Tadpole-like Conformations of Huntingtin Exon 1 Are Characterized by Conformational Heterogeneity that Persists regardless of Polyglutamine Length

Soluble huntingtin exon 1 (Httex1) with expanded polyglutamine (polyQ) engenders neurotoxicity in Huntington's disease. To uncover the physical basis of this toxicity, we performed structural studies of soluble Httex1 for wild-type and mutant polyQ lengths. Nuclear magnetic resonance experiments show evidence for conformational rigidity across the polyQ region. In contrast, hydrogen–deuterium exchange shows absence of backbone amide protection, suggesting negligible persistence of hydrogen bonds. The seemingly conflicting results are explained by all-atom simulations, which show that Httex1 adopts tadpole-like structures with a globular head encompassing the N-terminal amphipathic and polyQ regions and the tail encompassing the C-terminal proline-rich region. The surface area of the globular domain increases monotonically with polyQ length. This stimulates sharp increases in gain-of-function interactions in cells for expanded polyQ, and one of these interactions is with the stress-granule protein Fus. Our results highlight plausible connections between Httex1 structure and routes to neurotoxicity.     

Health risk assessment of heavy metals and metalloids via dietary intake of a potential vegetable (Coriandrum sativum L.) grown in contaminated water irrigated agricultural sites of Sargodha,Pakistan

Levels of Mo, As, Se, Fe, Cu, Zn, Ni, and Pb were determined in a vegetable commonly consumed in Pakistan. Samples were collected from three different sites (Ratokala, Phularwan, and Mailowal) supplied with three different water sources. Mo and Pb in water and Mo and As in C. sativum were higher than the suggested standards at the international level. Lower bioconcentration factor and pollution load index were seen at Site-I than at other sites. A positive association was found between the rhizosphere metals and those of the vegetable at all three sites. Enrichment factor at three sites ranged from 0.28 to 10.39. The highest value of daily intake of metals was found for Fe (0.245 mg/kg/day) and of the health risk index for As (70.41) at the wastewater inundated site. It is inferred that uptake of Mo, As, Cu, Ni, and Pb through C. sativum represents a high health danger to the individuals using this vegetable.     

Accumulation of metals and metalloids in radish (Raphanus sativus L.) and spinach (Spinacea oleracea L.) irrigated with domestic wastewater in the peri-urban areas of Khushab City,Pakistan

Accumulation of different metals and metalloids was assessed in two vegetables radish (Raphanus sativus L.) and spinach (Spinacea oleracea L.) irrigated with domestic wastewater in the peri-urban areas of Khushab City, Pakistan. In general, the metal and metalloid concentrations in radish and spinach were higher at site-II treated with sewage water than those found at site-I treated with canal water. In case of radish at both sites the levels of metals (Cr, Mn, Fe, Co, Ni, Cu, Zn, As, Se, Mo, Cd, and Pb) were below the permissible level except those of Mn, Ni, Mo, Cd, and Pb. At both sites, the transfer factor ranged from 0.047–228.3 mg kg^?1 with Cr having the highest transfer factor. The metal pollution index in soil was in the following order: As > Fe > Ni > Zn > Cd > Mo > Se > Co > Pb > Mn > Cr > Cu, respectively. While in case of spinach at both sites, the concentrations of metals and metalloids in vegetable samples irrigated with canal and sewage water were observed below the permissible level except Mn, Ni, Zn, Mo, and Pb. At both sites, the transfer factor ranged from 0.038–245.4 mg kg^?1 with Cr having the highest transfer factor. The metal pollution index in soil was in the following order: Cd > Ni > Co > Se > Mn > Zn > Mo > Pb > Fe > Cr > As > Cu, respectively.     

Deciphering role of technical bioprocess parameters for bioethanol production using microalgae

Microalgae biomass is considered an important feedstock for biofuels and other bioactive compounds due to its faster growth rate, high biomass production and high biomolecules accumulation over first and second-generation feedstock. This research aimed to maximize the specific growth rate of fresh water green microalgae Closteriopsis acicularis, a member of family Chlorellaceae under the effect of pH and phosphate concentration to attain enhanced biomass productivity. This study investigates the individual and cumulative effect of phosphate concentration and pH on specific growth characteristics of Closteriopsis acicularis in autotrophic mode of cultivation for bioethanol production. Central-Composite Design (CCD) strategy and Response Surface Methodology (RSM) was used for the optimization of microalga growth and ethanol production under laboratory conditions. The results showed that high specific growth rate and biomass productivity of 0.342 day⁻¹ and 0.497 g L⁻¹ day⁻¹ respectively, were achieved at high concentration of phosphate (0.115 g L⁻¹) and pH (9) at 21st day of cultivation. The elemental composition of optimized biomass has shown enhanced elemental accumulation of certain macro (C, O, P) and micronutrients (Na, Mg, Al, K, Ca and Fe) except for nitrogen and sulfur. The Fourier transform infrared spectroscopic analysis has revealed spectral peaks and high absorbance in spectral range of carbohydrates, lipids and proteins, in optimized biomass. The carbohydrates content of optimized biomass was observed as 58%, with 29.3 g L⁻¹ of fermentable sugars after acid catalyzed saccharification. The bioethanol yield was estimated as 51 % g ethanol/g glucose with maximum of 14.9 g/L of bioethanol production. In conclusion, it can be inferred that high specific growth rate and biomass productivity can be achieved by varying levels of phosphate concentration and pH during cultivation of Closteriopsis acicularis for improved yield of microbial growth, biomass and bioethanol production.     

Characterization of fusarium wilt-resistant and fusarium wilt-susceptible somaclones of banana cultivar rastali (Musa AAB) by random amplified polymorphic DNA and retrotransposon markers

Two DNA fingerprinting techniques, random amplified polymorphic DNA (RAPD) and inter-retrotransposon amplified polymorphism (IRAP), were used to characterize somaclonal variants of banana. IRAP primers were designed on the basis of repetitive and genome-wide dispersed long terminal repeat (LTR) retrotransposon families for assessing the somaclonal variation in 2Musa clones resistant and susceptible toFusarium oxysporum f. sp.cubense race 4. RAPD markers successfully detected genetic variation within and between individuals of the clones. IRAP makers amplified either by a single primer or a combination of primers based on LTR orientation successfully amplified different retrotransposons dispersed in theMusa genome and detected new events of insertions. RAPD markers proved more polymorphic than IRAP markers. Somaclonal variation seems to be the result of numerous indels occurring genome-wide accompanied by the activation of retroelements, as a result of stress caused by micropropagation. It is concluded that characterization of the somaclonal variants requires more than one DNA marker system to detect variation in diverse components of the genome.     

Differential inter- and intra-specific defense induction in Lupinus by Myzus persicae feeding

Experiments were conducted to investigate the potential induction of plant defenses by Myzus persicae Sulzer (Homoptera: Aphididae) feeding on five lupin, Lupinus spp. (Leguminosae), varieties with well‐characterized levels of aphid resistance. Myzus persicae feeding on L. angustifolius and L. luteus varieties induced genotype‐specific changes in their host that were not consistent with the level of aphid resistance or the plant species. The plant responses were systemically detected by apterous and alate forms of the aphids. Chemical assays revealed no induction of oxidizing enzyme (catalase, peroxidase, or polyphenol oxidase) activity, serine or cystein proteinase inhibitors, or soluble phenolics in any of the five varieties tested following 3 days of feeding by 10 or 30 aphids. However, there were significant differences among the five lupin varieties in the levels of peroxidase and polyphenol oxidase activity, proteinase inhibitors, and soluble phenolics.     

High-yield rapid production of hepatitis B surface antigen in plant leaf by a viral expression system

Recombinant hepatitis B surface antigen (HBsAg) constitutes currently used vaccines against hepatitis B virus, and has been successfully employed as a carrier for foreign epitopes. With the aim of developing an inexpensive, easily administered vaccine source for global immunization, several groups have expressed HBsAg in plant systems. Transgenic plant-derived HBsAg assembles into virus-like particles (VLPs) and is immunogenic in both mice and humans. However, HBsAg expression is relatively low in transgenic plant systems. The time-consuming and labour-intensive process of generating transgenic plants also significantly limits high-throughput analyses of various HBsAg fusion antigens. In this paper, the high-yield rapid production of HBsAg in plant leaf using a novel viral transient expression system is described. Nicotiana benthamiana leaves infiltrated with the MagnICON viral vectors produced HBsAg at high levels, averaging 295 µg/g leaf fresh weight at 10 days post-infection, as measured by a polyclonal enzyme-linked immunosorbent assay. Transiently expressed HBsAg accumulated as the full-length product, formed disulphide-linked dimers, displayed the conformational 'a' antigenic determinant and assembled into VLPs. Immunization of mice with partially purified HBsAg elicited HBsAg-specific antibodies. Furthermore, it was found that transient production of HBsAg using vacuum infiltration of whole plants, rather than syringe infiltration of leaves, was readily scalable, and greatly improved the accumulation of correctly folded HBsAg that displays the protective 'a' determinant.     

Survey and phylogenetic analysis of culturable microbes in the oral secretions of three bark beetle species

In a recent study, we reported a previously undescribed behavior in which a bark beetle exuded oral secretions containing bacteria that have antifungal properties, and hence defend their galleries against pervasive antagonistic Hyphomycete fungi. Actinobacteria, a group known for their antibiotic properties, were the most effective against fungi that invade the spruce beetle galleries. In the present study, we describe the isolation and identification of microorganisms from oral secretions of three bark beetles (Coleoptera: Curculionidae: Scolytinae): the spruce beetle, Dendroctonus rufipennis Kirby, the mountain pine beetle, Dendroctonus ponderosae Hopkins, and the pine engraver, Ips pini Say. Bacteria isolated from these three species span the major bacterial classes α-, β-, and γ-Proteobacteria, Firmicutes, Bacteroidetes, and Actinobacteria, except for D. ponderosae , which yielded no α-proteobacteria or Bacteroidetes isolates. Spruce beetles and pine engraver beetles had similar numbers of α-proteobacteria isolates, but pine engravers yielded twice as many Bacteroidetes isolates as spruce beetles. In contrast, mountain pine beetles yielded more isolates in the β- and γ-proteobacteria than spruce beetles and pine engravers. The highest percentage of Actinobacteria was obtained from spruce beetles, followed by pine engravers and mountain pine beetles. All of the fungal isolates obtained from the three beetle species were Ascomycetes. The greatest fungal diversity was obtained in spruce beetles, which had nine species, followed by pine engravers with five, and mountain pine beetles with one.     

Plasma proteome analysis of cervical intraepithelial neoplasia and cervical squamous cell carcinoma

Although cervical cancer is preventable with early detection, it remains the second most common malignancy among women. An understanding of how proteins change in their expression during a particular diseased state such as cervical cancer will contribute to an understanding of how the disease develops and progresses. Potentially, it may also lead to the ability to predict the occurrence of the disease. With this in mind, we aimed to identify differentially expressed proteins in the plasma of cervical cancer patients. Plasma from control, cervical intraepithelial neoplasia (CIN) grade 3 and squamous cell carcinoma (SCC) stage IV subjects was resolved by two-dimensional gel electrophoresis and the resulting proteome profiles compared. Differentially expressed protein spots were then identified by mass spectrometry. Eighteen proteins were found to be differentially expressed in the plasma of CIN 3 and SCC stage IV samples when compared with that of controls. Competitive ELISA further validated the expression of cytokeratin 19 and tetranectin. Functional analyses of these differentially expressed proteins will provide further insight into their potential role(s) in cervical cancer-specific monitoring and therapeutics.