Purification of Intracellular Dextranases and D-Glucosidases from Pseudomonas UQM 733

The intracellular enzymes of Pseudomonas UQM 733 which act on dextran have been reinvestigated mainly by isoelectric focusing. At least three dextranases are present, and one of them (D₄) has been purified and shown to be very similar to one of the extracellular endo-dextranases (D₁). Three different α-D-glucosidases have also been purified.     

Making gene drive biodegradable

Gene drive systems have long been sought to modify mosquito populations and thus combat malaria and dengue. Powerful gene drive systems have been developed in laboratory experiments, but may never be used in practice unless they can be shown to be acceptable through rigorous field-based testing. Such testing is complicated by the anticipated difficulty in removing gene drive transgenes from nature. Here, we consider the inclusion of self-elimination mechanisms into the design of homing-based gene drive transgenes. This approach not only caused the excision of the gene drive transgene, but also generates a transgene-free allele resistant to further action by the gene drive. Strikingly, our models suggest that this mechanism, acting at a modest rate (10%) as part of a single-component system, would be sufficient to cause the rapid reversion of even the most robust homing-based gene drive transgenes, without the need for further remediation. Modelling also suggests that unlike gene drive transgenes themselves, self-eliminating transgene approaches are expected to tolerate substantial rates of failure. Thus, self-elimination technology may permit rigorous field-based testing of gene drives by establishing strict time limits on the existence of gene drive transgenes in nature, rendering them essentially biodegradable.This article is part of the theme issue ‘Novel control strategies for mosquito-borne diseases''.     

Contractability of a Chorionic Opercular Corolla on Eggs of the Assassin bug Sinea rileyi Montandon (Heteroptera: Reduviidae)

Summary

A chorionic opercular rim corolla contracts in response to submersion of eggs in water and dilates again when eggs dry. This “behavior” ceases in hatched and dead unhatched eggs. We suggest that contraction serves a group respiratory function. Egg clutches are sibships of high relatedness in close spatial proximity with shared susceptibilities. These factors suggest the possibility of various cooperative interactions especially in insects with elaborate chorionic structures.     

α-(15-Hydroxyhexadecyl)itaconic acid from Usnea aliphatica

The structure of a new aliphatic acid from Usnea aliphatica was established by spectroscopic and chemical methods as α- (15-hydroxyhexadecyl)itaconic acid.     

Aedes aegypti dyspepsia encodes a novel member of the SLC16 family of transporters and is critical for reproductive fitness

As a key vector for major arthropod-borne viruses (arboviruses) such as dengue, Zika and chikungunya, control of Aedes aegypti represents a major challenge in public health. Bloodmeal acquisition is necessary for the reproduction of vector mosquitoes and pathogen transmission. Blood contains potentially toxic amounts of iron while it provides nutrients for mosquito offspring; disruption of iron homeostasis in the mosquito may therefore lead to novel control strategies. We previously described a potential iron exporter in Ae. aegypti after a targeted functional screen of ZIP (zinc-regulated transporter/Iron-regulated transporter-like) and ZnT (zinc transporter) family genes. In this study, we performed an RNAseq-based screen in an Ae. aegypti cell line cultured under iron-deficient and iron-excess conditions. A subset of differentially expressed genes were analyzed via a cytosolic iron-sensitive dual-luciferase reporter assay with several gene candidates potentially involved in iron transport. In vivo gene silencing resulted in significant reduction of fecundity (egg number) and fertility (hatch rate) for one gene, termed dyspepsia. Silencing of dyspepsia reduced the induction of ferritin expression in the midgut and also resulted in delayed/impaired excretion and digestion. Further characterization of this gene, including a more direct confirmation of its substrate (iron or otherwise), could inform vector control strategies as well as to contribute to the field of metal biology.     

Species specificity of hybridoma antibodies to surface antigens of guinea pig sperm

The species specificity of hybridoma antibodies to sperm surface antigens was studied. A collection of over 50 hybridoma antibodies that bind to the guinea pig sperm surface was tested for binding to mouse, rat, hamster, and human sperm by indirect immunofluorescence. None of the antibodies bind to mouse sperm. rat sperm, or human sperm. All but three of the antibodies also fail to bind to hamster sperm. AH-30, AH-31, and AH-1032, the three antibodies that crossreact with hamster sperm, show a different topographical localization on hamster sperm from that seen on guinea pig sperm. The three antibodies do not precipitate a ¹²⁵I surface-labeled antigen from hamster sperm extracts. However, from guinea pig sperm extracts, all three antibodies precipitate ¹²⁵I surface-labeled polypeptides with molecular weights (M_r) of 62,000, 52,000, and 38,000. This result suggests that the crossreacting antibodies may be recognizing different antigens on hamster and guinea pig sperm.     

Macrophytic primary production and nutrient concentrations in a deltaic floodplain marsh of the Lower Paraná River

Biomass changes across an annual cycle were followed at two sampling sites in the floodplain marsh of the Lower Paraná River: close to the river-shore and 800 m inside the floodplain marsh, both dominated by Scirpus californicus and Cyperus giganteus. Tidal influence determines a daily exchange of water between the river and the floodplain marsh.Estimated net primary production was higher in the river (2820 against 1770 g m^–2). Contents of nitrogen and phosphorus in plant tissue decreased from the river to the floodplain (0.62 to 0.45% N and 0.18 to 0.14% P). In spite of the important water exchange between the river and the floodplain, a decrease in nitrate, oxygen and suspended matter, and an increase in soluble reactive phosphorus in the water were observed from the river towards the floodplain marsh.A primary production gradient exists from the river to the inner floodplain marsh, where production is nitrogen-limited, sustained mainly on nutrients supplied by the river. Floodplain marshes are nitrate sinks, probably through denitrification losses and macrophyte uptake.     

Ceramide glycosylation potentiates cellular multidrug resistance.

Ceramide glycosylation, through glucosylceramide synthase (GCS), allows cellular escape from ceramide-induced programmed cell death. This glycosylation event confers cancer cell resistance to cytotoxic anticancer agents [Liu, Y. Y., Han, T. Y., Giuliano, A. E., and M. C. Cabot. (1999) J. Biol. Chem. 274, 1140-1146]. We previously found that glucosylceramide, the glycosylated form of ceramide, accumulates in adriamycin-resistant breast carcinoma cells, in vinblastine-resistant epithelioid carcinoma cells, and in tumor specimens from patients showing poor response to chemotherapy. Here we show that multidrug resistance can be increased over baseline and then totally reversed in human breast cancer cells by GCS gene targeting. In adriamycin-resistant MCF-7-AdrR cells, transfection of GCS upgraded multidrug resistance, whereas transfection of GCS antisense markedly restored cellular sensitivity to anthracyclines, Vinca alkaloids, taxanes, and other anticancer drugs. Sensitivity to the various drugs by GCS antisense transfection increased 7- to 240-fold and was consistent with the resumption of ceramide-caspase-apoptotic signaling. GCS targeting had little influence on cellular sensitivity to either 5-FU or cisplatin, nor did it modify P-glycoprotein expression or rhodamine-123 efflux. GCS antisense transfection did enhance rhodamine-123 uptake compared with parent MCF-7-AdrR cells. This study reveals that GCS is a novel mechanism of multidrug resistance and positions GCS antisense as an innovative force to overcome multidrug resistance in cancer chemotherapy.