Double-stranded RNAs targeting inhibitor of apoptosis gene show no significant cross-species activity

RNA interference (RNAi) has become an integral part of mainstream research due to its versatility and ease of use. However, the potential nontarget effects associated with double-stranded RNAs (dsRNA) are poorly understood. To explore this, we used dsRNAs targeting the inhibitor of apoptosis (iap) gene from nine insect species and assayed their possible nontarget effects. For each assay, we used a control (dsRNA targeting the gene coding for green fluorescent protein, GFP) and a species-specific dsRNA targeting nine iap genes in insect species to evaluate target gene knockdown efficiency, apoptosis phenotype in cells and mortality in insects. Our results revealed that dsIAP efficiently knocks down iap gene expression and induces apoptosis phenotype and mortality in target insect species. In contrast, no significant knockdown of the iap gene expression, apoptosis phenotypes, or mortality were detected in cell lines developed from nontarget insects or nontarget insects treated with dsIAPs. Interestingly, even among closely related insects such as stink bugs, Nezara viridula, Halyomorpha halys, and Murgantia histrionica, with substantial sequence similarity among iap genes from these insects, no significant nontarget effects of dsIAP were observed under the conditions tested. These data demonstrate no significant nontarget effects for dsIAPs and suggest that the threat of nontarget effects of RNAi technology may not be substantial.     

Development and quantification of biodiesel production from chicken feather meal as a cost-effective feedstock by using green technology

Increased urbanization and increase in population has led to an increased demand for fuels. The result is the prices of fuels are reaching new heights every day. Using low-cost feedstocks such as rendered animal fats in biodiesel production will reduce biodiesel expenditures. One of the low-cost feedstocks for biodiesel production from poultry feathers. This paper describes a new and environmentally friendly process for developing biodiesel production technology from feather waste produced in poultry industry. Transesterification is one of the well-known processes by which fats and oils are converted into biodiesel. The reaction often makes use of acid/base catalyst. If the material possesses high free fatty acid then acid catalyst gives better results. The data resulted from gas chromatography (GC) revealed these percentages for fatty acid compositions: myristic acid, palmitic acid, stearic acid, oleic acid, linoleic acid and arachidonic acid. The biodiesel function group was analyzed by using FTIR. This study concluded that the rooster feathers have superior potential to process them into biodiesel than broiler chicken feathers fat because of fatty acid composition values and it has important properties of biodiesel.     

Chitosan nanoparticles help double-stranded RNA escape from endosomes and improve RNA interference in the fall armyworm,Spodoptera frugiperda

RNA interference (RNAi) is a promising technology for the development of next-generation insect pest control products. Though RNAi is efficient and systemic in coleopteran insects, it is inefficient and variable in lepidopteron insects. In this study, we explored the possibility of improving RNAi in the fall armyworm (FAW), Spodoptera frugiperda by conjugating double-stranded RNA (dsRNA) with biodegradable chitosan (Chi). dsRNA conjugated with chitosan was protected from degradation by endonucleases present in Sf9 cell-conditioned medium, hemolymph, and midgut lumen contents collected from the FAW larvae. Chi–dsRNA complexes showed reduced accumulation in the endosomes of Sf9 cells and FAW tissues. Exposing chitosan formulated dsRNA in Sf9 cells and the tissues induced a significant knockdown of endogenous genes. Chi–dsIAP fed to FAW larvae induced knockdown of iap gene, growth retardation, and mortality. Processing of dsRNA into small interfering RNA was detected with chitosan-conjugated ³²P-UTP-labeled ds green fluorescent protein in Sf9 cells and FAW larval tissues. Overall, these data suggest that dsRNA conjugated with chitosan helps dsRNA escape from the endosomes and improves RNAi efficiency in FAW cells and tissues.     

Isolation of sterols from the marine fungus Corollospora lacera

Several marine fungi collected from the waters of Prince Edward Island, Canada, were screened for the presence of natural products exhibiting antibacterial activity. Both broths and mycelia of these fungi were studied using the bioassay-guided chromatographic separation. The 4 fractions from the extract of mycelia of Corollospora lacera exhibited weak antibacterial activity and were analyzed further. From these fractions, 2 sterols (5 alpha,8 alpha-epidioxyergosterol and 22E,24R-ergosta-7,22-diene-3beta,5 alpha,6 beta-triol) and a 3:1 mixture of linoleic and oleic acids were isolated. The presence of ergosterol was confirmed in dichloromethane extracts of mycelia of every fungus in this study and this sterol was isolated from the extract of mycelium of Corollospora lacera. Two other known compounds (5-hydroxymethylfuran-2-carbaldehyde and bis(2-ethylhexyl) phthalate), were isolated from the dichloromethane extract of mycelium of Monodictys pelagica. The phthalate was reported in the literature as a metabolite isolated from the fungi, but in our study it was proven to be an artifact of the culturing and (or) extraction procedures rather than a true fungal metabolite.     

A comparison of aquaporin function in mediating stomatal aperture gating among drought-tolerant and sensitive varieties of rice (Oryza sativa L.)

Protoplasma - Climate change drastically affects the cultivation of rice, and its production is affected significantly by water stress. Adaptation of a plant to water deficit conditions is...     

Hypertrophic Stimulation Increases β-actin Dynamics in Adult Feline Cardiomyocytes

The myocardium responds to hemodynamic stress through cellular growth and organ hypertrophy. The impact of cytoskeletal elements on this process, however, is not fully understood. While α-actin in cardiomyocytes governs muscle contraction in combination with the myosin motor, the exact role of β-actin has not been established. We hypothesized that in adult cardiomyocytes, as in non-myocytes, β-actin can facilitate cytoskeletal rearrangement within cytoskeletal structures such as Z-discs. Using a feline right ventricular pressure overload (RVPO) model, we measured the level and distribution of β-actin in normal and pressure overloaded myocardium. Resulting data demonstrated enriched levels of β-actin and enhanced translocation to the Triton-insoluble cytoskeletal and membrane skeletal complexes. In addition, RVPO in vivo and in vitro hypertrophic stimulation with endothelin (ET) or insulin in isolated adult cardiomyocytes enhanced the content of polymerized fraction (F-actin) of β-actin. To determine the localization and dynamics of β-actin, we adenovirally expressed GFP-tagged β-actin in isolated adult cardiomyocytes. The ectopically expressed β-actin-GFP localized to the Z-discs, costameres, and cell termini. Fluorescence recovery after photobleaching (FRAP) measurements of β-actin dynamics revealed that β-actin at the Z-discs is constantly being exchanged with β-actin from cytoplasmic pools and that this exchange is faster upon hypertrophic stimulation with ET or insulin. In addition, in electrically stimulated isolated adult cardiomyocytes, while β-actin overexpression improved cardiomyocyte contractility, immunoneutralization of β-actin resulted in a reduced contractility suggesting that β-actin could be important for the contractile function of adult cardiomyocytes. These studies demonstrate the presence and dynamics of β-actin in the adult cardiomyocyte and reinforce its usefulness in measuring cardiac cytoskeletal rearrangement during hypertrophic stimulation.