A synthetic biology approach for consistent production of plant-made recombinant polyclonal antibodies against snake venom toxins.

Antivenoms developed from the plasma of hyperimmunized animals are the only effective treatment available against snakebite envenomation but shortage of supply contributes to the high morbidity and mortality toll of this tropical disease. We describe a synthetic biology approach to affordable and cost-effective antivenom production based on plant-made recombinant polyclonal antibodies (termed pluribodies). The strategy takes advantage of virus superinfection exclusion to induce the formation of somatic expression mosaics in agroinfiltrated plants, which enables the expression of complex antibody repertoires in a highly reproducible manner. Pluribodies developed using toxin-binding genetic information captured from peripheral blood lymphocytes of hyperimmunized camels recapitulated the overall binding activity of the immune response. Furthermore, an improved plant-made antivenom (plantivenom) was formulated using an in vitro selected pluribody against Bothrops asper snake venom toxins and has been shown to neutralize a wide range of toxin activities and provide protection against lethal venom doses in mice.

Microbial community composition and cooccurrence patterns driven by co-contamination of arsenic and antimony in antimony-mining area.

In the current study, a typical Sb mine was selected to explore the microbial community composition and assembly driven by the cocontamination of As/Sb with geographic distance. Our results showed that environmental parameters, especially pH, TOC, nitrate, total and bioavailable As/Sb contents largely affected the microbial community diversity and composition. The total and bioavailable As/Sb levels were significantly positively correlated with the relative abundance of Zavarzinella, Thermosporothrix and Holophaga, while the pH presented a significant negative correlation with the three genera, potentially implying they are important taxonomic groups in acid-mining soils. The cooccurrence network analysis indicated the environmental stress dominated by pH and As/Sb co-contamination affected the microbial modularity and interaction. Meanwhile, Homogeneous selection (HoS, 26.4-49.3%), and drift and others (DR, 27.1∼40.2%) were the most important assembly processes for soil bacterial, and the importance of HoS decreased and the DR increased with geographic distance to the contamination source respectively. Soil pH, nutrient availability, total and bioavailable As/Sb contents significantly affected the HoS and DR processes. This study provides theoretical support for microbial remediation in metal(loid)-contaminated soils.

CARM1 activates myogenin gene via PCAF in the early differentiation of TPA-induced rhabdomyosarcoma-derived cells.

CARM1/PRMT4 is a member of the protein arginine methyltransferase (PRMT) family. CARM1 as a transcriptional coactivator plays an active role on mammalian genes. Here, we show that CARM1 can be recruited to the promoter of myogenin gene to enhance its transcriptional activation via PCAF at the early stage of TPA-induced RD cell differentiation. By adding adenosine dialdehyde, AdOx, to inhibit the PRMT in RD cells, the TPA-induced recruiting of p300, PCAF and the Brg1 at the myogenin promoter is abolished and myogenic differentiation is blocked. More specifically, the expression of PCAF and its nucleation are prohibited when CARM1 is knockdown by its specific siRNA. We suggest that the physical interaction of CARM1 and PCAF is likely pivotal for the activation of PCAF in the downstream of CARM1 pathway for inducing myogenin under TPA-induced differentiation. The findings shed lights on novel therapeutic targets in the treatment of rhabdomyosarcoma patients.

Oxic and anoxic conditions affect arsenic (As) accumulation and arsenite transporter expression in rice.

Arsenic (As) exposure from rice consumption has now become a global health issue. This study aimed to investigate the effects of rice rhizosphere oxic conditions on silicate transporter (responsible for arsenite transportation) expressions, and on As accumulation and speciation in four rice genotypes, including two hybrid genotypes (Xiangfengyou9, Shenyou9586) and two indica subspecies (Xiangwanxian17, Xiangwanxian12). Oxic and anoxic treatments have different effects on root length (p < 0.001) and weight (p < 0.05). Total As concentrations in roots were dramatically lower in oxic treatments (88.8-218 mg/kg), compared to anoxic treatments (147-243 mg/kg) (p < 0.001). Moreover, root and shoot arsenite concentrations in oxic treatments were lower than that in anoxic treatments in arsenite treatments. The relative abundance of silicate transporter expressions displayed a trend of down-regulation in oxic treatments compared to anoxic treatments, especially significantly different for Xiangwanxian17, Xiangwanxian12 in Lsi1 expressions (p < 0.05), Xiangfengyou9, Shenyou9586, Xiangwanxian17 in Lsi2 expressions (p < 0.05). However, there were no significant differences of transporter expressions in different As treatments and genotypes. It may be a possible reason for low As accumulation in rice growing aerobically compared to flooded condition and a potential route to reduce the health risk of As in rice.

Effect of silicate on arsenic fractionation in soils and its accumulation in rice plants.

Four rice genotypes, two hybrid and two indica, were selected to investigate the effects of silicate (Si) application on arsenic (As) accumulation and speciation in rice and As fractionation in soil. There were significant differences in root, straw and grain biomass among genotypes (p < 0.05), and Si application significantly increased root (p < 0.05) and grain biomass (p < 0.001). Silicate addition reduced the proportion of As associated with well-crystallized hydrous oxides of Fe and Al and residual phases, whilst increasing the proportions of specifically-sorbed As and As associated with amorphous and poorly-crystalline Fe and Al hydrous oxides. Furthermore, the results indicated that the fraction proportions of non-specifically sorbed, specifically-sorbed, and associated with amorphous and poorly-crystalline hydrous oxides of Fe and Al in rhizosphere soils, were greater than non-rhizosphere soils. Silicate application had a significant effect decreasing total As concentrations in root (p < 0.005), straw (p < 0.05) and husk (p < 0.001) of rice plants. The effect of Si on reducing As accumulation in rice leaves was revealed by SXRF. Indica genotypes transported and accumulated less As than hybrid genotypes. Both percentage and concentration of iAs were lower in indica genotype XFY-9 than in hybrid genotype XWX-12. Silicate reduced iAs and DMA by 21% and 58% in grain (polished) respectively. DMA may have a greater translocation capacity from straw to grain (polished) than inorganic As. The study provides the potential for understanding As uptake mechanisms in rice and mitigating the health risks posed by As contamination in paddy fields.

The effect of silicon on iron plaque formation and arsenic accumulation in rice genotypes with different radial oxygen loss (ROL).

Rice is one of the major pathways of arsenic (As) exposure in human food chain, threatening over half of the global population. Greenhouse pot experiments were conducted to examine the effects of Si application on iron (Fe) plaque formation, As uptake and rice grain As speciation in indica and hybrid rice genotypes with different radial oxygen loss (ROL) ability. The results demonstrated that Si significantly increased root and grain biomass. Indica genotypes with higher ROL induced greater Fe plaque formation, compared to hybrid genotypes and sequestered more As in Fe plaque. Silicon applications significantly increased Fe concentrations in iron plaque of different genotypes, but it decreased As concentrations in the roots, straws and husks by 28-35%, 15-35% and 32-57% respectively. In addition, it significantly reduced DMA accumulation in rice grains but not inorganic As accumulation. Rice of indica genotypes with higher ROL accumulated lower concentrations of inorganic As in grains than hybrid genotypes with lower ROL.

Transcriptome responses involved in artemisinin production in Artemisia annua L. under UV-B radiation.

Artemisinin, an endoperoxide sesquiterpene lactone, is an effective antimalarial drug isolated from Artemisia annua L. In this study, a low dose (1.44 kJm(-2)d(-1)) of UV-B radiation (280-320 nm) for short-term (1h per day for 10 days) was applied to A. annua seedlings to stimulate artemisinin production. UV-B treatment not only induced the generation of reactive oxygen species (ROS), enhanced peroxidase activity and endogenous content of abscisic acid (ABA), but stimulated the biosynthesis of artemisinin in the seedlings. Here, transcriptomic changes during UV-B radiation in A. annua were detected using an Agilent GeneChip with 43,692 probe sets. In total, 358 transcripts were identified as differentially expressed under UV-B stress, of which 172 transcripts increased and 186 transcripts decreased in abundance. In terms of biological processes, gene ontology (GO) terms including primary carbohydrate and nitrogen compound metabolic processes were enriched in UV-B-repressed genes. The up-regulated genes were enriched in response to stress, ROS generation, hormone (ethylene, ABA) stimulus and cell cycle control. The expression of key enzymes such as amorpha-4,11-diene synthase (ADS) and cytochrome P450 dependent monooxygenase/hydroxylase (CYP71AV1), and related WRKY transcription factors was up-regulated significantly for artemisinin biosynthesis. This profile of global gene expression patterns during UV-B stress will be valuable for further identification of the enzymes involved in artemisinin biosynthesis.

Observation of magnetic field effects on transient fluorescence spectra of cryptochrome 1 from homing pigeons.

Cryptochromes are suggested to be involved in the bird magnetoreception based on the radical pair mechanism (RPM), a well established theory of weak magnetic field effects on chemical reactions. Two members of cryptochrome/photolyase family were found to respond to magnetic field, however, no direct responses of bird cryptochrome to magnetic field as weak as the Earth's magnetic field have been obtained so far. In this study, we used transient fluorescence spectroscopy to characterize the weak magnetic field effects of bird cryptochromes. To do this, we cloned the cryptochrome 1 gene (clCRY1) from the retina of homing pigeons (Columba livia), expressed it in insect Sf9 cells and analyzed the transient fluorescence of purified clCRY1 by application of 45-300 µT magnetic fields. The flavin adenine dinucleotide (FADox ) and glucose oxidase (GOD) in PBS buffer were set as controls which could be excited by light to generate radicals, but would not be sensitive to magnetic field. We observed that the transient fluorescence spectra of clCRY1 were sensitive to the applied magnetic field at room temperature. Our result provides a new proof of the cryptochrome-based model of avian magnetoreception in vitro.