A physical map of the 1-gigabase bread wheat chromosome 3B

As the staple food for 35% of the world's population, wheat is one of the most important crop species. To date, sequence-based tools to accelerate wheat improvement are lacking. As part of the international effort to sequence the 17-billion-base-pair hexaploid bread wheat genome (2n = 6x = 42 chromosomes), we constructed a bacterial artificial chromosome (BAC)-based integrated physical map of the largest chromosome, 3B, that alone is 995 megabases. A chromosome-specific BAC library was used to assemble 82% of the chromosome into 1036 contigs that were anchored with 1443 molecular markers, providing a major resource for genetic and genomic studies. This physical map establishes a template for the remaining wheat chromosomes and demonstrates the feasibility of constructing physical maps in large, complex, polyploid genomes with a chromosome-based approach.     

Interactions between a non glycosylated human proline-rich protein and flavan-3-ols are affected by protein concentration and polyphenol/protein ratio

Interactions between salivary proline-rich proteins and tannins are involved in astringency, which is one of the most important organoleptic sensations perceived when drinking wine or tea. This work aimed to study interactions between a recombinant human salivary proline-rich protein, IB-5, and a flavan-3-ol monomer, epigallocatechin gallate (EGCG). IB-5 presented the characteristics of natively unfolded proteins. Interactions were studied by dynamic light scattering, isothermal titration microcalorimetry, and circular dichroism. The interaction mechanism was dependent on protein concentration. At low concentrations, a three-stage mechanism was evidenced. Saturation of the interaction sites (first stage) was followed by protein aggregation into metastable colloids at higher EGCG/protein ratios (second stage). Further increasing this ratio led to haze formation (third stage). At low ratios, a disorder-to-order transition of IB-5 structure upon binding was evidenced. At high protein concentrations, direct bridging between proteins and EGCG was observed, resulting in significantly lower aggregation and turbidity thresholds.     

Geographical variation of solanidane aglycone glycoalkaloids in the wild potato species Solanum chacoense Bitter

The Colorado Potato Beetle is a serious pest of the cultivated potato. Natural resistance has been found in a few wild species, including Solanum chacoense Bitter, in which the resistance is attributed to the presence of leptine glycoalkaloids. Production and accumulation of these compounds within S. chacoense varies widely and appears to be inherited in a quantitative fashion, but high leptine-producing clones occur rarely. In the present study, 15 different accessions from various locations and altitudes of origin within central to northern Argentina and Paraguay were analyzed for foliar glycoalkaloid (leptine, leptinine, solanine, chaconine) content. The objective was to infer the frequency of leptine production in ecogeographically distinct S. chacoense accessions, and to ascertain any possible association between leptine levels/concentrations and ecogeographical location. Leptines were detected in 8 of the 15 accessions, and the amounts within each accession varied widely. Most of the leptine-containing accessions originated from western Argentina except two in province Córdoba in central Argentina. There was no relationship between elevational level and leptine, but there was a negative trend with total glycoalkaloids (TGA) and elevation, due to solanine and chaconine levels which decreased with increasing elevation. In addition, nine unidentified, putative glycoalkaloids were detected, in very high proportions in some individuals and accessions. This study raises interesting questions about glycoalkaloid distribution, helps provide direction for new avenues of leptine and glycoalkaloid research, and proposes a systematic, ecogeographically based method for bioprospecting genes controlling rare plant secondary compounds.     

Methanolic extract of Verbascum macrurum as a source of natural preservatives against oxidative rancidity

The antioxidant properties of various fractions of a methanolic extract obtained from the aerial parts of Verbascum macrurum have been determined by monitoring their capacity to scavenge the stable free-radical DPPH. They were also evaluated as natural preservatives against oxidative rancidity using the accelerated Rancimat method. Their activities expressed as protection factor (PF(r)) indicated that the fractions rich with phenylpropanoid glycosides were more potent compared to alpha-tocopherol and of the same magnitude as BHT, which were used as reference standards. Ten natural compounds were identified as components of this methanolic extract and were isolated by medium-pressure liquid chromatography (MPLC). Assessment of their antioxidant activities established that acteoside, a polyhydroxylated phenylpropanoid glycoside derivative, is the most potent free radical scavenger and showed the highest protection factor (PF(r)) against sunflower-oil-induced oxidative rancidity. Its activity is comparable to the synthetic antioxidant BHT and clearly superior to natural alpha-tocopherol. This compound therefore represents a very interesting candidate for use in food preservation as natural protecting agent against oxidative rancidity.     

Role of endothelium and nitric oxide in modulating in vitro responses of colonic arterial and venous rings to vasodilatory neuropeptides in horses

The objective of this study was to determine and compare the in vitro responses of equine large colon arterial and venous rings to vasodilatory neuropeptides; calcitonin gene-related peptide (CGRP); substance P (SP); vasoactive intestinal polypeptide (VIP); and acetylcholine (ACh), a standard nonpeptide endothelium-dependent vasodilator. Responses of vessel rings to graded concentrations (10(-11) M to 10(-5) M) of each drug were determined in endothelium-intact, denuded, and Nomega-nitro-L-arginine methyl ester (L-NAME, 10(-5) M)-treated rings that were pre-contracted with norepinephrine. Percentage maximal relaxation (PMR), defined as the % decrease from the contracted state, was determined. Because all rings did not relax at least 50%, EC50 values could not be consistently calculated. Arterial rings with intact endothelium were more sensitive to CGRP, compared with VIP and SP, and venous rings of all conditions were more sensitive to VIP than CGRP or SP. Overall, arteries had a greater PMR for ACh compared with SP and VIP. Intact and L-NAME treated arteries had a greater PMR than denuded arteries; there were no differences in PMR of intact and L-NAME treated arteries. Veins had a greater PMR for VIP than CGRP, SP, or ACh. Calcitonin gene-related peptide caused greater relaxation in intact arteries, whereas VIP causes greater relaxation in veins. Arterial relaxation was dependent upon the presence of intact endothelium. The response of veins to VIP among the conditions tested was not different, suggesting VIP has direct actions on venous smooth muscle. These neuropeptides modulate vasomotor tone via vasorelaxation in colonic arteries and veins.     

Epiphytic life is the main characteristic of the life cycle ofPseudomonas syringae pv.pisi,pea bacterial blight agent

Pseudomonas syringae pv.pisi, pea bacterial blight agent, is seed-transmitted. Some aspects of its life cycle and its biology were investigated. The colonization of pea plants obtained from naturally infected seeds was studied in natural conditions while high populations of bacteria developed on plants showing no symptoms. Two streptomycin-resistant mutants were used to study the epiphytic life of the pathogen. Populations were monitored in different host-parasite compatibilities. When race 2 or race 6 of the pathogen was surface-inoculated on susceptible cultivars, a decrease of population size was observed during the following one to three days but was followed by an increase to levels 1000 times greater than the initial number detected, without symptoms for most of the plants. When race 2 was surface-inoculated on resistant genotypes or race 6 on non-host plants, bacteria did not multiply but population levels slightly decreased.Pseudomonas syringae pv.pisi shows a resident phase and its development is race-specific. Weeds collected in naturally contaminated pea fields, diseased or not, often harboured the pathogen but with levels smaller than those observed on peas. Pea crop debris and volunteers kept high levels of bacteria for at least eight months after the harvest of a diseased crop. As long as two pea crops are not grown one after the other in the same field, it is unlikely that debris and volunteers will act as an important inoculum source. The development of this pathogen during the growing season is considered as an important parameter to take into account for controlling the disease through seed health testing.     

Detection of Salmonella organisms and assessment of a protocol for removal of contamination in horse stalls at a veterinary teaching hospital

OBJECTIVES: To assess methods of detecting environmental contamination with Salmonella organisms and evaluate a cleaning and disinfection protocol for horse stalls in a veterinary teaching hospital. DESIGN: Original study. SAMPLE POPULATION: 37 horses with diarrhea likely to be caused by Salmonella infection and their stall environments. PROCEDURES: Fecal samples were collected from horses daily during hospitalization; samples were obtained from stall sites after cleaning and application of disinfectants. Fecal and environmental samples were cultured for Salmonella spp and tested via polymerase chain reaction (PCR) assay to detect Salmonella DNA. RESULTS: 1 horse died and 2 were discharged prior to sample collection. Fecal samples from 9 of 34 horses yielded growth of Salmonella organisms on bacteriologic culture, and 23 yielded positive results via PCR assay on > or = 1 occasion. Among environmental samples from 21 stalls, salmonellae were detected at > or = 1 stall site on 6 of 78 occasions, and > or = 1 stall site yielded positive results via PCR assay on 69 of 77 occasions. Salmonella DNA was detected more frequently in samples of stall drains, cracks, and corners. Salmonella spp were cultured from samples of 3 stalls after both initial and second cleaning and disinfection cycles, but no organisms were detected in samples obtained after use of a peroxygen disinfectant. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that stalls in which horses with salmonellosis were housed should only be used to accommodate newly hospitalized horses after samples (collected after 2 cycles of cleaning and disinfection) from drains, cracks, and corners yield negative results on bacteriologic culture.     

A comparison of the cytologic and histologic features of meningiomas in four dogs

Abstract: The cytologic and histologic features of 2 intracranial and 2 spinal (extramedullary cervical) canine meningiomas were compared. Cerebrospinal fluid analysis in 2 cases revealed mild, mixed cell pleocytosis, primarily composed of small lymphocytes and monocytoid cells, with a moderate increase in total protein concentration. Cytologic features suggestive of meningioma included cells with both epithelial and mes-enchymal characteristics and a tendency towards cell clustering. Tumor location also was useful in making a diagnosis. The 4 meningiomas differed histologically from one another, and included angioblastic, psam-momatous, meningotheliomatous, and microcystic anaplastic types, which conformed to a classification scheme for human meningiomas. The classification scheme could not be applied to cytologic specimens.