Similar regulation of cell surface human T-cell leukemia virus type 1 (HTLV-1) surface binding proteins in cells highly and poorly transduced by HTLV-1-pseudotyped virions

Little is known about the requirements for human T-cell leukemia virus type 1 (HTLV-1) entry, including the identity of the cellular receptor(s). Previous studies have shown that although the HTLV receptor(s) are widely expressed on cell lines of various cell types from different species, cell lines differ dramatically in their susceptibility to HTLV-Env-mediated fusion. Human cells (293, HeLa, and primary CD4(+) T cells) showed higher levels of binding at saturation than rodent (NIH 3T3 and NRK) cells to an HTLV-1 SU immunoadhesin. A direct comparison of the binding of the HTLV-1 surface glycoprotein (SU) immunoadhesin and transduction by HTLV-1 pseudotyped virus revealed parallels between the level of binding and the titer for various cell lines. When cells were treated with phorbol myristate acetate (PMA), which down-modulates a number of cell surface molecules, the level of SU binding was markedly reduced. However, PMA treatment only slightly reduced the titer of murine leukemia virus(HTLV-1) on both highly susceptible and poorly susceptible cells. Treatment of target cells with trypsin greatly reduced binding, indicating that the majority of HTLV SU binding is to proteins. Polycations, which enhance the infectivity of several other retroviruses, inhibited HTLV-1 Env-mediated binding and entry on both human and rodent cells. These results suggest that factors other than the number of primary binding receptors are responsible for the differences in the titers of HTLV-1 pseudotypes between highly susceptible cells and poorly susceptible cells.     

Ecological characteristics of plant species and associations along two Dinaric karstic rivers (Bosnia and Herzegovina, the Balkans)

This paper describes the plant communities of two oligotrophic karstic rivers with a slight anthropogenic influence — the Trebi?at and the Li?tica — in South Bosnia and Herzegovina, their sinecology, and the relationship between vegetation and plant species and environmental parameters. According to 87 relevés, a total of 26 plant associations, using Braun-Blanquet methods, were found in the rivers and nearby surveyed area. Only nine associations were common to both rivers. Eight associations were recorded for the territory of Bosnia and Herzegovina for the first time. Associations from both rivers differed according to following parameters measured at the sampling site: water temperature, distance from the mouth (river kilometers), river width, water depth, flow, pH and slope. There were no differences in nutrient concentrations among the associations. According to Canonical Correspondence Analysis (CCA), three environmental variables out of the 11 initially considered were retained as being related to plant distribution. The first two axes explained 56.8% and 35.2% of variance of species-environment relationship in the Trebi?at and Li?tica rivers, respectively. Water depth in the Trebi?at River, and pH and river kilometers in the Li?tica River were found to be the most influential, while nutrients and other physico-chemical parameters were not significant in either river. Water depth vector was identified as an underlying environmental factor determining distribution of Potamogeton lucens and Myriophyllum verticillatum (Potamion), and Nymphaea alba (Nymphaeion albae). According to in situ measurements of light intensity, the highest coefficients of light attenuation, radiance and reflection were found at a station with dense populations of Potamogeton lucens and Nuphar luteum (Potametum lucentis). The vegetation and floristic value of the rivers is discussed as a basis for biodiversity conservation programmes and evaluating the state of these ecosystems in the future.     

The Cedar Project: risk factors for transition to injection drug use among young, urban Aboriginal people

Background:

Studies suggest that Aboriginal people in Canada are over-represented among people using injection drugs. The factors associated with transitioning to the use of injection drugs among young Aboriginal people in Canada are not well understood.

Methods:

The Cedar Project is a prospective cohort study (2003–2007) involving young Aboriginal people in Vancouver and Prince George, British Columbia, who use illicit drugs. Participants’ venous blood samples were tested for antibodies to HIV and the hepatitis C virus, and drug use was confirmed using saliva screens. The primary outcomes were use of injection drugs at baseline and tranisition to injection drug use in the six months before each follow-up interview.

Results:

Of 605 participants, 335 (55.4%) reported using injection drugs at baseline. Young people who used injection drugs tended to be older than those who did not, female and in a relationship. Participants who injected drugs were also more likely than those who did not to have been denied shelter because of their drug use, to have been incarcerated, to have a mental illness and to have been involved in sex work. Transition to injection drug use occurred among 39 (14.4%) participants, yielding a crude incidence rate of 19.8% and an incidence density of 11.5 participants per 100 person-years. In unadjusted analysis, transition to injection drug use was associated with being female (odds ratio [OR] 1.98, 95% confidence interval (CI) 1.06–3.72), involved in sex work (OR 3.35, 95% CI 1.75–6.40), having a history of sexually transmitted infection (OR 2.01, 95% CI 1.07–3.78) and using drugs with sex-work clients (OR 2.51, 95% CI 1.19–5.32). In adjusted analysis, transition to injection drug use remained associated with involvement in sex work (adjusted OR 3.94, 95% CI 1.45–10.71).

Interpretation:

The initiation rate for injection drug use of 11.5 participants per 100 person-years among participants in the Cedar Project is distressing. Young Aboriginal women in our study were twice as likely to inject drugs as men, and participants who injected drugs at baseline were more than twice as likely as those who did not to be involved in sex work.Aboriginal leadership in Canada is deeply concerned about substance use, more specifically injection drug use and its association with the spread of HIV and the hepatitis C virus among Aboriginal young people.^1,2 Recent studies in Canada suggest that Aboriginal people are over-represented among people who use injection drugs.^3,4 For Aboriginal young people in Canada under the age of 24 years, injection drug use accounts for the majority of infections with the hepatitis C virus (70%–80%)^5,6 and over half (59%) of HIV infections.⁷Indigenous scholars have stated that research on substance use within Aboriginal communities must consider the context of colonization, including the intergenerational impacts of the residential school and child welfare systems.^8–11 It is now well documented that Aboriginal children experienced extensive psychological, sexual, physical and emotional abuses within those systems.^12,13 As former students of residential schools raise children and grandchildren, the intergenerational effects of abuse and familial fragmentation are evident in communities where interpersonal violence and drug dependence are pervasive.^14–16A priority for preventing infections with HIV and hepatitis C among young Aboriginal people is the development of programs and rights-based,^18,19 youth-informed¹⁷ policies aimed at preventing the use of injection drugs. However, research to date has not provided sufficient evidence to inform such development.^2,19 Concerns over this paucity of information led to the launch of a two-city cohort study in 2003 to address HIV-related vulnerabilities among young Aboriginal people in British Columbia — a unique study centred on at-risk youth and supported by and partnered with Aboriginal investigators and collaborators.We report here baseline and longitudinal data on the factors associated with injection drug use and the transition to injection drug use to inform the development of prevention programs and policies.     

Antimicrobial activity of new phorbins from Jatropha curcas Linn. (Euphorbiaceae) leaves

The crude methanol extract of Jatropha curcas leaves exhibited activity against Staphylococcus aureus, Bacillus subtilis, Mycobacterium phlei, Candida albicans, and Trichophyton mentagrophytes but was inactive against Escherichia coli, Pseudomonas aeruginosa, and Saccharomyces cerevisiae. In a bioassay-directed fractionation, two new phorbins were isolated and analysed by spectroscopic methods. Isolate 1 was characterized as an analogue of pheophytin b with a phytyl moiety containing three double bonds which are at positions P2/P3, P6/P7, and P10/P11. Compound 2 was characterized as methyl pheophorbide a with 132-OH and 17- and 17(1)-CH3 moieties. It is active against Serratia marcescens.     

Potentials for win-win alliances among animal agriculture and forest products industries: Application of the principles of industrial ecology and sustainable development

Commercial forests in many parts of the world are deficient in nitrogen and phosphorus. These nutrient-deficient forests often exist in close proximity to large animal feeding operations, meat processing and other food, textile, or other biomass-processing plants, and municipal waste treatment facilities. Many of these facilities produce large surpluses of nitrogen, phosphorus, and organic matter as gaseous ammonia, urea, uric acid, phosphorus compounds, bacterial sludges, and partially treated municipal wastewaters. These co-existing and substantial nutrient deficiencies and surpluses offer ready-made opportunities for discovery, demonstration, and commercial development of science-based, technology-facilitated, environmentally sound, economically viable, and socially acceptable "win-win alliances" among these major industries based on the principles of industrial ecology and sustainable development. The major challenge is to discover practical means to capture the surplus nutrients and put them to work in forest stands from which value-added products can be produced and sold at a profit.     

Identification and characterization of human apurinic/apyrimidinic endonuclease-1 inhibitors

The repair of abasic sites that arise in DNA from hydrolytic depurination/depyrimidination of the nitrogenous bases from the sugar-phosphate backbone and the action of DNA glycosylases on deaminated, oxidized, and alkylated bases are critical to cell survival. Apurinic/apyrimidinic endonuclease-1/redox effector factor-1 (APE-1; aka APE1/ref-1) is responsible for the initial removal of abasic lesions as part of the base excision repair pathway. Deletion of APE-1 activity is embryonic lethal in animals and is lethal in cells. Potential inhibitors of the repair function of APE-1 were identified based upon molecular modeling of the crystal structure of the APE-1 protein. We describe the characterization of several unique nanomolar inhibitors using two complementary biochemical screens. The most active molecules all contain a 2-methyl-4-amino-6,7-dioxolo-quinoline structure that is predicted from the modeling to anchor the compounds in the endonuclease site of the protein. The mechanism of action of the selected compounds was probed by fluorescence and competition studies, which indicate, in a specific case, direct interaction between the inhibitor and the active site of the protein. It is demonstrated that the inhibitors induce time-dependent increases in the accumulation of abasic sites in cells at levels that correlate with their potency to inhibit APE-1 endonuclease excision. The inhibitor molecules also potentiate by 5-fold the toxicity of a DNA methylating agent that creates abasic sites. The molecules represent a new class of APE-1 inhibitors that can be used to probe the biology of this critical enzyme and to sensitize resistant tumor cells to the cytotoxicity of clinically used DNA damaging anticancer drugs.     

Cell-free HTLV-1 infects dendritic cells leading to transmission and transformation of CD4(+) T cells

Cell-free human T-lymphotropic virus type 1 (HTLV-1) virions are poorly infectious in vitro for their primary target cells, CD4(+) T cells. Here, we show that HTLV-1 can efficiently infect myeloid and plasmacytoid dendritic cells (DCs). Moreover, DCs exposed to HTLV-1, both before and after being productively infected, can rapidly, efficiently and reproducibly transfer virus to autologous primary CD4(+) T cells. This DC-mediated transfer of HTLV-1 involves heparan sulfate proteoglycans and neuropilin-1 and results in long-term productive infection and interleukin-2-independent transformation of the CD4(+) T cells. These studies, along with observations of HTLV-1-infected DCs in the peripheral blood of infected individuals, indicate that DCs have a central role in HTLV-1 transmission, dissemination and persistence in vivo. In addition to altering the current paradigm concerning how HTLV-1 transmission occurs, these studies suggest that impairment of DC function after HTLV-1 infection plays a part in pathogenesis.