High-throughput bioluminescence screening of ubiquitin-proteasome pathway inhibitors from chemical and natural sources

To discover original inhibitors of the ubiquitin-proteasome pathway, the authors have developed a cell-based bioluminescent assay and used it to screen collections of plant extracts and chemical compounds. They first established a DLD-1 human colon cancer cell line that stably expresses a 4Ubiquitin-Luciferase (4Ub-Luc) reporter protein, efficiently targeted to the ubiquitin-proteasome degradation pathway. The assay was then adapted to 96- and 384-well plate formats and calibrated with reference proteasome inhibitors. Assay robustness was carefully assessed, particularly cell toxicity, and the statistical Z factor value was calculated to 0.83, demonstrating a good performance level of the assay. A total of 18,239 molecules and 15,744 plant extracts and fractions thereof were screened for their capacity to increase the luciferase activity in DLD-1 4Ub-Luc cells, and 21 molecules and 66 extracts inhibiting the ubiquitin-proteasome pathway were identified. The fractionation of an active methanol extract of Physalis angulata L. aerial parts was performed to isolate 2 secosteroids known as physalin B and C. In a cell-based Western blot assay, the ubiquitinated protein accumulation was confirmed after a physalin treatment confirming the accuracy of the screening process. The method reported here thus provides a robust approach to identify novel ubiquitin-proteasome pathway inhibitors in large collections of chemical compounds and natural products.     

Maternal immunity and mother-to-child transmission of HCV and HIV-1: challenges and recent advances

Human immunodeficiency virus type 1 (HIV-1) and hepatitis C virus (HCV) are two viral pathogens that establish chronic infections in their hosts and that are at present responsible for serious public health problems on a pandemic scale. HIV-1 and HCV can be transmitted from person to person by contact with bodily fluids. Both can also be transmitted from mother to child during the course of pregnancy and childbirth. There are currently no vaccines available to immunize against HIV-1 and HCV infection or to prevent mother-to-child transmission (MTCT), and accessible treatments have significant yet limited efficacy. However, important progresses have been made since the discovery of HCV and HIV-1 : (a) sensitive screening and detection methods have been perfected ; (b) risk factors for acquisition, replicative cycles, pathogenesis, and mechanisms of transmission have been better characterized ; (c) specific treatments, immunotherapy, and antiretroviral prophylaxis regimen were developed ; (d) immune correlates of protection are better understood ; and (e) vaccine design was undertaken. In addition, co-infection with HCV and HIV-1, which is common among high-risk groups including injection drug users, significantly increases the incidence of MTCT of both viruses. The mechanisms by which this facilitation occurs are still under investigation and may involve direct replicative facilitation, enhancement of placental transfer, and/or interference with host immune responses. Taken together, these developments could lead to the implementation of global scale strategies to prevent MTCT of HCV and HIV-1.     

Soil sampling for <Emphasis Type="Italic">Metarhizium anisopliae</Emphasis> spores in Queensland sugarcane fields

Three sugarcane fields in Bundaberg and four fields in each of the Burdekin, Tully and Innisfail (Queensland, Australia) were sampled for spores of Metarhizium anisopliae (Metchnikoff) (Deuteromycotina: Hyphomycetes). This entomopathogenic fungus is the active ingredient in the biocide “BioCane^TM”, which was developed for the management of the greyback canegrub Dermolepida albohirtum (Waterhosue) (Coleoptera: Scarabaeidae) and other scarabs in cane fields. Fields sampled were of different crop ages and all had a history of BioCane^TM treatment in Plant Cane in past years. Soil samples were taken in each field from four depths (0–10, 10–20, 20–30 and 30–40 cm below soil level) with the use of an auger. Spore levels were highest at the depths of 10–20 and 20–30 cm. Spore levels differed between locations with Innisfail and Tully recording the highest spore counts. Spores were also found in the inter-row space in plots sampled in Tully. Sampling statistics were determined for M. anisopliae spores at the four soil depths with 0.1 and 0.25 precision levels. Three sampling methods were compared (use of marker beads; use of 100 mm auger and 150 mm auger). Samples that relied on marker beads resulted in higher spore counts, however, an auger can still be used since BioCane^TM does not normally contain coloured markers. Results obtained demonstrate the ability of the pathogen to translocate in soil profile and across rows, most likely due to grub movements and other soil fauna. Sampling for M. anisopliae spores provides good monitoring of their levels in soil. The implications of this on grub management decisions are discussed.     

Estimation of linear and mass attenuation coefficients of soy–lignin bonded Rhizophora spp. particleboard as a potential phantom material using caesium-137 and cobalt-60

In this study, linear and mass attenuation coefficients of fabricated particleboards intended for use as phantom material were estimated using ¹³⁷Cs and ⁶⁰Co radiation sources. Particleboards made of Rhizophora spp. wood trunk bonded with soy flour and lignin were fabricated at a target density of 1.0 g cm^?3, with and without gloss finish coating. Elemental composition of the particleboards was obtained by means of energy dispersive X-ray (EDX) spectroscopy. Experimental setups were simulated via the GATE Monte Carlo (MC) package, with particle histories of 1?×?10⁶–1?×?10⁷. Linear and mass attenuation coefficients obtained from measurements and GATE simulations were compared and discussed. The percentage differences between the measured and simulated linear and mass attenuation coefficients of the samples were reasonably small (2.05–4.88% for ¹³⁷Cs and 3.24–5.38% for ⁶⁰Co). It is shown that all the particleboards have the potential to be used as phantom materials as the attenuation coefficients measured were in good agreement with those of water (calculated with XCOM) and with those simulated with the GATE toolkit. The use of gloss finish coating also did not show any significant effect on the attenuation coefficient of the phantom material. Verification of experimental results via GATE simulations has been shown crucial in providing reliable data for energy transmission studies. Based on the results achieved in this study, it is concluded that the studied material—Rhizophora spp. wood trunk bonded with soy flour and lignin including gloss finish coating—can be used in radiation dosimetry studies.

     

Tetrahydrobiopterin and Its Multiple Roles in Neuropsychological Disorders

Neurochemical Research - Tetrahydrobiopterin (BH4) is a multifunctional co-factor of various enzymes and a substantial amount of studies have shown BH4 as a key regulator in the synthesis of...     

Peridontitis as a Risk Factor for Attention Deficit Hyperactivity Disorder: Possible Neuro-inflammatory Mechanisms

Periodontitis is a condition caused mostly by the creation of a biofilm by the bacterium P. gingivalis, which releases toxins and damages the tooth structure. Recent research studies have reported association between dental health and neuropsychiatric illnesses. Neuroinflammation triggered by the first systemic inflammation caused by the bacterium present in the oral cavities is a plausible explanation for such a relationship. Substantial amount of evidence supports the role of neuroinflammation and dysfunction of the dopaminergic system in the pathology of ADHD (Attention deficit hyperactivity disorders). Recent epidemiological, microbiological and inflammatory findings strengthen that, periodontal bacteria, which cause systemic inflammation can contribute to neuroinflammation and finally ADHD. Although both diseases are characterized by inflammation, the specific pathways and crosslink’s between periodontitis and ADHD remain unknown. Here, the authors describe the inflammatory elements of periodontitis, how this dental illness causes systemic inflammation, and how this systemic inflammation contributes to deteriorating neuroinflammation in the evolution of ADHD. Therefore, the aim of this review is to present possible links and mechanisms that could confirm the evidence of this association.

     

Untargeted metabolomics reveals differences between commercial and non-commercial Camellia sinensis cultivars used in black tea production

Journal of Plant Biochemistry and Biotechnology - Tea (Camellia sinensis) has enthralled both consumers and researchers, due to its taste, aroma and its medicinal attributes. Tea consumers concern...