A host-independent role for Fasciola hepatica transforming growth factor-like molecule in parasite development

The trematode parasite Fasciola hepatica causes chronic infection in hosts, enabled by an immunosuppressed environment. Both host and parasite factors are known to contribute to this suggesting that avoidance of immunopathology is beneficial to both parties. We have previously characterised a parasite transforming growth factor (TGF)-like molecule, FhTLM, that interacts with host macrophages to prevent antibody-dependent cell cytotoxicity (ADCC). FhTLM is one of many described helminth TGF homologues and multiple helminths are now known to utilise host immune responses as developmental cues. To test whether, or how, F. hepatica uses FhTLM to manipulate host immunity, we initially examined its effects on the CD4 T-cell phenotype. Despite inducing IL-10, there was no induction of FoxP3 within the CD4 T-cell compartment. In addition to inducing IL-10, a wide range of chemokines were elicited from both CD4 T-cells and macrophages. However, no growth or survival advantage was conferred on F. hepatica in our co-culture system when CD4 T-cells, macrophages, or eosinophils were tested. Finally, using RNA interference we were able to verify a host-independent role for FhTLM in parasite growth. Despite the similarities of FhTLM with other described helminth TGF homologues, here we demonstrate species-specific divergence.     

A Novel W1999S Mutation and Non-Target Site Resistance Impact on Acetyl-CoA Carboxylase Inhibiting Herbicides to Varying Degrees in a UK Lolium multiflorum Population

Background

Acetyl-CoA carboxylase (ACCase) inhibiting herbicides are important products for the post-emergence control of grass weed species in small grain cereal crops. However, the appearance of resistance to ACCase herbicides over time has resulted in limited options for effective weed control of key species such as Lolium spp. In this study, we have used an integrated biological and molecular biology approach to investigate the mechanism of resistance to ACCase herbicides in a Lolium multiflorum Lam. from the UK (UK21).

Methodology/Principal Findings

The study revealed a novel tryptophan to serine mutation at ACCase codon position 1999 impacting on ACCase inhibiting herbicides to varying degrees. The W1999S mutation confers dominant resistance to pinoxaden and partially recessive resistance to cycloxydim and sethoxydim. On the other hand, plants containing the W1999S mutation were sensitive to clethodim and tepraloxydim. Additionally population UK21 is characterised by other resistance mechanisms, very likely non non-target site based, affecting several aryloxyphenoxyproprionate (FOP) herbicides but not the practical field rate of pinoxaden. The positive identification of wild type tryptophan and mutant serine alleles at ACCase position 1999 could be readily achieved with an original DNA based derived cleaved amplified polymorphic sequence (dCAPS) assay that uses the same PCR product but two different enzymes for positively identifying the wild type tryptophan and mutant serine alleles identified here.

Conclusion/Significance

This paper highlights intrinsic differences between ACCase inhibiting herbicides that could be exploited for controlling ryegrass populations such as UK21 characterised by compound-specific target site and non-target site resistance.     

THE DARK ADAPTATION OF RETINAL FIELDS OF DIFFERENT SIZE AND LOCATION

The decrease in threshold shown by the eye during dark adaptation proceeds in two steps. The first is rapid, short in duration, and small in extent. The second is slow, prolonged, and large. The first is probably due to cone function; the second to rod function. In centrally located fields the two parts of adaptation change differently with area. With small, foveal fields the first part dominates and only traces of the second part appear. As the area increases the first part changes a little, while the second part covers an increasing range of intensities and appears sooner in time. Measurements with an annulus field covering only the circumference of a 20° circle show most of the characteristics of a 20° whole field centrally located. Similarly a 2° field located at different distances from the center shows dark adaptation characteristics essentially like those of large centrally located fields whose edges correspond to the position of the central field. Evidently the behavior in dark adaptation of centrally located fields of different size is determined in the main not by area as area, but by the fact that the retina gradually changes in sensitivity from center to periphery, and therefore the larger the field the farther it reaches into peripheral regions of permanently greater sensibility.     

THE ABSORPTION SPECTRUM OF VISUAL PURPLE

The absorption spectra of visual purple solutions extracted by various means were measured with a sensitive photoelectric spectrophotometer and compared with the classical visual purple absorption spectrum. Hardening the retinas in alum before extraction yielded visual purple solutions of much higher light transmission in the blue and violet, probably because of the removal of light-dispersing substances. Re-extraction indicated that visual purple is more soluble in the extractive than are the other colored retinal components. However, the concentration of the extractive did not affect the color purity of the extraction but did influence the keeping power. This suggests a chemical combination between the extractive and visual purple. The pH of the extractive affected the color purity of the resulting solution. Over the pH range from 5.5 to 10.0, the visual purple color purity was greatest at the low pH. Temperature during extraction was also effective, the color purity being greater the higher the temperature, up to 40°C. Drying and subsequent re-dissolving of visual purple solutions extracted with digitalin freed the solution of some protein impurities and increased its keeping power. Dialysis against distilled water seemed to precipitate visual purple from solution irreversibly. None of the treatments described improved the symmetry of the unbleached visual purple absorption spectrum sufficiently for it to resemble the classical absorption spectrum. Therefore it is very likely that the classical absorption spectrum is that of the light-sensitive group only and that the absorption spectra of our purest unbleached visual purple solutions represent the molecule as a whole.     

THE COURSE OF ROD DARK ADAPTATION AS INFLUENCED BY THE INTENSITY AND DURATION OF PRE-ADAPTATION TO LIGHT

An increase in the degree of light adaptation causes a decrease in the slope of the subsequent rod dark adaptation function and a displacement of the function to the right on the time axis. Over a wide range, these changes occur to the same extent whether the increase in the degree of light adaptation is produced by raising the intensity or by prolonging the exposure. Within these limits, the Bunsen-Roscoe reciprocity law applies to the intensity and duration of pre-exposure. Over a still wider range, dark adaptation has the same course following brief exposure to a bright light as it has following prolonged exposure to a dim light, provided the degree of light adaptation is the same in both instances (as indicated by identical initial dark adaptation thresholds).