Delta-9 tetrahydrocannabinol (THC) inhibits lytic replication of gamma oncogenic herpesviruses in vitro

BACKGROUND: The major psychoactive cannabinoid compound of marijuana, delta-9 tetrahydrocannabinol (THC), has been shown to modulate immune responses and lymphocyte function. After primary infection the viral DNA genome of gamma herpesviruses persists in lymphoid cell nuclei in a latent episomal circular form. In response to extracellular signals, the latent virus can be activated, which leads to production of infectious virus progeny. Therefore, we evaluated the potential effects of THC on gamma herpesvirus replication. METHODS: Tissue cultures infected with various gamma herpesviruses were cultured in the presence of increasing concentrations of THC and the amount of viral DNA or infectious virus yield was compared to those of control cultures. The effect of THC on Kaposi's Sarcoma Associated Herpesvirus (KSHV) and Epstein-Barr virus (EBV) replication was measured by the Gardella method and replication of herpesvirus saimiri (HVS) of monkeys, murine gamma herpesvirus 68 (MHV 68), and herpes simplex type 1 (HSV-1) was measured by yield reduction assays. Inhibition of the immediate early ORF 50 gene promoter activity was measured by the dual luciferase method. RESULTS: Micromolar concentrations of THC inhibit KSHV and EBV reactivation in virus infected/immortalized B cells. THC also strongly inhibits lytic replication of MHV 68 and HVS in vitro. Importantly, concentrations of THC that inhibit virus replication of gamma herpesviruses have no effect on cell growth or HSV-1 replication, indicating selectivity. THC was shown to selectively inhibit the immediate early ORF 50 gene promoter of KSHV and MHV 68. CONCLUSIONS: THC specifically targets viral and/or cellular mechanisms required for replication and possibly shared by these gamma herpesviruses, and the endocannabinoid system is possibly involved in regulating gamma herpesvirus latency and lytic replication. The immediate early gene ORF 50 promoter activity was specifically inhibited by THC. These studies may also provide the foundation for the development of antiviral strategies utilizing non-psychoactive derivatives of THC

The JCMT Legacy Survey of the Gould Belt: Mapping 13CO and C 18O in Orion A

The Gould Belt Legacy Survey will map star-forming regions within 500 pc, using Heterodyne Array Receiver Programme (HARP), Submillimetre Common-User Bolometer Array 2 (SCUBA-2) and Polarimeter 2 (POL-2) on the James Clerk Maxwell Telescope (JCMT). This paper describes HARP observations of the J= 3 → 2 transitions of 13CO and C18O towards Orion A. The 15 arcsec resolution observations cover 5 pc of the Orion filament, including OMC 1 (including BN–KL and Orion bar), OMC 2/3 and OMC 4, and allow a comparative study of the molecular gas properties throughout the star-forming cloud. The filament shows a velocity gradient of ∼1 km s−1 pc−1 between OMC 1, 2 and 3, and high-velocity emission is detected in both isotopologues. The Orion Nebula and Bar have the largest masses and linewidths, and dominate the mass and energetics of the high-velocity material. Compact, spatially resolved emission from CH3CN, 13CH3OH, SO, HCOOCH3, CH3CHO and CH3OCHO is detected towards the Orion Hot Core. The cloud is warm, with a median excitation temperature of ∼24 K; the Orion Bar has the highest excitation temperature gas, at >80 K. The C18O excitation temperature correlates well with the dust temperature (to within 40 per cent). The C18O emission is optically thin, and the 13CO emission is marginally optically thick; despite its high mass, OMC 1 shows the lowest opacities. A virial analysis indicates that Orion A is too massive for thermal or turbulent support, but is consistent with a model of a filamentary cloud that is threaded by helical magnetic fields. The variation of physical conditions across the cloud is reflected in the physical characteristics of the dust cores. We find similar core properties between starless and protostellar cores, but variations in core properties with position in the filament. The OMC 1 cores have the highest velocity dispersions and masses, followed by OMC 2/3 and OMC 4. The differing fragmentation of these cores may explain why OMC 1 has formed clusters of high-mass stars, whereas OMC 4 produces fewer, predominantly low-mass stars

An ALMA study of hub-filament systems – I. On the clump mass concentration within the most massive cores

The physical processes behind the transfer of mass from parsec-scale clumps to massive star-forming cores remain elusive. We investigate the relation between the clump morphology and the mass fraction that ends up in its most massive core (MMC) as a function of infrared brightness, i.e. a clump evolutionary tracer. Using Atacama Large Millimeter/submillimeter Array (ALMA) 12 m and Atacama Compact Array, we surveyed six infrared dark hubs in 2.9 mm continuum at ∼3 arcsec resolution. To put our sample into context, we also re-analysed published ALMA data from a sample of 29 high-mass surface density ATLASGAL sources. We characterize the size, mass, morphology, and infrared brightness of the clumps using Herschel and Spitzer data. Within the six newly observed hubs, we identify 67 cores, and find that the MMCs have masses between 15 and 911 M⊙ within a radius of 0.018–0.156 pc. The MMC of each hub contains 3–24 per cent of the clump mass (fMMC), becoming 5–36 per cent once core masses are normalized to the median core radius. Across the 35 clumps, we find no significant difference in the median fMMC values of hub and non-hub systems, likely the consequence of a sample bias. However, we find that fMMC is ∼7.9 times larger for infrared dark clumps compared to infrared bright ones. This factor increases up to ∼14.5 when comparing our sample of six infrared dark hubs to infrared bright clumps. We speculate that hub-filament systems efficiently concentrate mass within their MMC early on during its evolution. As clumps evolve, they grow in mass, but such growth does not lead to the formation of more massive MMCs

Changes in Channel Trafficking and Protein Stability Caused by LQT2 Mutations in the PAS Domain of the HERG Channel

Inherited human long-QT2 syndrome (LQTS) results from mutations in the gene encoding the HERG channel. Several LQT2-associated mutations have been mapped to the amino terminal cytoplasmic Per-Arnt-Sim (PAS) domain of the HERG1a channel subunit. Here we have characterized the trafficking properties of some LQT2-associated PAS domain mutants and analyzed rescue of the trafficking mutants by low temperature (27°C) or by the pore blocker drug E4031. We show that the LQT2-associated mutations in the PAS domain of the HERG channel display molecular properties that are distinct from the properties of LQT2-associated mutations in the trans-membrane region. Unlike the latter, many of the tested PAS domain LQT2-associated mutations do not result in trafficking deficiency of the channel. Moreover, the majority of the PAS domain mutations that cause trafficking deficiencies are not rescued by a pore blocking drug. We have also explored the in vitro folding stability properties of isolated mutant PAS domain proteins using a thermal unfolding fluorescence assay and a chemical unfolding assay

Calmodulin Interaction with hEAG1 Visualized by FRET Microscopy

BACKGROUND: Ca(2+)-mediated regulation of ion channels provides a link between intracellular signaling pathways and membrane electrical activity. Intracellular Ca(2+) inhibits the voltage-gated potassium channel EAG1 through the direct binding of calmodulin (CaM). Three CaM binding sites (BD-C1: 674-683, BD-C2: 711-721, BD-N: 151-165) have been identified in a peptide screen and were proposed to mediate binding. The participation of the three sites in CaM binding to the native channel, however, remains unclear. METHODOLOGY/PRINCIPAL FINDINGS: Here we studied the binding of Ca(2+)/CaM to the EAG channel by visualizing the interaction between YFP-labeled CaM and Cerulean-labeled hEAG1 in mammalian cells by FRET. The results of our cellular approach substantiate that two CaM binding sites are predominantly involved; the high-affinity 1-8-14 based CaM binding domain in the N-terminus and the second C-terminal binding domain BD-C2. Mutations at these sites completely abolished CaM binding to hEAG1. CONCLUSIONS/SIGNIFICANCE: We demonstrated that the BD-N and BD-C2 binding domains are sufficient for CaM binding to the native channel, and, therefore, that BD-C1 is unable to bind CaM independently

HIV infection and drugs of abuse: role of acute phase proteins

Background HIV infection and drugs of abuse such as methamphetamine (METH), cocaine, and alcohol use have been identified as risk factors for triggering inflammation. Acute phase proteins such as C-reactive protein (CRP) and serum amyloid A (SAA) are the biomarkers of inflammation. Hence, the interactive effect of drugs of abuse with acute phase proteins in HIV-positive subjects was investigated. Methods Plasma samples were utilized from 75 subjects with METH use, cocaine use, alcohol use, and HIV-positive alone and HIV-positive METH, cocaine, and alcohol users, and age-matched control subjects. The plasma CRP and SAA levels were measured by ELISA and western blot respectively and the CD4 counts were also measured. Results Observed results indicated that the CRP and SAA levels in HIV-positive subjects who are METH, cocaine and alcohol users were significantly higher when compared with either drugs of abuse or HIV-positive alone. The CD4 counts were also dramatically reduced in HIV-positive with drugs of abuse subjects compared with only HIV-positive subjects. Conclusions These results suggest that, in HIV-positive subjects, drugs of abuse increase the levels of CRP and SAA, which may impact on the HIV infection and disease progression

Lineage Analysis of Circulating Trypanosoma cruzi Parasites and Their Association with Clinical Forms of Chagas Disease in Bolivia

Around 30–50% of Trypanosoma cruzi infections in Latin America cause chronic Chagas disease 10–30 years after the primary infection due to lack of effective treatment. The major clinical complications associated with chronic Chagas disease are cardiac myositis (leading to cardiac failure), and autonomous neuroplexus degeneration of the digestive tract that can cause megacolon or megaesophagus. Therefore, there are three major clinical forms of Chagas disease; cardiac, digestive and indeterminate (asymptomatic). The parasites, which can infect humans as well as other mammals, are transmitted by species of triatomines commonly found in the Americas. The parasite is divided in at least six discrete typing units: TcI, TcIIa–e. In humans, the TcI is mainly observed in Central America and northern parts of South America while the TcIIb/d/e is confined mainly to the southern cone of Latin America. We determined which DTU were prevalent in chronic patients in Bolivia, where the three clinical forms and several DTUs of the parasites are present, in order to determine whether there was a link between a particular parasite DTU and a particular clinical outcome. We found a vast majority of TcIId but its kDNA polymorphism showed no association with any of the clinical manifestations of chronic Chagas

Recreational and occupational field exposure to freshwater cyanobacteria – a review of anecdotal and case reports, epidemiological studies and the challenges for epidemiologic assessment

Cyanobacteria are common inhabitants of freshwater lakes and reservoirs throughout the world. Under favourable conditions, certain cyanobacteria can dominate the phytoplankton within a waterbody and form nuisance blooms. Case reports and anecdotal references dating from 1949 describe a range of illnesses associated with recreational exposure to cyanobacteria: hay fever-like symptoms, pruritic skin rashes and gastro-intestinal symptoms are most frequently reported. Some papers give convincing descriptions of allergic reactions while others describe more serious acute illnesses, with symptoms such as severe headache, pneumonia, fever, myalgia, vertigo and blistering in the mouth. A coroner in the United States found that a teenage boy died as a result of accidentally ingesting a neurotoxic cyanotoxin from a golf course pond. This death is the first recorded human fatality attributed to recreational exposure to cyanobacteria, although uncertainties surround the forensic identification of the suspected cyanotoxin in this case. We systematically reviewed the literature on recreational exposure to freshwater cyanobacteria. Epidemiological data are limited, with six studies conducted since 1990. Statistically significant increases in symptoms were reported in individuals exposed to cyanobacteria compared to unexposed counterparts in two Australian cohort studies, though minor morbidity appeared to be the main finding. The four other small studies (three from the UK, one Australian) did not report any significant association. However, the potential for serious injury or death remains, as freshwater cyanobacteria under bloom conditions are capable of producing potent toxins that cause specific and severe dysfunction to hepatic or central nervous systems. The exposure route for these toxins is oral, from ingestion of recreational water, and possibly by inhalation. A range of freshwater microbial agents may cause acute conditions that present with features that resemble illnesses attributed to contact with cyanobacteria and, conversely, acute illness resulting from exposure to cyanobacteria or cyanotoxins in recreational waters could be misdiagnosed. Accurately assessing exposure to cyanobacteria in recreational waters is difficult and unreliable at present, as specific biomarkers are unavailable. However, diagnosis of cyanobacteria-related illness should be considered for individuals presenting with acute illness following freshwater contact if a description is given of a waterbody visibly affected by planktonic mass development

Morphine Induces Expression of Platelet-Derived Growth Factor in Human Brain Microvascular Endothelial Cells: Implication for Vascular Permeability

Despite the advent of antiretroviral therapy, complications of HIV-1 infection with concurrent drug abuse are an emerging problem. Morphine, often abused by HIV-infected patients, is known to accelerate neuroinflammation associated with HIV-1 infection. Detailed molecular mechanisms of morphine action however, remain poorly understood. Platelet-derived growth factor (PDGF) has been implicated in a number of pathological conditions, primarily due to its potent mitogenic and permeability effects. Whether morphine exposure results in enhanced vascular permeability in brain endothelial cells, likely via induction of PDGF, remains to be established. In the present study, we demonstrated morphine-mediated induction of PDGF-BB in human brain microvascular endothelial cells, an effect that was abrogated by the opioid receptor antagonist-naltrexone. Pharmacological blockade (cell signaling) and loss-of-function (Egr-1) approaches demonstrated the role of mitogen-activated protein kinases (MAPKs), PI3K/Akt and the downstream transcription factor Egr-1 respectively, in morphine-mediated induction of PDGF-BB. Functional significance of increased PDGF-BB manifested as increased breach of the endothelial barrier as evidenced by decreased expression of the tight junction protein ZO-1 in an in vitro model system. Understanding the regulation of PDGF expression may provide insights into the development of potential therapeutic targets for intervention of morphine-mediated neuroinflammation

The Alchemist

JOURThis is the final version of the article. It was first published by IOP Publishing for the Royal Astronomical Society via http://dx.doi.org/10.3847/0004-637X/817/2/167We present a first look at the SCUBA-2 observations of three sub-regions of the Orion B molecular cloud: LDN 1622, NGC 2023/2024, and NGC 2068/2071, from the JCMT Gould Belt Legacy Survey. We identify 29, 564, and 322 dense cores in L1622, NGC 2023/2024, and NGC 2068/2071 respectively, using the SCUBA-2 850 μm map, and present their basic properties, including their peak fluxes, total fluxes, and sizes, and an estimate of the corresponding 450 μm peak fluxes and total fluxes, using the FellWalker source extraction algorithm. Assuming a constant temperature of 20 K, the starless dense cores have a mass function similar to that found in previous dense core analyses, with a Salpeter-like slope at the high-mass end. The majority of cores appear stable to gravitational collapse when considering only thermal pressure; indeed, most of the cores which have masses above the thermal Jeans mass are already associated with at least one protostar. At higher cloud column densities, above 1–2 × 1023 cm‑2, most of the mass is found within dense cores, while at lower cloud column densities, below 1 × 1023 cm‑2, this fraction drops to 10% or lower. Overall, the fraction of dense cores associated with a protostar is quite small (<8%), but becomes larger for the densest and most centrally concentrated cores. NGC 2023/2024 and NGC 2068/2071 appear to be on the path to forming a significant number of stars in the future, while L1622 has little additional mass in dense cores to form many new stars