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Effect of autoclaving of a lactose-containing diet on cholesterol and bile acid metabolism of conventional and germ-free rats.

Feeding of lactose in amounts comparable to the adult human intake in developed countries (6% of diet, and in later studies 10%) had no major effect on cholesterol and bile acid metabolism of germ-free and conventional rats. However, when lactose-containing casein-starch diets were sterilized by autoclaving, changes in intestinal and/or fecal bile acids were found. Both germ-free and conventional rats demonstrated some increase in intestinal beta-muricholic acid concentrations ascribable to the mere presence of lactose in the diet. Autoclaving of the diet produced additional changes, especially in the fecal bile acid pattern of conventional rats. Here the ratio between the beta-muricholic-derived secondary bile acids hyodeoxycholic and omega-muricholic acids changed from the usual 5:3 to approximately 1:10, with omega-muricholic acid becoming the major fecal bile acid. These changes point to a notable effect of lactose-derived products, formed during steam-sterilization, on the microbial modification of intestinal bile acids in the lower gut. Similar changes have been observed after oral administration of aureomycin and other, unrelated antibiotics that inhibit growth of gram positive organisms.

Animals↗

Effect of acetylsalicylic acid on symptoms and hydrogen excretion in the disaccharide tolerance test with lactose or lactulose.

Prostaglandins play a role in the pathogenesis of symptoms of food intolerance. An alleviation of lactose intolerance by premedication with the prostaglandin-synthesis inhibitor acetylsalicylic acid (ASA) has been reported. We studied the effect of 900 mg of ASA and of a glucose placebo on disaccharide intolerance symptoms and on breath hydrogen (H2) excretion in 16 healthy adult males 12 of whom were lactose malabsorbers and received 50 g of lactose 30 to 40 min after ASA; the remaining four were lactose absorbers and received 30 g of lactulose. Premedication with ASA had no significant effect on the severity and duration of intolerance symptoms and on the timing of H2 excretion. In contrast, the maximal breath H2 concentration and the total H2 excretion were significantly increased after ASA administration. We conclude that prostaglandins are of minor importance in causing the usually moderate symptoms of disaccharide intolerance. Prostaglandin-synthesis inhibition by ASA may reduce the motility of the colon thereby permitting more colonic gas (derived from unabsorbed carbohydrates) to be absorbed and excreted.

Aspirin↗

Lactose malabsorption in Australian Aborigines.

The aim of this study was to determine the incidence of lactose malabsorption in healthy, adult Australian Aborigines. Malabsorption of lactose was measured in 45 full-blood Aboriginal subjects and 37 non-Aboriginal, multiracial controls using the breath hydrogen method. Eighty-four percent of the Aboriginal subjects were found to be lactose malabsorbers and 64% developed abdominal pain or diarrhea. In the control subjects, 20% were found to be lactose malabsorbers and all of these developed symptoms of diarrhea. The results provide strong evidence that Australian Aborigines, in common with most human adults, are lactase deficient.

Australia↗

Cyclic AMP-mediated jejunal secretion in lactose-fed malnourished rats.

We evaluated the effects of a hyperosmolar lactose load on the functional properties of the jejunal mucosa of protein-energy malnourished rats. Malnourished animals exposed to an oral lactose load showed an enhanced loss of DNA, protein, and sodium into the intestinal lumen as compared to well nourished controls exposed to the same lactose load. Only the jejunum of lactose-fed malnourished rats showed increased levels of 3'-5' cyclic adenosine monophosphate (cAMP), that appeared principally due to an enhancement of adenyl cyclase. Overall, our data strongly suggest that the intestinal mucosa in malnutrition is more sensitive and responds to an osmotic stress by enhanced secretion.

3',5'-Cyclic-AMP Phosphodiesterases↗

Intestinal-transit and lactose intolerance in chronic alcoholics.

Intestinal transit time of lactulose and malabsorption of lactose were measured by hydrogen excretion in breath following ingestion of 20 g lactulose or 25 g lactose. Patients were 20 recently drinking, adequately nourished, chronic, male alcoholics. Many (65%) complained of diarrhea while drinking actively. Small bowel transit was significantly shorter in alcoholics with diarrhea (62 +/- 8.6 min) than in normal controls (93 +/- 10.4 min, p less than 0.05). It increased significantly after 8-10 days abstinence, to a value of 101.5 +/- 11 min, not significantly different from normal controls. Following lactulose 75% of alcoholics developed diarrhea after 1-2 days compared with only 15% after 8-10 days of abstinence; 40% had diarrhea after lactose in the initial study, whereas no subjects had diarrhea from the same dose after a period of sobriety. These data suggest that drinking alcoholics have an increased sensitivity to osmotic loads, which is associated with diarrhea, shortened transit time, and lactose intolerance. Abnormalities disappeared 8-10 days following cessation of drinking and normal diet.

Adult↗

Milk and lactose intakes and ovarian cancer risk in the Swedish Mammography Cohort.

BACKGROUND: High intakes of dairy products and of the milk sugar lactose have been hypothesized to increase ovarian cancer risk, but prospective data are scarce. OBJECTIVE: We examined the association between intakes of dairy products and lactose and the risk of total epithelial ovarian cancer and its subtypes. DESIGN: This was a prospective population-based cohort study of 61 084 women aged 38-76 y who were enrolled in the Swedish Mammography Cohort. Diet was assessed in 1987-1990 with the use of a self-administered food-frequency questionnaire. During an average follow-up of 13.5 y, 266 women were diagnosed with invasive epithelial ovarian cancer; 125 of those women had serous ovarian cancer. RESULTS: After adjustment for potential confounders, women who consumed >/=4 servings of total dairy products/d had a risk of serous ovarian cancer (rate ratio: 2.0; 95% CI: 1.1, 3.7; P for trend = 0.06) twice that of women who consumed <2 servings/d. No significant association was found for other subtypes of ovarian cancer. Milk was the dairy product with the strongest positive association with serous ovarian cancer (rate ratio comparing consuming >/=2 glasses milk/d with consuming milk never or seldom: 2.0; 95% CI: 1.1, 3.7; P for trend = 0.04). We observed a positive association between lactose intake and serous ovarian cancer risk (P for trend = 0.006). CONCLUSIONS: Our data indicate that high intakes of lactose and dairy products, particularly milk, are associated with an increased risk of serous ovarian cancer but not of other subtypes of ovarian cancer. Future studies should consider ovarian cancer subtypes separately.

Adult↗

Evidence for a lactose-mediated association between two nuclear carbohydrate-binding proteins.

Nuclear proteins were extracted in 2 M NaCl from membrane-depleted nuclei isolated from HL60 cells. Extracted proteins were submitted to affinity chromatography columns containing immobilized glucose, galactose or lactose. The polypeptides present in the different eluted fractions were resolved by SDS-PAGE and were either silver stained or analysed by immunoblotting with monoclonal or polyclonal antibodies, respectively, raised against the glucose-binding protein CBP67 and the galactose-binding proteins CBP35 and L14. The results presented here show that HL60 cell nuclei contain CBP35 and a glucose-binding lectin of 70 kDa (CBP70). These data account for the previously reported binding of neoglycoproteins containing glucosyl and galactosyl residues to HL60 cell nuclei. Furthermore, the present study provides the new information that CBP35 can associate with CBP70 by interactions dependent on the binding of CBP35 to lactose, and the results of some affinity chromatography experiments strongly suggest that CBP35 and CBP70 associate by protein-protein interactions. The potential function of this lactose-mediated interaction is discussed with respect to data recently reported by others showing that CBP35 is involved in in vitro mRNA splicing and that lactose inhibits the processing of the pre-RNA substrate.

Antigens, Differentiation↗

Autoregulation of GAL4 transcription is essential for rapid growth of Kluyveromyces lactis on lactose and galactose.

Transcriptional induction of genes in the lactose-galactose regulon of the yeast Kluyveromyces lactis requires the GAL4 transcription activator protein. Previous data indicated that the concentration of GAL4 was tightly regulated under basal, inducing, and glucose repressing conditions but the mechanisms were unknown. In this paper we demonstrate that transcription of the GAL4 gene (KI-GAL4) increases 3- to 4-fold during induction of the regulon. This increase requires a KI-GAL4 binding site, UASG, in front of the KI-GAL4 gene, indicating that the KI-GAL4 protein autoregulates transcription of its own gene. Our data demonstrate that the autoregulatory circuit is essential for full induction of the lactose-galactose regulon and, hence, for rapid growth on lactose or galactose. Other data indicate that basal transcription of the KI-GAL4 gene is governed by unidentified promoter elements. The existence of the autoregulatory circuit reveals an important difference between the lactose-galactose regulon and its homologue in Saccharomyces cerevisiae, the melibiose-galactose regulon. This difference may have evolved in response to different selective pressures encountered by the two organisms.

Base Sequence↗

Membrane assembly of lactose permease of Escherichia coli.

Lactose permease, the lacY gene product in Escherichia coli, is an integral membrane protein. Its induction was examined in secAts and secYts mutants by measuring o-nitrophenyl-beta-galactoside uptake activity. In contrast to the synthesis of the maltose binding protein, the malE gene product, which is dependent on the secA and secY gene products, lactose permease seemed to be produced and integrated functionally into membrane independently of SecA or SecY. Gene fusion of the lamB signal sequence to the N-terminal part of the lactose permease gene resulted in production of active fused permease in the E. coli membrane. The signal sequence did not seem to be processed, judging from its mobility on SDS polyacrylamide gel electrophoresis. E. coli cell growth was super-sensitive to induction of production of the fused permease with the signal sequence in contrast to induction of the normal lactose permease. These results are consistent with the above observation that production and integration of LacY protein into membrane is relatively independent of the SecY protein that may have a certain specificity for the signal sequence or, more generally, membrane translocation intermediates.

Amino Acid Sequence↗

Dietary regulation of intestinal lactase and sucrase in adult rats: quantitative comparison of effect of lactose and sucrose.

To characterize the mechanisms leading to dietary evoked increases of lactase and sucrase activities by carbohydrates, we performed a quantitative comparison of the effects of lactose and sucrose on the corresponding disaccharidases in the jejunum of 2-month-old rats. For 7-10 days the rats were fed a low-starch (5 cal%), high-fat (73 cal%) diet, and for various periods of time (3-72 h) were given an isocaloric sucrose or lactose (20, 40, or 70 cal%) diet. Lactase and sucrase activities in jejunal homogenates were significantly increased within 3 h after the initial feeding of the sucrose (40 cal%) diet. After 3 h of feeding the sucrose diet, sucrase activity gradually increased and reached its maximum at 24 h, whereas lactase activity did not exhibit further change. Increased intake of sucrose led to an increase of lactase and sucrase activity. Within a range of doses of digestible amounts of lactose, the effect of diet containing lactose on these disaccharidase activities was similar to the effect of the diet containing sucrose. This similarity suggests the important role of the common constituent sugar, i.e., glucose. Further, analyses of response to these disaccharides along the villus-crypt axis revealed that the increase of lactase activity occurs at a more apical and broader locus of cohort of epithelial cells along the height of the villus than that of sucrase, suggesting that different mechanisms are involved in dietary regulation of lactase and sucrase.

Animals↗

Feeding lactose-intolerant children with a powdered fermented milk.

Fresh yogurt has been proposed as a milk substitute for lactase-deficient patients. We investigated the possibility that a dried, low-fat milk fermented by yogurt microorganisms would be effective. Processing of a fermented milk by spray-drying led to a powder with persistence of lactase activity (11.7 units/g) without storage at 4 degrees C and stable at room temperature (20-23 degrees C). The tolerance and absorption of 10.5 g of lactose in a volume of 150 ml was studied in 25 Gabonese lactase-deficient children aged 5-14 years in the form of a standard humanized milk formula followed 1 week later by the powdered fermented milk formula. Results of the breath hydrogen test showed that in 24 of 25 cases, lactose absorption was normal with a maximal rise of hydrogen over baseline of under 13 ppm after ingestion of the powdered fermented milk. The rise was above 20 ppm in the 25 children with the same load of lactose in the form of the standard formula and one-third had symptoms of lactose intolerance. This powdered fermented milk preparation should be considered as a valid approach in programs of nutritional support targeted to countries with a high prevalence of lactase deficiency.

Adolescent↗

Lactose in blood in nonpregnant, pregnant, and lactating women.

Lactose synthesized in the mammary gland can pass into the bloodstream by either a paracellular or transcellular pathway. In nonpregnant, nonlactating women, the concentration of lactose in the blood plasma was 1.5 +/- 0.1 microM (mean +/- SEM) in 9 women and undetectable in another 11 women. During pregnancy, this concentration was 3.7 +/- 0.4 microM at 10-21 weeks of gestation, with an increase to 8.7 +/- 1.8 microM by 38-40 weeks of gestation. At the initiation of lactation, the concentration of lactose peaked 3-5 days after birth, with a mean peak concentration of 75 +/- 18 microM, and then decreased to 30 +/- 8 microM when lactation was well established at 6 weeks after birth. These findings suggest that the mammary glands are synthetically active by the beginning of the second trimester of pregnancy and reach maximum synthetic capacity soon after birth. Measurement of concentrations of lactose in the blood plasma during pregnancy and lactation may allow an assessment of the successful initiation of both lactogenesis I and II.

Female↗

The use of bacterial luciferase genes as reporter genes in Lactococcus: regulation of the Lactococcus lactis subsp. lactis lactose genes.

Lactose metabolism is an important industrial trait in dairy lactococci. In Lactococcus lactis, lactose is taken up via the phosphoenolpyruvate-dependent phosphotransferase system (PEP-PTS) and is subsequently metabolized via the glycolytic and tagatose 6-phosphate pathways. Genes for the lactose-specific PEP-PTS proteins, phospho-beta-galactosidase and tagatose 6-phosphate pathway enzymes are encoded by a single 8 kb operon, lacABCDFEGX, and there is a divergently transcribed lacR repressor gene. Transcriptional fusions of both the lac operon promoter and the lacR promoter to the luxAB genes of Vibrio fischeri were used to investigate the regulation of expression of both promoters. In vivo bioluminescence assays demonstrated that lacR negatively regulates the lac operon and also autoregulates itself. Induction of transcription occurred for both promoters during growth on lactose: sevenfold for lacR and fivefold for the lac operon. The lacR promoter was demonstrated to be a particularly strong promoter, being approximately four times more efficient than the lac operon promoter. Both promoters provide good potential for the inducible expression of foreign proteins in Lactococcus.

Base Sequence↗

Intestinal permeability and lactose hydrolysis in human rotaviral gastroenteritis assessed simultaneously by non-invasive differential sugar permeation.

Changes in intestinal permeability and lactose hydrolysis have been investigated in three adults and fifteen infants with acute rotaviral gastroenteritis by differential sugar absorption. The method involves chromatographic measurement of urinary lactose, lactulose and L-rhamnose excretion following combined ingestion in an iso-osmolar test solution. All patients had abnormal intestinal permeability indicated by raised urine lactulose/L-rhamnose excretion, ratio of percentages recovered in 5 h, of 0.462 (0.100-1.227) mean and range, compared with 0.027 (0.008-0.052) for healthy controls (P less than 0.001). Ten patients also had urinary lactose/lactulose excretion ratios raised above the normal range (0.014-0.41, mean 0.258) during their acute illness, indicating impaired intestinal lactose hydrolysis. Both indices had become normal 4 weeks after the acute illness, serial investigation of five patients showing that improvement was complete much earlier. Except for the short duration these changes are similar to those associated with villous atrophy in coeliac disease. The test procedure was verified with respect to intestinal lactose hydrolysis by demonstrating a linear relationship between lactose/lactulose excretion and log jejunal mucosal lactase activity by in vitro assay (R2 = 0.95) in a further group of subjects. Differential lactose/lactulose/L-rhamnose absorption provides a non-invasive and sensitive index of small intestinal integrity of value for the interpretation of prolonged or otherwise complicated enteritis and the distinction of primary secondary intestinal lactase deficiency.

Adult↗

Sequence, organization, transcription and regulation of lactose and galactose operons in Lactobacillus rhamnosus TCELL-1.

AIMS: Understanding the metabolism of lactose and galactose and their regulation in Lactobacillus rhamnosus. METHODS AND RESULTS: A gene cluster containing nine open reading frames (ORFs) involved in the metabolism of lactose and galactose in Lact. rhamnosus TCELL-1 was sequenced and characterized. The order of the ORFs was lacTEGF and galKETRM. Northern blotting experiments revealed that the gene cluster could be transcribed as one lacTEGF-galKETRM mRNA though three major transcripts (lacTEGF, galKETRM and galETRM) were detected for the gene cluster. The transcription of the lac or gal operon was independently induced in the presence of lactose or galactose. Northern blotting and primer extension experiments found the presence of four putative promoters upstream from the ORFs lacT (lacTp), galK (galKp1 and galKp2) and galE (galEp). The measurements of enzymatic activities of GalK, GalE and GalT suggested that the expression of the gal operon was subjected to a galactose activation and glucose repression mechanism. CONCLUSIONS: In Lact. rhamnosus TCELL-1, the galactose moiety of lactose could be metabolized by two alternative pathways (the Leloir and the tagatose 6-phosphate pathways) whereas galactose metabolism could be mediated by the Leloir pathway. SIGNIFICANCE AND IMPACT OF THE STUDY: This work provides important information about sugar metabolism in Lact. rhamnosus.

Amino Acid Sequence↗

Molecular rearrangement of lactose plasmid DNA associated with high-frequency transfer and cell aggregation in Lactococcus lactis 712.

High-frequency conjugation of the lactose plasmid pLP712 is associated with a constitutive cell aggregation phenotype and is facilitated by cointegration with a sex factor. Analysis of 23 independently derived enlarged lactose plasmids revealed that the sex factor DNA present in cointegrates varied in size. This suggested that more than simple cointegration with a sex factor plasmid was involved. Further analysis led to the discovery of a chromosomally located sex factor that could excise and be lost or exist as labile plasmid DNA. Cointegration with this sex factor was shown to be promoted by transposition of a copy of ISSI present on the lactose plasmid, and models are presented to account for the complex and variable structures of the resulting enlarged lactose plasmids.

Base Sequence↗

Evaluation of additional acetone and urea analyses, and of the fat-lactose-quotient in cow milk samples in the herd recording system in Norway.

The purpose of this study was to evaluate the use of additional acetone and urea analyses to the established herd recording system (protein, fat, lactose, and somatic cell count) in cow milk samples every second month. Samples were obtained from 43/82 herds (800/1368 cows) in a lowland/highland area. Acetone concentration (mmol/l) were classified into acetone classes 1 (<0.7); 2 (0.7-1.4); and 3 (>1.4). There was an association between increasing acetone class and increasing ketosis incidence in herd (Kendall's Tau-b = 0.13). The highest percentage frequencies of acetone classes 2 and 3 were in weeks 4-6 postpartum. Milk yield, urea, and lactose were significantly lower; milk protein not different, and milk fat was significantly higher in herds with high ketosis incidence and in cows in acetone classes 2 and 3. General linear models, obtained by backward elimination of non-significant variables, showed that milk protein was negatively associated to milk yield and positively associated to milk fat. Milk yield decreased, and milk fat concentration increased with increasing acetone level. Milk fat and milk lactose were negatively associated, and a fat-lactose-quotient might be promising in a model to rule out subclinical ketosis cows.

Acetone↗

Evaluation of bulk milk analyses of acetone, urea, and the fat-lactose-quotient as diagnostic aids in preventive veterinary medicine.

The purpose of this study was to evaluate the use of additional acetone and urea analyses to the established milk quality system (protein, fat, lactose, and somatic cell count) in bulk milk samples twice a month. Samples were obtained from 44/93 herds in a lowland/highland area. When ranging herds with most acetone values over 0.24mmol/1 (90th percentile), only two out of 10 herds would be on a corresponding list when ranged for highest ketosis treatment incidence. Most high acetone values occur after periods with concentrated calvings, but also in periods without calvings, presumably because of feeding with inferior silage high in butyric acid. Milk fat concentration, milk lactose concentration and the interaction term between these, the fat-lactose-quotient, show the largest absolute values in all three models with ketosis treatment incidence, milk protein concentration, and milk yield as dependent variables in general linear analyses on standardized variables. Acetone and urea concentrations in bulk milk show only small absolute values or are omitted from the models. The practical conclusion from these analyses on bulk milk samples to be used by advisory personnel is small, but it seems that high bulk milk fat concentrations and low lactose concentrations are unfavourable when it comes to ketosis treatment incidence, but favourable when it comes to milk protein concentration. Bulk milk acetone concentrations as a measure of the energy supply in herds must only be interpreted after calving intensity and silage quality are assessed.

Acetone↗