Interactions between bovine beta-lactoglobulin A and various bioactive peptides as studied by front-face fluorescence spectroscopy

Front-face fluorescence spectroscopy was used for the first time to study the interactions between bovine beta-lactoglobulin variant A (beta-Lg A) and various beta-Lg-derived bioactive peptides. Fluorescence spectra were recorded for beta-Lg A-peptide mixtures at 25 degrees C and pH 6.8 with an excitation wavelength of 290 nm to characterize the molecular environment of tryptophan (Trp) residues present in the protein but absent in the peptides. Spectra remained unchanged following addition of peptides beta-Lg f92-100 and beta-Lg f125-135, while Phe-Phe interaction between beta-Lg f69-83 molecules interfered with analysis. Addition of beta-Lg f102-105 produced a blue shift (3 nm) and a significant increase in fluorescence intensity, while addition of beta-Lg f142-148 also caused a significant increase in fluorescence intensity but accompanied by a red shift (3 nm). These results indicate that the polarity of the Trp environment in the beta-Lg A structure may be modified differently depending on the peptide added.     

Possible parachordoma in a dog

Abstract

CASE HISTORY: A 15-mm diameter mass developed in the S/C tissue overlying the right lateral gluteal muscle of a 6½-year-old female Flat-coated Retriever.

PATHOLOGICAL FINDINGS: Cytological preparations following aspiration of the mass were highly cellular and consisted of a population of large polygonal cells containing single to multiple nuclei, large prominent nucleoli, and intracytoplasmic vacuoles. Histologically, the neoplasm consisted of similar large cells surrounded by thick fibrous connective tissue trabeculae. The large polygonal cells reacted positively with antibodies against vimentin, low- and high-molecular-weight variants of cytokeratin (AE1/AE3), but not with antibodies to desmin or glial fibrillary acidic protein (GFAP).

DIAGNOSIS: The clinical, gross, histological and immunohistochemical findings are similar to those reported for parachordomas in humans. Neither recurrence nor metastases were noted 18 months after surgical excision of the mass.

CLINICAL RELEVANCE: This is the first reported case of a possible parachordoma in a dog, a benign tumour with cytological features of malignancy.     

Pseudogout in two dogs

Extract

Pseudogout is a common condition in elderly people in which sudden onset of intense joint pain and synovitis develop. Peri-synovial calcium crystal deposition is the cause of the synovitis rather than the uric acid deposition in common gout. It is a rare condition in dogs, having only been reported twice before. Two dogs presenting with acute lameness and pyrexia showed many macrophages and occasional neutrophils containing intracellular crystals on cytological examination of synovial fluid. The crystals were single or multiple, small to large, clear, square or rhomboid-shaped, weakly birefringent and resembled those described for pseudogout. Occasional crystals were also present free in the matrix. Non-steroidal anti-inflammatory drugs that are effective in reducing pain in people with pseudogout provided no relief to either patient, but twice daily oral prednisolone therapy was successful. Dosage was gradually tapered as clinical signs improved over a period of 6 weeks.     

Mycobacterial infections in striped bass from Delaware Bay

Eighty striped bass Morone saxatilis were obtained from Delaware Bay using commercial gill nets set adjacent to Woodland Beach (n = 70) and Bowers Beach (n = 10) in December 2003. Fish were examined for gross lesions. Total lengths (TLs) and eviscerated weights were determined to calculate condition factors (K). Portions of spleens were aseptically harvested for bacterial culture, and portions of spleens, kidneys (anterior and posterior), livers, and gonads were obtained for histological examination. The size distribution of the striped bass was relatively homogeneous; the mean TL was about 600 mm for all samples. Mean K exceeded 0.95 in all samples and was not significantly different (P > 0.05) among samples. Significant differences in mycobacterial infection prevalence (P < or = 0.05) were observed among samples; samples obtained at Woodland Beach (WB) on December 10 (53.8%, n = 13) and December 17 (7.1%, n = 42) exhibited the most striking differences in prevalence. Mycobacterial infection intensity ranged from 1 X 10(2) to 1 X 10(7) colony-forming units per gram of spleen. Acanthocephalan infection prevalence and intensity, non-acid-fast bacterial infection prevalence, and fish sex ratio were also significantly different among the samples (P < or = 0.05). Similar to the mycobacterial infections, differences in sex ratio, acanthocephalan infection, and non-acid-fast bacterial infection were observed between the WB samples taken on December 10 and 17. However, no significant associations (P > 0.05) were observed between sex ratio or these infections and mycobacterial infection. The differences in bacterial and parasite infection prevalence and intensity and fish sex ratio in some samples indicate that these fish had a different history and that the epizootiology of mycobacterial infection in striped bass from Delaware Bay may be relatively complex.     

A 100,000-year periodicity in the accretion rate of interplanetary dust

Numerical modeling of the orbital evolution of interplanetary dust particles revealed that, over the past 1.2 million years, the rate of accretion of dust by Earth has varied by a factor of 2 to 3. These variations display a 100,000-year periodicity and are anticorrelated with Earth's changing orbital eccentricity. Extraterrestrial helium-3 concentrations in a deep-sea sediment core display a similar periodicity but are 50,000 years out of phase with the predicted variations. Also, because collisions between large bodies in the asteroid belt are inevitable, it is expected that large-amplitude stochastic variations on 10(7)- to 10(8)-year time scales would be superimposed on the 10(5)-year periodic variations.     

A comparison of aggressiveness and deoxynivalenol production between Canadian <Emphasis Type="Italic">Fusarium graminearum</Emphasis> isolates with 3-acetyl and 15-acetyldeoxynivalenol chemotypes in field-grown spring wheat

Twenty four isolates of Fusarium graminearum, half of which were 3-acetyldeoxynivalenol (3-ADON) and half 15-acetyldeoxynivalenol (15-ADON) chemotypes, were tested for their ability to produce deoxynivalenol and to cause Fusarium head blight (FHB) in spring wheat cultivars. The objectives of this study were to determine (1) whether 3-ADON isolates differ in aggressiveness, as measured by the FHB index, and DON production from 15-ADON isolates under field conditions, and (2) whether the performance of resistant host cultivars was stable across isolates. Field tests of all isolates were conducted with three replicates at each of two locations in Canada and Germany in 2008 with three host genotypes differing in FHB resistance level. The resistant host genotype showed resistance regardless of the chemotype or location. The differences between mean FHB indices of 3-ADON and 15-ADON isolates were not significant for any wheat genotype. In contrast, average DON production by the 3-ADON isolates (10.44 mg kg⁻¹) was significantly (P < 0.05) higher than for the 15-ADON isolates (6.95 mg kg⁻¹) at three of the four locations where moderately resistant lines were tested, and at both locations where susceptible lines were evaluated. These results indicate that 3-ADON isolates could pose a greater risk to food safety. However, as the mean aggressiveness and DON production of 3-ADON and 15-ADON chemotypes was similar on highly resistant lines, breeding and use of highly resistant lines is still the most effective measure of reducing the risks associated with DON in wheat.     

A high-throughput seed germination assay for root parasitic plants

Background

Some root-parasitic plants belonging to the Orobanche, Phelipanche or Striga genus represent one of the most destructive and intractable weed problems to agricultural production in both developed and developing countries. Compared with most of the other weeds, parasitic weeds are difficult to control by conventional methods because of their life style. The main difficulties that currently limit the development of successful control methods are the ability of the parasite to produce a tremendous number of tiny seeds that may remain viable in the soil for more than 15 years. Seed germination requires induction by stimulants present in root exudates of host plants. Researches performed on these minute seeds are until now tedious and time-consuming because germination rate is usually evaluated in Petri-dish by counting germinated seeds under a binocular microscope.

Results

We developed an easy and fast method for germination rate determination based on a standardized 96-well plate test coupled with spectrophotometric reading of tetrazolium salt (MTT) reduction. We adapted the Mosmann’s protocol for cell cultures to germinating seeds and determined the conditions of seed stimulation and germination, MTT staining and formazan salt solubilization required to obtain a linear relationship between absorbance and germination rate. Dose–response analyses were presented as applications of interest for assessing half maximal effective or inhibitory concentrations of germination stimulants (strigolactones) or inhibitors (ABA), respectively, using four parameter logistic curves.

Conclusion

The developed MTT system is simple and accurate. It yields reproducible results for germination bioassays of parasitic plant seeds. This method is adapted to high-throughput screenings of allelochemicals (stimulants, inhibitors) or biological extracts on parasitic plant seed germination, and strengthens the investigations of distinctive features of parasitic plant germination.