Characterization of black locust (Robinia pseudoacacia L.) heartwood extractives: identification of resveratrol and piceatannol

Robinia pseudoacacia L. heartwood is characterized by a very high natural durability. However, a significant difference was observed between the mature and juvenile heartwood, the latter presenting less durability against fungi decay, which could be attributed to lower extractive content. In order to elucidate this idea, extractives from mature and juvenile heartwoods of black locust trees were investigated. Results showed that extractive and phenolic contents were higher in mature than in juvenile heartwoods. The identification of phenolic compounds by UPLC–DAD–MS/MS revealed, for the first time, the presence of resveratrol and piceatannol. These two stilbenes as well as the flavonoid dihydrorobinetin were present at the highest level in mature heartwood, and as they are known antifungals, they could account for the great durability of mature heartwood. The stilbenes were detected in significant amounts particularly in mature heartwood where piceatannol reached a level tenfold higher than that reported for Japanese knotweed roots, the primary natural source of these stilbenes, whereas resveratrol level was comparable with reported values. As resveratrol and piceatannol receive increasing demand for nutraceutical, cosmetic and, possibly, pharmaceutical purposes, due to their beneficial health effects, this study underlines the use of R. pseudoacacia as a promising sustainable and economical source of resveratrol and piceatannol.     

Evaluation of the arterial blood pressure of dogs by two noninvasive methods.

The determination of the arterial blood pressure was done on 12 healthy mixed breed dogs in both the anesthetized and the conscious state, to evaluate two instruments (Doppler flow detector and infrasonde D4000), in their ability to indirectly determine arterial blood pressure. The coefficients of variation were higher with indirect methods when compared with the results obtained by cannulation. These coefficients were lower with the Doppler flow detector. The correlation study showed that both apparatuses were reliable in most situations. The infrasonde D4000 was more accurate than the Doppler in the conscious animals. However the results showed a lack of precision in hypertensive conscious dogs. The diastolic arterial blood pressure was particularly precise in the case of the anesthetized hypotensive dogs. Its sensitivity allowed it to register muscle movement artifacts. The Doppler flow detector showed less variation and was particularly accurate in both anesthetized and conscious hypertensive dogs. Its sensitivity allowed artifact movement sounds to be detected. The Doppler should be used in quiet surroundings or earphones should be worn by the evaluator. Some form of restraint is needed with the use of both instruments. Even if the correlations with the direct arterial blood pressure values were better with the infrasonde D4000, greater variations were found in the individual readings. The Doppler instrument represents in the hands of the investigators a better instrument for routine monitoring of blood pressure in the dog.     

Ecosystem health as a clinical rotation for senior students in Canadian veterinary schools.

We describe 4 years of an experimental rotation in ecosystem health offered to senior veterinary students in Canada. Faculty from the 4 Canadian veterinary colleges collaborated in offering the rotation once annually at 1 of the colleges. The 1st rotation was held in Guelph in 1993, followed in successive years by rotations at Saskatoon, Saint-Hyacinthe, and Charlottetown. The rotation is a predominantly field-based experience that allows students to work with veterinary and other role models who are actively engaged in clinical research related to ecosystem health. Five specific field studies that worked particularly well during the rotations are presented. These studies involved investigating mortality in wildlife due to botulism, designing an environmental surveillance system around herds of beef cattle, using belugas to evaluate the health of the St. Lawrence River, dealing with competition for water use by aquaculture and agriculture, and exploring the role of veterinarians during major coastal oil spills. The experience has resulted in our developing the subject matter, field examples, teaching approach, and confidence necessary to make ecosystem health the focus of a productive clinical rotation for senior year veterinary students.     

Influence of litter size on metabolic status and reproductive axis in primiparous sows

This study on primiparous sows was designed to 1) determine the impact of nursing a large litter on LH secretion and follicular development, and 2) investigate the metabolic adaptations by which milk yield increases with litter size. At farrowing, crossbred, primiparous sows were assigned to 1 of 3 experimental groups differing in litter size and feed allowance. Sows with 13 or 14 piglets (13AL, n = 7) were fed ad libitum. Sows with 7 piglets were fed ad libitum (7AL, n = 6) or were feed-restricted (7R, n = 8). The restriction was based on the estimated energy deficiency for the 13AL sows. On d 9 +/- 1 of lactation, a jugular catheter was surgically implanted. Serial blood samplings and glucose tolerance tests were performed in mid- and late lactation. Sows were slaughtered 3 d after weaning, and ovarian characteristics were recorded. During lactation, the 7AL sows lost no or little body reserves, and their estimated energy balance was near zero. The 13AL and 7R sows exhibited similar negative energy balances and similar losses of backfat and estimated lipid content. Litter growth rate was greater (P < 0.05) in the 13AL than in the 7AL and 7R groups. After weaning, the volume of the largest 14 follicles was smaller (P < 0.05) in sows nursing 13 or 14 piglets than in sows with 7 piglets. Plasma concentrations of LH and LH pulse frequency did not differ between groups (P > 0.1). The longer glucose half-life on d 16 than on d 27 of lactation (22.5 vs. 18.8 min; P < 0.05) indicated a lower glucose tolerance in mid- than in late lactation. The area under the insulin curve was greater in the 7AL than in the 13AL sows (P = 0.08) and intermediate in the 7R group, with no differences in glucose profiles. This led to the suggestion that the 7AL sows were more resistant to insulin than the 13AL sows. In all groups of sows, follicular development after weaning was correlated with LH secretion in midlactation. Active follicular development was associated with prolonged secretion of insulin in response to glucose challenge. Our results show that besides litter size, a sow's metabolic status in lactation influences follicular maturation after weaning and also indicate that the metabolic adaptations by which primiparous sows nursing large litters increase litter growth rate and body reserve mobilization do not involve an accentuated peripheral insulin resistance.     

Risk assessment of outcrossing of transgenic rapessed to related species:

Summary The risk for a gene dispersal is reported for reciprocal crosses between a transgenic rapeseed variety resistant to the herbicide phosphinotricin and five related species. The first stages after pollination were cytologically observed and fertilized ovaries were established in in vitro culture for the production of interspecific hybrids. A similar classification was observed for the index of pollination compatibility and embryo yield. From the 243 embryos produced, 109 plantlets were obtained in a greenhouse. All the interspecific combinations tested were able to produce hybrid plants. A higher number of hybrids was obtained when rapeseed was used as the female parent. The hybrids had the expected triploid structure except for two amphidiploid, B. napus × B. oleracea, and one amphidiploid, B. napus × S. arvensis, plants with 56 chromosomes. The triploid hybrids were sterile or partially fertile but two of the amphidiploid plants, B. napus × B. oleracea, were fully fertile. The cytoplasm source did not seem to affect the fertility of the hybrids.     

Characterization of dinucleotide and trinucleotide EST-derived microsatellites in the wheat genome

Over the past decade microsatellites or simple sequence repeats (SSRs) have attracted a considerable amount of attention from researchers. The aim of the present paper was to analyse expressed sequence tag-derived SSR (EST-SSR) marker variability in wheat and to investigate the relationships between the number and type of repeat units and the level of microsatellite polymorphism. Two hundred and forty-one new EST-SSR markers available in a public database () were characterized in eight durum wheat cultivars (Svevo, Ciccio, Primadur, Duilio, Meridiano, Claudio, Latino, Messapia), two accessions of Triticum turgidum var. dicoccoides (MG4343, MG29896), one accession of T. turgidum var. dicoccum (MG5323) and in the common wheat cv. Chinese Spring. Of these, 201 primer pairs (83.4%) amplified PCR products successfully, while the remaining 40 (16.6%) failed to amplify any product. Of the EST-SSRs analysed, 45.2% of the primer pairs amplified one or two PCR products. Multiple discrete PCR products were observed among both di- and trinucleotide EST-SSR markers (31.2 and 40.5%, respectively). Markers based on dinucleotide microsatellites were more polymorphic than those based on trinucleotide SSRs in the 12 wheat genotypes tested (68.9 and 52.7%, respectively). An average of 2.5 alleles for dinucleotide and 2.0 alleles for trinucleotide SSRs was observed. The data reported in the present work indicate the presence of a significant relationship between motif sequence types and polymorphism. The primer set based on the AG repeat motif showed the lowest percentage of polymorphism (55.0%), while the primer set based on the AC repeat motif showed t he highest percentage (85.0%). Among trinucleotide SSRs, the AGG microsatellite markers showed the highest percentage of polymorphism (70.0%), and the ACG motif the lowest value (25.0%). The characterization of these new EST-SSR markers and the results of our studyon the effect of repeat number and type of motifs could have important applications in the genetic analysis of agronomically important traits, quantitative trait locus discovery and marker-assisted selection.     

Heredity of seventeen isozyme loci in cassava (Manihot esculenta Crantz)

Summary Starch gel electrophoresis was used to assess isozyme polymorphism in two Manihot species. Crude extracts were obtained from leaves and pollen. Ten enzymes were examined for their polymorphism in a germplasm collection of 365 cultivated plus 109 wild accessions, mainly from Africa. The inheritance of these enzymes was examined using 13 intra and interspecific progenies. Seventeen polymorphic loci were found for the ten enzyme systems, with 59 alleles. All the markers showed disomic heredity and three linkage groups were identified.     

Assessing success in gene transfer betweenLolium multiflorum andFestuca arundinacea

Summary AFestuca arundinacea (2n=6x-42) plant with three PGI/2 homoeoalleles marking three homoeologous chromosomes was crossed with aLolium multiflorum plant (2n=4x=28) with a different PGI/2 phenotype to give a pentaploid hybrid (2n=5x=35) with five chromosomes each marked by a different PGI/2 allele. This hybrid plant was backcrossed twice with diploidL. multiflorum (2n=2x=14) with a different PGI/2 phenotype. Numbers of interspecific recombinants involving chromosomes marked by PGI/2 were then determined in both backcross generations. In the BC1, recombinants involving only one PGI/2 allele were found but in the BC2, all three homoeologousF. arundinacea chromosomes carrying the PGI/2 locus recombined withLolium with one in greater frequency and the others in equal but lower frequency. The evidence supports claims made forF pratensis (2n=2x=14) andF. arundinacea var.glaucescens (2n=4x=28) being progenitors forF. arundinacea.     

Pressure-induced unfolding and aggregation of the proteins in whey protein concentrate solutions

Whey protein concentrate solutions (12% w/v, pH 6.65 +/- 0.05) were pressure treated at 800 MPa for 20-120 min and then examined using size exclusion chromatography (SEC), small deformation rheology, transmission electron microscopy, and various types of one-dimensional (1D) and two-dimensional (2D) polyacrylamide gel electrophoresis (PAGE). The pressure-treated samples showed a time-dependent loss of native whey proteins by SEC and 1D PAGE and a corresponding increase in non-native proteins and protein aggregates of different sizes. These aggregates altered the viscosity and opacity of the samples and were shown to be cross-linked by intermolecular disulfide bonds and by noncovalent interactions using 1D PAGE [alkaline (or native), sodium dodecyl sulfate (SDS), and SDS of reduced samples (SDS(R))] and 2D PAGE (native:SDS and SDS:SDS(R)). The sensitivity of the major whey proteins to pressure was in the order beta-lactoglobulin B (beta-LG B) > beta-LG A > bovine serum albumin (BSA) > alpha-lactalbumin (alpha-LA), and the large internal hydrophobic cavity of beta-LG may have been partially responsible for its sensitivity to high-pressure treatments. It seemed likely that, at 800 MPa, the formation of a beta-LG disulfide-bonded network preceded the formation of disulfide bonds between alpha-LA or BSA and beta-LG to form multiprotein aggregates, possibly because the disulfide bonds of alpha-LA and BSA are less exposed than those of beta-LG either during or after pressure treatment. It may be possible that intermolecular disulfide bond formation occurred at high pressure and that hydrophobic association became important after the high-pressure treatment.