Isolation and characterization of the viscous, high-molecular-mass microbial carbohydrate fraction from faeces of healthy subjects and patients with Crohn's disease and the consequences for a therapeutic approach

1998 ◽  
Vol 95 (4) ◽  
pp. 425-433
Author(s):  
Johanneke G. H. RUSELER-VAN EMBDEN ◽  
Leo M. C. VAN LIESHOUT ◽  
Doede J. BINNEMA ◽  
Maarten P. HAZENBERG

1.An earlier study by our group revealed that the viscosity of faeces from patients with Crohn's disease is significantly lower than that of healthy subjects. This is due to low concentrations of a high-molecular-mass carbohydrate, probably of bacterial origin. The cause of this phenomenon might be the impaired barrier function of the gut mucosa. Low viscosity may allow close contact of intestinal contents (bacterial products and toxins) with the intestinal wall. This could play a role in the maintenance of the disease. 2.The first aim of this study was to investigate the high-molecular-mass carbohydrate fraction, responsible for viscosity, in detail. We also tried (in a pilot study) to raise the intestinal viscosity of patients with Crohn's disease with the undegradable food additive hydroxypropylcellulose (E463), in an attempt to alleviate clinical symptoms. 3.The high-molecular-mass fraction (> 300 ;kDa) responsible for faecal viscosity was sensitive to lysozyme and contained high levels of muramic acid. It was concluded that this material consisted mainly of peptidoglycan polysaccharides and was consequently of bacterial origin. The muramic acid in material from patients with Crohn's disease was 7.5 (1.5–13.9)%, which was less than in healthy subjects [11.4 (8.5–24.1)%; P = 0.0004]. Furthermore, viscosity in material from patients with Crohn's disease was found to be half [14.9 (1.0–33.6) cP] of that found in healthy subjects [35.0 (2.7–90.7) cP; P = 0.004]. 4.A daily dose of 1 ;g of hydroxypropylcellulose caused an increase in faecal viscosity in patients with Crohn's disease (from 1.4 to 2.3 cP) and in healthy subjects (from 4.9 to 7.5 cP). Faecal consistency improved in patients with Crohn's disease (from watery and loose to formed) and the defecation frequency decreased from 3–4 to about 2 times a day. No changes in defecation patterns were found in healthy subjects. 5.These data indicate that the high-molecular-mass fraction that is responsible for faecal viscosity is peptidoglycan. Furthermore, a daily dose of a hydroxypropylcellulose solution to increase the viscosity of the intestinal contents of patients with Crohn's disease might be beneficial. This approach merits further study.

2008 ◽  
Vol 24 (2) ◽  
pp. 111-117 ◽  
Author(s):  
N. Rodríguez-Pérez ◽  
A. Aguinaga-Barrilero ◽  
Marina B. Gorroño-Echebarría ◽  
Mercedes Pérez-Blas ◽  
J. M. Martín-Villa

We wished to analyse the frequency of Crohn’s disease-linked CARD15 polymorphisms (P268S, R702W, G908R and 1007fs) in a group of Spanish patients with idiopathic uveitis. To this aim, DNA samples were obtained from 111 unrelated patients. P268S, R702W and G908R polymorphisms were detected using TaqMan Genotyping kits (Applied Biosystems), and the 1007fs variation by direct DNA sequencing. Control group consisted of 105 healthy subjects.None of the polymorphisms studied revealed a significant increase in the groups of patients, when compared to the control group. Thus, P268S is found in 50% of patients (gene frequency 0.284) vs 44% of control individuals (gene frequency 0.245); R702W in 7% of patients (0.036) vs 7% (0.033); G908R in 2% of patients (0.009) vs 4% (0.019) and, finally, 1007fsin 2% of uveitis patients (0.008) vs 4% (0.021). Moreover, DNA sequencing has allowed us to define two new intronic polymorphisms in phase, in the 5' and 3' boundaries of the exon 11 (GenBank accession number #DQ 869189).Altogether, our results suggest that the Crohn’s disease-linked CARD15 polymorphisms do not seem to predispose to idiopathic uveitis in the Spanish population.


1993 ◽  
Vol 291 (3) ◽  
pp. 793-798 ◽  
Author(s):  
A Thewles ◽  
R A Parslow ◽  
R Coleman

Biliary cholesterol output in rats was stimulated over 3-fold by feeding diosgenin for 5 days, whereas biliary outputs of phospholipid and bile salts were not changed by diosgenin feeding. Isolating and perfusing the liver without bile salts resulted in a rapid and substantial decrease in biliary bile salt output; bile salt depletion abolished the diosgenin-induced increment in biliary cholesterol output, showing that the diosgenin-elevated biliary cholesterol output was bile-salt-dependent. Diosgenin treatment also produced a significant decrease in biliary alkaline phosphodiesterase I. Fresh bile obtained from control and diosgenin-fed rats was subjected to gel-permeation chromatography in order to separate different-sized biliary cholesterol carriers. Two major peaks of cholesterol were eluted, with cholesterol also being eluted between the peaks. The cholesterol peak eluted at the lower molecular mass (20-30 kDa) was observed in all bile samples. The higher-molecular-mass peak, which was eluted at the void volume, was not observed in all biles; control biles contained very little high-molecular-mass form of cholesterol, whereas biles from the diosgenin group contained up to 47% of cholesterol in the high-molecular-mass fraction. Diosgenin treatment produced a range of elevated biliary cholesterol values which positively correlated with the proportion of cholesterol contained in the high-molecular-mass fraction (r = 0.98). The results show that diosgenin induced a marked bile-salt-dependent increase in biliary cholesterol output and a shift in biliary cholesterol transport to higher-molecular-mass structures.


2006 ◽  
Vol 400 (3) ◽  
pp. 477-484 ◽  
Author(s):  
Nick Sirijovski ◽  
Ulf Olsson ◽  
Joakim Lundqvist ◽  
Salam Al-Karadaghi ◽  
Robert D. Willows ◽  
...  

Magnesium chelatase inserts Mg2+ into protoporphyrin IX and is the first unique enzyme of the chlorophyll biosynthetic pathway. It is a heterotrimeric enzyme, composed of I- (40 kDa), D- (70 kDa) and H- (140 kDa) subunits. The I- and D-proteins belong to the family of AAA+ (ATPases associated with various cellular activities), but only I-subunit hydrolyses ATP to ADP. The D-subunits provide a platform for the assembly of the I-subunits, which results in a two-tiered hexameric ring complex. However, the D-subunits are unstable in the chloroplast unless ATPase active I-subunits are present. The H-subunit binds protoporphyrin and is suggested to be the catalytic subunit. Previous studies have indicated that the H-subunit also has ATPase activity, which is in accordance with an earlier suggested two-stage mechanism of the reaction. In the present study, we demonstrate that gel filtration chromatography of affinity-purified Rhodobacter capsulatus H-subunit produced in Escherichia coli generates a high- and a low-molecular-mass fraction. Both fractions were dominated by the H-subunit, but the ATPase activity was only found in the high-molecular-mass fraction and magnesium chelatase activity was only associated with the low-molecular-mass fraction. We demonstrated that light converted monomeric low-molecular-mass H-subunit into high-molecular-mass aggregates. We conclude that ATP utilization by magnesium chelatase is solely connected to the I-subunit and suggest that a contaminating E. coli protein, which binds to aggregates of the H-subunit, caused the previously reported ATPase activity of the H-subunit.


1981 ◽  
Vol 87 (1) ◽  
pp. 25-33 ◽  
Author(s):  
J. P. Van de Merwe ◽  
P. I. M. Schmitz ◽  
F. Wensinck

SummaryAnaerobic coccoid rods belonging to species ofEubacteriumandPeptostreptococcusagglutinate more frequently with sera from patients with Crohn's disease than with sera from patients suffering from other diseases and from healthy subjects. Results of agglutination tests with four strains of coccoid anaerobes were used to estimate the probability that a patient suffers from Crohn's disease. The data on healthy subjects and patients with Crohn's disease were subjected to logistic discriminant analysis. With the methods and interpretation described, 52% of the patients with Crohn's disease were recognized as ‘definite’ or ‘probable’ Crohn's disease and 14% as ‘suspected’. Only 1% of the healthy subjects were classified as ‘suspected’ and none as ‘definite’ or ‘probable’ Crohn's disease.


1998 ◽  
Vol 114 (2) ◽  
pp. 262-267 ◽  
Author(s):  
Geert R. D'Haens ◽  
Karel Geboes ◽  
Mark Peeters ◽  
Filip Baert ◽  
Freddy Penninckx ◽  
...  

PLoS ONE ◽  
2019 ◽  
Vol 14 (1) ◽  
pp. e0211328 ◽  
Author(s):  
Valerie Collij ◽  
Floris Imhann ◽  
Arnau Vich Vila ◽  
Jingyuan Fu ◽  
Gerard Dijkstra ◽  
...  

1989 ◽  
Vol 17 (6) ◽  
pp. 526-531 ◽  
Author(s):  
O. Sangaletti ◽  
M. Petrillo ◽  
G. Bianchi Porro

Urinary oxalate concentrations were measured in 45 patients with quiescent Crohn's disease, four patients with chronic pancreatitis and five healthy subjects after a normal oxalate (150 g/day) diet, after a high-fat (150 g/day), normal oxalate diet and after a high-oxalate (500 mg/day) diet. Urinary oxalate concentrations were significantly ( P<0.05) higher in patients with Crohn's disease and steatorrhoea, but not in those with chronic pancreatitis, after administrating a high-oxalate diet compared with healthy subjects. Mean oxalate values were 19.1 mg/24 h in controls compared with 65.8 mg/24 h in Crohn's disease patients. A direct correlation ( r=0.37, P<0.01) was established between faecal fats and urinary oxalate after oral oxalate load; this correlation ( r=0.43, P<0.01) is closer when only patients with Crohn's disease are considered. The study, therefore, confirmed a correlation between steatorrhoea and hyperoxaluria in patients with Crohn's disease; however, the high percentage of false positive results limits the use of urinary oxalate concentrations as a reliable indicator of lipid malabsorption. It is concluded that, at present, measurement of urinary oxalate cannot be recommended as a valid alternative to the Van de Kamer method for diagnosing lipid malabsorption.


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