Delivery of Nucleic Acids, Proteins, and Nanoparticles by Arginine-Rich Cell-Penetrating Peptides in Rotifers

Cell-penetrating peptides (CPPs) are a group of short, membrane-permeable cationic peptides that represent a nonviral technology for delivering nanomaterials and macromolecules into live cells. In this study, two arginine-rich CPPs, HR9 and IR9, were found to be capable of entering rotifers. CPPs were able to efficiently deliver noncovalently associated with cargoes, including plasmid DNAs, red fluorescent proteins (RFPs), and semiconductor quantum dots, into rotifers. The functional reporter gene assay demonstrated that HR9-delivered plasmid DNAs containing the enhanced green fluorescent protein and RFP coding sequences could be actively expressed in rotifers. The 1-(4,5-dimethylthiazol-2-yl)-3,5-diphenylformazan assay further confirmed that CPP-mediated cargo delivery was not toxic to rotifers. Thus, these two CPPs hold a great potential for the delivery of exogenous genes, proteins, and nanoparticles in rotifers.     

Short- and long-term diets of the threatened longhorned pygmy devil ray,Mobula eregoodoo determined using stable isotopes

Most mobulids are listed as near threatened to endangered. Nonetheless, effective conservation measures are hindered by knowledge gaps in their ecology and behaviour. In particular, few studies have assessed diets and trophic ecologies that could inform methods to avoid fishing mortality. Here, a shortfall in data for the longhorned pygmy devil ray, Mobula eregoodoo was addressed by describing temporal variability in dietary preferences using stable isotope analysis. During summer and autumn in 2017, five bather-protection gillnets were deployed off eastern Australia (29° S, 153.5° E). From the catches of these gillnets, 35 adult M. eregoodoo had liver, muscle and stomach contents sampled to determine δ¹³C and δ¹⁵N profiles. Analyses revealed that surface zooplankton and zooplanktivorous teleosts were important dietary components across short- and long-term temporal scales. Large quantities of undigested sandy sprat, Hyperlophus vittatus, in the stomachs of some specimens unequivocally confirm feeding on teleosts. A narrow isotopic niche and minimal isotopic overlap with reef manta rays, Mobula alfredi from the same geographic region in eastern Australia implies M. eregoodoo has unique and highly specialised resource use relative to other mobulids in the area. The species is clearly vulnerable to capture during inshore migrations, presumably where they feed on shallow-water shoaling teleosts. Female M. eregoodoo likely have a low annual reproductive output, so population recoveries from fishing-induced declines are likely to be slow. Measures to reduce the by catch of M. eregoodoo in local bather-protection gillnets, and artisanal fisheries more broadly, should be given priority.     

Efficacy of neem compost on root knot nematode pest of Lagos spinach,Celosia argentea

The effects of neem compost on root knot nematode, Meloidogyne incognita, pest of Lagos spinach, Celosia argentea, cv. TLV 8, was studied during 2010 and 2011 planting seasons on the field. The trial was conducted at the Teaching and Research Farm of Ladoke Akintola University of Technology, Ogbomoso, Nigeria. There were four treatments, replicated five times fitted into randomised complete block design. Neem compost was applied at 1.0, 1.5 and 2.0?tonnes/ha. Experimental plots, where no compost was applied served as the control. The result obtained shows that Lagos spinach treated with neem compost significantly (p?<?0.05) reduced the final soil nematode population and root infections (damage), with resultant improvement on crop growth and yield. The result of the chemical analysis of the neem compost revealed the presence of flavonoids, sterols, glycosides, alkaloids and saponins.     

Subunit Secondary Structure in Filamentous Viruses: Predictions and Observations

Abstract

The algorithm of Gamier, Osguthorpe and Robson (J. Mol. Biol. 120, 97–120, 1978) for prediction of protein secondary structure has been applied to the coat protein sequences of six filamentous bacteriophages: fd, Ifl, IKe, Pfl, Xf and Pf3. For subunits of Class I virions (fd, Ifl, IKe), the algorithm predicts a very high percentage of helix in comparison to other structure types, which is in accord with the results of laser Raman and circular dichroism measurements. For subunits of the Class II virions (Pfl, Xf, Pf3), the algorithm consistently predicts a predominance of β structure, which is compatible with the demonstrated facility for conversion of Class II subunits from α-helix to β-strand under appropriate experimental conditions (Thomas, Prescott and Day, J. Mol. Biol. 165, 321–356, 1983). Even when the algorithm is biased to favor helix, the Class II virion subunits are predicted to contain considerably more strand than helix. Qualitatively similar results are obtained using the algorithm of Chou and Fasman {Adv. Enzym. 47, 45–148,45-148). Therefore, both predictive and experimental methods indicate a distinction between Gass I and II subunits, which is reflected in a greater tendency of the latter to adopt other than uniform β-helical conformation. The results suggest a possible model for the disassembly of filamentous viruses which may involve the unraveling of coat protein helices at the N terminus.     

Isolation and distribution of iridescent Cellulophaga and other iridescent marine bacteria from the Charente-Maritime coast,French Atlantic

An intense colored marine bacterium, identified as Cellulophaga lytica, was isolated previously from a sea anemone surface on the Charente-Maritime rocky shore (Atlantic Coast, France), and iridescence of its colonies under direct light was recently described. In addition, iridescence intensities were found to differ strongly between C. lytica strains from different culture collections. However, importantly, the occurrence and distribution of iridescent bacteria in the marine environment were still unknown. Therefore, in this study, a search was undertaken for marine iridescent bacterial strains in different biotopes of the Charente-Maritime coast. Various marine samples (water, sediment, macroalgae, other macroorganisms and detritus) were collected from seven biotopes using a direct plate inoculation method. As a result, 34 iridescent strains related to the genus Cellulophaga, as well as 63 iridescent strains affiliated to the genera Tenacibaculum and Aquimarina, were isolated. Iridescent colors were different according to the genera but iridescent marine bacteria were widely distributed. However, a majority of strains were isolated from rocky shores and, in particular, red seaweed surfaces and mollusks. The data from the study suggested that isolates with iridescent properties were well conserved in stressful environments such as the coastal shoreline. This origin may provide an insight into the ecological and biological functions of iridescence.     

Humanization of Antibodies Using Heavy Chain Complementarity-determining Region 3 Grafting Coupled with in Vitro Somatic Hypermutation

A method for simultaneous humanization and affinity maturation of monoclonal antibodies has been developed using heavy chain complementarity-determining region (CDR) 3 grafting combined with somatic hypermutation in vitro. To minimize the amount of murine antibody-derived antibody sequence used during humanization, only the CDR3 region from a murine antibody that recognizes the cytokine hβNGF was grafted into a nonhomologous human germ line V region. The resulting CDR3-grafted HC was paired with a CDR-grafted light chain, displayed on the surface of HEK293 cells, and matured using in vitro somatic hypermutation. A high affinity humanized antibody was derived that was considerably more potent than the parental antibody, possessed a low pm dissociation constant, and demonstrated potent inhibition of hβNGF activity in vitro. The resulting antibody contained half the heavy chain murine donor sequence compared with the same antibody humanized using traditional methods.     

Dual effect of chemokine CCL7/MCP-3 in the development of renal tubulointerstitial fibrosis

Most end-stage renal disease kidneys display accumulation of extracellular matrix (ECM) in the renal tubular compartment (tubular interstitial fibrosis – TIF) which is strongly correlated with the future loss of renal function. Although inflammation is a key event in the development of TIF, it can also have a beneficial anti-fibrotic role depending in particular on the stage of the pathology. Chemokines play an important role in monocyte extravasation in the inflammatory process. CCL2 has already been shown to be involved in the development of TIF but CCL7, a close relative of CCL2 and able to bind to similar receptors, has not been studied in renal disease. We therefore studied chemokine CCL7 in a model of unilateral ureteral obstruction (UUO)-induced TIF. We observed that the role of CCL7 differs depending on the stage of the pathology. In early stages (0–8 days), CCL7 deficient (CCL7-KO) mice displayed attenuated TIF potentially involving two mechanisms: an early (0–3 days) decrease of inflammatory cell infiltration followed (3–8 days) by a decrease in tubular ECM production independent of inflammation. In contrast, during later stages of obstruction (10–14 days), CCL7-KO mice displayed increased TIF which was again associated with reduced inflammation. Interestingly, the switch between this anti- to profibrotic effect was accompanied by an increased influx of immunosuppressive regulatory T cells. In conclusion, these results highlight for the first time a dual role for CCL7 in the development of renal TIF, deleterious in early stages but beneficial during later stages.     

Discovery of TAK-960: An orally available small molecule inhibitor of polo-like kinase 1 (PLK1)

Using structure-based drug design, we identified and optimized a novel series of pyrimidodiazepinone PLK1 inhibitors resulting in the selection of the development candidate TAK-960. TAK-960 is currently undergoing Phase I evaluation in adult patients with advanced solid malignancies.     

Modulation of adipose tissue inflammation by bioactive food compounds

Adipose tissue has an important endocrine function in the regulation of whole-body metabolism. Obesity leads to a chronic low-grade inflammation of the adipose tissue, which disrupts this endocrine function and results in metabolic derangements, such as type-2 diabetes. Dietary bioactive compounds, such as polyphenols and certain fatty acids, are known to suppress both systemic and adipose tissue inflammation and have the potential to improve these obesity-associated metabolic disorders. Mechanistically, polyphenolic compounds including non-flavonoids, such as curcumin and resveratrol, and flavonoids, such as catechins (tea-polyphenols), quercetin and isoflavones, suppress nuclear factor-κB (NF-κB) and mitogen-activated protein (MAP) kinases (MAPK) pathways while activating the 5′ adenosine monophosphate-activated protein kinase (AMPK) pathway in adipose tissue. Dietary polyunsaturated fatty acids, such as eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), conjugated linoleic acid (CLA) and monounsaturated fatty acids (MUFA), such as oleic acid, also impart anti-inflammatory effects through several mechanisms. These include activation of AMPK and peroxisome proliferator-activated receptor gamma (PPAR-γ), as well as suppression of toll-like receptors (TLRs) and NF-κB pathway. This review discusses the major molecular mechanisms of dietary polyphenols and fatty acids, alone or in combination, which are responsible for adipose tissue-associated anti-inflammatory effects.