CONTAMINATION BY PERSISTENT ORGANIC POLLUTANTS AND ENDOCRINE DISRUPTING CHEMICALS IN VIETNAM : PATTERNS, BEHAVIOR, TRENDS AND TOXIC POTENTIAL

Joint Research on Environmental Science and Technology for the Eart

In vitro bioactivities of Codonopsis javanica root extract from Kon Tum province, Vietnam

Dangshen Codonopsis javanica exhibits invaluable medicinal properties in herbal remedies; however, there has currently not been much specific analysis of the phytochemicals and bioactivities of this plant. The root ethanol extract of C. javanica contains substances such as saponins, phenolic acids, terpenoids, and alkaloids. It displays an antibacterial effect against Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Bacillus cereus with the IC50 values of 150, 100, 150, and 90 μg/mL, respectively. The antioxidant capacity of the root extract was also observed with an IC50 value of 46.8 ± 6.8 μg/mL. Furthermore, the extract exhibits activity on human cancer cell lines HepG2 (IC50 = 83.6 ± 2.7 μg/mL) and MCF-7 (IC50 = 95.3 ± 2.3 μg/mL). Hence, this study provides the basic data for further research on the bioactivities of natural compounds of Dangshen C. javanica for the first time

Mechanisms Underlying Stage-1 TRPL Channel Translocation in Drosophila Photoreceptors

Background: TRP channels function as key mediators of sensory transduction and other cellular signaling pathways. In Drosophila, TRP and TRPL are the light-activated channels in photoreceptors. While TRP is statically localized in the signaling compartment of the cell (the rhabdomere), TRPL localization is regulated by light. TRPL channels translocate out of the rhabdomere in two distinct stages, returning to the rhabdomere with dark-incubation. Translocation of TRPL channels regulates their availability, and thereby the gain of the signal. Little, however, is known about the mechanisms underlying this trafficking of TRPL channels. Methodology/Principal Findings: We first examine the involvement of de novo protein synthesis in TRPL translocation. We feed flies cycloheximide, verify inhibition of protein synthesis, and test for TRPL translocation in photoreceptors. We find that protein synthesis is not involved in either stage of TRPL translocation out of the rhabdomere, but that re-localization to the rhabdomere from stage-1, but not stage-2, depends on protein synthesis. We also characterize an ex vivo eye preparation that is amenable to biochemical and genetic manipulation. We use this preparation to examine mechanisms of stage-1 TRPL translocation. We find that stage-1 translocation is: induced with ATP depletion, unaltered with perturbation of the actin cytoskeleton or inhibition of endocytosis, and slowed with increased membrane sterol content. Conclusions/Significance: Our results indicate that translocation of TRPL out of the rhabdomere is likely due to protei

Stage-1 TRPL Translocation is Not Regulated by <i>shibire</i>-Mediated Endocytosis.

<p>Shown are representative retinal sections of single ommatidia from wild-type and <i>shibire^ts1</i> mutants. Dark-raised flies were first incubated at 30°C to block endocytosis in <i>shi^ts1</i>, indicated by paralysis. Flies continued to be incubated at 30°C but either remained in the dark or were light-exposed for 2 hours. Tissue sections were immunostained for TRPL. TRPL displayed normal light-induced translocation from the rhabdomere to the neighboring stalk membrane. Multiple tissue sections were taken from 14 eyes from nine flies (wild-type, Dark), 10 eyes from 8 flies (wild-type, Light), 10 eyes from 7 flies (<i>shibire^ts1</i>, Dark), 15 eyes from 10 flies (<i>shibire^ts1</i>, Light).</p

Testing the Requirement of Protein Synthesis for TRPL Channel Translocation.

<p>(A) <i>hs-inaD</i> transgenic flies (in an <i>inaD¹</i> null background) express INAD protein only when heat-shocked at 37°C. One hour of heat-shock induces INAD expression that can be easily detected by immunoblot analysis (see -CHX). Flies fed green-colored cycloheximide (CHX) for 30 minutes were selected with for abdomens with various degrees of greenness (degree green), heat-shocked, and analyzed by immunoblot analysis (three heads per lane). Degree greenness had an inverse relationship with inducible expression of INAD, indicating that CHX treatment did indeed block protein synthesis. Anti-syntaxin was used as a loading control. (B) Dark-raised wild-type flies were fed +/− CHX (colored green) in the dark for at least 30 minutes, then light-exposed for either 30 minutes (Stage-1) or 12 hours (Stage-2), while remaining on the +/− CHX food throughout the light-exposure. Only flies with dark-green abdomens were selected for sectioning. Shown are representative retinal sections immunostained for TRPL. TRPL channels translocated to both stage-1 and -2 with CHX treatment. Multiple tissue sections were taken from 11 eyes from 8 flies (Dark), 8 eyes from 7 flies (Stage 1, -CHX), 9 eyes from 7 flies (Stage 1, +CHX). (C) Left, Wild-type flies were fed +/− CHX in the dark for at least 30 minutes to inhibit protein synthesis, light-exposed for 30 minutes to induce stage-1 translocation, followed by 6 hours of dark incubation. Only flies with dark-green abdomens were selected for sectioning and immunostaining. TRPL channels relocalized to the rhabdomeres in –CHX flies, but were unable to translocate back to the rhabdomere in +CHX fed flies. Right, Wild-type flies were light-exposed for 12 hours to induce stage-2 translocation, then transferred to +/− CHX food in continuous light for 1 additional hour, followed by dark incubation for 10 hours. Only flies with dark-green abdomens were selected for sectioning and immunostaining. Representative sections show that TRPL channels relocalized to the rhabdomere from stage-2 in both +/− CHX. Multiple sections were taken from 13 eyes from 9 flies (Stage 1), 7 eyes from 5 flies (Dark 6 hrs, -CHX), 12 eyes from 9 flies (Dark 6 hrs, +CHX), 13 eyes from 11 flies (Stage 2), 11 eyes from 9 flies (Dark 10 hrs, -CHX), 16 eyes from 10 flies (Dark 10 hrs, +CHX).</p

Increased Dietary Ergosterol Slowed the Rate of TRPL Translocation.

<p>(A) Wild-type flies were fed a defined medium containing either wild-type or <i>mot3Δ</i> mutant yeast for 10, 15, 20 or 30 days in the dark. Sterols were extracted from whole fly membranes and then scanned spectrophotometrically. Left, shown are representative spectral profiles (250–300 nm) of sterols from flies raised for 30 days on food made with wild-type yeast (wt-fed) or <i>mot3Δ</i> mutant yeast (<i>mot3Δ</i>-fed). This four-peaked profile is characteristic of ergosterol, the major sterol present in <i>Drosophila</i>. <i>mot3Δ</i>-fed flies consistently displayed an increased absorption spectrum, compared to wt-fed flies. Right, quantification of ergosterol increase in flies fed <i>mot3Δ</i>-food, relative to wt-food, for 10, 15, 20, and 30 days. (B) Left, shown are representative retinal sections immunostained for TRPL from wt-fed or <i>mot3Δ</i>-fed flies. Dark-raised flies showed rhabdomeric localization of TRPL. With <u>17</u> minutes of light-exposure, wt-fed flies displayed a more evident stage-1 TRPL translocation, while flies fed <i>mot3Δ</i>-based food displayed a more rhabdomeric-like TRPL localization. After 30 minutes of light-exposure, both wt-fed and <i>mot3Δ</i>-fed flies appeared to display stage-1 translocation. Right, Box-and-Whisker-Plots showing the relative rhabdomeric TRPL signal between wt-fed and <i>mot3Δ</i>-fed flies when exposed to 17 and 30 min of light. Multiple tissue sections were taken from multiple eyes/flies (across different experiments), as follows. For wild-type, 5 eyes from 4 flies (Dark), <u>10</u> eyes from <u>5</u> flies (LEX, <u>17</u> min), <u>2</u> eyes from <u>2</u> flies (LEX, 30 min); for <i>mot3Δ</i>: <u>5</u> eyes from <u>4</u> flies (dark), <u>7</u> eyes from <u>5</u> flies (LEX, <u>17</u> min), <u>3</u> eyes from <u>3</u> flies (LEX, 30 min). ** t-test analysis indicates a significant difference (P<0.01).</p

A novel, high-performance random array platform for quantitative gene expression profiling

We have developed a new microarray technology for quantitative gene-expression profiling on the basis of randomly assembled arrays of beads. Each bead carries a gene-specific probe sequence. There are multiple copies of each sequence-specific bead in an array, which contributes to measurement precision and reliability. We optimized the system for specific and sensitive analysis of mammalian RNA, and using RNA controls of defined concentration, obtained the following estimates of system performance: specificity of 1:250,000 in mammalian poly(A(+)) mRNA; limit of detection 0.13 pM; dynamic range 3.2 logs; and sufficient precision to detect 1.3-fold differences with 95% confidence within the dynamic range. Measurements of expression differences between human brain and liver were validated by concordance with quantitative real-time PCR (R(2) = 0.98 for log-transformed ratios, and slope of the best-fit line = 1.04, for 20 genes). Quantitative performance was further verified using a mouse B- and T-cell model system. We found published reports of B- or T-cell-specific expression for 42 of 59 genes that showed the greatest differential expression between B- and T-cells in our system. All of the literature observations were concordant with our results. Our experiments were carried out on a 96-array matrix system that requires only 100 ng of input RNA and uses standard microtiter plates to process samples in parallel. Our technology has advantages for analyzing multiple samples, is scalable to all known genes in a genome, and is flexible, allowing the use of standard or custom probes in an array

Trinary Component Adsorption of Methylene Blue, Methyl Orange, and Methyl Red from Aqueous Solution Using TiO2/Activated Carbon

Porous TiO2/activated carbon (AC) material was synthesized by grafting peroxo-hydro titanium complexes to rice husk-derived activated carbon. It was found that the morphology of TiO2/AC consists of TiO2 fine particles highly dispersed on the AC matrix. The obtained TiO2/AC composites with high surface area and a red shift exhibit an excellent adsorption performance in both single and trinary system toward methylene blue (MB), methyl orange (MO), and methyl red (MR). The isotherm models including extended Langmuir, P-factor, ideal adsorbed solution theory (IAST) for Langmuir, Freundlich, and Sips models were applied to study the adsorption equilibrium data of trinary solutions. It was found that IAST for Freundlich and Langmuir models were the most suitable one to describe the adsorption of the three dyes on TiO2/AC material. The high maximum adsorption capacities (mmol g-1) in single/trinary mixture were found as 0.452/0.340 for MB; 0.329/0.321 for MO; and 0.806/2.04 for MR. Moreover, the recyclability experiments showed that the adsorbent could be reused through photocatalytic self-cleaning for at least three cycles with stable capacity. Thus, the TiO2/AC can be effectively employed for the removal of dyes from industrial textile wastewater

Human versus equine intramuscular antitoxin, with or without human intrathecal antitoxin, for the treatment of adults with tetanus: a 2 × 2 factorial randomised controlled trial

Background Intramuscular antitoxin is recommended in tetanus treatment, but there are few data comparing human and equine preparations. Tetanus toxin acts within the CNS, where there is limited penetration of peripherally administered antitoxin; thus, intrathecal antitoxin administration might improve clinical outcomes compared with intramuscular injection. Methods In a 2  × 2 factorial trial, all patients aged 16 years or older with a clinical diagnosis of generalised tetanus admitted to the intensive care unit of the Hospital for Tropical Diseases, Ho Chi Minh City, Vietnam, were eligible for study entry. Participants were randomly assigned first to 3000 IU human or 21 000 U equine intramuscular antitoxin, then to either 500 IU intrathecal human antitoxin or sham procedure. Interventions were delivered by independent clinicians, with attending clinicians and study staff masked to treatment allocations. The primary outcome was requirement for mechanical ventilation. The analysis was done in the intention-to-treat population. The study is registered at ClinicalTrials.gov, NCT02999815; recruitment is completed. Findings 272 adults were randomly assigned to interventions between Jan 8, 2017, and Sept 29, 2019, and followed up until May, 2020. In the intrathecal allocation, 136 individuals were randomly assigned to sham procedure and 136 to antitoxin; in the intramuscular allocation, 109 individuals were randomly assigned to equine antitoxin and 109 to human antitoxin. 54 patients received antitoxin at a previous hospital, excluding them from the intramuscular antitoxin groups. Mechanical ventilation was given to 56 (43%) of 130 patients allocated to intrathecal antitoxin and 65 (50%) of 131 allocated to sham procedure (relative risk [RR] 0·87, 95% CI 0·66–1·13; p=0·29). For the intramuscular allocation, 48 (45%) of 107 patients allocated to human antitoxin received mechanical ventilation compared with 48 (44%) of 108 patients allocated to equine antitoxin (RR 1·01, 95% CI 0·75–1·36, p=0·95). No clinically relevant difference in adverse events was reported. 22 (16%) of 136 individuals allocated to the intrathecal group and 22 (11%) of 136 allocated to the sham procedure experienced adverse events related or possibly related to the intervention. 16 (15%) of 108 individuals allocated to equine intramuscular antitoxin and 17 (16%) of 109 allocated to human antitoxin experienced adverse events related or possibly related to the intervention. There were no intervention-related deaths. Interpretation We found no advantage of intramuscular human antitoxin over intramuscular equine antitoxin in tetanus treatment. Intrathecal antitoxin administration was safe, but did not provide overall benefit in addition to intramuscular antitoxin administration