Immunohistochemical detection of p53 tumor-suppressor protein is a poor indicator of prognosis for canine cutaneous mast cell tumors

Eighty-three canine cutaneous mast cell tumors were graded histologically and evaluated immunohistochemically for p53 tumor-suppressor protein expression. An avidin-biotin immunohistochemical protocol incorporated a rabbit polyclonal antibody (CM-1) directed against normal and mutant p53 protein. Positive staining was observed in 44.6% (37/83) of tumors and included 50% (12/24) of grade I (well differentiated) tumors, 46.9% (23/49) of grade II (intermediate differentiation) tumors, and 20% (2/10) of grade III (poorly differentiated) tumors. A statistically significantly higher proportion (P < 0.019) of tumors from the head and neck (83.3%, 10/12), stained positive for p53 than tumors from the thorax, back, abdomen, and axilla (39.4%, 13/33), legs (35.7%, 10/28), or prepuce, scrotal, or inguinal areas (44.4%, 4/9). No statistically significant difference between p53 labeling and histologic grade, breed, or tumor size was present. Survival data were available for 53/83 (63.9%) of dogs. Positive reactivity for p53 was observed in 47% (25/53) of tumors within this group, with 57.9% (11/19) of grade I, 43.3% (13/30) of grade II, and 25% (1/4) of grade III tumors labeled. Mean survival time for the 53 dogs was 12.1 months. The median survival time for dogs with grade III tumors or tumors >5 cm was statistically significantly shorter (P < 0.0001) than for dogs with grades I and II or smaller tumors. Although p53 protein abnormalities may play a role in tumor development or behavior in some canine cutaneous mast cell tumors, immunoreactivity was not associated with lack of tumor differentiation, tumor locations previously shown to demonstrate aggressive biological behavior, breed predisposition, or survival times.     

Predicting non‐native fish dispersal under conditions of climate change: case study in England of dispersal and establishment of pumpkinseed Lepomis gibbosus in a floodplain pond

Predictions of future climate change include shifts in patterns of precipitation, evapotranspiration and water run‐off, resulting in increased periods of drought as well as variability and intensity of rainfall events. In the United Kingdom, the non‐native North American sunfish, pumpkinseed Lepomis gibbosus (L.), is expected to benefit from these changes. We examine how hydrological variability induced by predicted changes in climate will affect the dispersal and spread of pumpkinseed in England by: (i) determining the relationship between discharge regime and pumpkinseed propagule pressure; (ii) examining a newly‐established pumpkinseed population following a flood event in 2007; and (iii) comparing the growth and life‐history traits of this new population with fish collected from the source population to demonstrate how the pumpkinseed's life‐history plasticity contributes to its success as a coloniser. Using Bayesian modelling, we determined that the number of pumpkinseed escapees is likely to increase with increasing discharge. The newly‐established pumpkinseed population showed fast juvenile growth, early age at maturity and small size at maturity. These traits differed significantly from the source population, specifically total length (TL) means at ages 1 and 2 were significantly greater in the new population, whereas TL at age 4 was significantly greater in the source population, and a significantly higher proportion of mature females were found at smaller size classes in the newly established pumpkinseed population. This study demonstrates the potential link between hydrological variability (current and future) and the dispersal of non‐native pumpkinseed, leading to the establishment of new populations.     

Growth-independent effects of a fluctuating thermal regime on the life-history traits of the Japanese medaka (Oryzias latipes)

Abstract –  Temperature fluctuation has been shown to affect somatic growth rates and metabolic functions of fish, but little is known about the effects of fluctuating temperatures on life-history traits. We examined these effects on female Japanese medaka ( Oryzias latipes ) reared from hatch to maturity at two mean temperatures (27 and 30 °C) with diel temperatures either held constant or allowed to fluctuate ±5 °C. Feeding regimes were manipulated to equilibrate medaka growth rates in all treatments. Age and length at maturity decreased as a result of thermal fluctuation, whereas there was no effect of temperature fluctuation on the proportion of fish attaining maturity or on the number of eggs produced by a mature female. We hypothesise that short-term high temperatures associated with temperature fluctuation decreases development time, whereas reduced energy conversion efficiency at higher temperatures does not allow for increased egg production.     

Alpha-amylase/subtilisin Inhibitor Levels in Australian Barleys

An enzyme-linked immunosorbent assay (ELISA) was used to determine the bifunctional alpha-amylase/subtilisin inhibitor (BASI) content of barley grain from 11 cultivars grown in six diverse locations in Australia. The inhibitor ranged from 119 to 254 μg/g in 57 barley samples. Genotype had a significant (P<0·05) effect on BASI content but there was no effect due to environment. Total protein varied independently of BASI and was influenced by environment and genotype. BASI content was higher (P<0·05) in malting barley than in feed barley and was correlated positively (r=0·29;P<0·05) with alpha-amylase activity in corresponding malts. The ELISA used monoclonal and polyclonal antibodies raised against purified BASI. In immunoblot analysis the monoclonal antibody showed high specificity for the inhibitor in barley and also detected the inhibitor in wheat. Low levels of inhibitor (mean 3·2 μg/g) were found in 12 Australian wheat cultivars using the ELISA developed for barley. The assay had a linear working range of 5–50 ng/mL with a detection limit of 2 ng/mL. Reproducibility between assays was good (CV=4·9%) but mean recoveries were high, ranging from 116–129% when purified inhibitor was added to barley extracts. The ELISA may have useful applications in brewing research and barley breeding programmes.