Recovery and fine structure variability of RGII sub-domains in wine (Vitis vinifera Merlot).

BACKGROUND AND AIMS: Rhamnogalacturonan II (RGII) is a structurally complex pectic sub-domain composed of more than 12 different sugars and 20 different linkages distributed in five side chains along a homogalacturonan backbone. Although RGII has long been described as highly conserved over plant evolution, recent studies have revealed variations in the structure of the polysaccharide. This study examines the fine structure variability of RGII in wine, focusing on the side chains A and B obtained after sequential mild acid hydrolysis. Specifically, this study aims to differentiate intrinsic structural variations in these RGII side chains from structural variations due to acid hydrolysis. METHODS: RGII from wine (Vitis vinifera Merlot) was sequentially hydrolysed with trifluoroacetic acid (TFA) and the hydrolysis products were separated by anion-exchange chromatography (AEC). AEC fractions or total hydrolysates were analysed by MALDI-TOF mass spectrometry. KEY RESULTS: The optimal conditions to recover non-degraded side chain B, side chain A and RGII backbone were 0·1 m TFA at 40 °C for 16 h, 0·48 m TFA at 40 °C for 16 h (or 0·1 m TFA at 60 °C for 8 h) and 0·1 m TFA at 60 °C for 16 h, respectively. Side chain B was particularly prone to acid degradation. Side chain A and the RGII GalA backbone were partly degraded by 0·1 m TFA at 80 °C for 1-4 h. AEC allowed separation of side chain B, methyl-esterified side chain A and non-methyl-esterified side chain A. The structure of side chain A and the GalA backbone were highly variable. CONCLUSIONS: Several modifications to the RGII structure of wine were identified. The observed dearabinosylation and deacetylation were primarily the consequence of acidic treatment, while variation in methyl-esterification, methyl-ether linkages and oxidation reflect natural diversity. The physiological significance of this variability, however, remains to be determined.

Monoclonal antibodies to rhamnogalacturonan I backbone.

Monoclonal antibodies were raised against rhamnogalacturonan I backbone, a pectin domain, using Arabidopsis thaliana seed mucilage-derived rhamnogalacturonan I oligosaccharides--BSA conjugates. Two monoclonal antibodies, designated INRA-RU1 and INRA-RU2, selected for further characterization, were specific for the backbone of rhamnogalacturonan I, displaying no binding activity against the other pectin domains i.e. homogalacturonans, galactans or arabinans. A range of oligosaccharides was prepared by enzymatic digestion of rhamnogalacturonan I isolated from Arabidopsis thaliana seed mucilage and from sugar beet pectin, purified by low-pressure chromatography and characterized by high-performance anion-exchange chromatography and mass spectrometry. These rhamnogalacturonan I oligomers were used to characterize the binding site of the two monoclonal antibodies by competitive inhibition. Both INRA-RU1 and INRA-RU2 showed maximal binding to the [-->2)-alpha-L-rhamnosep-(1-->4)-alpha-D-galacturonic acid p-(1-->](7) structural motif but differed in their minimum binding requirement. INRA-RU2 required at least two disaccharide (rhamnose-galacturonic acid) repeats for the antibody to bind, while INRA-RU1 required a minimum of six disaccharide repeats. Furthermore, the binding capacity of INRA-RU1 decreased steeply as the number of disaccharide repeats go beyond seven. Each of these antibodies reacted with hairy regions isolated from sugar beet pectin. Immunofluorescence microscopy indicated that both antibodies can be readily used to detect rhamnogalacturonan I epitopes in various cell wall samples.

Aleurone cell walls of wheat grain: high spatial resolution investigation using synchrotron infrared microspectroscopy.

Infrared microspectroscopy and immunolabeling techniques were employed in order to obtain deeper insight into the biochemical nature of aleurone cell walls of wheat grain. The use of a synchrotron source, thanks to its intrinsic brightness, has provided unprecedented information at the level of a few micrometers and has allowed the discrimination of various polysaccharides in cell walls. The high spectral quality obtained in the small analyzed domain has been beneficial in estimating the relative proportions of beta-glucan and arabinoxylan, through the use of principal component analysis (PCA). The highest amount of beta-glucan is found in periclinal cell walls close to the starchy endosperm. The junction regions between aleurone cells are enriched in arabinoxylan. At the early stage of wheat grain development (271 degrees D), the chemical composition along the cell walls is more heterogeneous than at the mature stage. Both synchrotron infrared microspectroscopy and immunolabeling experiments made it possible to reveal the spatial heterogeneity of the various chemical compositions of aleurone cell walls.

Carbohydrate fractions of legumes: uses in human nutrition and potential for health.

Starch and fibre can be extracted, using wet or dry processes, from a variety of grain legumes and used as ingredients for food. alpha-Galactosides can be isolated during wet processes from the soluble extract. Starch isolates or concentrates are mostly produced from peas, whereas dietary fibre fractions from peas and soyabean are commercially available. The physico-chemical characteristics of fibre fractions very much depend on their origin, outer fibres being very cellulosic whereas inner fibres contain a majority of pectic substances. Inner fibres are often used as texturing agents whereas outer fibres find their main uses in bakery and extruded products, where they can be introduced to increase the fibre content of the food. Most investigations on impacts on health have been performed on soyabean fibres. When positive observations were made on lipaemia, glucose tolerance or faecal excretion, they were unfortunately often obtained after non-realistic daily doses of fibres. Legume starches contain a higher amount of amylose than most cereal or tuber starches. This confers these starches a lower bioavailability than that of most starches, when raw or retrograded. Their low glycaemic index can be considered as beneficial for health and especially for the prevention of diseases related to insulin resistance. When partly retrograded, these starches can provide significant amount of butyrate to the colonic epithelium and may help in colon cancer prevention. alpha-Galactosides are usually considered as responsible for flatus but their apparent prebiotic effects may be an opportunity to valorize these oligosaccharides.

Highlighting of intervertebral movements and variations of intradiskal pressure during lumbar spine manipulation: a feasibility study.

OBJECTIVES: To demonstrate relative movement of the vertebrae and variations in intradiskal pressure during 2 different lumbar spinal manipulations, in flexion or extension, in 2 unembalmed cadavers. DESIGN: A pressure sensor was inserted into the L3-4 disk in cadaver 1 and into the L1-2 to L4-5 disks in cadaver 2. Two adjacent vertebrae (L3 and L4 in cadaver 1, and L4 and L5 in cadaver 2) were each equipped with 2 monoaxial accelerometers to record acceleration in the caudocranial axis and a biaxial accelerometer to record acceleration in the "horizontal" anatomic plane. SETTING: Laboratory study. RESULTS: During the thrust, relative intervertebral movements were demonstrated; movements differed with the type of manipulation (in flexion or extension). Intradiskal pressure initially increased, then decreased. CONCLUSIONS: Lumbar spinal manipulations have a biomechanical effect on the intervertebral disks, producing a brief but marked change in intradiskal pressure. This effect, which differs slightly with the different types of manipulation studied, is the consequence of movements of the adjacent vertebrae.

Digestion of carbohydrate from white beans (Phaseolus vulgaris L.) in healthy humans.

Resistant starch (RS) is thought to be present in large amounts in legume seeds; however, it has never been quantified in healthy humans. RS from cooked (atmospheric pressure) white beans was quantified in humans and pigs, and characterized to explain its low digestibility. Six human volunteers were intubated to collect ileal digesta after an experimental meal composed of orange juice, butter and 167 g beans (dry matter basis). The reliability of the intubation method was examined in a pig study in which it was compared with another collection method, ileal cannulation. Chemical analyses, microscopy and size exclusion chromatography were performed on human and pig digesta. The pig study showed that the intubation method may underestimate the quantity of RS. However, no chemical/physical difference was observed between the RS collected by the two techniques. In the human study, 16.5 +/- 1.3% (11.3 g) of the ingested starch was recovered as RS. The microscopy of the digesta showed that part of the RS was enclosed in the cell walls. Although the RS was composed mainly of alpha-glucan molecules with a degree of polymerization (DP) 40 to 60, oligosaccharides and large molecules of DP > 400 were also present. Retrogradation was not found to be the main factor responsible for starch malabsorption. We conclude that white beans may contain a large amount of RS formed mainly by partially degraded molecules protected by the cell walls during their transit through the gut.

Emulsification and lipolysis of triacylglycerols are altered by viscous soluble dietary fibres in acidic gastric medium in vitro.

This in vitro study was designed to test the hypothesis that soluble dietary fibres can alter the process of intragastric lipid emulsification and possibly subsequent triacylglycerol lipolysis. Three guar gums, two pectins and gum arabic were dissolved in acidic gastric medium in the concentration range 0.3-2.0% (w/v). Viscosities of fibre solutions were measured and apparent viscosities varied over a wide range (0.7-77 mPa/s). Emulsification of a lipid mixture (triolein/phosphatidylcholine/cholesterol) was performed under mild conditions in the presence of increasing concentrations of soluble fibres. The amount of emulsified lipid was not affected whereas the size of the emulsified droplets was increased by raising the concentration of viscous fibres only. The droplet size (r=0.75, P=0.006) and overall droplet surface area (r=-0.69, P=0.009) were strongly correlated with the medium viscosity in the range 0-20 mPa/s. The addition of solutions of viscous fibres to a preformed standard emulsion did not change the initial velocity of human gastric lipase reaction. Conversely, when emulsions prepared in the presence of fibres (i.e. with different droplet sizes) were incubated with excess gastric enzyme for 2 h, the high-viscosity guar gum significantly reduced the extent of triacylglycerol lipolysis, as compared with control and low- or medium-viscosity fibres. In conclusion, the data obtained show that reducing emulsification of dietary lipids in the mildly acid medium found in the stomach is a mechanism by which soluble viscous fibres can alter lipid assimilation.

Urethral Syndrome in the female ("irritable bladder"): the expression of fantasies about the urogenital area.

The urethral syndrome is a frequently encountered psychosomatic disorder; it constitutes the main complaint of some 20% of all women who consult a urologist. After a brief survey of the history of this syndrome, the authors examine 55 case reports of women with the urethral syndrome. The authors particularly emphasize the significance of the manner in which patients describe their perceptions of the pain, which is a valuable diagnostic sign. A follow-up study of patients for 3 to 21 years after the first interview demonstrated the releative efficiency of the various forms of treatment. A discussion of the therapeutic possibilities and their limitations follows.