Method for the detection of synthetic cry3A in transgenic potatoes

All transgenic cultivars of potatoes registered in Canada and the United States have been modified to express a synthetic cry3A gene as a means of conferring resistance against the Colorado potato beetle, an important economic pest of potatoes. A PCR method was developed to amplify a 499 bp region of the synthetic cry3A gene. Using this method, synthetic cry3A could be detected in six different transgenic cultivars. Positive results could be confirmed with PvuII restriction digestion of the PCR-generated amplicon, which resulted in two fragments that were 283 and 216 bp in size. Of the 52 tuber extracts tested with this method, no false positive or false negative results were obtained, suggesting the method could be used with a high degree of accuracy. The absolute limit of detection was the number of cry3A copies present in one or perhaps two haploid copies of the potato genome. The practical limit of detection in tubers on a fresh weight basis was 0.02% for the NL 10-SUP and 0.01% for the remaining cultivars. Synthetic cry3A could also be detected in processed food products such as potato chips, shoestring potatoes, and frozen French fries. The method was suitable for screening potato tuber lots and some processed foods for the presence of synthetic cry3A.     

A comparison of constructed and natural habitat for frog conservation in an Australian agricultural landscape

Constructed ponds are an important consideration in the conservation of wetland biota in agricultural landscapes. Twenty-two natural ponds and 22 adjacent constructed ponds (farm dams) were surveyed on the Southern Tablelands of New South Wales to compare patterns of use by frogs and develop frog conservation recommendations. Farm dams supported similar numbers of frog species to natural ponds, although differences in frog assemblage were observed between the pond types. Limnodynastes tasmaniensis and Uperolia laevigata were significantly more likely to occur at farm dams while L. peronii was more likely to occur at natural ponds. Results suggest waterbodies with high levels of emergent vegetation cover that lack fish are likely to support a high number of frog species, regardless of origin (i.e. natural or constructed). However, it is important for landholders to conserve natural waterbodies as these environments appear likely to support frog species that do not use farm dams.     

Effects of meat consumption and vegetarian diet on risk of wrist fracture over 25 years in a cohort of peri- and postmenopausal women

BACKGROUND: Evidence suggesting that a diet high in fruits and vegetables may be beneficial to bone health has sparked interest in the potential benefit of a vegetarian diet. However, other studies have raised a question regarding the adequacy of protein in such a diet. OBJECTIVE: The aim of the present study was to take a whole foods approach in examining the effects of foods high in protein on the risk of wrist fracture (WF) in a cohort with a significant proportion consuming a meat-free diet. DESIGN: A cohort study of women who completed two lifestyle surveys 25 years apart. SUBJECTS: One thousand eight hundred and sixty-five peri- and postmenopausal women at the time of the first survey. RESULTS: There was a significant interaction between meat consumption and foods high in vegetable protein. Among vegetarians, those who consumed the least vegetable protein intake were at highest risk for fracture. However, increasing levels of plant-based high-protein foods decreased WF risk, with a 68% reduction in risk (hazard ratio (HR) = 0.32, 95% confidence interval (CI) 0.13-0.79) in the highest intake group. Among those with lowest vegetable protein consumption, increasing meat intake decreased the risk of WF, with the highest consumption decreasing risk by 80% (HR = 0.20, 95% CI 0.06-0.66). CONCLUSIONS: The finding that higher consumption frequencies of foods rich in protein were associated with reduced WF supports the importance of adequate protein for bone health. The similarity in risk reduction by vegetable protein foods compared with meat intake suggests that adequate protein intake is attainable in a vegetarian diet.     

Assay of the set of all Sudan azodye (I, II, III, IV, and Para-Red) contaminating agents by liquid chromatography-tandem mass spectrometry and isotope dilution methodology

A high-throughput mass spectrometric method is presented for the simultaneous detection of Sudan I, II, III, IV, and Para-Red azodyes in foodstuff. The method is based on the use of deuterium-labeled internal standards and atmospheric pressure chemical ionization (APCI) in a triple-quadrupole instrument. The gas-phase breakdown pattern of each labeled and unlabeled analogue displays the naphthoic moiety as a common fragment. The search for the parents of this common species (parent ion scans) allows, by flow injection and in a single run, the evaluation of the presence of each polluting species spiked in typical foodstuffs. A detailed assay of each azodye was performed by LC-APCI and isotope dilution method, through the multiple reaction monitoring approach, using deuterium-labeled internal standards. Sudan dyes can be quantified above the threshold of 10 ppb except for Sudan Para-Red, for which the limit of quantification was 20 ppb, likely due to the different ionization efficiency.     

Developmental biology. Rocks that roll zebrafish

The vestibular organs of the inner ear of higher vertebrates control balance, and their counterparts in fish control both balance and hearing. Essential to the operation of these sensory organs are the biomineralized structures--otoconia in higher vertebrates or otoliths in fish--that deflect the sensory hair bundles situated beneath them. In her Perspective, Fekete explores the fascinating world of otolith biomineralization in zebrafish; revealing the importance of a protein called Starmaker for coordinating the shape and type of crystal in fish otoliths ( S?llner et al.).     

Rehydration fluid temperature affects voluntary drinking in horses dehydrated by furosemide administration and endurance exercise

To determine whether temperature of rehydration fluid influences voluntary rehydration by horses, six 2-3-year-old horses were dehydrated (4-5% body weight loss) by a combination of furosemide administration and 30 km of treadmill exercise. For the initial 5 min following exercise, horses were offered a 0.9% NaCl solution at 10, 20, or 30 degrees C. Subsequently, after washing and cooling out, voluntary intake of water at 10, 20, or 30 degrees C from 20 to 60 min after exercise was measured. Fluid intake (FI) during the first 5 min of recovery was 9.8+/-2.5,12.3+/-2.1 and 9.7+/-2.0L (p>0.05) for saline at 10, 20, and 30 degrees C, respectively. Although not a significant finding, horses offered 0.9% NaCl at 20 degrees C tended to take fewer (p=0.07), longer drinks than when saline at either 10 or 30 degrees C was offered. Between 20 and 60 min of recovery, intake of water at 20 degrees C (7.7+/-0.8L) and 30 degrees C (6.6+/-1.2L) was greater (p<0.05) than that at 10 degrees C (4.9+/-0.5L). Thus, total FI was 14.7+/-2.5,19.9+/-2.5, and 16.3+/-2.4L for rehydration fluids at 10, 20, and 30 degrees C, respectively (p<0.05, value for 20 degrees C water greater than that for 10 degrees C water). Although the amount of metabolic heat transferred to the initial saline drink was correlated with the decrease in core temperature during the initial 5 min of recovery, heat transfer to ingested fluid was most likely responsible for the dissipation of, at most, 5% of the heat generated during endurance exercise. In conclusion, following exercise these dehydrated-normothermic horses voluntary drank the greatest amount of fluid at near ambient (20 degrees C) temperature. Although not determined in this study, greater satiation of thirst by oropharyngeal cooling may have contributed to lesser intake of colder (10 degrees C) fluid.     

Cloning, expression and characterization of monkey (Macaca fascicularis) CD137

CD137 plays an important role as a co-stimulatory molecule in activated T cells. Agonistic CD137 specific antibodies have been investigated as therapeutic agents to promote tumor-specific immune responses by direct activation of T cells. As part of the pre-clinical pharmacological evaluation of cynomolgus monkeys, monkey CD137 was cloned and characterized. The deduced amino acid sequence encoded a full-length gene of 254 amino acids 95% identical to human CD137. Sequence variants identified in monkey CD137 include four splice variants lacking the transmembrane domain. These variants were detectable in human including two previously unreported variants. Two missense single nucleotide polymorphisms were detected present in 42 and 50% of 36 monkeys tested. In both monkey and human, mRNA expression of full-length CD137 and splice variants were significantly increased in peripheral blood mononuclear cells (PBMCs) upon stimulation by anti-CD3 antibodies. Recombinant monkey CD137 protein was bound with high affinity by an agonistic anti-human CD137 antibody but not by an anti-mouse CD137 antibody. In summary, compared to human, monkey CD137 showed distinct extracellular domain amino acid sequence and sequence polymorphisms. Thus, antibodies directed against epitopes in this extracellular domain could have differences in pharmacologic activity between cynomolgus monkeys and human or across individual cynomolgus monkeys.     

Recent human‐induced salinity changes in Ramsar‐listed Orielton Lagoon,south‐east Tasmania,Australia: a new approach for coastal lagoon conservation and management

• 1. Many coastal lagoons in eastern Australia have changed as a result of recent human activities. However, it is often difficult to determine the extent of change and the ecological implications. A palaeoecological approach allows a quantitative assessment of how an aquatic ecosystem changes and responds to human impacts beyond what is possible with historical data or monitoring programmes.
• 2. Orielton Lagoon (south‐east Tasmania, Australia) is a Ramsar‐listed coastal wetland of international importance for conservation. This study was undertaken at the site to determine whether recent anthropogenic hydrological modifications to the lagoon had influenced its ecology, particularly salinity, and compromised its Ramsar status.
• 3. A diatom–salinity transfer function was constructed from a training data set of 96 diatom taxa from 34 sampling sites in 19 lagoons along the east coast of Tasmania.
• 4. The salinity of Orielton Lagoon has changed in response to a causeway constructed across its mouth, which has transformed the lagoon from an open marine environment to an enclosed, virtually stagnant, brackish water body.
• 5. These changes have compromised the protected coastal wetland status of Orielton Lagoon. Environmental remediation attempts have since partially restored the natural hydrology of the lagoon and it is now returning towards the state it was in prior to causeway construction.
• 6. A palaeoecological approach using diatoms was found to be successful in reconstructing recent salinity changes and investigating human impacts on Orielton Lagoon over the last 50–55 years.

Copyright © 2006 John Wiley & Sons, Ltd.