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Intestinal glycosidase activities in one adult and two suckling echidnas: absence of a neutral lactase (beta-D-galactosidase).

The activities of various glycosidases in homogenates of the small-intestinal mucosa of one adult and two suckling echidnas, Tachyglossus aculeatus, were investigated. The activities of lactase (beta-D-galactosidase), beta-N-acetylglucosaminidase, neuraminidase and alpha-L-fucosidase were higher in the sucklings than in the adult animal. Maltase and isomaltase activities were lower. Sucrase and cellobiase activities were absent or present in trace amounts only. The lactase activity had a pH optimum of 4.0-4.5, was predominantly in the soluble fraction following ultracentrifugation and was inhibited by p-chloromercuribenzene sulfonate, suggesting that it was due to a lysosomal acid beta-galactosidase and not a brush-border neutral lactase. The maltase activity of the sucklings also had the characteristics predominantly of a lysosomal acid hydrolase. It is proposed that in suckling echidnas, the oligosaccharides (mainly neuraminyllactose and fucosyllactose) of the mother's milk are digested intracellularly by lysosomal enzymes, rather than at the brush border, of the epithelial cells of the small-intestinal mucosa.

Animals↗

Multiple glycosidase deficiencies in a case of juvenile (type 3) Gaucher disease.

Biochemical investigations were performed on autopsy tissues obtained from an 11-year-old girl who died with the juvenile, subacute neuropathic form of Gaucher disease. In addition to the expected deficiency of glucocerebrosidase activity, extracts of both liver and kidney from this individual displayed a profound (greater than or equal to 90%) deficiency of "soluble" beta-glucosidase, beta-xylosidase, and beta-galactosidase activities. Fibroblasts obtained from this individual also contained markedly reduced levels of beta-xylosidase activity but normal levels of beta-D-fucosidase and beta-galactosidase activity. Because the soluble beta-glucosidase, beta-xylosidase, and a portion of the beta-galactosidase activities from control human liver all cochromatographed on a gel filtration column of Sephadex G-200, it is suggested that these activities all reside in a single enzyme, analogous to the situation described in a number of nonhuman, mammalian tissues. This demonstration of multiple glycosidase deficiencies in addition to the deficiency of glucocerebrosidase in a case of subacute neuropathic Gaucher disease suggests that other biochemical aberrations, in addition to a deficiency of glucocerebrosidase, might contribute to pathology in some cases of Gaucher disease.

Child↗

Molecular defect in combined beta-galactosidase and neuraminidase deficiency in man.

In normal human fibroblasts, an enzymically active 85,000-dalton precursor form of beta-galactosidase is processed, via a number of intermediates, into a mature 64,000-dalton form. In addition there is an enzymically inactive 32,000-dalton component and its 54,000-dalton precursor. In fibroblasts from patients with a combined deficiency of beta-galactosidase and neuraminidase these last two components are absent and hardly any mature beta-galactosidase can be demonstrated. Nevertheless, in the mutant fibroblasts, precursor beta-galactosidase is synthesized and processed normally. The excessive intralysosomal degradation that is responsible for the deficiency of mature beta-galactosidase can be partially corrected by addition of the protease inhibitor leupeptin, which results in the accumulation of 85,000-dalton precursor beta-galactosidase and of a partially processed 66,000-dalton form. When mutant cells were grown in the presence of a "corrective factor" purified from the medium of NH4Cl-stimulated cell cultures, both beta-galactosidase and neuraminidase activities were restored to low control levels. The immunoprecipitation pattern was completely normal after addition of the corrective factor, and mature 64,000-dalton beta-galactosidase accumulated in the mutant fibroblasts. We propose that the combined beta-galactosidase/neuraminidase deficiency is caused by a defective 32,000-dalton glycoprotein which is normally required to protect beta-galactosidase and neuraminidase against excessive intralysosomal degradation and to give these enzymes their full hydrolytic activity.

Cells, Cultured↗

Reclassification of Bacteroides putredinis (Weinberg et al., 1937) in a new genus Alistipes gen. nov., as Alistipes putredinis comb. nov., and description of Alistipes finegoldii sp. nov., from human sources.

During studies on the bacteriology of appendicitis in children, we often isolated from inflamed and non-inflamed tissue samples, an unusual bile-resistant pigment-producing strictly anaerobic gram-negative rod. Phenotypically this organism resembles members of Bacteroides fragilis group of species, as it is resistant to bile and exhibits a special-potency-disk pattern (resistance to vancomycin, kanamycin and colistin) typical for the B. fragilis group. However, the production of brown pigment on media containing haemolysed blood and a cellular fatty acid composition dominated by iso-C15:0, suggests that the organism most closely resembles species of the genus Porphyromonas. However, the unidentified organism differs from porphyromonads by being bile-resistant and by not producing butyrate as a metabolic end-product. Comparative 16S ribosomal RNA gene sequencing studies show the unidentified organism represents a distinct sub-line, associated with but distinct from, the miss-classified species Bacteroides putredinis. The clustering of the unidentified bacterium with Bacteroides putredinis was statistically significant, but they displayed > 4% sequence divergence with each other. Chromosomal DNA-DNA pairing studies further confirmed the separateness of the unidentified bacterium and Bacteroides putredinis. Based on phenotypic and phylogenetic considerations, it is proposed that Bacteroides putredinis and the unidentified bacterium from human sources be classified in a new genus Alistipes, as Alistipes putredinis comb. nov. and Alistipes finegoldii sp. nov., respectively. The type strain of Alistipes finegoldii is CCUG 46020(T) (= AHN243(T)).

Appendicitis↗

Prospective evaluation of lactose malabsorption by lactose hydrogen breath test in individuals infected with Entamoeba histolytica and passing cysts.

The aim of the present prospective study was to detect lactose malabsorption in subjects in northern India infected with Entamoeba histolytica and passing cysts. The study group included forty-one patients with E. histolytica cysts in at least one of three consecutive faecal samples. Lactose malabsorption was detected by a lactose H2 breath test. The results were compared with those of forty controls subjects. Thirty-two of forty-one (78.0 %) subjects passing E. histolytica cysts had lactose malabsorption compared with seventeen of forty (42.5 %) control subjects (P<0.01). In conclusion, the present study shows that lactose malabsorption is significantly more common in individuals infected with E. histolytica and passing cysts compared with control subjects.

Adult↗

Beta-galactosidase production by Streptococcus thermophilus is higher in the small intestine than in the caecum of human-microbiota-associated mice after lactose supplementation.

Transit kinetics and survival rates of a bacterial species from yoghurt (i.e. Streptococcus thermophilus strain FBI3) were examined in different digestive compartments of gnotoxenic and human-microbiota-associated mice. The production of the lactose-hydrolysing enzyme (i.e. beta-galactosidase) was also investigated within the digestive tract, using a chromosomal reporter system based on luciferase genes from Photorhabdus luminescens under the control of the plac promoter. In both mice models, S. thermophilus cells transited within 2 h from the stomach to the caecum-colon compartment of the digestive tract where they displayed a survival rate of nearly 100 %. In gnotoxenic mice, luciferase activity was found to increase in the second half of the small intestine and in the caecum-colon compartment when lactose was added to the drinking water provided to the animals. In human-microbiota-associated mice drinking lactose, luciferase activity was similarly increased in the second half of the small intestine but was drastically reduced in the caecum-colon compartment. This feature could be ascribed to the presence of the resident human microbiota.

Animals↗

Intestinal gas.

Patients with functional bowel disease commonly complain of abdominal pain, bloating, and excessive flatulence and eructation. Pain and bloating may be primarily caused by abnormal intestinal motility rather than by excessive intestinal gas. As yet there are no data available that prove excessive flatulence is actually caused by the presence of excessive intestinal gas. A study of the composition of intestinal gas provides insight into whether it is derived from swallowed air or from intraluminal metabolism. Therapy aims primarily at excluding the presence of organic disease as a cause and reassuring the patient that the disorder is functional in nature. Dietary manipulation, changing the habit of aerophagia, exercise, and pressure and heat applied to be abdominal area are all possibilities to be tried.

Aerophagy↗

Coffee, sugars, and chronic diarrhea. Why a dietary history is important.

Patients with chronic diarrhea should be carefully questioned about their diet. A correlation may be found between gastrointestinal symptoms and the ingestion of coffee, milk, or sugars such as sorbitol and fructose. If the offending agent can be identified and withdrawn from the diet, the diarrhea will stop and further evaluation can be avoided.

Chronic Disease↗

Comparison of a portable breath hydrogen analyser (Micro H2) with a Quintron MicroLyzer in measuring lactose maldigestion, and the evaluation of a Micro H2 for diagnosing hypolactasia.

The measurement of hydrogen in exhaled air and changes in the concentration of blood glucose and urine galactose excretion are indirect methods of diagnosing hypolactasia. The aim of this study was to compare a portable breath hydrogen analyser (Micro H2) with a widely used model (Quintron MicroLyzer) and to compare them with the blood glucose, urine galactose, and gastrointestinal symptoms in the lactose tolerance test. After an overnight fast, 44 volunteers (18-66 y) ingested 50 g lactose in a single oral dose. Changes in exhaled breath hydrogen concentrations were measured with the two analysers, and changes in blood glucose and urinary galactose were assayed for 4 h and used as a reference. Eighteen subjects were diagnosed as maldigesters according to our gold standard of at least two positive tests out of the three: breath hydrogen by Quintron, blood glucose concentration, and urine galactose excretion. The highest increase in the breath hydrogen concentration over the baseline was highly variable: 44-366 ppm (Micro H2) or 27-187 ppm (Quintron MicroLyzer). The sensitivity, specificity, and positive and negative predictive values of the Micro H2 compared to the gold standard were 83%, 96%, 94% and 89%, respectively. Overall agreement was 91% (95% CI 78-97%). Compared to the Quintron, the diagnoses were identical in 100% of the cases (92-100%). Thus, for diagnosing hypolactasia, the Micro H2 appeared as reliable for measuring breath hydrogen concentrations as Quintron MicroLyzer commonly used in oral lactose tolerance tests.

Adolescent↗

A novel method for diagnosis of adult hypolactasia by genotyping of the -13910 C/T polymorphism with Pyrosequencing technology.

The lactose-loading test and other functional tests that have been the most widely used clinically in the diagnosis of adult hypolactasia up to now are labour intensive and costly, and suffer from low sensitivity as well as low specificity. In addition, lactose-loading tests may be painful to the patient. Here, a new genotyping method for the diagnosis of adult hypolactasia is described. The method utilizes Pyrosequencing technology, which gives the DNA sequence around the recently identified C/T polymorphic site in the MCM6 gene. Among the advantages compared to the other genotyping methods published are less staff hands-on time than for example RFLP analyses, and the avoidance of radioactivity, as in the originally described isotope-minisequencing. Most importantly, Pyrosequencing, which is a direct DNA sequencing technique, gives unambiguous genotyping results as well as some redundant sequence information beyond the SNP position, which serves as a valuable internal control, obtained for each sample.

Adult↗

Probiotics in gastroenterology: indications and future perspectives.

Nowadays. there is a growing interest in probiotics as a safe way of changing the intestinal bacterial flora. Probiotics may have potential in several gastroenterological conditions, especially when the intestinal flora has been disturbed. Most scientific evidence is available for diarrhoea patients treated with Lactobacillus GG, Lactobacillus reuteri or Saccharomyces boulardii. Meta-analyses have shown an overall reduction in the risk of antibiotic-associated diarrhoea during treatment with probiotics, and benefits have also been demonstrated for patients with rota-virus-associated diarrhoea. Patients with inflammatory bowel disease, an inflammatory disorder characterized by a change in the intestinal flora, are another important target group for which probiotics may be beneficial. It has been claimed that in ulcerative colitis and Crohn disease patients, lactobacilli, S. boulardii and Escherichia coli reduce relapses. but most studies were not placebo-controlled. A reduction in relapses has also been demonstrated in pouchitis patients treated with a multispecies probiotic. Irritable bowel syndrome might be another clinical indication for probiotic therapy, but results of clinical trials performed in these patients are inconsistent. Additionally, probiotics may improve lactose absorption. Helicobacter pylori eradication and constipation. Finally, in animal models of colorectal cancer, treatment with probiotics reduces the prevalence of this disease, and in humans the amount of genotoxic substances in faeces has been reduced. In conclusion, the results of studies on the effects of probiotics in gastrointestinal conditions are encouraging. but well-designed placebo-controlled studies are warranted before recommendations for therapeutic or preventive use can be given. Many issues still have to be resolved, including optimal dose and duration of treatment, selection of and differences between the several available probiotic strains, and, importantly, their mechanisms of actions have to be elucidated.

Colorectal Neoplasms↗

Is cow's milk suitable for the dietary supplementation of rural Gambian children? 1. Prevalence of lactose maldigestion.

To define the age-related prevalence of lactose maldigestion (LM), 218 children aged 13-72 months from a rural Gambian village were studied using a lactose breath hydrogen test. There was a significant rise in LM from 21% to 76% between the 2nd and 3rd-5th years of life (p < 0.001). Only eight children had diarrhoea within 2 weeks before the test. Diarrhoea and abdominal discomfort followed in only seven children (3%). There was no difference in mean time of introduction of supplementary diet or cessation of breastfeeding between children aged 12-36 months with lactose digestion and LM. There was a greater proportion of children with normal lactose digestion than with LM in those still receiving breast milk (85% vs 15%), and a greater proportion of LM in the fully weaned group (63% vs 37%) (p < 0.001). There was no relation between weight-for-age or weight-for-height and the ability to digest lactose. The ability to digest lactose is lost in the majority of children during the 2nd year, coincident with the cessation of breastfeeding, but is not a major factor associated with growth failure thereafter.

Age Factors↗

Intestinal lactase status of adults in Papua New Guinea.

Fifty hospital patients in Port Moresby (mean age 30 yr) without evidence of malnutrition or gastro-intestinal disease, who came from many parts of Papua New Guinea (P.N.G.), were given 50 g lactose orally after an overnight fast. In 49 blood glucose rise was less than 1.1 mmol 1(-1), indicating hypolactasia. Thirty-three gave a clear history of symptoms, usually diarrhoea, after lactose. Adult hypolactasia approaches 100% in P.N.G. There is no indication that the genetic structure of any part of the population of P,N.G. with regard to lactase has been influenced by genetic drift from peoples with persistence of lactase into adult life (PL).

Adolescent↗

The effect of the digestion products of lactose (glucose and galactose) on its intraintestinal, in vivo hydrolysis by exogenous microbial beta-D-galactosidase.

Exogenous, microbial beta-D-galactosidases are capable of effecting hydrolysis of lactose in situ in the gastrointestinal tract of lactase-deficient subjects when given as replacement therapy at mealtime. As its digestion products-glucose and galactose-are known to inhibit lactose hydrolysis in vitro, the effect of adding excess monosaccharide to milk on the hydrolytic efficiency of a beta-galactosidase from Aspergillus niger in adult lactose-malabsorbers was tested. Subjects were studied with 360-ml volumes of milk containing 18 g of carbohydrate. This was administered as intact milk, as lactose-prehydrolyzed milk, and as milk to which 399 mg of Lactase N was added within 5 minutes of consumption. This latter Lactase N-treated milk was administered alone and with graded levels of glucose-9, 18, and 36 g-and with similar doses of galactose. The Lactase N enzyme alone at mealtime reduced breath H2 production by 68% as compared to intact milk. The addition of monosaccharides produced no change in the apparent hydrolytic efficiency of the Lactase N in situ. Thus, product inhibition is unlikely to be the basis for the limited efficiency of intraintestinal hydrolysis of milk lactose by the enzyme from A niger.

Adult↗