conditionally immortalized macrophages (CIMs) retain characteristics of primary macrophages derived from the bone marrow yet allow for easy genetic manipulation and a virtually unlimited supply of cells. We demonstrate the utility of this system for dissection of host genetics during intracellular bacterial infection using two important human pathogens: Listeria monocytogenes and Mycobacterium tuberculosis

The CDC Revised Recommendations for HIV Testing: Reactions of Women Attending Community Health Clinics

The purpose of this study was to examine reactions to the Centers for Disease Control and Prevention revised recommendations for HIV testing by women attending community health clinics. A total of 30 women attending three community clinics completed semistructured individual interviews containing three questions about the recommendations. Thematic content analysis of responses was conducted. Results were that all agreed with the recommendation for universal testing. Most viewed opt-out screening as an acceptable approach to HIV testing. Many emphasized the importance of provision of explicit verbal informed consent. The majority strongly opposed the elimination of the requirement for pretest prevention counseling and spontaneously talked about the ongoing importance of posttest counseling. The conclusion was that there was strong support for universal testing of all persons 13 to 64 years old but scant support for the elimination of pretest prevention counseling. In general, respondents believed that verbal informed consent for testing as well as provision of HIV-related information before and after testing were crucial

Is mindfulness Buddhist? (and why it matters).

Modern exponents of mindfulness meditation promote the therapeutic effects of "bare attention"--a sort of non-judgmental, non-discursive attending to the moment-to-moment flow of consciousness. This approach to Buddhist meditation can be traced to Burmese Buddhist reform movements of the first half of the 20th century, and is arguably at odds with more traditional Theravāda Buddhist doctrine and meditative practices. But the cultivation of present-centered awareness is not without precedent in Buddhist history; similar innovations arose in medieval Chinese Zen (Chan) and Tibetan Dzogchen. These movements have several things in common. In each case the reforms were, in part, attempts to render Buddhist practice and insight accessible to laypersons unfamiliar with Buddhist philosophy and/or unwilling to adopt a renunciatory lifestyle. In addition, these movements all promised astonishingly quick results. And finally, the innovations in practice were met with suspicion and criticism from traditional Buddhist quarters. Those interested in the therapeutic effects of mindfulness and bare attention are often not aware of the existence, much less the content, of the controversies surrounding these practices in Asian Buddhist history

Hydrogen-like nitrogen radio line from hot interstellar and warm-hot intergalactic gas

Hyperfine structure lines of highly-charged ions may open a new window in observations of hot rarefied astrophysical plasmas. In this paper we discuss spectral lines of isotopes and ions abundant at temperatures 10^5-10^7 K, characteristic for warm-hot intergalactic medium, hot interstellar medium, starburst galaxies, their superwinds and young supernova remnants. Observations of these lines will allow to study bulk and turbulent motions of the observed target and will broaden the information about the gas ionization state, chemical and isotopic composition. The most prospective is the line of the major nitrogen isotope having wavelength 5.65 mm (Sunyaev and Churazov 1084). Wavelength of this line is well-suited for observation of objects at z=0.15-0.6 when it is redshifted to 6.5-9 mm spectral band widely-used in ground-based radio observations, and, for example, for z>=1.3, when the line can be observed in 1.3 cm band and at lower frequencies. Modern and future radio telescopes and interferometers are able to observe the absorption by 14-N VII in the warm-hot intergalactic medium at redshifts above z=0.15 in spectra of brightest mm-band sources. Sub-millimeter emission lines of several most abundant isotopes having hyperfine splitting might also be detected in spectra of young supernova remnants.Comment: 12 pages, 5 figures, accepted by Astronomy Letters; v3: details added; error fixe

The Pesticide Risk Beliefs Inventory: A Quantitative Instrument for the Assessment of Beliefs about Pesticide Risks

Recent media attention has focused on the risks that agricultural pesticides pose to the environment and human health; thus, these topics provide focal areas for scientists and science educators to enhance public understanding of basic toxicology concepts. This study details the development of a quantitative inventory to gauge pesticide risk beliefs. The goal of the inventory was to characterize misconceptions and knowledge gaps, as well as expert-like beliefs, concerning pesticide risk. This study describes the development and field testing of the Pesticide Risk Beliefs Inventory with an important target audience: pesticide educators in a southeastern U.S. state. The 19-item, Likert-type inventory was found to be psychometrically sound with a Cronbach’s alpha of 0.780 and to be a valuable tool in capturing pesticide educators’ beliefs about pesticide risk, assessing beliefs in four key categories. The Pesticide Risk Beliefs Inventory could be useful in exploring beliefs about pesticide risks and in guiding efforts to address misconceptions held by a variety of formal and informal science learners, educators, practitioners, the agricultural labor force, and the general public

Probiotics, Anticipation Stress, and the Acute Immune Response to Night Shift

IntroductionSleep disturbance and sleep disruption are associated with chronic, low grade inflammation and may underpin a range of chronic diseases in night shift workers. Through modulation of the intestinal microbiota, probiotic supplements may moderate the effects of sleep disruption on the immune system. The aim of this study was to examine 14 days of daily probiotic supplementation on the acute response of acute phase proteins and immune markers to sleep disruption associated with night shift work (Australia and New Zealand Clinical Trials Registry: 12617001552370).MethodsIndividuals (mean age 41 ± 11 yrs; 74% female) performing routine night shift were randomly assigned to a probiotic group (1 × 1010 colony forming units (CFU) Lactobacillus acidophilus DDS-1 or 1 × 1010 CFU Bifidobacterium animalis subsp. lactis UABla-12) or placebo (n= 29 per group). Participants undertook a 14-day supplementation period that coincided with a period of no night shifts followed by two consecutive night shifts. Blood samples were collected prior to the start of supplementation (V1), prior to commencing the first night shift (V2), after the first night shift (V3) and after the second night shift (V4). Serum was assessed for markers of stress (cortisol), acute phase response (C reactive protein (CRP), erythrocyte sedimentation rate, pentraxin), adhesion markers (serum E-selectin, mucosal vascular addressin cell adhesion molecule 1 (MAdCAM-1), and serum cytokines (interleukin (IL)-1ra, IL-1β, IL-6, tumor necrosis factor (TNF)-α, IL-10). Sleep quality was assessed with the Pittsburgh Sleep Quality Index (PSQI) and a Fitbit activity tracker.ResultsThe groups were well balanced on key markers and the probiotic strains were well tolerated. The 14-day supplementation period that coincided with typical night-day sleep-wake cycles leading up to night shift (V1 to V2) was associated with significant changes in the placebo group in the concentration of serum cortisol (p = 0.01), pentraxin (p = 0.001), MAdCAM-1 (p = 0.001), and IL-1ra (p=0.03). In contrast, probiotic supplementation moderated changes in these serum markers from V1 to V2. No significant interaction effects (time by group) were observed for the serum markers prior to and after night shift work following probiotic supplementation due to the substantial changes in the serum markers that occurred during the normal sleep period from V1 to V2.ConclusionsProbiotics may moderate the effects of anticipatory stress on the immune system in the lead up to night shift

A Human IgSF Cell-Surface Interactome Reveals a Complex Network of Protein-Protein Interactions

Cell-surface protein-protein interactions (PPIs) mediate cell-cell communication, recognition, and responses. We executed an interactome screen of 564 human cell-surface and secreted proteins, most of which are immunoglobulin superfamily (IgSF) proteins, using a high-throughput, automated ELISA-based screening platform employing a pooled-protein strategy to test all 318,096 PPI combinations. Screen results, augmented by phylogenetic homology analysis, revealed ∼380 previously unreported PPIs. We validated a subset using surface plasmon resonance and cell binding assays. Observed PPIs reveal a large and complex network of interactions both within and across biological systems. We identified new PPIs for receptors with well-characterized ligands and binding partners for “orphan” receptors. New PPIs include proteins expressed on multiple cell types and involved in diverse processes including immune and nervous system development and function, differentiation/proliferation, metabolism, vascularization, and reproduction. These PPIs provide a resource for further biological investigation into their functional relevance and may offer new therapeutic drug targets

A Human IgSF Cell-Surface Interactome Reveals a Complex Network of Protein-Protein Interactions

Cell-surface protein-protein interactions (PPIs) mediate cell-cell communication, recognition, and responses. We executed an interactome screen of 564 human cell-surface and secreted proteins, most of which are immunoglobulin superfamily (IgSF) proteins, using a high-throughput, automated ELISA-based screening platform employing a pooled-protein strategy to test all 318,096 PPI combinations. Screen results, augmented by phylogenetic homology analysis, revealed ∼380 previously unreported PPIs. We validated a subset using surface plasmon resonance and cell binding assays. Observed PPIs reveal a large and complex network of interactions both within and across biological systems. We identified new PPIs for receptors with well-characterized ligands and binding partners for “orphan” receptors. New PPIs include proteins expressed on multiple cell types and involved in diverse processes including immune and nervous system development and function, differentiation/proliferation, metabolism, vascularization, and reproduction. These PPIs provide a resource for further biological investigation into their functional relevance and may offer new therapeutic drug targets